PRIYANKA S Thesis On Probiotic Shrikhand Very Importand PDF

DEVELOPMENT OF FIBER FORTIFIED
PROBIOTIC SHRIKHAND
A
THESIS
SUBMITTED TO THE
ANAND AGRICULTURAL UNIVERSITY
IN PARTIAL FULFILMENT OF THE REQUIREMENTS
FOR THE AWARD OF THE DEGREE
OF
Master of Technology
IN
DAIRY TECHNOLOGY
BY
PRIYANKA SUVERA
B. Tech (DAIRY & FOOD TECHNOLOGY)
DEPARTMENT OF DAIRY TECHNOLOGY

S. M C. COLLEGE OF DAIRY SCIENCE
ANAND-388 110, INDIA
2014
REGISTRATION NO: 04-2024-2012

DEVELOPMENT OF FIBER FORTIFIED
PRIYANKA SUVERA
PROBIOTIC SHRIKHAND
M. Tech
BY
PRIYANKA SUVERA
DAIRY TECHNOLOGY

2014
2014

DEVELOPMENT OF FIBER FORTIFIED PROBIOTIC
SHRIKHAND
Name of Student M ajor Advisor

Priyanka Suvera Dr. P. S. Prajapati

S. M . C. COLLEGE OF DAIRY SCIENCE
ANAND – 388 110, INDIA
ABSTRACT
The present investigation was planned with the objective of developing

fiber-fortified probiotic Shrikhand using the probiotic culture thermophilic
lactic acid freeze-dried DVS (Direct Vat Set) culture (ABY-3).For preparation of
Shrikhand, skim milk was heated to 90ºC for 10 min and then cooled to 39ºC.
Starter culture ABY-3 was added at the rate of 0.1g/liter skim milk. Then it
was incubated at 39ºCfor 7 h to get desired acidity (0.7-0.8 percent LA) and
firmness in dahi. The chakka was prepared by hanging the dahi in muslin
cloth for 8-9 h. The chakka was mixed with calculated amount of pasteurized
cream (60-62 per cent fat) to obtained 6.2 per cent milk fat in Shrikhand. The
fiber containing powders were added based on expected yield of Shrikhand. The
sugar was added at the rate of 72.5 per cent of dry matter on the (w/w) of
chakka. It was mixed properly and passing through wire mesh for smooth and
uniform texture in Shrikhand.
Abstract
Three fiber containing powder were incorporated in Shrikhand at three

rates viz., banana powder (1.0, 3.0, 5.0 per cent for B1, B2, B3 respectively), oat
powder (1.0, 2.0, 3.0 per cent for O1, O2, O3 respectively), inulin powder (1.0,
2.0, 3.0 per cent for I1, I2, I3 respectively) on w/w basis of expected yield of
Shrikhand. The sample of Shrikhand without addition of fiber containing
powder was used as a control (C). All the samples were evaluated for the
composition, physico-chemical, rheological property i.e. consistency, sensory
properties and microbial quality. The data generated during the investigations
was statistically analyzed by using Completely Randomized Design.
Among the three levels studied for each fiber containing powder, B3, O2
and I1 were selected as the most acceptable rates of addition for fiber
fortification in Shrikhand.
In the next part of the study comparison of Shrikhand, containing

selected level of each fiber containing powder viz. B3, O2 I1and C was done and
O2 was selected as the best supplementation of fiber followed by I1 and B3 and
C.
Comparison of cardamom and rajbhog as the flavoring substances for

the fiber-fortified probiotic Shrikhand with control containing no added
flavouring substances revealed that Shrikhand containing cardamom flavor had
significantly the highest total score i.e. 91.72, while control had the lowest total
score i.e.85.89.
The gross composition of the developed cardamom flavor probiotic fiber

fortified Shrikhand were total solids 59.01 per cent, milk fat 6.37 per cent,
protein 5.72 per cent, carbohydrate 44.76 per cent, sucrose 41.47 per cent,
ash 0.68 per cent and fiber 0.24 per cent. The physico chemical properties and
consistency of the developed cardamom flavor probiotic fiber fortified
Shrikhand were acidity 0.98 per cent LA, free fatty acids 0.58 µ.eq/g, soluble
nitrogen 0.27 per cent, water activity 0.938, pH 4.36 and consistency 8.65 N.
the total sensory score obtained by the product was 93.28. Hence, the
ii
Abstract
developed product can be considered excellent as per Indian standards (IS

15348: 2003) grading system. The probiotic count of the developed product was
8.46-log cfu/g, which was much higher than minimum count (6.00 log cfu/g)
to state that product is probiotic.The estimated cost of the developed fiber
fortified probiotic Shrikhand was found to be Rs. 131.84/kg compared to
current market rate of Rs. 160/kg for cardamom flavour Shrikhand without
fiber.
The fiber fortified Shrikhand was packaged in PS cups and stored at

7±2ºC to study its shelf life. The fiber fortified Shrikhand packed in PS cups
was stored at 7±2ºC up to 35d. The packed product was monitored for its
physico chemical, microbiological and a sensory property at predetermined
time intervals after every 7 d of refrigerated storage or until the product was
rejected on sensory basis. Fresh Shrikhand had the acidity value of 0.98per
cent LA. Which increased significantly (P<0.05) to 1.33per cent LA on 35 d of
storage. The FFA content increased from 0.64(0 d) to 1.13 (35 d) during
storage. Fresh Shrikhand had soluble nitrogen content 0.24 per cent, which
increased significantly (P<0.05) to 0.44 per cent upon storage of 35 d. During
storage, the water activity of Shrikhand decreased significantly from 0.9391(0
d) to 0.9330(35 d).Fresh Shrikhand had the pH value 4.81 with decreased
significantly (P<0.05) to 4.37 on 35 d of storage. The consistency of Shrikhand
ranged from 7.81 (at 0 d) to 9.23 (at 35 d).
Storage period had significant (P<0.05) effect on flavor of Shrikhand at

refrigeration temperature. The flavor score of the product was 45.66 at 0 day.
There was significant (P<0.05) decreased in flavor score of Shrikhand at each
interval of storage. On 35 d of storage there was significantly (P<0.05) lower
flavor score i.e. 33.79. Body and texture score of Shrikhand during storage
declined significant (P<0.05) with the advancement of storage period. At 0 day
the highest score (33.26) was observed, it was significantly (P<0.05) higher
than all the storage intervals while at 35 d of storage the lowest score (28.36)
was observed which was significantly (P<0.05) lower than all the storage
iii
Abstract
intervals. The color score of Shrikhand during storage decreased with the
increased in storage period. Fresh Shrikhand had a score of 9.61 on 0 day
which was found to decrease significantly (P<0.05) to 7.08 at 35 d of storage at
refrigeration temperature. At 35 d score of color was significantly (P<0.05) lower
than all other storage intervals. The total score of Shrikhand stored at
refrigeration temperature was found to decrease significantly (P<0.05) with the
increase in storage period. Fresh Shrikhand has a total score of 93.53, which
was significantly (P<0.05) the highest score but decreased significantly at each
interval of storage. The overall sensory scores gave a shelf life of more than 35
d at refrigeration temperature and found acceptable on sensory basis.
Highest probiotic count was recorded in Shrikhand prepared (8.73 log

cfu/g) on 0 day and their after it were gradually decreased with increasing of
storage days. The significantly (P<0.05) lowest score was recorded in Shrikhand
prepared (7.50 log cfu/g) on 35 d of storage. There was decline in probiotic
count that can be observed clearly from the graph. Nevertheless, in the
Shrikhand sample probiotic count remained well above 10 7 even after the end
of refrigerated storage for 35 d. Fresh Shrikhand sample had a SPC count of
5.29 log cfu/g but increased significantly (P<0.05) to 5.85 log cfu/g after 35 d
of storage. The average values of YMC for stored Shrikhand ranged from (7
d)1.53 to (35 d) 2.06 log cfu/g. There was significant effect (P<0.05) of storage
period independently on yeast and mold count of product sample on each
interval of storage at refrigerated temperature. Fresh Shrikhand samples found
to free from yeast and mold at refrigeration temperature.
The fiber containing oat powder (2 per cent) fortified probiotic Shrikhand
had the shelf life up to 35 d of refrigerated storage temperature (7±2ºC).
iv
SHETH M. C. COLLEGE OF DAIRY SCIENCE
ANAND – 388 110 (GUJARAT)
Dr. P. S. Prajapati
Professor and Head,
Department of Dairy Technology,
S. M. C. College of Dairy Science,
Anand Agricultural University,
Anand – 388 110, INDIA
This is to certify that the thesis entitled “DEVELOPMENT OF
FIBER FORTIFIED PROBIOTIC SHRIKHAND” submitted by
Ms. Priyanka Suvera (Registration No. : 04-2024-2012) in partial
fulfillment of the requirements for the degree of Master of
Technology in Dairy Technology of Anand Agricultural University
is a record of bonafide research work carried out by him under my
guidance and supervision and the thesis has not previously formed
the basis for the award of any degree, diploma or other similar title.
Place: Anand P. S. Prajapati

Date: /10/2014 (M ajor Advisor)
ACKNOWLEDGEMENT
“I would maintain that thanks are the highest form of thought; and that gratitude is happiness
doubled by wonder.”
I would like to present my first greatest honour for Jesus Christ for His incredibly and
miraculously awesome blessing through the process of writing this thesis. Thank you Jesus for
accompanying me every single second of my life, to lift me up when I am down, to cheer me up
when I lose my confidence. Thank you for sending these great people to help me in writing my
thesis.
I would like to express my sincere gratitude to my Major Advisor Dr. P S Prajapati,
Professor and Head, Department of Dairy Technology, Sheth M.C. College of Dairy Science,
Anand Agricultural University, Anand, for the continuous support of my M.Tech study and
research, for his patience, motivation, enthusiasm, and immense knowledge. His guidance helped
me in all the time of research and writing of this thesis. I could not have imagined having a
better advisor and mentor for my M.Tech study.Thank you for keeping me in track and being
very patient while mentoring me.
I would like to express the deepest appreciation to my committee members
Dr. A G Bhadania, Minor Advisor, Professor and Head, Dairy Engineering Dept; Dr. A H
Jana, Professor, Dairy Technology Dept; Dr. V B Darji, Associate Professor, Department of
Agricultural Statistics, I.T. College of Agriculture, AAU, Anand. Who has shown the attitude and
the substance of a genius, they continually and persuasively conveyed a spirit of adventure in
regard to research, and an excitement in regard to teaching. Without them supervision and
constant help this dissertation would not have been possible.
I wish to endow my thanks to Dr. B P Shah, Principal and Dean, Sheth M. C. College of
Dairy Science, AAU, Anand, for providing the required infrastructure facilities provided to carry
out the present investigation.
I am also thankful to my wonderful Dairy Technology Department where I share and

study and consult for these 2 years. I am grateful to Dr. Suneeta Pinto for her encouragement
and practical advice. I am also thankful to her for reading my reports; commenting on my views
and helping me understand and enrich my ideas. I would like to acknowledge Dr. Sunil Patel,
Dr. J P Prajapati, Dr. H G Patel, Dr. J B Prajapati, Shri Hiral Modha, Shri Amit Patel,
Mr. A J Gokhle, Ms.Jarita Malik, Mr Chetan Dharaiya, Shri Kunal Kadiya, Ms. Komal Patel,
Mr. Dhinal Patel, Mr. Kunal Gawai for numerous discussions and lectures on related topics
that helped me improve my knowledge in the area. The help rendered by all the peons of Dairy
Technology dept. Is worth appreciating, so I wish to thank all of them.
Most importantly, none of this would have been possible without the love and patience of
my “Mother and father”. My immediate family to whom this dissertation is dedicated to, has
been a constant source of love, concern, support and strength all these years. I would like to
express my heart-felt gratitude to my family. My extended family has aided and encouraged me
throughout this endeavour. Word are not enough to thank to my Sister Maulika Suvera and
Brother Rohan Suvera for providing me constant support, boosting my moral. I specially thank
to Dr. Ashish for all the support and encouragement.
I would also like to extend huge, warm thanks to my roommates, Lalita and Binjan. I am
indebted to some of my good colleagues Vishal, Hitendra, Ronak, Amit, Denzil, Montu, Sharad,
Hetal, Krupa, Moumita, Rohit, Tejas, Vaishali, Rashmi, Janki, Kinjal,Divyta, Hetal, Nikita for
their valuable help and support.
Last but not least, I want to thank you all who have helped me directly or indirectly,
academically or spiritually. Forgive me if I cannot mention your name one by one here but your
help will remain in my heart forever. I know the phrase „Thank you‟ cannot represent my big
gratitude, but at least I want you to know that you are awesome. Thank you.
Place : Anand (PriyankaSuvera)

Date: /10 / 2014
CONTENTS
No. Title Page

NO.
1 INTRODUCTION 01-05
2 REVIEW OF LITERATURE 06-48
2.1 Shrikhand 07
2.1.1 Definition and Standards of Shrikhand 08
2.1.2 Composition of Shrikhand 09-10
2.1.3 Traditional method for Shrikhand making 11
2.1.3.1Quality of milk 11
2.1.3.2 Types of milk 12
2.1.3.3 Standardization of milk 13
2.1.3.4 Heat treatments and homogenization to Milk 14
2.1.3.5 Starter culture and their incubation temperature 16
for preparation of dahi
2.1.3.6 Removal of whey 18
2.1.3.7 Quality and composition of chakka 19
2.1.3.8 Blending and mixing of chakka with additives 22
2.1.4 Physico-chemical properties of Shrikhand 23
2.1.5 Sensory Quality of Shrikhand 25
2.1.6 Microbiological Quality of Shrikhand 26
2.1.7 Rheological Quality of Shrikhand 27
2.1.8 Packaging of Shrikhand 28
2.1.9 Shelf-life of Shrikhand 28
2.2 Technological innovations in commercial production 30
2.2.1Mechanization in manufacture of Shrikhand 30
2.2.2 Manufacture of Shrikhand using membrane Technology 31

No. Title Page
NO.
2.2.3 Manufacture of Direct Acidified Shrikhand 33
2.3 Value addition in Shrikhand 34
2.4 Probiotics and their Potential Use 36
2.4.1 Probiotics in Dairy Products 37
2.4.1.1 Probiotics Shrikhand 37
2.4.1.2 Probiotics in Yoghurt 38
2.1.1.3 Probiotics in Ice cream 39
2.4.1.4 Probiotics in Cheese 40
2.5 Dietary Fibers: Types, Availabilities and Requirements 41
2.5.1 Dietary fiber components 42
2.5.2 Mechanism of dietary fiber 43
2.5.3 Inulin, Banana and Oat as Different Dietary Fiber 44
2.5.4 Nutrition Claim 45
2.5.5 Dietary fiber in dairy products 46
3 MATERIALS AND METHODS 49-68
3.0 Introduction 49
3.1 Materials used in Shrikhand making 49
3.1.1 Skim Milk 50
3.1.2 Cream 50
3.1.3 Probiotic culture 50
3.1.4 Sugar 50
3.1.5 Powder containing fiber ingredients 50
3.1.6 Packaging material 51
3.1.7 Culture media and reagents 51
3.2 Preparation of Shrikhand 51
3.2.1 Preparation of dahi 52
3.2.2 Preparation of chakka 52
No. Title Page
NO.
3.2.3 Mixing of chakka with other ingredients 52
3.2.4 Packaging and storage of Shrikhand 52
3.3 Analysis 52
3.3.1 Composition analysis 54
3.3.2 Physico chemical analysis 59
3.3.3 Microbiological analysis 66
3.3.3.1 Preparation of samples for microbiological 66
analysis of Shrikhand
3.3.4 Sensory evaluation 67
3.4 Statistical analysis 68
4 RESULTS AND DISCUSSION 69-141
4.1 PHASE-I: PREPARATION OF SHRIKHAND USING
70
DIFFERENT FIBER CONTAININGPOWDERS
4.2PHASE-II: OPTIMIZATION OF THE RATE OF ADDITION OF
71
DIFFERENT FIBER CONTAINING POWDER
4.2.1 Optimization of rate of addition of banana powder 72
4.2.1.1 Effect of varying levels of banana powder on
72
the composition of Shrikhand
the physico-chemical properties and consistency of 74
Shrikhand
77
the sensory characteristics of Shrikhand
4.2.2 Optimization of rate of addition of oat powder 80
4.2.2.1 Effect of varying levels of oat powder on the
80
composition of Shrikhand
physico-chemical properties and consistency 83
of Shrikhand
4.2.2.3 Effect of varying levels of fiber containing
oat powder on the sensory attributes of 85
Shrikhand
No. Title Page
NO.
4.2.3 Optimization of rate of addition of inulin powder 88
inulin powder on the composition of Shrikhand 88

inulin powder on the physico-chemical 91
properties and consistency of Shrikhand
4.2.3.3 Effect of varying levels of inulin powder on
94
the sensory attributes of Shrikhand
4.2.4 Fiber contents in different experimental Shrikhand 96
4.3 PHASE-III: COMPARISON OF SHRIKHAND CONTAINING
OPTIMIZED LEVEL OF EACH FIBER CONTAINING
97
POWDER TO SELECT THE BEST FIBER CONTAINING
POWDER IN FIBER FORTIFIED SHRIKHAND
4.3.1 Chemical composition 98
4.3.2 Physico -chemical properties and Consistency 101
4.3.3 Sensory attributes 103
4.4 PHASE - IV: SELECTION OF FLAVOURING INGRDIENTS
109
FOR THE FINAL PREPARED SHRIKHAND
4.4.1 Preparation of optimized fiber fortified probiotic Shrikhand 141
4.4.2 Estimation of cost of production of fiber fortified probiotic
115
Shrikhand
4.5 PHASE – V: STUDY OF THE SHELF LIFE OF PREPARED
SHRIKHAND AT REFRIGERATION TEMPERATURE 116
(7 ±20 C)
4.5.1 Effect of storage on physico chemical and consistency of
117
Shrikhand
4.5.2 Effect of storage on sensory properties of Shrikhand 127
4.5.3 Effect of storage on microbiological quality of Shrikhand 135
5 SUMMARY AND CONCLUSIONS 142-155
BIBLIOGRAPHY i-xx
APPENDICES i-vi
LIST OF TABLES
Table No. Title Page No.
2.1 FSSA standards for Shrikhand 09

2.2 Composition of Shrikhand 10
2.3 New nomenclature of lactic acid bacteria 18
2.4 FSSA standards for Chakka 21
2.5 Composition of Chakka 22
2.6 Composition of raw banana powder 45
2.7 Composition of oat 45
4.1 Code name of experimental Shrikhand 71
Composition of fresh banana powder containing
4.2 73
probiotic Shrikhand
Physico chemical properties and consistency of
4.3 fresh banana powder containing probiotic 75
Shrikhand
Sensory characteristics of fresh banana powder
4.4 78
containing probiotic Shrikhand
Composition of fresh oat powder containing
4.5 81
probiotic Shrikhand
Physico chemical and consistency of fresh oat
4.6 84
powder containing probiotic Shrikhand
Sensory characteristics of fresh oat powder
4.7 87
Composition of fresh inulin powder containing
4.8 89
probiotic Shrikhand
Physico chemical and consistency of fresh inulin
4.9 92
Sensory characteristics of fresh inulin powder
4.10 95
4.11 Content of fiber in different samples 97
Average composition of fiber fortified probiotic
4.12 99
Shrikhand
Physico chemical and consistency of fiber
4.13 101
fortified probiotic Shrikhand
i
Sensory characteristics offiber fortified probiotic
4.14 104
Shrikhand
Microbiological quality of fiber fortified probiotic
4.15 107
Shrikhand
Selection of flavour substance for the final
4.16 110
prepared Shrikhand
4.17 Gross composition of final product Shrikhand 114
Estimated cost of production of fiber fortified
4.18 115
probiotic Shrikhand
Effect of storage period on physico-chemical
4.19 properties and consistency of Shrikhandat 117
refrigeration temperature (7±20C)
Effect of storage period on sensory attributes of
4.20 127
Shrikhand
Effect of storage period on microbial quality
4.21 136
ofShrikhandat refrigeration temperature (7±20C)
ii
LIST OF FIGURES
Fig. No. Title Page No.

Flow-chart of standardized process for Shrikhand
3.1 51
manufacture
A representative two-bite force distance texture
3.2 profile curve of banana powderShrikhandsample 62
sample
3.3 63
profile curve of oat powderShrikhandsample sample
3.4 profile curve of inulin powderShrikhandsample 64
sample
3.5 65
profile curve of controlShrikhandsample sample
4.1 Method for fiber fortified probiotic Shrikhand 112
Influence of storage period on titratable
4.2 acidity(%LA) of Shrikhandat refrigerated 118
temperature (7±2ºC)
Influence of storage period on free fatty acid(µ.eq/g)
4.3 120
of Shrikhandat refrigerated temperature (7±2ºC)
Influence of storage period on soluble nitrogen(%)
4.4 122
Influence of storage period on water activity of
4.5 123
Shrikhandat refrigerated temperature (7±2ºC)
Influence of storage period on pH of Shrikhandat
4.6 125
refrigerated temperature (7±2ºC)
Influence of storage period on consistency of
4.7 126
Influence of storage period on flavour of
4.8 128
Influence of storage period on body and texture of
4.9 130
Influence of storage period on colour and
appearance of Shrikhandat refrigerated
4.10 132
temperature (7±2ºC)properties of Burfi at room
temperature (37±20C)
Influence of storage period on overall acceptability
4.11 134
Influence of storage period on probiotic count of
4.12 137
Influence of storage period on standard plate count
4.13 139
Influence of storage period on yeast and mold
4.14 count of Shrikhandat refrigerated temperature 141
(7±2ºC)
LIST OF PLATES
Plate
Title
No.
I Water Activity Meter
II Instrumental Texture Analyzer
III Fiber fortified probiotic Shrikhand
LIST OF APPENDICES
Sr. Page
Title
No. No.
I Sensory Score card for Shrikhand i
II Buffer and Media for Microbiological Analysis ii
III Sharp’s extraction solution v
IV Composition of banana, oat and inulin powder vi
LIST OF ABBREVIATIONS
% Per cent
@ At the rate of
± Plus or minus
≤ Less than or equal to
≥ More than or equal to
0C Degree Celsius
µm Micrometer
µM Micromole
aw Water activity
ANOVA Analysis of Variance
BIS Bureau of Indian Standards
C.D. Critical difference
C.V. Co-efficient of variance
Cfu Colony forming units
Cm Centimeter
CRD Completely Randomized Design
e.g. For example
et al., And co-workers
etc. Etcetera
FDM Fat on Dry Matter
FSSAI Food Safety and Standards Authority of India
g Gram
H2SO4 Sulphuric acid
h Hour
IS Indian Standards
i.e. That is
Kg Kilogram
Kcal/g Kilo-calories per gram
L.A. Lactic acid
Lit. Litre
Log Logarithm
Max. Maximum
Min. Minimum
Ml Milliliter
Mm Millimeter
MSNF Milk solids-not-fat
N Newton
NaOH Sodium Hydroxide
N/mm Newton per millimeter
NLT Not Less Than
NMT Not More Than
NS Non-significant
ppm Parts per million
pH Negative logarithm of hydrogen ion concentration
RO Reverse Osmosis
PS Polystyrene
S Second
SCBM Sweet cream buttermilk
S.Em. Standard error of mean
SNF Solid Not Fat
SPC Standard Plate Count
UF Ultra filtration
viz. Vide licet (Namely)
w/w Weight by weight
YMC Yeast and Mould
Plate- III: - Fiber fortified probiotic Shrikhand
Plate- II: Water Activity Meter
Plate- I: - Texture Analyzer
REVIEW
OF
Introduction
LITERATU
RE
Materials
&
Methods
Results
And
Discussion
Summary
and
Conclusion
Review
of
Literature
SHRIKHAND
A
THESIS
SUBMITTED TO THE
IN PARTIAL FULFILMENT OF THE REQUIREMENTS
FOR THE AWARD OF THE DEGREE
OF
Master of Technology
IN
DAIRY TECHNOLOGY
BY
PRIYANKA SUVERA

2014

PRIYANKA SUVERA
PRIYANKA SUVERA
SHRIKHAND
M. Tech
M. Tech
BY
PRIYANKA SUVERA
DAIRY TECHNOLOGY
DAIRY TECHNOLOGY

S. M. C. COLLEGE OF DAIRY SCIENCE
2014
2014

2014
SHRIKHAND
Name of Student Major Advisor

Priyanka Suvera Dr. P. S. Prajapati

S. M. C. COLLEGE OF DAIRY SCIENCE
ANAND – 388 110, INDIA
ABSTRACT
The present investigation was planned with the objective of developing

fiber-fortified probiotic Shrikhand using the probiotic culture thermophilic
lactic acid freeze-dried DVS (Direct Vat Set) culture (ABY-3).For preparation of
Shrikhand, skim milk was heated to 90ºC for 10 min and then cooled to 39ºC.
Starter culture ABY-3 was added at the rate of 0.1g/liter skim milk. Then it
was incubated at 39ºCfor 7 h to get desired acidity (0.7-0.8 percent LA) and
firmness in dahi. The chakka was prepared by hanging the dahi in muslin
cloth for 8-9 h. The chakka was mixed with calculated amount of pasteurized
cream (60-62 per cent fat) to obtained 6.2 per cent milk fat in Shrikhand. The
fiber containing powders were added based on expected yield of Shrikhand. The
sugar was added at the rate of 72.5 per cent of dry matter on the (w/w) of
chakka. It was mixed properly and passing through wire mesh for smooth and
uniform texture in Shrikhand.
Abstract

rates viz., banana powder (1.0, 3.0, 5.0 per cent for B1, B2, B3 respectively), oat
powder (1.0, 2.0, 3.0 per cent for O1, O2, O3 respectively), inulin powder (1.0,
2.0, 3.0 per cent for I1, I2, I3 respectively) on w/w basis of expected yield of
Shrikhand. The sample of Shrikhand without addition of fiber containing
powder was used as a control (C). All the samples were evaluated for the
composition, physico-chemical, rheological property i.e. consistency, sensory
properties and microbial quality. The data generated during the investigations
was statistically analyzed by using Completely Randomized Design.
Among the three levels studied for each fiber containing powder, B3, O2
and I1 were selected as the most acceptable rates of addition for fiber
fortification in Shrikhand.
In the next part of the study comparison of Shrikhand, containing

selected level of each fiber containing powder viz. B3, O2 I1and C was done and
O2 was selected as the best supplementation of fiber followed by I 1 and B3 and
C.
Comparison of cardamom and rajbhog as the flavoring substances for

the fiber-fortified probiotic Shrikhand with control containing no added
flavouring substances revealed that Shrikhand containing cardamom flavor had
significantly the highest total score i.e. 91.72, while control had the lowest total
score i.e.85.89.
The gross composition of the developed cardamom flavor probiotic fiber

fortified Shrikhand were total solids 59.01 per cent, milk fat 6.37 per cent,
consistency of the developed cardamom flavor probiotic fiber fortified
ii
Abstract

to state that product is probiotic.The estimated cost of the developed fiber
current market rate of Rs. 160/kg for cardamom flavour Shrikhand without
fiber.
The fiber fortified Shrikhand was packaged in PS cups and stored at

7±2ºC to study its shelf life. The fiber fortified Shrikhand packed in PS cups
was stored at 7±2ºC up to 35d. The packed product was monitored for its
rejected on sensory basis. Fresh Shrikhand had the acidity value of 0.98per
cent LA. Which increased significantly (P<0.05) to 1.33per cent LA on 35 d of
storage. The FFA content increased from 0.64(0 d) to 1.13 (35 d) during
storage. Fresh Shrikhand had soluble nitrogen content 0.24 per cent, which
increased significantly (P<0.05) to 0.44 per cent upon storage of 35 d. During
storage, the water activity of Shrikhand decreased significantly from 0.9391(0
d) to 0.9330(35 d).Fresh Shrikhand had the pH value 4.81 with decreased
significantly (P<0.05) to 4.37 on 35 d of storage. The consistency of Shrikhand
ranged from 7.81 (at 0 d) to 9.23 (at 35 d).
Storage period had significant (P<0.05) effect on flavor of Shrikhand at

refrigeration temperature. The flavor score of the product was 45.66 at 0 day.
There was significant (P<0.05) decreased in flavor score of Shrikhand at each
interval of storage. On 35 d of storage there was significantly (P<0.05) lower
flavor score i.e. 33.79. Body and texture score of Shrikhand during storage
the highest score (33.26) was observed, it was significantly (P<0.05) higher
than all the storage intervals while at 35 d of storage the lowest score (28.36)
was observed which was significantly (P<0.05) lower than all the storage
iii
Abstract
intervals. The color score of Shrikhand during storage decreased with the
increased in storage period. Fresh Shrikhand had a score of 9.61 on 0 day
which was found to decrease significantly (P<0.05) to 7.08 at 35 d of storage at
refrigeration temperature. At 35 d score of color was significantly (P<0.05) lower
than all other storage intervals. The total score of Shrikhand stored at
refrigeration temperature was found to decrease significantly (P<0.05) with the
increase in storage period. Fresh Shrikhand has a total score of 93.53, which
was significantly (P<0.05) the highest score but decreased significantly at each
interval of storage. The overall sensory scores gave a shelf life of more than 35
d at refrigeration temperature and found acceptable on sensory basis.

prepared (7.50 log cfu/g) on 35 d of storage. There was decline in probiotic
count that can be observed clearly from the graph. Nevertheless, in the
Shrikhand sample probiotic count remained well above 10 7 even after the end
of refrigerated storage for 35 d. Fresh Shrikhand sample had a SPC count of
5.29 log cfu/g but increased significantly (P<0.05) to 5.85 log cfu/g after 35 d
of storage. The average values of YMC for stored Shrikhand ranged from (7
d)1.53 to (35 d) 2.06 log cfu/g. There was significant effect (P<0.05) of storage
interval of storage at refrigerated temperature. Fresh Shrikhand samples found
to free from yeast and mold at refrigeration temperature.
The fiber containing oat powder (2 per cent) fortified probiotic Shrikhand
had the shelf life up to 35 d of refrigerated storage temperature (7±2ºC).
iv
SHETH M. C. COLLEGE OF DAIRY SCIENCE
ANAND – 388 110 (GUJARAT)
Dr. P. S. Prajapati
Professor and Head,
Department of Dairy Technology,
S. M. C. College of Dairy Science,
Anand Agricultural University,
Anand – 388 110, INDIA
This is to certify that the thesis entitled “DEVELOPMENT OF
FIBER FORTIFIED PROBIOTIC SHRIKHAND” submitted by
Ms. Priyanka Suvera (Registration No. : 04-2024-2012) in partial
fulfillment of the requirements for the degree of Master of
Technology in Dairy Technology of Anand Agricultural University
is a record of bonafide research work carried out by him under my
guidance and supervision and the thesis has not previously formed
the basis for the award of any degree, diploma or other similar title.
Place: Anand P. S. Prajapati

Date: /10/2014 (Major Advisor)
ACKNOWLEDGEMENT
“I would maintain that thanks are the highest form of thought; and that gratitude is happiness
doubled by wonder.”
I would like to present my first greatest honour for Jesus Christ for His incredibly and
miraculously awesome blessing through the process of writing this thesis. Thank you Jesus for
accompanying me every single second of my life, to lift me up when I am down, to cheer me up
when I lose my confidence. Thank you for sending these great people to help me in writing my
thesis.
I would like to express my sincere gratitude to my Major Advisor Dr. P S Prajapati,
Professor and Head, Department of Dairy Technology, Sheth M.C. College of Dairy Science,
Anand Agricultural University, Anand, for the continuous support of my M.Tech study and
research, for his patience, motivation, enthusiasm, and immense knowledge. His guidance helped
me in all the time of research and writing of this thesis. I could not have imagined having a
better advisor and mentor for my M.Tech study.Thank you for keeping me in track and being
very patient while mentoring me.
I would like to express the deepest appreciation to my committee members
Dr. A G Bhadania, Minor Advisor, Professor and Head, Dairy Engineering Dept; Dr. A H
Jana, Professor, Dairy Technology Dept; Dr. V B Darji, Associate Professor, Department of
Agricultural Statistics, I.T. College of Agriculture, AAU, Anand. Who has shown the attitude and
the substance of a genius, they continually and persuasively conveyed a spirit of adventure in
regard to research, and an excitement in regard to teaching. Without them supervision and
constant help this dissertation would not have been possible.
I wish to endow my thanks to Dr. B P Shah, Principal and Dean, Sheth M. C. College of
Dairy Science, AAU, Anand, for providing the required infrastructure facilities provided to carry
out the present investigation.
I am also thankful to my wonderful Dairy Technology Department where I share and

study and consult for these 2 years. I am grateful to Dr. Suneeta Pinto for her encouragement
and practical advice. I am also thankful to her for reading my reports; commenting on my views
and helping me understand and enrich my ideas. I would like to acknowledge Dr. Sunil Patel,
Dr. J P Prajapati, Dr. H G Patel, Dr. J B Prajapati, Shri Hiral Modha, Shri Amit Patel,
Mr. A J Gokhle, Ms.Jarita Malik, Mr Chetan Dharaiya, Shri Kunal Kadiya, Ms. Komal Patel,
Mr. Dhinal Patel, Mr. Kunal Gawai for numerous discussions and lectures on related topics
that helped me improve my knowledge in the area. The help rendered by all the peons of Dairy
Technology dept. Is worth appreciating, so I wish to thank all of them.
Most importantly, none of this would have been possible without the love and patience of
my “Mother and father”. My immediate family to whom this dissertation is dedicated to, has
been a constant source of love, concern, support and strength all these years. I would like to
express my heart-felt gratitude to my family. My extended family has aided and encouraged me
throughout this endeavour. Word are not enough to thank to my Sister Maulika Suvera and
Brother Rohan Suvera for providing me constant support, boosting my moral. I specially thank
to Dr. Ashish for all the support and encouragement.
I would also like to extend huge, warm thanks to my roommates, Lalita and Binjan. I am
indebted to some of my good colleagues Vishal, Hitendra, Ronak, Amit, Denzil, Montu, Sharad,
Hetal, Krupa, Moumita, Rohit, Tejas, Vaishali, Rashmi, Janki, Kinjal,Divyta, Hetal, Nikita for
their valuable help and support.
Last but not least, I want to thank you all who have helped me directly or indirectly,
academically or spiritually. Forgive me if I cannot mention your name one by one here but your
help will remain in my heart forever. I know the phrase ‘Thank you’ cannot represent my big
gratitude, but at least I want you to know that you are awesome. Thank you.
Place : Anand (PriyankaSuvera)

Date: /10 / 2014
CONTENTS
No. Title Page

NO.
1 INTRODUCTION 01-05
2 REVIEW OF LITERATURE 06-48
2.1 Shrikhand 07
2.1.1 Definition and Standards of Shrikhand 08
2.1.2 Composition of Shrikhand 09-10
2.1.3 Traditional method for Shrikhand making 11
2.1.3.1Quality of milk 11
2.1.3.2 Types of milk 12
2.1.3.3 Standardization of milk 13
2.1.3.4 Heat treatments and homogenization to Milk 14
2.1.3.5 Starter culture and their incubation temperature 16
for preparation of dahi
2.1.3.6 Removal of whey 18
2.1.3.7 Quality and composition of chakka 19
2.1.3.8 Blending and mixing of chakka with additives 22
2.1.4 Physico-chemical properties of Shrikhand 23
2.1.5 Sensory Quality of Shrikhand 25
2.1.6 Microbiological Quality of Shrikhand 26
2.1.7 Rheological Quality of Shrikhand 27
2.1.8 Packaging of Shrikhand 28
2.1.9 Shelf-life of Shrikhand 28
2.2 Technological innovations in commercial production 30
2.2.1Mechanization in manufacture of Shrikhand 30
2.2.2 Manufacture of Shrikhand using membrane Technology 31

No. Title Page
NO.
2.2.3 Manufacture of Direct Acidified Shrikhand 33
2.3 Value addition in Shrikhand 34
2.4 Probiotics and their Potential Use 36
2.4.1 Probiotics in Dairy Products 37
2.4.1.1 Probiotics Shrikhand 37
2.4.1.2 Probiotics in Yoghurt 38
2.1.1.3 Probiotics in Ice cream 39
2.4.1.4 Probiotics in Cheese 40
2.5 Dietary Fibers: Types, Availabilities and Requirements 41
2.5.1 Dietary fiber components 42
2.5.2 Mechanism of dietary fiber 43
2.5.3 Inulin, Banana and Oat as Different Dietary Fiber 44
2.5.4 Nutrition Claim 45
2.5.5 Dietary fiber in dairy products 46
3 MATERIALS AND METHODS 49-68
3.0 Introduction 49
3.1 Materials used in Shrikhand making 49
3.1.1 Skim Milk 50
3.1.2 Cream 50
3.1.3 Probiotic culture 50
3.1.4 Sugar 50
3.1.5 Powder containing fiber ingredients 50
3.1.6 Packaging material 51
3.1.7 Culture media and reagents 51
3.2 Preparation of Shrikhand 51
3.2.1 Preparation of dahi 52
3.2.2 Preparation of chakka 52
No. Title Page
NO.
3.2.3 Mixing of chakka with other ingredients 52
3.2.4 Packaging and storage of Shrikhand 52
3.3 Analysis 52
3.3.1 Composition analysis 54
3.3.2 Physico chemical analysis 59
3.3.3.1 Preparation of samples for microbiological 66
analysis of Shrikhand
3.3.4 Sensory evaluation 67
3.4 Statistical analysis 68
4 RESULTS AND DISCUSSION 69-141
4.1 PHASE-I: PREPARATION OF SHRIKHAND USING
70
DIFFERENT FIBER CONTAININGPOWDERS
4.2PHASE-II: OPTIMIZATION OF THE RATE OF ADDITION OF
71
4.2.1 Optimization of rate of addition of banana powder 72
72
the composition of Shrikhand
the physico-chemical properties and consistency of 74
Shrikhand
77
the sensory characteristics of Shrikhand
4.2.2 Optimization of rate of addition of oat powder 80
80
physico-chemical properties and consistency 83
of Shrikhand
oat powder on the sensory attributes of 85
Shrikhand
No. Title Page
NO.
4.2.3 Optimization of rate of addition of inulin powder 88
inulin powder on the composition of Shrikhand 88

inulin powder on the physico-chemical 91
4.2.3.3 Effect of varying levels of inulin powder on
94
the sensory attributes of Shrikhand
4.2.4 Fiber contents in different experimental Shrikhand 96
4.3 PHASE-III: COMPARISON OF SHRIKHAND CONTAINING
OPTIMIZED LEVEL OF EACH FIBER CONTAINING
97
POWDER TO SELECT THE BEST FIBER CONTAINING
POWDER IN FIBER FORTIFIED SHRIKHAND
4.3.1 Chemical composition 98
4.3.2 Physico -chemical properties and Consistency 101
4.3.3 Sensory attributes 103
4.4 PHASE - IV: SELECTION OF FLAVOURING INGRDIENTS
109
FOR THE FINAL PREPARED SHRIKHAND
4.4.1 Preparation of optimized fiber fortified probiotic Shrikhand 141
4.4.2 Estimation of cost of production of fiber fortified probiotic
115
Shrikhand
4.5 PHASE – V: STUDY OF THE SHELF LIFE OF PREPARED
SHRIKHAND AT REFRIGERATION TEMPERATURE 116
(7 ±20C)
4.5.1 Effect of storage on physico chemical and consistency of
117
Shrikhand
4.5.2 Effect of storage on sensory properties of Shrikhand 127
4.5.3 Effect of storage on microbiological quality of Shrikhand 135
5 SUMMARY AND CONCLUSIONS 142-155
BIBLIOGRAPHY i-xx
APPENDICES i-vi
LIST OF TABLES
Table No. Title Page No.
2.1 FSSA standards for Shrikhand 09

2.2 Composition of Shrikhand 10
2.3 New nomenclature of lactic acid bacteria 18
2.4 FSSA standards for Chakka 21
2.5 Composition of Chakka 22
2.6 Composition of raw banana powder 45
2.7 Composition of oat 45
4.1 Code name of experimental Shrikhand 71
Composition of fresh banana powder containing
4.2 73
probiotic Shrikhand
Physico chemical properties and consistency of
4.3 fresh banana powder containing probiotic 75
Shrikhand
Sensory characteristics of fresh banana powder
4.4 78
Composition of fresh oat powder containing
4.5 81
probiotic Shrikhand
Physico chemical and consistency of fresh oat
4.6 84
Sensory characteristics of fresh oat powder
4.7 87
Composition of fresh inulin powder containing
4.8 89
probiotic Shrikhand
Physico chemical and consistency of fresh inulin
4.9 92
Sensory characteristics of fresh inulin powder
4.10 95
4.11 Content of fiber in different samples 97
Average composition of fiber fortified probiotic
4.12 99
Shrikhand
Physico chemical and consistency of fiber
4.13 101
i
Sensory characteristics offiber fortified probiotic
4.14 104
Shrikhand
Microbiological quality of fiber fortified probiotic
4.15 107
Shrikhand
Selection of flavour substance for the final
4.16 110
prepared Shrikhand
4.17 Gross composition of final product Shrikhand 114
Estimated cost of production of fiber fortified
4.18 115
probiotic Shrikhand
Effect of storage period on physico-chemical
4.19 properties and consistency of Shrikhandat 117
refrigeration temperature (7±20C)
Effect of storage period on sensory attributes of
4.20 127
Shrikhand
Effect of storage period on microbial quality
4.21 136
ofShrikhandat refrigeration temperature (7±20C)
ii
LIST OF FIGURES
Fig. No. Title Page No.

Flow-chart of standardized process for Shrikhand
3.1 51
manufacture
3.2 profile curve of banana powderShrikhandsample 62
sample
3.3 63
profile curve of oat powderShrikhandsample sample
3.4 profile curve of inulin powderShrikhandsample 64
sample
3.5 65
profile curve of controlShrikhandsample sample
4.1 Method for fiber fortified probiotic Shrikhand 112
Influence of storage period on titratable
4.2 acidity(%LA) of Shrikhandat refrigerated 118
temperature (7±2ºC)
Influence of storage period on free fatty acid(µ.eq/g)
4.3 120
Influence of storage period on soluble nitrogen(%)
4.4 122
Influence of storage period on water activity of
4.5 123
Influence of storage period on pH of Shrikhandat
4.6 125
Influence of storage period on consistency of
4.7 126
Influence of storage period on flavour of
4.8 128
Influence of storage period on body and texture of
4.9 130
Influence of storage period on colour and
appearance of Shrikhandat refrigerated
4.10 132
temperature (7±2ºC)properties of Burfi at room
temperature (37±20C)
Influence of storage period on overall acceptability
4.11 134
Influence of storage period on probiotic count of
4.12 137
Influence of storage period on standard plate count
4.13 139
Influence of storage period on yeast and mold
4.14 count of Shrikhandat refrigerated temperature 141
(7±2ºC)
LIST OF PLATES
Plate
Title
No.
I Instrumental Texture Analyzer
II Water Activity Meter
III Fiber fortified probiotic Shrikhand
LIST OF APPENDICES
Sr. Page
Title
No. No.
I Sensory Score card for Shrikhand i
II Buffer and Media for Microbiological Analysis ii
III Sharp’s extraction solution v
IV Composition of banana, oat and inulin powder vi
LIST OF ABBREVIATIONS
% Per cent
@ At the rate of
± Plus or minus
≤ Less than or equal to
≥ More than or equal to
0C Degree Celsius
µm Micrometer
µM Micromole
aw Water activity
ANOVA Analysis of Variance
BIS Bureau of Indian Standards
C.D. Critical difference
C.V. Co-efficient of variance
Cfu Colony forming units
Cm Centimeter
CRD Completely Randomized Design
e.g. For example
et al., And co-workers
etc. Etcetera
FDM Fat on Dry Matter
FSSAI Food Safety and Standards Authority of India
g Gram
H2SO4 Sulphuric acid
h Hour
IS Indian Standards
i.e. That is
Kg Kilogram
Kcal/g Kilo-calories per gram
L.A. Lactic acid
Lit. Litre
Log Logarithm
Max. Maximum
Min. Minimum
Ml Milliliter
Mm Millimeter
MSNF Milk solids-not-fat
N Newton
NaOH Sodium Hydroxide
N/mm Newton per millimeter
NLT Not Less Than
NMT Not More Than
NS Non-significant
ppm Parts per million
pH Negative logarithm of hydrogen ion concentration
RO Reverse Osmosis
PS Polystyrene
S Second
SCBM Sweet cream buttermilk
S.Em. Standard error of mean
SNF Solid Not Fat
SPC Standard Plate Count
UF Ultra filtration
viz. Vide licet (Namely)
w/w Weight by weight
YMC Yeast and Mould
Chapter 1
INTRODUCTION
Fermented milk and milk products occupy a place in satisfying

nutritional requirements of human being since the time antiquity. Fermented
milk products have been well recognized to have therapeutic,
anticholeterolemic, anticarcinogenic properties (Devshete et al., 2012).
Amongst the various fermented milk products, dahi a well-known indigenous
fermented milk products prepared by lactic acid fermentation is being
converted in to Shrikhand because of its better shelf life.
Shrikhand in Sanskrit means “Sandal”. Shrikhand, the product might

have been so named because of its flavour, the cooling effect and the colour,
the qualities, which it shares with sandal. However, the fact is that this word is
originated from the Sanskrit word “Shrikharini” as mentioned in “Bhava-
Prakash”, the old classic. With the advent of time, the word “Shikharan” was
derived from “Shikhrini”. Subsequently, the derivation of the words,
“Shrikhand” and “Shikhand”. The later term is used for this product in Gujarat
State. “Shrikhand” was later on modified as “Shrikhand” giving it Sanskrit
touch, thereby suggesting that it might be a Sanskrit word, Hence in
Maharashtra, Mysore and other states this product is known as “Shrikhand”.
Shrikhand is an indigenous fermented and sweetened milk product

having a typical pleasant sweet-sour taste. It is prepared by blending Chakka,
a semi-solid mass obtained after draining whey from dahi, with sugar, cream
and other ingredients like fruit pulp, nut, flavour, spices and colour to achieve
the finished product of desired composition, consistency and sensory
attributes. Shrikhand has a typical semi-solid consistency with a characteristic
smoothness, firmness and pliability that makes it suitable for consumption
directly after meal or with poori (made of a dough of whole-meal wheat, rolled
Introduction
out and deep-fried) or bread. Although largely produced on small scale

adopting age-old traditional methods, Shrikhand is now commercially
manufactured in organized dairy sector to cater to the growing demand.
Shrikhand is served as special delicacy during festivals and ceremonial

occasions. Consumption of Shrikhand is reported to be effective in treatment
of many diseases like diarrhoea, acidity, gastro enteritis (Devshete et al., 2012).
The (FAO/WHO, 2002) definition of probiotic is that they are “live

microorganisms which when administered in adequate amounts confer a
health benefit on the host”. Fermented dairy products have long been an
important component of nutritional diet. In the commercial manufacture of
fermented products, starters containing specific genera, species and strains of
cultures are used to generate desired flavor, body and texture characteristics.
Lactic acid bacteria used as starters in the industrial production of dahi,
Shrikhand, mishit doi, lassi, buttermilk and yoghurt. The lactic acid bacteria
are naturally accepted as GRAS (Generally regarded as safe) for human
consumption. The probiotic bacteria used in commercial products today are
mainly members of the genera Lactobacillus and Bifidobacterium.
Probiotic have been therapeutically, to modulate immunity, improve

digestive process, prevent cancer, improve lactose intolerance, etc. (Makhal et
al., 2005). Lactic acid bacteria decreases serum cholesterol levels and increases
vitamin B content in the product (Grill et al., 2000).
Foods containing probiotic microorganisms come within the category of

functional foods, i.e., foods, which have a positive effect on health.
Furthermore, increased commercial interest in exploiting the proposed health
attributes of probiotic has contributed significantly to the rapid growth and
expansion of this sector of the market (Stanton et al., 2001). Food
manufacturers are enthusiastic about developing such products because the
added ingredients give increased value to food. The global market for probiotic
foods in the coming years is growing rapidly. Indian probiotic industry is
Page | 2
Introduction
achieving its pace at steady rate with opportunities for rapid growth in
near future. Indian probiotic market is valued at $12 million in 2011, is
expected to witness a CAGR (Compound Annual Growth Rate) of 11 per cent by
2016.
Since the awareness about the probiotic and its health benefits has
grown tremendously among Indian population, demand for probiotic foods has
gained much popularity. Indian and multinational companies are coming in
to the picture since they entered the Indian food industry in 2007.
Indian probiotic market was valued at $2 million as per 2010 estimates.
According to figures released by US-based research firm, Frost and Sullivan,
in August 2012, probiotic products in India generated revenue of US
$310 million in 2011. The market value supposed to increase up to US
$522.8 million by 2018. Indian probiotic market is estimated worth Rs 20
million with annual growth rate of 22.6 per cent till 2015 (ICMR-DBT, 2011).
In Indian Probiotic industry different brands are exist like Nestle, Amul,
Yakult Danone and Mother Dairy along with other minor players operating in
different regions in their own capacities. Amul is the leader with nearly 70%
market share in 2011 and is likely to be at the top in the coming years as well.
In 2011, Amul Probiotic share accounted for more than double than the
combined share of its competitors like Mother Dairy, Nestle and Yakult. Nestle
and Mother Dairy is holding second and third position in the Indian Probiotic
Market respectively (www.renub.com). Amul has entered in to the probiotic
segment with the introduction of ice cream and lassi. Nestle has launched
Active Plus dahi and Yakult-Danone, a probiotic drink Yakult (Raja and
Arunachalam, 2011).
It is important for food materials to be delicious as well as natural.

Rapidly increasing of human population of world, environmental pollution
caused by consistently developing technology, insufficient education and
problems caused by wrong nutrition are making supplying of natural elements
Page | 3
Introduction
(lipids, carbohydrates, proteins, vitamins, minerals) for body cells to work

smoothly. Nevertheless, oil content present in the structure of some food
materials is a problematic situation for consumers. In order to solve this
problem, dietary fiber can be used, which can improve the textural and sensual
qualities of products in addition to being functional. An excessive interest has
been observed over the last few years in fibrous nutrients in developed
countries.
An Australian scientist, Eben Hipsley, in 1951, describe lignin, cellulose,

and hemicelluloses in food (Hipsley,1953) first used the term dietary fiber.
Dietary Fibre means carbohydrate polymers with a degree of polymerisation
(DP) not less than three which are not hydrolysed by the endogenous enzymes
in the small intestine of humans (FSSA, 2010). It includes polysaccharides,
oligosaccharides, lignin and associated plant substances. Fiber is found in
varying quantities in all plant foods including legumes, cereals, some fruits
(particularly apples and bananas) and berries, certain vegetables (such as
broccoli, carrots and root vegetables, potatoes, sweet potatoes, and onions),
and seed husk and bran. The dietary fibers can be divided into two groups:
water-soluble and water insoluble fibers. Depending on the source, plants can
contain both types of fibers in varying content. Fibers have different functions
in food products. They have technical functionality relating to texture, as well
as nutritional functionality relating to health (Ramirez et al., 2010; Ajila and
Prasada Rao, 2013).
Dietary fibres can provide a multitude of functional properties when they

are incorporated in food systems. Thus, fibres addition contributes to the
modification and improvement of the texture, sensory characteristics and shelf-
life of foods due to their water-binding capacity, gel-forming ability, fat mimetic,
anti-sticking, anti-clumping, texturising and thickening effects (Thebaudin et
al., 1997; Gelroth and Ranhotra, 2001; Dello et al.,2004). Dietary fiber have
beneficial physiological effect like improved laxation, attenuation of blood
cholesterol, attenuation of post prandial blood glucose, influence of immune
Page | 4
Introduction
function, fermentability and production of SCFAS (short chain fatty acids),

Decreasing of intestinal transits time, increasing of stool bulk (EU, 2008).
Milk is an excellent medium to carry or generate live and active cultured

dairy products. The technology of application of probiotic organisms in
fermented dairy products aims to combine the potential health benefits of the
bacteria with their ability to grow in milk resulting in a nutritional healthy and
desirable product for the consumers. Milk and milk products considered as a
vehicle for dietary fiber would not only take care of their own role in human
health but could also enhance the heath fullness of the diet as a whole.
Therefore, the present study was undertaken with objective to study the
development of traditional dairy product with an addition of probiotic culture
as well as dietary fiber containing ingredients.
Objectives of Research work:
1) To develop fiber fortified probiotic Shrikhand.
2) To study the physico – chemical, rheological, microbiological and

sensory properties of fiber fortified Shrikhand.
3) To evaluate viable count of probiotic in developed product
4) To study shelf life of developed fiber fortified Shrikhand at refrigerated

temperature (7± 2˚C).
5) To analyze of fiber content in the final product.
Page | 5
Chapter 2
REVIEW OF LITERATURE
Cultured dairy products are the vital component of the human diet in
India. Apart from imparting nutrition and novelty, these products help in
preserving the precious nutrients in milk that tend to quick deterioration. Curd
(yoghurt), makhan (cultured butter), Mishti Doi (sweet yoghurt), cheese
analogous, buttermilk, lassi (sweet buttermilk) prominently used as
supplementary food in different parts of India, due to high nutritive
characteristics flavour, taste, palatable nature and possible therapeutic value.
Shrikhand is one amongst the most preferred dairy product in western India.
The product has undergone constant modification to suit the changing needs of
consumer and the present day market. Shrikhand available in various parts of
country has considerable variations with respect to its composition, sensory
quality, microbiological attributes and textural parameters. Several reports are
available in the literature on Shrikhand but scanty information is available on
its market quality. In this chapter, literature is reviewed and collated in the
form of tables for comprehensive understanding.
In the present context, the available literature on Shrikhand is reviewed
under the following groups.
 Shrikhand - Definition and standards of Shrikhand
 Composition of Shrikhand
 Traditional method for Shrikhand making
 Physico-chemical properties of Shrikhand
 Sensory quality of Shrikhand
 Microbiological quality of Shrikhand
 Rheological Quality of Shrikhand
 Packaging of Shrikhand
 Shelf-life of Shrikhand
Review of literature
 Technological innovations in commercial production

 Value addition in Shrikhand
 Probiotics - Dairy Products
 Dietary Fibers: types, availabilities and requirements
 Mechanism of dietary fiber
 Dietary fiber in dairy products
2.1 Shrikhand
Shrikhand is an indigenous fermented and sweetened milk product of
Indian origin and regularly consumed in Gujarat, Maharashtra and certain
parts of Karnataka, Madhya Pradesh and Rajasthan. Shrikhand is sweetened
dewatered dahi. In addition to these places, because of its typical sweet-sour
taste, it is becoming popular in other parts of the country. Shrikhand is
prepared by blending chakka with sugar, cream and other ingredients like
fruit pulp, nut, flavor, spices, color, etc. to achieve the finished product
of desired composition, consistency and sensory attributes. The popularity
of the product is ascribed to its uniqueness with respect to taste, richness,
diversity, cooling effect and method of its manufacture (Upadhyay and Dave,
1977).
Shrikhand has a typical semi- solid consistency with a
characteristic smoothness, firmness and pliability that make it suitable for
consumption directly after meal or with “puree” or bread. It resembles
sweetened Quarg or Quark produced in Germany (Patel and Abd-EL-
Salam, 1986). Although largely produced on small scale adopting age old
traditional methods, with the growing demand, Shrikhand is now
commercially manufactured in organized dairy sector.Shrikhand has
distinctive rich flavour and long shelf life. Shrikhand is served as special
delicacy during festivals and ceremonial occasions. Consumption of Shrikhand
is reported to be effective in treatment of many diseases like diarrhoea, acidity,
gastro enteritis (Devshete et al.,2012).
Page | 7
Shrikhand originated in Persia using Frasi-shir (milk) and khand (sugar),

and was later brought to the shores of Gujarat by the Parsi Zohrastrian settlers
(Narayanan and Lingam, 2013).
Shrikhand is an indigenous semi-soft whole milk product prepared using
Chakka (strained dahi/curd). The curd (dahi) is partially strained through a
cloth to remove the whey and thus produce solid mass called Chakka. Chakka
is finely mixed with sugar and flavouring agents (Kulkarni et al., 2006).
Shrikhand available in various parts of country has considerable
variations with respect to its composition, sensory quality, microbiological
attributes and also textural parameters (Sharma and Zariwala, 1980;
Upadhyay et al.,1984; Salunke et al., 2005; Salunke et al.,2006). Consumers
in Gujarat prefer its sweet variety while those in Maharashtra prefer the
tangy / sour variety ( Aneja et al.,2002).
2.1.1 Definition and Standards of Shrikhand
According to FSSA (2011), Shrikhand-means the product obtained from
Chakka or Skimmed Milk Chakka to which milk fat is added. It may contain
fruits, nuts, sugar, cardamom, saffron and other spices. It shall not contain
any added colour and artificial flavor substances. It shall conform to the
following specifications, namely listed in table 2.1.
Page | 8
Table 2.1: FSSA standards for Shrikhand
Requirement Shrikhand Full Cream Shrikhand
Total Solids, % by wt Not less than 58 Not less than 58
Milk Fat (on dry matter basis), % by wt Not less than 8.5 Not less than 10
Milk protein (on dry matter basis), % by wt Not less than 9 Not less than 7
(for Fruit Shrikhand – not less than 6.0)
Titrable acidity (as lactic acid), % by wt Not more than 1.4 Not less than 1.4
Sugar (sucrose) (on dry matter basis), % by Not more than Not more than 72.5
wt 72.5
Total Ash (on dry matter basis), % by wt Not more than Not more than 0.9
0.9
The Bureau of Indian Standards (BIS) and Prevention of Food

Adulteration Act (PFA) has prescribed a minimum of 58 per cent total solids,
minimum of 8.5 per cent fat (in dry matter), minimum of 10.5 per cent protein
(in dry matter), maximum of 1.4 per cent lactic acid, maximum of 72.5 per cent
sugar (in dry matter), maximum of 0.9 per cent total ash (in dry matter).
2.1.2 Composition of Shrikhand

Wide variations are observed by several workers in the composition of
Shrikhand as shown in Table 2.2.
Page | 9
Table 2.2
Page | 10
2.1.3 Traditional method for Shrikhand making
The traditional method of making Shrikhand involves the preparation of
curd or dahi by culturing cow or buffalo milk with natural starter (curd of the
previous batch). After a firm curd is formed, it is transferred in a muslin cloth
and hung for 12 - 18 hr to remove free whey. The semi- solid mass obtained is
called as “chakka”. The chakka is mixed with required amount of sugar, color,
flavoring materials and spices and blended to smooth and homogenous
consistency. Finally cooling it to a low temperature (≤10ºC) before it is ready for
consumption (Bhattacharya et al., 1972; Rangappa and Achaya, 1974; Parikh
1977; Upadhyay and Dave, 1977; Patel and Chakraborty, 1988). Aneja et al.
(1977a) developed a semi – mechanized method for Shrikhand making which
led to its production on commercial scale. The published literature on the
various aspects of Shrikhand manufacture is presented under the following sub
– groups:
a) Quality of milk
b) Types of milk use
c) Standardized of milk
d) Heat treatment of milk
e) Preparation of Dahi
f) Removal of whey
g) Quality and composition of chakka, and
h) Blending and mixing of chakka with additives
2.1.3.1 Quality of milk
Good-quality raw milk is required to make good-quality dairy products.

Once raw milk is defective, it cannot be improved during processing, and
defects often become more pronounced. Therefore, it is important that raw milk
be produced and handled from farm to plant under conditions that do not
reduce its quality or, consequently, the quality of the product. Many factors
can influence the quality of raw milk.
Page | 11
The raw milk intended for fermented milk manufacture should be free
from antibiotics as well as from any off-flavours, and should have lower
standard plate count and psychotropic count (Riber, 1989).
Milk differing from their normal composition, i.e. colostrums, late

lactation or mastitis, etc., is ales undesirable for dahi making. These milks
contain certain inhibitory components’ such as high somatic cell counts ,
enzymes , and have altered milk constituents, alkaline ph as well as low heat
stability, which adversely affects the growth of starter organisms resulting in
undesirable characteristics of dahi (Singh and Lakhminarayan, 1948;
Anantaramaiah and Iya,1952; Moniz et al.,1976; Singh and Singh, 1980;
Saraswat and Agrawal,1982;). Milk with high somatic count negatively
influences the microbiological quality of dahi (Singh and Singh, 1980).
During the manufacture of cheese and cultured dairy products,

antibiotics can inhibit dairy starter cultures used to develop acid (e.g., lactic
acid bacteria), which can result in the loss of significant amounts of product
and milk (Lundstedt, 1979). Hamdan et al. (1971) found that potassium
sorbate (0.05 and .10 per cent w/v) residue affects production of acid and
acetaldehyde as well as growth of L. bulgaricus and S. thermophilus, the higher
concentration being more hazardous.
2.1.3.2 Types of milk
In India, more than 50 per cent of available milk is from buffalo (Rao et
al., 1989). Aneja et al. (1977) developed an industrial process for the
manufacture of Shrikhand, for that buffalo, skim milk was used for making
dahi for the manufacture of Shrikhand.
Different sources of milk solids were used for Shrikhand making.

Reconstituted skim milk, diluted concentrated milk, concentrated skim milk
(40 to 43 per cent TS) and butter milk, were used for making curd.
Page | 12
Reconstituted milk was found to be an effective alternative to fresh skim milk

for Shrikhand making (Anon,1982; Patel and Chakraborty,1985b).
By using skim milk for preparation of Shrikhand, not only fat losses are
eliminated, but also faster moisture expulsion and less moisture retention in
the curd were achieved (Patel, 1982; Patel and Chakraborty,1985a).
Buffalo milk prepared for making Shrikhand due to higher yield and
better quality of the finished product. Buffalo milk Shrikhand is rich in
minerals including calcium, magnesium, phosphorus, copper, iron and zinc
compared to cow milk Shrikhand. Similarly, higher proportions of dissolved
proportions of calcium, magnesium, phosphorus were found in buffalo milk
Shrikhand than cow milk Shrikhand (Kadan et al., 1984; Boghra, 1998).
Shrikhand prepared using skim milk from buffaloes, cow and goats was
examined. There were no differences in the total sensory scores of Shrikhand
prepared using buffalo, cow and goat milk (Subramonian et al., 1995).
Shrikhand was made from mix cow milk: buffalo milk in ratio of 1:3, 1:1
and 3:1. The best quality Shrikhand was made from cow milk: buffalo milk
ratio 1:1, which imparted smooth texture and firm, soft body (Ghatak and
Dutta, 1998).
Shrikhand was prepared from buffalo skim milk and sweet cream
buttermilk (SCBM) with 9, 12, 15 or 18per cent TS. The percentage of TS in
SCBM affected chemical, physical and sensory qualities of Shrikhand.
Shrikhand prepared from SCBM with 15per cent TS were similar to those of
control samples prepared from buffalo skim milk and superior to those of other
SCBM Shrikhand samples (Karthikeyan et al., 1999).
2.1.3.3 Standardization of milk
Standardization of milk for fat and/or SNF is essential for production of

uniform quality of any dairy product. The same is true in this case for
Page | 13
production of a uniform quality chakka suitable for Shrikhand preparation.

Standardization of skim milk to 9per cent SNF and 0.05per cent fat was found
to be useful for making satisfactory quality of Dahi to prepared Shrikhand
(Aneja et al., 1977a; Patel, 1982). Shrikhand of superior quality was produced
from milk with 6per cent fat and 9per cent SNF and sodium citrate treatment
(Reddy et al., 1984). Chakka prepared from 4.0per cent fat milk produced
Shrikhand of the best sensory quality (De and Patel, 1989).
Standardized buffalo milk containing 4 per cent fat and 9.8 to 10.0per
cent solids· not-fat was prepared from fresh buffalo whole milk containing
about 7 to 8per cent fat and 9.8 to 10.0per cent solids-not-fat. Chakka
obtained from whole milk is much superior in both consistency and aroma
(Desai et al., 1985).
2.1.3.4 Heat treatments and homogenization to Milk
Heat treatment is essential processing step in dahi manufacture for

obtaining desirable quality of the finished product with respect to flavour as
well as body and textural properties. Heat treatment of milk for dahi making
have several advantages e.g. it reduces bacterial load , inactivates naturally
occurring enzymes and increases shelf-life if final product, also higher
temperatures are believed to result in the alteration of proteins in milk that
favours the growth and activity of starter culture (Rangappa,1947;Ray,1970;
Madan Mohan,1980; Patel, 1982). It also influences the culture activity
(Sreenivasan and Ranganathan,1972; Kohk et al., 1980; Madan Mohan, 1980;
Maurya and Saraswat, 1982; Saraswat and Agrawal, 1982).
Traditionally dahi is made from simmered or boiled milk (Madhok and

Kapoor, 1942). Heat treatment of dahi at 63ºC for 30 minutes resulted in
maximum titratable acidity of dahi when S.lactis and S.thermophilus were used
as a starter cultures, whereas, in case of the cultures S.cremoris,
S.diacetylactis and L.balgaricus; milk heated at 85ºC for 10 minutes and gave
maximum acidity (Dutta et al.,1973; Singh,1983).
Page | 14
In 1977b, Aneja et al. reported that increasing the heating temperature

(70-100°C for 16 sec) of the skim milk increase the yield of Chakka. The
optimum conditions are heating temperature of milk 90°C for 10 sec for
preparation of Chakka.
Shrikhand was prepared from cow and buffalo milks, spray-dried at an

inlet air temperature of 180-200ºC and outlet temperature of 100ºC,
reconstituting capacity and flavour of cow and buffalo milks were satisfactory
up to 45 days (Mahajan et al., 1979).
Garg and Jain (1980) found that 82.2 ºC temperatures for 26 to 20

seconds holding time was the best heat treatment of milk for dahi making. In
1987b, Rao et al. whole milk were heated to 85°C for 15 min and cool to 30°C,
for prepared Chakka used to make Shrikhand.
Shrikhand is manufacture by ultra filtration technique using ceramic

membrane module for ultra filtration of milk. Ultra filtration, approx. 50°C, 4
bars pressure yielded, 23 per cent more Chakka from milk than traditional
method probably due to the recovery of whey proteins (Sharma and Reuter,
1989).
Manufacture of fermented milk high temperature short time

pasteurization process is generally used , wherein temperatures ranges from 85
to 98ºC and corresponding time between 20 sec and 7 min were employed
(Lucey, 2004).
Homogenization is one of the important processing of parameters

employed in the manufacture of dairy products. Homogenization of milk affects
the body and texture, moisture content and yield of Chakka. Homogenization of
standardized milk was carried out at 100 kg/cm 2 pressure at 6O-65ºC, using
'Rannie' single stage homogenizer. Both, homogenized and unhomogenized lots
of milk were heated to 90'C for 5 min and then cooled to incubation
temperature of 42 to 43°C. Homogenization of milk was found to improve the
Page | 15
appearance, consistency and texture of chakka. This treatment also gives

higher yield and higher fat recovery. Use of homogenized milk is recommended
for improving organoleptic quality and yield of Shrikhand (Desai et al., 1985).
2.1.3.5 Starter culture and their incubation temperature for

preparation of dahi
The use of right type of culture is an essential pre-requisite for the

manufacture of Dahi. Dahi making is an essential pre-requisite for Shrikhand
manufacture as the quality of curd greatly influences the quality of chakka and
Shrikhand. Quality attributes are largely dependent on number of factors such
as type and composition of milk used, treatments given to milk, starter culture
used, rate of inoculation and conditions of incubation etc. (Miyani,1982; Patel,
1982; Desai, 1983).
Puntamberkar (1968) made use of Lactococcus lactis or

Lactococcuscremoris at the rate of 1-2 per cent and incubated room
temperature for preparing dahi to be used for Shrikhand making.
In1974, Ingle and Joglekar recommended for Shrikhand preparation,

buffalo milk was inoculated with 2 per cent mixed culture of Str. lactis,
Leucinostoc citrovorum and L. bulgaricus and incubated at 34ºC for 16 h to get
desirable texture and pleasing aroma in curd.
Aneja et al. (1977a) prepared curd from skim milk was inoculated with
0.25 to 0.50 per cent dahi culture, obtained from NDRI Karnal. After 8 h of
incubation the curd had acidity between 0.8 to 1.1 per cent lactic acid.
Gandhi and Jain (1977) reported the use of mixed culture containing Str.
Lactis,Str. diacetylactis, Leucinostoc citrovorumin a ratio of 1:1:1 incubated
30ºC for 12-14 h till sufficient acidity developed to give pH 4.6 to 4.8 for
preparing dahi to be used for Shrikhand making.
Page | 16
Dahi intended for Shrikhand making should have optimum firmness, and
0.8 to 1.0 per cent acidity (as LA) to ensure satisfactory rate of whey drainage.
It should also have desire aroma and flavour (Upadhyay and Dave, 1977; Patel
and Chakraborty,1988).
Shrikhand was prepared from buffalo skim milk inoculated with a

yoghurt starter cultures Streptococcus thermophilus, Lactobacillus bulgaricus
gave a pH of 4.6 after only 4 h at 42°C, thus reducing the normal curd setting
time by from 12 to 16 h to 4h. (Anon, 1982; Patel and Chakroborty,1985).
Miyani (1982) prepared curd for Shrikhand making by employing skim

milk inoculated with 1 per cent active mixed starter culture LF - 40 (NDRI,
Karnal) and incubated for 12-15 h at 29-32ºC until the curd developed the
desired acidity of 0.8 per cent.
Suryawanshi et al. (1993) reported that the use of Streptococcus

themophillus and Lactobacillus acidophilus for making dahi for Shrikhand
preparation, at the rate of different 2, 4 and 6 per cent levels with active starter
culture, it has been observed that higher levels of inoculums of curd
significantly reduced the time required for Shrikhand manufacture.
Rao et al. (1986) reported that dahi prepared from boild milk using Str.
cremoris as starter culture ( @ 2 per cent inoculums and incubated 30- 37ºC
for 16 h), produced best organoleptic quality with highest yield of chakka as
compared to other starter cultures used as (Str. lactis, Str. lactis subsp.
diacetilactis and Str. thermophilus + l. Bulgaricus).
In 1992, Prajapati reported that dahi prepared from buffalo skim milk for
making Shrikhand inoculated with active starter cultures of Str. salivarious
subsp. thrmophilus and L. delbrukii subsp. bulgaricus were added in equal
proportation at the rate of 2.5 per cent to get desired curd in about 6-7 h.
Patel (2013) reported that dahi prepared from skim milk using str.
thermophilus, L.helveticus and L.rhamnosus were added at the rate of 2 per
Page | 17
cent, while Acidophilus Bifidobacterium yoghurt culture (ABY) and yoghurt

culture (Yo-flex) were added at the rate of 0.2 g/ litre, it was incubated at 39ºC
for 12 h to get the desired acidity and firmness in dahi. The new as well as the
old nomenclature for various starter cultures, which are used in dahi making is
given table 2.3
Table-2.3: New nomenclature of lactic acid bacteria.
Old New
Streptococcus Lactococcus
S.lactis ssp. Lactis L.lactis ssp. Lactis
S.lactis ssp. Diacetylactis L.lactis biovar. Diacetylactis
S.lactis ssp. Cremoris S.lactis ssp. Cremoris
Lactobacillus Lactobacillus
L.delbrueckii L.delbrueckii ssp. Delbrueckii
L.leichmanii L.delbrueckii ssp. Leichmanii
L.bulgaricus L.delbrueckii ssp. Bulgaricus
L.lactis L.delbrueckii ssp. Lactis
L.caucasicus L.kefir
Leuconostoc Leuconostock
L.mesenteroids L.mesenteroids ssp. Mesenteroides
L.cremoris L.mesenteroids ssp. Cremoris
Streptococcus Streptococcus
S.thermophilus S. salivarius ssp. Thermophilus
(Marshall, 1987)
2.1.3.6 Removal of Whey
The removal of whey from curd is very important step in Shrikhand

making since it influence the body and texture characteristics of the finished
product.
Page | 18
Traditionally, drainage of whey from curd to obtain chakka, is done by

hanging/ tying curd in muslin cloth for about 6 – 8 h and more until the
draining has apparently ceased (Puntamberkar, 1968; Ingle and Joglekar,
1974; Gandhi and Jain,1977; Upadhyay and Dave, 1977; Kadan et al., 1984;
Reddy et al., 1984; Rao et al.,1986).
A mechanical means of whey removal from curd i.e. basket centrifuge

(revolving at 1100 to 1200 r.p.m) to obtain chakka was employed by Aneja et
al.,1977; Patel and Chakraborty, 1985 a, b, c; Patel and Abd-El- Salam,1986).
A batch type vacuum whey drainer was developed by Shah and Sharma
(unpublished work) was used for rapid expulsion of whey from curd to obtain
chakka of satisfactory quality in about 20 to 30 min at 350 to 400 millimetre
mercury vacuum (Upadhyay, 1981; Miyani, 1982; Desai, 1983).
2.1.3.7 Quality and composition of chakka
As the chakka is the base material for Shrikhand making, its chemical
composition, acidity, body and texture and flavour greatly influence the quality
attributes of the Shrikhand.
Chakka is the semi-solid curd mass obtained by the removal of whey

from dahi (curd) is commonly known as chakka that is the base material for
making Shrikhand. Chakka has milky white colour, smooth texture and mild
acidic flavour. Increasing temperature of heating of milk before coagulation led
to an increase in yield due to complete denaturation of whey protein and
precipitation with casein (Aneja et al., 1977a).
It has been experienced by Upadhyay and Dave (1977) and Desai (1983)
that standardized milk yields chakka with pleasant flavour and smooth body
but had high fat losses in whey and adversely affected recovery of fat in
chakka. In contrast, skim milk chakka was found to be rough and dry.
However when calculated amount of cream (70 to 80 per cent fat) was added to
such chakka at kneading stage, the finished product became equally
Page | 19
acceptable. They also reported that the quantity of sugar to be added depends
on the acidity of chakka.
Highest yield and best organoleptic properties were obtained with chakka
prepared from milk added with Str.cremoris. Drainage over night for 16 h
caused an increase in acidity, decrease in yield and dry texture of chakka (Rao
et al., 1986).
Use of sodium alginate if used at higher proportion (0.3 per cent)

produced inferior quality chakka. Similar observations were also made by
(Kadan et al.,1984; Desai et al.,1987).
Sachdeva et al. (1994) reported manufacture of ‘Chakka’ from milk

concentrated by reverse- osmosis (RO). Increased yield, higher solids recovery,
reduced processing time, increased through put, access to mechanisation and
alleviation of whey disposal problem are claimed as major advantages of this
process.
Shrikhand prepared using skim milk from buffaloes, cow and goat was
examined. The yield of Chakka was greatest with buffalo milk (26.2 per cent)
and lowest with goat milk (24 per cent). Suggested that the lower yield in goat
milk resulted from its smaller micelles and higher water content (Subramonian
et al., 1995).
The effect of coagulation pH (4.6, 4.8 and 5) and addition of disodium

hydrogen phosphate at 0.0, 0.05 and 0.1per cent on sensory quality of directly
acidified Chakka was studied. Coagulation pH did not have a significant effect
on sensory attributes of Chakka. However, use of disodium hydrogen
phosphate significantly improved body, texture and flavour scores. Coagulation
pH of 4.8 and disodium hydrogen phosphate at 0.1 per cent produced Chakka
with most acceptable texture and this combination was used for Shrikhand
manufacture (Biyabani et al., 1998).
Page | 20
Sharma and Verma (2005) reported to develop ‘Chakka’ by direct

acidification process. The sensory evaluation of Chakka for flavour, body &
texture and colour & appearance showed that optimum quality of product
could be obtained by using a combination of lactic acid (1:9 dilution) and
glucono delta lactone (2.5 per cent), coagulated at pH 4.8 and incubation
temperature of 35°C. The Chakka obtained from conventional and direct
acidification was found to have similar sensory characteristics.
According to FSSA (2011), Chakka means a white pale yellow semi-solid

product of good texture and uniform consistency obtained by draining off the
whey from the yoghurt obtained by lactic fermentation of cow’s milk, buffalo
milk, skimmed milk and recombined milk that has been subjected to minimum
heat treatment equivalent to that of pasteurization.
Table 2.4: FSSA standard for Chakka
Requirements Chakka Skimmed milk Full cream

Chakka Chakka
Total solids, percent by weight Min.30 Min.20 Min.28
Milk fat (on dry basis) percent by

Min.33 Max.5 Min.38
weight
Milk protein(on dry basis) percent by Min.30 Min.60 Min.30

weight
Titratable acidity (as lactic acid)

Max.2.5 Max.2.5 Max.2.5
percent by weight
Total ash (on dry basis) percent by

Max.3.5 Max.5.0 Max.3.5
weight
Page | 21
Wide variations are observed by several workers in the composition of

Chakka as shown in table 2.5.
Table 2.5 - Composition of chakka
Titrable
Moisture Protein Ash
Author Sources Fat (%) acidity
(%) (%) (%)
(%LA)
Ganguly et al. Laboratory 58 – 59 22.4 10.30 2.34 1.03
(1959)
Sharma and Laboratory 52.7 8.5 13-15.6 - 0.67-0.78
Zariwal (1978)
Sharma and Market 59.8 14.4-20.8 10.40-18.4 - 0.68-0.97
Zariwal (1978) sample
Kadanet al. Buffalo 66.63 17.94 - 1.39 -
(1984) milk
Kadanet al. Cow milk 67.95 11.88 - 1.24 -
(1984)
Patel and Buffalo 76.5 - 14.1 2.2 -
Chakraborty skim milk
(1988)
2.1.3.8 Blending and mixing of chakka with additives
Shrikhand is prepared by blending and mixing of chakka with sugar,

cream and flavouring materials such as condiments, spices fruits etc. Quantity
and quality of these blending materials, method of blending etc. determine
compositional, microbiological and body and texture characteristics of the
resultant product.
Puntambekar (1968) prepared Shrikhand by blending skim milk chakka

with unsalted butter granules and sugar.
Ingle and Joglekar (1974) obtained Shrikhand by mixing chakka with

adequate amount of sugar (36 per cent by weight of chakka), nuts, cardamom,
charoli etc. This was mixed properly by working it through the hand operator
Shrikhand machine.
Page | 22
Upadhyay and Dave (1977) suggested that best quality of Shrikhand

having homogeneous consistency can be prepared by addition of equal quantity
of sugar along with plastic cream (70-80 per cent fat) or unsalted white butter
(calculated amount) to skim milk cahkka and kneading on S.S wiremesh. The
product so obtained may be blended with desired flavour, colour, fruits and
nuts.
Miyani (1982) kneaded chakka over a cleaned and sanitized S.S

wiremesh to obtained uniformly, then it was mixed with crystalline sugar and
pasteurized cream (70 per cent fat). The admixture was then mechanically
mixed by application of 60 (to and fro) strokes with the help of S.S. ladle.
Desai (1983) prepared Shrikhand by kneading chakka over a cleaned and

sanitized S.S. wiremesh screen (30 mesh) to obtain uniformity and then it was
mixed with sugar (80 per cent by weight of chakka), allowed to stand for about
30 min to ensure proper dissolution of sugar. The mixture was then manually
stirred using a S.S. ladle and giving about 60 (to and fro) strokes.
Use of planetary mixer has been recommended for industrial method of

Shrikhand manufacture for mixing and blending of chakka and other
ingredients (Aneja et al., 1977; Patel and Chakraborty, 1985 a, b, c; Patel and
Abd-El- Salam, 1986).
Prajapati (1989) prepared Shrikhand like product from buffalo skim milk
chakka by blending with cream. The chakka-cream mixture was passed
through a colloidal mill to get fine and homogenous consistency. To this sugar
was added at the rate of 55 per cent (w/w) of chaka-cream mixture and the
contents were blended in a planetary mixture for 30 min. Ground cardamom
also added at the rate of 0.2 per cent (w/w) of Shrikhand.
2.1.4 Physico-chemical Properties of Shrikhand
Shrikhand collected from 5 shops of Anand, Gujarat, India, showed wide

variation in TS, fat, protein, carbohydrates, ash and pH values. Fat varied from
Page | 23
2.0 to 5.0per cent. Not all samples, except one under an organized dairy
undertaking, conformed to the min. limit of 8.5per cent as prescribed by
Bureau of Indian Standards. Wide variations were also observed for different
minerals in Shrikhand from various shops. Market samples also showed wide
variations in citrate, copper and iron levels (Boghra and Mathur, 1992).
In 2006, Salunke et al. investigated physicochemical properties of

Shrikhand made and sold by 5 major manufactures in 4 cities in Maharashtra,
India. TS, fat, protein, reducing sugar (lactose), non reducing sugar (sucrose)
and ash contents were 57 -67.8, 3-8.9, 4.6-6.7, 1.5-6.5, 44.5-51 and 0.3-045
per cent respectively. Acidity, pH and contents of soluble nitrogen and free fatty
acids were 1.04-1.54 per cent lactic acid, 3.9-4.3, 0.1-0.29 per cent and 0.68-
0.78 per cent oleic acid, respectively. Comparision of Shrikhand made in the
4cities indicated variation in composition between the FFA and titratable
acidity. The majority of Shrikhand samples did not meet the protein or sucrose
in DM requirements of the Indian Prevention of Food Adulteration Act and
Bureau of Indian Standards.
Fortification of sour whey concentrate in Chakka for preparation of

Shrikhand was studied. Addition of 5per cent sour whey concentrate to Chakka
(both from cows’ milk) increased the yield of Shrikhand by 5per cent over that
obtained with the traditional method; addition of whey concentrate at this level
had no detrimental effect on physicochemical and sensory properties of
Shrikhand. Addition of whey concentrate at level >5per cent caused
deterioration in quality of Shrikhand in terms of flavour, consistency and
texture (Giram et al., 2001).
Changes in physicochemical properties and mineral composition of

buffalo and cow’s milk during preparation of Shrikhand were studied.
Progressive fermentation of milk in to dahi brought about little or no changes
in total solids, fat, proteins and milk salts, but significant decrease in lactose
content and pH value. Mineral composition showed little change, with the
Page | 24
exception of citrate which disappeared completely the dahi stage in both milk.
Although solubilised minerals drained along with whey, considerable amounts
of all minerals remained aggregated in to chakka from both milk types. The
fermentation of milk brought about highly significant changes in mineral
content during transition from the colloidal to the soluble phase; this
transition occurred more rapidly and to greater extent with buffalo milk
(Boghra and Mathur, 2000).
2.1.5 Sensory Quality of Shrikhand
In 1985c, Patel and Chakraborty studied the factors affecting sensory

properties of Shrikhand. Moisture, fat and sugar content of Shrikhand made
from buffalo milk affected its organoleptic properties, the most desirable
product having 6 per cent fat, 41 per cent sugar and 40 per cent moisture.
In 1998, Nalawade et al. studied the effects of compositional variables on

sensory quality and consistency of Shrikhand in order to optimize
compositional parameters for maximum acceptability. The most desirable
combination of fat and sugar levels in Shrikhand that can give maximum
acceptability, were found to be 8 per cent fat and 36 per cent sugar.
Effect of using sugar substitutes on the sensory properties of Shrikhand

was investigated. Combinations of different amount of raftilose (3, 4 and 5 per
cent) and sugar (8, 10, 12.5 and 25 per cent) were used in Shrikhand made
with separated buffalo milk and yoghurt culture. Samples were analysed for
sensory properties using a 9-point Hedonic scale to evaluate its storage ability
at 7 ± 1°C. Among the various combination tried, Shrikhand prepared with
raftilose (4 per cent) and sugar (12.5 per cent) was rated as most acceptable by
sensory panels (Singh and Jha, 2005).
Shrikhand was prepared from dahi with a constant level of sugar (40 per
cent) and a supplementing with banana pulp at 10 per cent (T1), 20 per cent
Page | 25
(T2) and 30 per cent (T3). T2 (20 per cent) supplementation of banana pulp to
Shrikhand was much preferred (Narayan and Lingam, 2013).
2.1.6 Microbiological Quality of Shrikhand
In 1975b, Upadhyay et al. examined 45 samples of market Shrikhand

from 15 different manufacturers in 4 different cities in India for bacteriological
quality. Viable counts are varied greatly between samples (from 25,000 to
1,65,000). 32 samples contained coli forms, 15 of which had >100/g and
yeasts and moulds were present in high numbers (≤ 1,83,000/g) suggesting
unhygienic practices. However, psychrotrops were low in number (0-10,000/g)
and enteric pathogens were absent.
Shrikhand was prepared using mixed cultures of Streptococcus lactis C-

10 and Streptococcus lactis subsp. diacetylactis DRC-1 and microbiological and
organoleptic changes during storage at 7±2°C were noted for 50 days. Total
bacterial count and counts of psychrotrops, acid-producing bacteria,
lactobacillus spp. and proteolytic and lipolytic bacteria and yeasts and moulds
increased steadily; the coli form count was negative in all cases. (Upadhyay et
al., 1984).
Shrikhand was prepared from cows’ milk inoculated, after steaming, with
both Staphylococcus aureus and 1 of 2 Lactobacillus spp. at 105 cfu/ml. S.
aureus strains K-283, K-192 and E-326 grew well and produced thermo stable
deoxyribonuclease (TDNase) in the milk in the presence of L. acidophilus;
counts of both organisms reached 106 cfu/ml in curd and Chakka, and then
declined 10-20 times in Shrikhand. The three strains of S. aureus grew only
poorly in the presence of L. bulgaricus and produced low levels of TDNase. S.
aureus A-100 also showed very little growth and TDNase production in the
presence of L. acidophilus and was completely inhibited by L. bulgaricus.
(Varadaraj and Ranganathan, 1988a).
Page | 26
The ability of Staphylococcus aureus strains to grow and produce

thermostable deoxyribonuclease (TDNase) in Shrikhand when used in
combination with Streptococcus lactis, Str. Cremoris, Str. thermophillus and Str.
Lactis subsp. diacetylactis was evaluated. Cultures of S. aureus, when
inoculated in milk at 1 X 105 cfu/ml produced detectable levels of TDNase.
Growth and production of TDNase by S. aureus was less in the presence of Str.
Lactis (Varadaraj and Ranganathan, 1988b).
Market samples of Shrikhand were collected from twenty popular

manufacturers, five from each of the four cities of Maharashtra and analysed
for microbiological quality. Microbiological differences among manufacturers of
different cities were too low to be significant. Almost no samples met the
requirements for YMC as laid down under BIS standards (Salunke et al., 2005).
2.1.7 Rheological Quality of Shrikhand
Consistency was measured at 30°C for freshly prepared samples and at

10°C for refrigerated samples (kept at 5°C/12h) using a cone penetrometer
under the effect of various levels of moisture (30-45 per cent), sugar (70-90 per
cent) and fat (4-8 per cent); values are reported in mm of penetration. At the
minimum levels of parameters studied, consistency was maximum in both
samples, and vice versa. Increasing levels resulted in higher penetration,
causing reduction in firmness. Moisture showed that the most pronounced
effect, followed by sugar and fat. An increase in these contents and in
temperature caused a reduction of curd tension and coming-up time and
increase in penetration values (Miyani et al., 1984; Patel and Chakraborty,
1985c).
Shrikhand exhibits a combination of several rheological properties such

as weak gel-like visco-elasticity, an apparent yield stress, thixotropy and long
structural recovery time scales. In an attempt to trace the origins of these
rheological properties in Shrikhand, the microstructure was characterized and
showed that there exist two different microstructures whose studies show that
Page | 27
while the temperature sensitivity of the visco-elastic parameters is dominated

by the semi-crystalline milk fat microstructure, the shear sensitivity is largely
dictated by the protein network (Kulkarni et al., 2006).
2.1.8 Packaging of Shrikhand
Shrikhand, being a semi-solid product, is packed in heat sealable

polystyrene containers of various sizes ranging from 100 g to 1.0 kg (Patel,
1998) . The use of pre-formed polystyrene cups for packing Shrikhand limited
the packaging capacity to 1000 cups/hr. By employing a form- fill- seal (FFS)
machine up to 6000 cups may be filled / packed hour (Aneja et al.,
2002).
2.1.9 Shelf-life of Shrikhand
Shrikhand was stored at (i) 7±2°C and (ii) -7±2°C for 50 days, and
chemical and organoleptic quality was assessed at 10-day intervals. Acidity
increased during storage at both temperatures, more so at 7±2°C. The study of
proteolytic changes increased on storage at both temperatures. Free fatty acid
levels increased faster at 7±2°C, while non-reducing sugars decreased. The
stored product became unacceptable after 40 and 50 days of storage at 7±2°C
and -7±2°C respectively, as judged by sensory scores. (Upadhyay et al., 1985).
The keeping quality of Shrikhand depends largely on its initial micro flora
like yeast, mould and other foreign organisms. Owing to both high acid and
sugar levels, Shrikhand has a long shelf life of 35 - 40 days at 8º C and 2 - 3
days at 30ºC. Deterioration of the product is usually associated with an
increased activity of yeast and mold, and proteolytic micro flora under
refrigeration conditions of storage. Addition of 0.5 per cent potassium
sorbate retards the growth of yeasts and molds (Patel, 1998). Prajapati,
et.al, (1992) reported that post-production heat treatment at 70ºC for 5 min
yielded Shrikhand with a superior overall quality and a shelf-life of 15 days
at 35-37ºC. However, to get best shelf life in the industrial process,
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thermization of Shrikhand at 65ºC / 10 min and its subsequent freezing is

most effective. The frozen product can be kept in good condition from six
months to twelve months at -18ºC (Aneja et al., 2002).
In 1980, Sharma and Zariwala, found that samples stored at 10°C kept
well for about 40 days, whereas those stored at 37°C became unacceptable
within 7 days due to development of off-flavours as milk fat deteriorated. No
marked change in fat content occurred during storage at 10°C. Acidity, volatile
acidity, soluble protein and reducing sugar increased at both storage
temperatures, sucrose content decreased, all changes being more marked at
37°C than at 10°C. Counts of coli forms and acid – producing organisms
tended to decreased, total and staphylococcal counts fluctuated and proteolytic
count increased during storage. The product generally became unfit for
consumption due to surface mould growth and developed rancidity.
In 1997, Agnihotri and Pal, examined changes in goat milk Shrikhand

stored at ambient (18.53±0.56°C) and refrigeration temperature (4±1°C). The
freshly prepared Shrikhand with had average log10 6.55 SPC, 3.40
psychrotrops, 5.14 coli forms, 4.88 lactobacilli and 4.93 yeast and moulds/g
and shelf-life of 3 days. Storage under refrigeration (4±1°C), except for
psychrotrops, retarded the growth of other microbes, as well as changes in
moisture, pH, T.A. and organoleptic traits, thereby extending product shelf-life
about 30 days.
Effect of addition of different levels of strawberry-pulp and sugar on

chemical composition of Shrikhand during storage was studied. The moisture
content in all the experimental samples was decreased during storage period.
The acidity of Shrikhand increased with pulp level, when samples stored at
room temperature. The average soluble ‘N’ fresh product was increased on 11 th
day of storage under refrigerated temperature (5±2°C) (Sonawane et al., 2007).
In 2011, Raghuwanshi et al., studied Shrikhand samples collected from

the markets of Akola and Amravati and prepared in the laboratory were stored
Page | 29
at room temperature (30±2°C) and at refrigerator temperature (5±1°C). The

samples stored at room temperature were analyzed daily until they were
spoiled i.e. for five days, while the refrigeration samples were analyzed at an
interval of seven days until they were spoiled i.e. thirty-five days, for the
determination of content of reducing and non-reducing sugar. Fresh Shrikhand
was having 2.96 per cent reducing sugar, which increased to 3.39 per cent
during five days of storage and finally reached to 4.70 per cent in 35 days of
storage. There was decrease in non-reducing sugar from 48.41 to 48.05 in 1st 5
days and thereafter to 40.80 in 35 days.
2.2 Technological innovations in commercial production
2.2.1 Mechanization in manufacture of Shrikhand
With a view to overcome the limitations of traditional method, successful

attempts have been made to develop an industrial process for the manufacture
of Shrikhand. Standardized the process for manufacturing Shrikhand on
industrial scale (100 kg/batch). It consists of preparing curd from skim milk,
centrifugation of curd at 110 rpm for 40 min in a basket centrifuge to separate
whey and Chakka (concentrated curd) and adding cream, sugar and flavouring
to Chakka (Aneja et al., 1977; Aneja et al., 1977b).
A fully mechanizes/continuous process has also been developed for

industrial production of Shrikhand. In this process, Chakka is prepared by
separating the whey from skim milk dahi employing 28” dia. basket centrifuge
at 1100 rpm. The resultant Chakka, sugar and plastic cream are then mixed in
a planetary mixer (Aneja and Vyas, 1983).
Patel and Abd-EL-Salman (1986) described the industrial method for

production of Shrikhand. It consists essentially of incubating heat treated (85°C
for 30 min) skim milk with a mixed culture of Streptococcus lactis + S. lactis
subsp. diacetylactis to form dahi from which the whey is separated by
centrifugation; the resulting ‘Chakka’ is then mixed with sugar, cream and in
Page | 30
some cases cardamom at 30°C to form Shrikhand, which is packaged in

polystyrene containers or paper board boxes. Keeping quality of the products is
35-40 days at 8°C and 2-3 days at 30°C and can be improved by heating the
product at 70°C for 2 min or by addition of 0.5per cent potassium sorbate.
Composition of Shrikhand prepared by this method was 57-60 per cent TS, 5-6
per cent fat, 6.5-7 per cent protein and 40-43 per cent sucrose with titratable
acidity of 1.05- 1.10 per cent and pH 4.4-4.6.
Though many dairy plants in Maharashtra and Gujarat states are

now manufacturing Shrikhand on commercial scale, the first modern plant
was established at the Baroda District Co-operative Milk Producers’ Union Ltd.
(Sugam unit in Gujarat) in 1980. Since then it has been producing and
marketing Shrikhand under the brand names of “Sugam” and “Amul”
(Aneja et.al.,2002).
2.2.2 Manufacture of Shrikhand using membrane Technology
Ultra filtration (UF) is a pressure-driven, cross-flow membrane system

that can simultaneously purify, concentrate, and fractionate organic molecules
of a feed stream (Pal, 2003). The applicability of ultra filtration increased
considerably in dairy industry and probably account for the largest share
of installed membrane capacity worldwide. In the traditional and industrial
methods developed for manufacturing Shrikhand there is consider able loss of
whey proteins through the expulsion of whey from chakka. These whey
proteins with a high biological value could be recovered in chakka by the
application of ultra filtration (UF) to make the so called UF-chakka. The UF
chakka could be handled in the same manner as traditional chakka to make
the final product, i.e., Shrikhand.
Shrikhand is manufacture by ultra filtration technique using ceramic

membrane module for ultra filtration of milk. Ultra filtration, at pH 4.5-4.6,
approx. 50°C, 4 bars pressure and velocity of 5 m/s yielded, 23 per cent more
Chakka from milk than traditional method (15.5 kg (26.3 per cent TS) vs. 11.91
Page | 31
kg (26 per cent TS) from 100 kg skim milk curd), probably due to the recovery
of whey proteins. It is concluded that Shrikhand of very good quality may be
manufacture by ultra filtration of Chakka (Sharma and Reuter, 1989).
Sharma and Reuter (1992) made a successful attempt to develop

UF-chakka for subsequent use in Shrikhand preparation using ceramic
membrane module. The UF setup was designed with a centrifugal pump of
5,000 liters /hr capacity to obtain a velocity of 5 m/sec in the module. The
temperature and trans membrane pressure employed during the UF process
were 50±2º C and 0.5 (p i + p0) = 4 bars, where p and p are inlet and
outlet pressures of the milk respectively. According to them, by UF process
recover all the whey proteins and increase the yield of the final product while
automating the process. The coagulum obtained was agitated slowly and
subjected to ultra filtration. Whey was removed in the form of permeate. The
Chakka thus obtained was mixed with 70 per cent fat cream and sugar to
manufacture Shrikhand that contained 6 per cent fat, 41 per cent sugar and 40
per cent moisture. The mixture was then kneaded in a planetary mixture at 25-
26°C in order to get smooth paste-like semi-solid consistency with no feeling of
sugar grains. It was reported that there was practically no difference between
traditional and UF- Shrikhand.
Sachdeva et al. (1994) attempted the manufacture of Chakka by reverse

osmosis which involved heat treatment (90°C for 5 min.) to RO concentrate,
cooling to 22°C, inoculation with 20 per cent mixed lactic culture, incubation
for 18 hrs and then removal of whey by filtration to get Chakka. Increased
yield, higher solid recovery, reduced processing time, access to mechanization
and alleviation of whey disposal problem were claimed as major advantages of
the process.
Ansari et al. (2006) also developed the technology for the

manufacture of Shrikhand using UF pre-concentrated skim milk It involved
pasteurization of skim milk (9 per cent TS) at 92ºC for 5 min followed by
Page | 32
cooling to 30ºC. The suspended solids were removed by cloth filtration and
the milk subjected to concentration using UF membrane. The UF-
concentrated skim milk with 11 to 12 per cent TS was used to make
chakka. This chakka after removal of whey had 18per cent TS. It was reported
that this UF-concentrated skim milk chakka when subsequently used for
making Shrikhand resembled conventional Shrikhand. However, the Milk
concentrate with 19 to 20 per cent TS, when directly was used for
Shrikhand making without whey drainage had a curdling time of 9 to 10 hr for
reaching 2 per cent acidity. Shrikhand made from this chakka was reported
to be poor in quality and showed formation of hard particles during
pasteurization. Hence, these recommended further studies to improve the
quality of Shrikhand using UF-concentrated skim milk.
In standardized UF process, skim milk coagulum obtained by yoghurt

culture was heated to 60°C for 5 min with continuous agitation and ultra
filtered to about 16.6 per cent TS concentration. Whey was then removed from
this concentrated coagulum by hanging it in a muslin cloth (eight layered)
room temperature followed by mild pressing to get Chakka. Chakka was then
kneaded with cream (70 per cent fat) and sugar to prepare Shrikhand of
smooth consistency. Higher amount of sugar was added in traditional/ UF
skim milk Chakka to meet PFA standards of total solids in Shrikhand. UF
process resulted in nil fat loss in whey and 20.70 per cent extra recovery of
total solids in Chakka. The protein content in skim milk Chakka through UF
process and in Shrikhand prepared from it was higher than in traditional
process (Shukla et al., 2007).
2.2.3 Manufacture of Direct Acidified Shrikhand
The traditional technology of Shrikhand making is cumbersome and time

consuming as about 12 to 15 hr are required for complete coagulation of milk
and another 6 to 8 hr for expulsion of whey. Use of direct acidification in
manufacture of Shrikhand would reduce the processing time considerably, save
Page | 33
the cost due to culture maintenance and propagation, and eliminate all the
problems related to starter culture.
Patel and Chakraborty (1985) have made the attempt manufacture

Shrikhand by direct acidification and found undesirable results as the
Shrikhand made by direct acidification had grainy texture. However, Biyabani
et.al., (1998) had attempted to develop directly acidified Shrikhand by
suitably modifying the earlier reported procedure to overcome the problem
of hard grainy texture. It involved standardization of buffalo milk to 6 per cent
fat and bringing to boil followed by addition of disodium hydrogen phosphate at
a level of 0.1 per cent. Then cooled to room temperature followed by chilling to
5ºC. Using 50 per cent (v/v) lactic acid solution, the pH of the chilled milk was
adjusted to 4.8. The acidified milk was then allowed to set in an incubator at
37ºC for 3 hr. Then the chakka was obtained from the well-set curd in the
similar manner to that of conventional chakka. The directly acidified chakka
was then added with 40 per cent ground sugar and 2 per cent cardamom
powder and mixed thoroughly. The mixture was then passed through
muslin cloth to obtain Shrikhand. However, it was reported that the overall
acceptability of directly acidified Shrikhand was significantly less than that of
conventional Shrikhand.
2.3Value addition in Shrikhand
With the growing interest in the diversification of food products to

attract wide consumers, in the recent past we have noticed that many
attempts have been made to incorporate different additives into Shrikhand.
Fruit pulps like apple, mango, papaya, banana, guava and sapota (Bardale et.
al, 1986; Dadarwal et al., 2005) and cocoa powder with and without papaya
pulp (Vagdalkar et al., 2002) have been tried in Shrikhand. However, in case of
post-fermentation addition of pulps, from the food safety point of view, it is
necessary that the fruit pulp intended for addition shall be subjected to
heat treatment equivalent to pasteurization.
Page | 34
Deshpande et at. (2008) studied consumer acceptability of Shrikhand

prepared from various combinations of soymilk and cow’s milk was compared.
The consumer acceptability decreased on increasing soymilk contents in
Shrikhand. The maximum acceptable soymilk content was 60 per cent. TS
content were >58 percentage in all products.
Shrikhand was prepared using papaya pulp incorporated in Chakka to

increase the nutritional quality and overall acceptability. Papaya was added at
20, 40 and 60 per cent levels. The fat, protein, lactose, sucrose, ash and total
solid contents significantly decreased with increase in the level of papaya pulp.
The Shrikhand prepared with 20 per cent level of papaya pulp was found most
acceptable (Nigam et al., 2009).
Shrikhand was prepared from cow milk Chakka with constant level of
sugar (40 per cent by wt. of Chakka) blended with varying levels of custard
apple by wt. of Chakka. The compositional and sensory attributes of Shrikhand
were significantly affected due to addition of custard apple pulp at different
levels. Increased level of custard pulp showed slight decrease in cost of
production of Shrikhand. A novel type of Shrikhand having acceptable quality
can be prepared by addition of 10 per cent custard apple pulp to it and its cost
of production can be reduced (Gavane et al., 2010).
In 2011, Landge et al. assessed the suitability of adding ashwagandha

powder in to Shrikhand. Ashwagandha powder @ 0.3, 0.5, and 0.7 per cent
with 40 per cent cane sugar (by wt. of Chakka) was mixed for manufacture of
Shrikhand. Shrikhand prepared by addition 0.5 per cent ashwagandha powder
(T2) was superior in organoleptic parameter.
Kumar et al. (2011) prepared Shrikhand using various levels of apple

pulp viz. 0, 10, 20 and 30 per cent. The protein, fat and ash percent of the
Shrikhand decreased significantly (p<0.05) where as moisture
content increased significantly (p<0.05) with increasing level of apple pulp.
Page | 35
Based on various sensory parameters, Shrikhand containing 20 per cent apple

pulp was selected as optimum.
Shrikhand was prepared by adding different volume (in gm) of gulkand

and rose petal powder in combinations viz. 1:1.5 (GRS 1), 1.5:1.5 (GRS 2),
2:1.5 (GRS 3) and 2.5:1.5 (GRS 4) in 100 gm Chakka. Based on various
sensory parameters, Shrikhand containing 2:1.5 gm of gulkand and dried rose
petal powder was most liked by panel members (Nadaf et al.,2012).
Soy fortified Shrikhand samples were prepared with various levels of

soymilk, cane sugar and yoghurt culture (Lactobacillus debrueckii subsp.
Bulgaricus and Streptococcus thermophlus in the ratio of 1:1) examined for
selective enumeration of health beneficial microorganisms. The formulated soy
fortified Shrikhand with 45 per cent soy milk, 30 per cent cane sugar and 3per
cent yoghurt culture was found highly suitable (P< 0.05) for obtaining the
maximum viable count of Lactobacillus delbrueckiisubsp. bulgaricus and
Streptococcus thermophilus where as the desirability index was 0.985 (Singh et
al., 2014).
2.4 Probiotics and their Potential Use
Probiotics are defined as single or mixed cultures of live microorganisms

that when applied to animals or humans in adequate amounts confer a health
benefit on the host by improving the properties of the indigenous micro flora.
Examples of probiotic microorganisms used in foods include Lactococcus lactis,
Lactobacillus sp., Streptococcus, Enterococcus, Bifidobacterium sp., Pediococcus,
Propionibacteria sp. (Tabbers and Benninga, 2007).
In addition, fermented functional foods with health benefits like Evolus®

and Calpis®, have been introduced in the market, which are based on bioactive
peptides released by probiotic organisms (Shah, 2007).
According to Crittenden et al. (2005) and Anonymous (2010) The main

targets of probiotic intervention are as following.
Page | 36
i. Increasing natural resistance to infectious disease in the

gastrointestinal tract and a first line of defense against disease;
ii. Preventing of dangerous fungal overgrowth and some allergic

reactions; Reducing putrefactive/toxic microbial metabolism in the gut;
iii. Promoting optimized digestive processes, allowing maximum

nutritional benefit from food;
iv. Improving resistance to toxic bowel problems and diarrhoea;
v. Stimulating of the immune system;
vi. Producting of needed nutrients, like vitamin K, one form of which

the body itself cannot make;
vii. Improving lactose intolerance conditions;
viii. Reducing of cholesterol levels;
ix. Acting as antioxidants;
x. Controlling diseases where components of the intestinal micro

biota have been implicated in aetiology.
2.4.1 Probiotics in Dairy Products
The probiotic often included in the probiotic group members of the L.

Acidophilus complex, L.Casei complex and Bifidobacterium spp. Certain dairy
products as sour cream, butter milk, yoghurt, powdered milk and frozen
desserts contains bifidus and acidophilus bacteria (Hilliam, 2000).
2.4.1.1 Probiotics Shrikhand
In order to harness the beneficial aspects of fermented foods with health

promoting culture organisms, prepared Shrikhand using three different
cultures: (1) Lactobacillus acidophilus, (2) yoghurt culture Lactobacillus
Page | 37
delbrueckii ssp. bulgaricus and Streptococcus salivarius ssp. thermophilus and

(3) Lactobacillus acidiphilus and Bifidobacterium bifidus. Shrikhand made with
yoghurt culture had a low pH and acidity with desirable flavour, body and
texture scores indicating higher acceptance by the consumer over the others
(Geetha et al., 2003). Optimized low calorie Shrikhand powder was prepared
using probiotic culture (S thermophilus, L bulgaricus, B. bifidum; 1:1:1) (Jha
and Rakhi, 2009).
Swapna and Suvarna, (2012) prepared of Shrikhand by the addition of

probiotics as well as lactic acid bacteria. They reported that the highest score
was recorded with combinations of probiotics i.e. Lctobacillus acidophilus +
Lctobacillus sporogens and the lowest score was recorded in Lctobacillus
rhamnosus in terms of colour, appearance, aroma, texture and overall
acceptability of the product and lactic acid bacteria isolates using cattle milk
isolates recorded highest score followed by dairy milk and buffalo milk.
Different combination of probiotic starter culture used for Shrikhand viz.,

Streptococcus thermophillus (NPC), Acidophilus Bifidobacterium yoghurt culture
(PC1), Yo-flex + Lactobacillus helveticus (PC2), Yo-flex + Lactobacillus rhamnosus
(PC3), Streptococcus thermophilus + Lactobacillus helveticus (PC4) and
Streptococcus thermophilus + Lactobacillus rhamnosus (PC5). Sample PC3 had
best quality of Shrikhand. (Patel, 2013).
2.4.1.2 Probiotics in Yoghurt
Viability of yoghurt and probiotic bacteria was assessed during

manufacture of yoghurt using probiotic cultures. The viability of L.
acidophilus was affected by the presence of L. delbrueckii ssp. bulgaricus,
whereas bifidobacteria exhibited better stability in the yoghurt prepared from
cultures that contained L. delbrueckii ssp. bulgaricus. The viability of both
probiotic organisms was improved when the dissolved oxygen concentration
was low in the product. The storage temperature of yoghurt affected the
viability of bifidobacteria, but not L. acidophilus. The variations in titratable
Page | 38
acidity, pH, dissolved oxygen and hydrogen peroxide were almost identical at
storage temperatures of 4 and 10 °C. In one of the commercial cultures, three
log cycles inhibition of bifidobacteria was observed from initial counts of
>106 mL−1 (Dave and Shah, 1997).
Donkor et al. (2007) had assessed the in vitro angiotensin-converting

enzyme (ACE)-inhibitory (ACE-I) activity of peptide fractions from different
yoghurt. Yoghurts were prepared either using a sole yoghurt culture
including Lactobacillus delbrueckii ssp. bulgaricus Lb1466 and Streptococcus
thermophilus St1342, or L. acidophilus L10, L. casei L26 and Bifidobacterium
lactis B94 in addition to yoghurt culture. Peptide fractions showing high ACE-I
activity. All probiotic yoghurts showed appreciable ACE-I activity during initial
stages of storage compared with the control yoghurt, with a significant (p<0.05)
decrease afterwards. The ACE-I activity ranged from IC50 of 103.30–
27.79 μg mL−1 with the greatest ACE inhibition achieved during first and third
week of storage.
Mango soy fortified probiotic yoghurt (MSFPY) was prepared by using

blends of 78.3per cent toned milk, 14.5 per cent soymilk and 7.2 per cent
mango pulp. Effect of yoghurt cultures Streptococcusthermophilus (ST),
Lactobacillus bulgaricus (LB) and probiotic culture Bifidobacterium bifidus (BB),
Lactobacillusacidophilus (LA) on physicochemical properties and sensory
properties were studied. The optimized concentration of cultures was found out
to be 1.75 per cent, 1.95 per cent, 2.44 per cent and 1.37 per cent for ST, LB,
BB and LA respectively that yielded an acceptable quality of MSFPY having
acidity 0.73 per cent, total solids 14.02 per cent, and syneresis 14.12 per cent
and scored 8.5 on hedonic rating (Shilpi and Kumar, 2013).
2.1.1.3 Probiotics in Ice cream
Probiotic ice cream was manufactured by mixing fortified milk fermented

with probiotic strains with an ice - cream mix, followed by freezing. Moussa et
al. (2005) they have noticed higher acidity and viscosity in the case of mixes
Page | 39
containing probiotic bacteria. There was no effect on specific gravity as well as

weight per gallon of bio-ice cream mixes. Freezing point of mixes decreased in
all bio-ice cream compared to control. All treatments showed slightly faster
melting than control. Initial freezing of ice cream mix followed by hardening
caused a reduction of less than one log cycle in viable counts of probiotics. The
viable counts decreased by 2.23, 1.68, 1.54, 1.23 and 1.77 log cfu/g for
Lactobacillus acidophilus, Bifidobacterium bifidum, Lactobacillus reuteri,
Lactobacillus gasseri and Lactobacillus rhamnosus, respectively, during 12
weeks of frozen storage. Although there was decrease in the number of viable
cells, the ice cream may be considered as a probiotic food during 12 weeks of
storage, since they remained was above the recommended minimum limit of
106 cfu/g.all the ice cream samples received a high score in the organoleptic
evaluation. Lactobacillus reuteri containing ice cream was judged sourer and
attained higher score for probitic flavour.
Probotic ice-cream mix was made by adding 25 and 50 per cent of

commercial cultured milks cultured with Lactobacillus acidophilus and
Bifidobacterium bifidum to control ice-cream mix (Christiansen et al., 1996).
The number of viable Lactobacillus acidophilus and Bifidobacterium bifidum was
0.5-1.0×107 cfu/ml. The control 25 per cent cultured milk added ice cream
samples were comparable in terms of overall melting properties; however, the
25 per cent addition level samples had the best melting resistance. Probiotic ice
cream showed decreased overrun and had mild sour, fresh and pleasant
flavour. Younis et al. (1998) prepared ice cream containing viable Lactobacillus
acidophilus and Bifidobacterium bifidum by direct acidification or by culturing
with starter.
2.4.1.4 Probiotics in Cheese
Hard cheese, such as cheddar, may offer certain advantages over

yoghurt-type products in terms of delivery of viable probiotics, such as the
reduced acidity of the cheese compared with yoghurt environments and the
Page | 40
high fat content and texture of cheddar cheese, which may offer protection to
the microorganism during passage through the GIT (Stanton et al., 1998). They
reported that Bifidobacterium spp.survived well in Cheddar and Gouda cheese.
Studies have involved the incorporation of a number of probiotic lactobacillus
strains into cheddar cheese and assessment of their performance during
ripening. These strains had previously been isolated from human upper GIT
during surgery. Two L.paracasei strains grew and sustained high viability in
cheese while L. salivarius strains did not survive during ripening.
Consequently, probiotic cheddar cheeses can be manufactured containing high
levels of L.paracasei strains (10 8cfu/g) at a relatively low cost by conventional
manufacturing procedures. Further they have studied that incorporation of
these strain did not influence negatively on cheese quality, including aroma,
flavour and texture. The cheese also compared very favourable with yoghurt
regarding delivery of viable cells to the GIT, despite the apparent age difference
of the products.
Cheddar cheese was manufactured with either Lactobacillus

salivarius NFBC 310, NFBC 321, or NFBC 348 or L. paracasei NFBC 338 or
NFBC 364 as the dairy starter adjunct. The results demonstrate that the
probiotic L. paracasei species used in this study are particularly suitable for
Cheddar cheese applications. They grow to numbers in excess of 10 8 CFU/g
and remain at this level even after 8 months of ripening, while their presence
has negligible effects on cheese composition, flavour, and aroma (Stantonet al.,
1998).
2.5Dietary Fibers: Types, Availabilities and Requirements
Eben Hipsley, an Australian scientist, in 1951 to described lignin,

cellulose, and hemicelluloses in food and first used the term dietary fiber
(Hipsley,1953). Dietary fiber is neither a single nor a well-defined chemical
entity, but rather corresponds to a complex mixture of compounds present in
plants, which strongly vary with regard to chemical structure as well as physic-
Page | 41
chemical properties (AACC, 2001; Rodriguez et al., 2006). Dietary fiber have
technical functionality relating to texture, as well as nutritional functionality
relating to health (Ramirez et al., 2010; Ajila and Prasada Rao, 2013).
In fact, dietary fiber is more a concept than a chemically entity. The

widely accepted reference is that of Trowel et al. (1976), who defined dietary
fiber as “remnants of plant cells, resistant to hydrolysis by alimentary enzymes
of man and plant polysaccharides plus lignin; that resist hydrolysis by human
digestive enzymes”.
The American Association of Cereal Chemist (AACC) and Food Safety and
Standards Authority of India (FSSA, 2009) have proposed new definitions,
which underline “resistant to digestion” as being the key characteristic of
dietary fiber (AACC, 2001). These include:
1. “Dietary fiber is an edible part of plants or analogous

carbohydrates that are resistance to digestion and absorption in the human
small intestine with complete or partial fermentation in the large intestine”
(AACC, 2001).
2. Dietary Fiber means carbohydrate polymers with a degree of
polymerization (DP) not lower than 3 which are not hydrolyzed by the
endogenous enzymes in the small intestine of humans (FSSA, 2010).
The different components of dietary fiber have miscellaneous

physiological effects also. It is therefore, not surprising that no definition is yet
universally accepted.
2.5.1 Dietary fiber components
The most widely accepted classification for dietary fibre has been to
differentiate dietary components on their solubility in a buffer at a defined pH,
and/or their ferment ability in an invitro system using an aqueous enzyme
solution representative of human alimentary enzymes. Thus, most
appropriately dietary fibre is classified into two categories (1) water-
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insoluble/less fermented fibres: cellulose, hemicellulose, lignin and (2) water-

soluble/well fermented fibres: pectin, gums and mucilages (Anita and
Abraham, 1997).
Dietary fiber are metabolized and completely fermented by the colonic

micro flora and are oxidized to produce gases and short chain fatty acids
(SCFAs) mainly butyric, propionic, acetic and succinic acids (Roberfroid,
2005a).Dietary fiber exhibits one or more of laxation (fecal bulking and
softening: increased frequency and/or regularity), blood cholesterol
attenuation, and blood glucose attenuation, preventing bowel inflammatory
diseases (IBD) and colorectal cancer (CRC) (Nugent, 2005). Dietary fiber acts as
protective agent against cardiovascular diseases, diverticulosis, constipation,
irritable colon, colon cancer and diabetes. The insoluble fraction of fiber seems
to be related to the intestinal regulation, whereas the soluble fiber is associated
to the decrease of cholesterol levels and absorption of intestinal glucose
(Rodriguez et al., 2006).
2.5.2 Mechanism of dietary fiber
Dietary fibers have three primary mechanisms: bulking, viscosity and

fermentation. The main action of dietary fiber is to change the nature of the
contents of the gastrointestinal tract, and to change how other nutrients and
chemicals are absorbed. Soluble fiber binds to bile acids in the small intestine,
making them less likely to enter the body; this in turn lowers cholesterol levels
in the blood. Soluble fibers also attenuates the absorption of sugar, reduces
sugar response after eating, and normalizes blood lipid levels and, once
fermented in colon, produce short chain fatty acids as by products with wide
raging physiological activities (FSSA, 2010; EU, 2008).
Authorities generally recommend that about 30 grams of fiber be

consumed daily (FSSA, 2010). World Health Organization has recommended
that total fiber intake be 25 g/day (WHO/FAO, 2003). National Institute of
Page | 43
Nutrition (NIN), Hyderabad recommends an intake of 25-40g total dietary fiber

per day or 12-14g total dietary fiber per 1000 kcal per day.
2.5.3 Inulin, Banana and Oat as Different Dietary Fiber
Inulin, a non digestible carbohydrate, is a fructan that is not only found

in many plants as a storage carbohydrate, but has also been part of man's
daily diet for several centuries. Inulins are a group of naturally
occurring polysaccharides produced by many types of plants, industrially most
often extracted from chicory. The inulins belong to a class of dietary
fibers known as fructans (Roberfroid, 2005b). Inulin enhances the growth and
activities of selected beneficial bacteria or inhibits growth or activities of certain
pathogenic bacteria, hence promoting colonic health (Saad et al., 2013). Inulin
have much lower solubility, improves the stability of foams and emulsions and
shows exceptional fat-like characteristics when used under the forms of gel in
water. Fat and carbohydrate replacement with chicory inulin and oligofructose
offers the advantage of not compromising on taste and texture, while delivering
nutritionally enhanced products (Franck, 2002).Inulin introduces body and
mouth feel in cheese analogues or ice cream, and reduces syneresis in yoghurt
and other fermented milk products (Blecker et al., 2001).
The word “banana” is a general term embracing a number of species or

hybrids in the genus Musa of the family Musaceae (Haque, 2008). In order to
increase the utilization of banana, some researchers have suggested converting
the green banana into flour and starch (Suntharalingam and Ravindran, 1993).
Green banana is rich in carbohydrate especially starch (20-25 per cent)
(Cordenunsi and Lajolo, 1995), dietary fiber especially hemi cellulose (6.08 per
cent) (Kayisu et al., 1981), essential minerals such as potassium, phosphorus
and calcium and various vitamins such as A, B1, B2 and C (Morton, 1987).
Besides, green banana has been found to reduce clinical severity of childhood
Shigellosis and is useful in treating diarrheal diseases (Rabbani et al., 2009).
Average chemical composition of raw banana powder is presented in Table 2.6.
Page | 44
Table 2.6 Average chemical composition of raw banana powder
Protein Fat Ash Calcium Phosphorus Iron Moisture Fiber

(%) (%) (%) mg/100gm mg/100gm mg/100gm (%) (%)
5.32 3.33 2.46 44 115.60 25.50 4.33 2.5
(Morton, 1987)
Oats are the seeds of the plant Avena sativa, a cereal grain that has been
used as a foodstuff for humans. Nutritionally oats are an excellent source of
soluble fiber in the form of beta-glucans, alpha tocopherols, B vitamins,
minerals, proteins, and plant fats. As a health food, oats have received
extensive attention in recent times for their ability to lower serum cholesterol
levels. Oat beta glucan isolates have been shown to shift serum cholesterol
profiles to contain less LDL and more of the desired HDL (Lazaridou et al.,
2004). Oat bran products have been shown to reduce postprandial glucose
spike after a glucose load in patients with type II diabetes. The average
chemical composition of oat is presented in Table 2.7.
Table 2.7 Average chemical composition of oat
Crude
Products Protein (%) Fat (%) Ash (%)
fiber (%)
Whole oats 12.1 5.1 12.1 3.4
Groats 15.8 7.2 2.8 1.9
(Coffman, 1961)
2.5.4 Nutrition Claim
Source of Dietary Fiber: A claim that a food is a source of fibre and any
claim likely to have the same meaning for the consumer, may only be made
where the product contains at least 3 g of fibre per 100 g. High in Dietary
Fiber - A claim that a food is high in fibre and any claim likely to have the
Page | 45
same meaning for the consumer, may only be made where the product
contains at least 6 g of fibre per 100 g or 100 ml (FSSA, 2009).
2.5.5 Dietary fiber in dairy products
Seven commercial dietary fibre preparations (maize, oat, soya, sugar beet
and rice) were used to fortify sweetened plain yoghurt by addition at 1.32per
cent prior to homogenization, heat treatment and addition of starter. Addition
of fibre accelerate the acidification rate, and increase viscosity use of sugar-
beet or one of the soya preparations was used, in decrease viscosity due to
partial syneresis of the whey. Fibre-fortified yoghurt had a grainy flavour and
gritty texture; fortification with oat fibre gave the best results, with flavour and
texture scores not being significantly lower than those of control yoghurt
(Fernandez et al., 1997).
Calorie-reduced yogurts that were fortified with 1.32per cent oat fiber
were prepared from lactose-hydrolyzed milk, alone and supplemented with 2
and 4 per cent sucrose or with 1.6, 3.6, and 5.5 per cent fructose. Fernandez et
al. (1998) according to them Addition of 5.5per cent fructose increased
fermentation time by 60 per cent, slowing down the production of lactic,
pyruvic, acetic, and propionic acids and the consumption of hippuric and
orotic acids. Fiber addition led to increases in concentrations of acetic and
propionic acid. Lactobacilli counts were lower in samples treated with fructose.
After 28 day of storage, lactobacilli counts were consistently higher in fiber-
fortified yogurts, but total bacteria counts were lower. Apparent viscosity
increased with the addition of sweetener and fiber. Fiber addition decreased
overall flavour quality. The lactose-hydrolyzed yogurts received the highest
flavour scores, independent of fiber fortification. Fiber addition improved the
body and texture of unsweetened yogurts but lowered overall scores for body
and texture in yogurts sweetened with sucrose.
In cheese making, β-glucan affects the curd formation by reducing the

curd cutting time and increasing curd yield. β-glucan has been used in the
Page | 46
manufacture of low fat yogurt and ice cream to act as a fat mimetic in low fat
dairy products by improving the mouth feel. However, β-glucan can have
negative effects on flavour and mouth feel in cheese. "Nutrim", a commercial β-
glucan reduced the firmness of cheddar cheese resulting in a paste rather than
a curd (Volikakis et al., 2004).
The use of inulin as a functional ingredient in ice cream resulted in

significant improvement in textural characteristics in terms of firmness,
melting properties (Akalin and Erisir, 2008). β-glucan in skim milk yogurt
decreased syneresis when compared to yogurt made with full fat milk. The
decreased in syneresis was attributed to β-glucan's ability to form a three-
dimensional network entrapping water. The sensory studies showed that
addition of β-glucan to yogurt improved mouth feel, scoopability, and sensory
attributes consistent with full fat yogurt (Brennan and Tudorica, 2008).
The effect of fortification with Date fiber (DF) on fresh yogurt quality was
investigated. Control yogurt (without fiber), yogurt fortified with 1.5, 3.0, and
4.5 per cent DF, and yogurt with 1.5 per cent wheat bran (WB) were prepared.
Fortification with DF did not cause significant changes in yogurt acidity,
although pH was increased. Yogurts fortified with DF had firmer texture (higher
hardness values) and darker color compared with control or WB yogurts.
Consumer test results indicated that the appearance, color, and flavor ratings
were significantly affected by fiber fortification. Yogurt fortified with up to 3 per
cent DF had similar sourness, sweetness, firmness, smoothness, and overall
acceptance ratings as the control yogurt. Sensory ratings and acceptability
of yogurt decreased significantly when increasing DF to 4.5 per cent or using
1.5 per cent WB. Flavoring yogurt fortified with 4.5 per cent DF with vanilla did
not improve flavor or overall acceptance ratings. Thus, fortifying yogurt with 3
per cent DF produced acceptable yogurt with beneficial health effects (Hashim
et al., 2009).
Page | 47
Pandiyan et al. (2012) carried out to prepare synbiotic ice cream

incorporating Lactobacillus acidophilus and inulin and viability of L.acidophilus
was analyzed on storage. Whey protein concentrate (WPC) was incorporated in
the ice cream mix to improve the textural and nutritional quality of ice cream.
A faster melting rate was noticed in the probiotic and synbiotic ice cream
samples. Incorporation of inulin in ice cream mix significantly (P<0.01)
improved the growth of Lactobacillus acidophilus. Freezing of the ice cream mix
caused a reduction of 0.61 to 0.77 log counts of L. acidophilus count. A
significant reduction (P<0.01) in the count of L. acidophilus was observed
during storage. It is concluded that incorporation of inulin increases count of L.
acidophilus and the organism could survive at therapeutic minimum probiotic
level of 106 cells /ml for 15 days of storage at -18 to -23°C in ice cream.
Apple, wheat and bamboo fibers were used in the production of strained
yogurt at different ratios (1 per cent, 2 per cent and 3 per cent). The type of
dietary fiber caused statistically significant changes in colour, texture values
and sensory evaluation scores. Apple fibrous strained yogurts weren’t
preferred by panellists because of their ragged structure, dominant apple
taste and strong odour. Panellists found bamboo and wheat fiber strained
yogurts acceptable (Kemal et al., 2012).
Various researches have been carried out on Shrikhand. Since Shrikhand

traditionally enjoys a huge popularity in our country, the present investigation
on Fiber fortified probiotic Shrikhand was carried out utilizing the existing
literature.
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Chapter 3
MATERIAL AND METHODS
3.0 Introduction
Traditionally, Shrikhand preparation involves production of curd

(dahi) by lactic fermentation of whole milk, either cows or buffalo’s milk,
followed by the draining of whey from the curd through a suspended
muslin cloth or bag. The resulting solid mass (known as Chakka) is
uniformly mixed with sugar, cream and other ingredients like fruits, nuts,
cardamom, saffron and other spices etc. to improve physico-chemical,
rheological and sensory attributes of the final product.
Milk is rightly considered a ‘nearly complete food’. However, with

rapidly changing lifestyle the diet has become much of a concern for
health. Life has greatly contributed to the association of heart and other
related ailments with saturated fat and cholesterol in the diet. Milk and
milk products come under the cloud because of this suspected role in
coronary heart disease. In this context, milk and milk products
considered as a vehicle for dietary fiber would not only take care of their
own role in human health but could also enhance the healthfulness of the
diet as a whole.
The present study derives its significance from the above stated
facts. The chapter deals with the materials and methods employed in this
investigation as delineated below.
3.1 Materials used in Shrikhand making
The raw materials utilized for the manufacture of Shrikhand are as

detailed below.
Materials and Methods
3.1.1 Skim Milk
Cow or buffalo skim milk used for manufacturing Shrikhand was

procured from “Anubhav Dairy”, SMC college of Dairy Science, Anand.
Fresh skim milk, after carried out, was strained through a muslin cloth
and chemical analysis of milk was done. It was free from any
objectionable flavor and contained on an average fat 0.1%, total protein
3.5%, total solids 9.33% and 0.14 to 0.16 % acidity (as % LA). Thereafter,
it was treated as per requirement and then used for Shrikhand
manufacture.
3.1.2 Cream
Buffalo milk cream was obtained from Anubhav Dairy of college.

The fat content of cream was 65 to 70%.
3.1.3 Probiotic Culture
Thermophilic lactic acid freeze dried DVS (Direct Vat Set) culture
(ABY-3) procured from the Christian Hansen ltd. Mumbai containing
Bifidobacteriumspp., Streptococcus thermophilus, Lactobacillus acidophilus,
Lactobacillus delbrueckii subsp. bulgaricus was used to manufacture
Shrikhand.
3.1.4 Sugar
For preparation of Shrikhand with sweetener of good quality,

granular form of sugar, free from impurities was purchased from local
market of Anand.
3.1.5 Powder containing fiber ingredients
Probiotic Shrikhand was fortified with three fiber containing

materials viz. banana, oat and inulinpowders. Banana powder was
procured from Aum fresh- AumAgri Freeze Foods, Baroda. Bagrrys India
Limited, Delhi, supplied oat. Inulin was supplied by Oraftiltd. Belgium.
Page | 50
3.1.6 Packaging material
Polystyrene cups of 200 g and 500 g capacity procured from local

market of Anand were used for filling Shrikhand.
3.1.7 Culture media and Reagents
The composition of media and reagents utilized during the course of

experimentation is given in appendix-III.
3.2 Preparation of Shrikhand
The experimental Shrikhand was prepared as per the procedure

described in flow diagram given in figure 3.1
Skim milk (0.1% fat)
Heat treatment (90˚C/ 10 min.)
Cooling to 39˚C
Inoculation with starter culture ABY-3
Incubation (39°C/12 hr)
Whey drainage (hanging overnight)
Chakka
Addition of sugar (72.5% on DM of chakka)
Addition of cream (6.2% fat in Shrikhand)
Mixing
Passing through mesh (no.30)
Shrikhand
Packing and Storage
Figure 3.1: Flow chart for Shrikhand preparation (Patel, 2013)
Page | 51
3.2.1 Preparation of Dahi
The skim milk was filtered through muslin cloth, heated indirectly
in boiling water-bath to 90°C for 10 min. and cooled to 39°C. The milk
was inoculated 0.1g/lit for ABY-3 culture. Thereafter milk was incubated
at 39°C until the desired firmness and acidity of 0.1% LA was obtained.
3.2.2 Preparation of Chakka
After having the desired acidity (1.0 % LA) and firmness in

dahi/curd, whey was drained from the curd through a suspended muslin
cloth. Then curd was hanged overnight to get solid mass known as
Chakka.
3.2.3 Mixing of chakka with other ingredients
Chakka was mixed with fine quality crystalline sugar at the @ of

72.5% (on dry matter basis) and calculated amount of pasteurized cream
(adjust about 6% fat content in the Shrikhand) also along with powder
containing fiber ingredients. The content was mixed with flavouring
substances uniformly, sugar was dissolved completely and thereafter the
mixture was passed through a Wire Mesh (mesh size 30) to get fine and
homogenous consistency in Shrikhand.
3.2.4 Packing and storage of Shrikhand

After mixing, Shrikhand packed in polystyrene cups (previously
sanitized in about 200-ppm chlorine solution for about 20 min and then
drained thoroughly). The samples of fiber fortified Shrikhand were stored
at refrigerated temperature (i.e. 7±2°C) to study changes taking place
during storage. The observations were taken at the interval of 7 d up to
35 d.
3.3 ANALYSIS
Skim milk, cream, whey, chakka and Shrikhand (fresh as well as
stored) were subjected to various analyses described below.
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Skim milk, cream and Shrikhand
Representative samples of skim milk used for Shrikhand making

were analysed for fat (IS: 1224, Part-I, 1997), total protein (IS: 1479, Part-
II, 1961) and titratable acidity (IS: 1479, Part-I, 1960). The total milk
solids were determined by Mojonnier milk tester, Model-D, as per
standard procedure (Laboratory Manual, Milk Industry Foundation,
1959).
The cream utilized for standardization of fat in the finished product

were analyzed for fat (IS: 1224, Part-II, 1997).
Whey
Fat, total solids and titratable acidity of whey were determined as

per methods outlined above for milk, but substituting whey for skim milk.
Chakka
Representative samples of chakka were drawn immediately after

preparation, analysed for total milk solids by Mojonnier method
(Laboratory Manual, Milk Industry Foundation, 1959), fat, and total
protein by standard methods suggested by BIS for chakka and Shrikhand
(IS: 9532:1980).
Titratable acidity of chakka was determined by taking 10g of sample

and diluting it with 30 ml distilled water. After through mixing it
wastitrated against 0.1 N NaOH, using phenolphthalein (1% in 75% ethyl
alcohol) as an indicator. The results were expressed as per cent lactic
acid.
Shrikhand
Representative samples of fresh Shrikhand were drawn after

through mixing. The fresh samples of Shrikhand were analyzed for
moisture, fat, total protein, ash, sugar, titratable acidity, free fatty acids,
soluble protein, water activity and consistency, where as the stored
samples were analyzed for moisture, titratable acidity, free fatty acids,
Page | 53
soluble protein, water activity and consistency at 0, 7, 14, 21, 28 and 35

d for product stored at 7±2°C. All the tests were carried out in duplicate.
3.3.1 Compositional Analysis
Total solids
The total solids of the Shrikhand were determined according to

procedure described in IS 1479 (II) (1961). To obtained percentage
moisture content of sample, the per cent T.S. was subtracted from 100.
Fat
Per cent fat in the Shrikhand was determined as per the Rose –
Gottlieb method prescribed in FSSA standards (2010).
Weigh accurately about 10 g of sample, transfer to extraction tube.
Add 1.25 mil of ammonia sp.gr. 0.8974 mix and shake thoroughly. Add
10 ml of ethyl alcohol and mix again. Add 25 ml of diethyl ether stopper
and shake vigorously for about a minute. Then add 25 ml petroleum ether
(boiling range 40-60°C and shake again vigorously for about half a
minute. Let it stand until the upper ethereal layer has separated
completely and is clear. If there is tendency to form emulsion, a little
alcohol may be added to help separation of the layers. Decant off the clear
ethereal layer in to a suitable vessel (aluminium dish). Wash the delivery
end of the extraction tube with a little ether and add the washing to the
flask. Repeat twice extraction of the liquid remaining in the extraction
tube using 15 ml of each solvent every time. Add the ethereal extract to
the same container and evaporate off completely. Dry the flask in an air
oven at 102±2°C for twohrs, cool in a desiccators and weigh. Repeat the
process of heating and cooling and weighing until the difference between
two successive weights does not exceed one mg. Wash out the fat from the
flask with petroleum ether carefully leaving any insoluble residue in the
flask. Dry the flask in the oven and reweigh. The difference in weights
represents the weight of fat extracted from the sample (Shrikhand).
Correct weight of extracted fat by blank determination on reagents used.
If reagent blank is more than 0.5 mg, purify or replace reagents.
Page | 54
Difference between duplicate determinations obtained simultaneously by

the same analyst should not be more than 0.03 g fat/100 g product.
Calculation
Fat % (W/W) = Weight of Extracted Fat × 100
Weight of sample
Total sugar
Total sugar (lactose + sucrose) in the Shrikhand was determined as

per the method prescribed in Indian Standards (IS: 2802, 1964).
Twenty grams of Shrikhand was accurately weighed in to 250 ml

conical flask and diluted with 250 ml lukewarm distilled water (35 to
40°C), the contents of the flask were mixed thoroughly. For precipitation
of protein of Shrikhand, neutral lead acetate, alumina cream and sodium
oxalate solution were used as per the procedure described for ice cream in
Indian Standards (IS: 2802, 1964). After precipitation, the contents were
filtered through a Whatman filter paper No.1 in to a 500 ml graduated
flask. The precipitates and paper were thoroughly washed with hot water,
collecting the washing in flask. The flask and contents were cooled and
the volume was made up to the mark. The filtrate so obtained was utilized
for determination of total carbohydrates (original reducing sugar +
sucrose) as per the method prescribed in Indian Standards (IS: 2802,
1964) using the following relationships:
I) 0.05 g glucose = 0.0678 g lactose,
II) Percentage of lactose = Per cent glucose × 0.0678/0.05,
III) Percentage of sucrose = (Reducing sugar after conversion) –

(Original reducing sugar) × 0.95
Page | 55
Ash
Ash content of the entire sample was determined by procedure

described in BIS (IS: 1547-1985). 2 to 3 grams of homogenous sample
was weighed in a silica crucible. The sample in crucible was heated on
naked-flame until it was completely evaporation of moisture. The sample
was then transferred to muffle furnace and held for 3.5 h at 550±2°C after
cooling in the desecrator, the crucible was weighed. The process was
repeated until constant weight. The total ash content in per cent was
calculated as follows.
Total ash (% by weight) = 100 (W2-W)
(W1-W)
Where, W = Weight in g of the empty crucible
W1 = Weight in g of the crucible with sample
W2 = Weight in g of the crucible with ash
Total protein
Total nitrogen/protein of Shrikhand was determined by Semi-
Microkjeldahl method (IS: 1479 – Part – II, 1961), using Kjel-plus
digestion system (Model- KPS 006L, M/s. Pelican Instruments, Chennai)
as follows.
In a digestion tube 0.5-1.0 g of the sample was accurately weighed
and then 2.4 g of digestion mixture (Potassium Sulphate: copper
sulphate: selenium dioxide; 1:0.1:0.1) was added. To the tube contents,
10 ml of nitrogen-free concentrated sulphuric acid was added. The tubes
were then transferred to the digestion block, where the contents were
digested for about 30 min at a final temperature of 350°C.
The cooled and digested contents were loaded in the Kje-plus
distillation unit and after the unit was ‘ready’, a fixed volume of alkali (20
ml of 40 per cent sodium hydroxide) was added automatically to the
sample. The distillation time was fixed for 3 min. The librated ammonia
Page | 56
was condensed and collected in 25 ml of saturated boric acid solution

containing three drops of mixed indicator [equal volume of saturated
solution of methyl red and 0.1 per cent methylene blue solution, both
made in 95% (v/v) ethanol]. The distillate in boric acid was titrated
against 0.05 N sulphuric acid. A reagent blank was simultaneously run
using all the above chemicals except the sample and its reading was
subtracted from the experimental reading.
The percent total protein calculated using formula:
% Total protein = 0.07×(Burette reading – Blank reading)
Where, W =Weight of sample
For converting the values of total nitrogen into percent total protein,
the values were multiplied by a factor of 6.3.
Soluble nitrogen
The soluble nitrogen content of milk or Shrikhand samples was

determined by the procedure outlined by Kosikowski (1982). Three grams
of sample was weighed and mixed with requisite (~ 40 ml) quantity of
warm (50°C) Sharp’s extraction solution (Appendix-III). Using additional
solution, the final volume was made up to 100 ml in a volumetric flask.
The contents, after through mixing, were tempered for 1 h at 50 ± 1°C in a
water-bath with intermittent shaking and then filtered through Whatman
filter paper No.40. Ten millilitres of this filtrate was used for estimation of
soluble nitrogen by semi—micro-Kjeldahl method as described in section
3.7.6.5. A reagent blank was run simultaneously. The soluble nitrogen
expressed in percent, was calculated using the formula.
% Soluble nitrogen = 0.07 × 10 × (X-B)
Page | 57
Where, W = Weight of sample in grams
X = volume of 0.05 N sulphuric acid required in sample titration
B = volume of 0.05 N sulphuric acid required in blank filtration
Crude fibre
Crude fiber was estimated by official method described in Analysis

of the Association of Official Analytical Chemist (AOAC, 1990).
The moisture content of the Shrikhand sample was reduced to

about 10 percent by evaporation in a hot air oven. The reduced moisture
sample was subjected to total solids determination and thereafter it was
extracted for about 1 hour with petroleum ether, using a Soxhlet
apparatus. About 1 g of the moisture cum fat free sample was weighed
accurately and transferred to a 1-litre flask. 200 ml of dilute sulphuric
acid was taken in a beaker and was brought to boil. The whole of the
boiling acid was transferred to the flask containing the fat-free material
and immediately the flask was connected with a water-cooled reflux
condenser and was heated so that the contents of the flask begin to boil
within 1 minute. The flask was rotated frequently, taking care not to allow
the material to stick to the sides of the flask, and not to keep the material
out of contact with the acid. The boiling was continued for exactly 30
minutes. The flask was removed and the content was filtered through fine
linen (about 18 threads to the centimetre) held in a funnel and washed
with boiling water until the washings were no longer acidic to litmus.
Some quantity of sodium hydroxide solution was brought to boil under a
reflux condenser. The residue was washed on the linen into the flask with
200 ml of the boiling sodium hydroxide solution. Immediately the flask
was connected with the reflux condenser and boiled for exactly 30
minutes. The flask was removed and immediately the content was filtered
through the filtering cloth. The residue was thoroughly washed with
boiling water and transferred to a Gooch crucible prepared with a thin but
compact layer of ignited asbestos. The residue was washed thoroughly
first with hot water and then with about 15 ml of ethyl alcohol, 95 per
Page | 58
cent by volume. The Gooch crucible with the contents was dried at
105±2°C in an air oven to a constant mass. It was cooled and weighed.
The contents of the Gooch crucible were incinerated in an electric muffle
furnace maintained at 600±20°C until all the carbonaceous matter was
burnt. The Gooch crucible containing the ash content was cooled
indesiccators and weighed.
Crude fiber(on dry basis), percent by mass =100 (M1 – M2) / M

Where,
M1 = Mass in g of Gooch crucible and contents before ashing,
M2= Mass in g of Gooch crucible containing asbestos and ash
M = mass in g of the moisture-free material taken for the test.
3.3.2 Physico-chemical analysis
The various physico-chemical properties of Shrikhand

weredeterminedusing standard procedure.
Titratable acidity
The acidity of Shrikhand was determined by method described in

BIS (IS: 1166-1968) for condensed milk.
Titratable acidity of all the samples was determined by titrating 10 g

of sample against 0.1 N sodium hydroxide using phenolphthalein (1% in
75% ethyl alcohol) as an indicator and the results expressed in terms of
percent lactic acid (LA).
Acidity (as % LA) = 9×N×V

W
Where, N = Normality of sodium hydroxide used
V = Volume in ml of NaOH used for titration
W = Weight of sample
Page | 59
pH
The pH of Shrikhand was measured using Systronic digital pH

meter, Model 335. The method described by Franklin and Sharpe (1963)
for cheese was used. The homogenate prepared by diluting 20 g of sample
in 20 ml of glass-distilled water was subjected to pH measurement.
Free fatty acids (FFA)
Lipolytic changes were measured in terms of free fatty acids (FFA).

The free fatty acids content was determined by the method suggested by
Deeth and Fitz-Gerald (1976).
Three grams of Shrikhand was weighed in 60 ml screw cap test

tube, stoppered and mixed with 10 ml of extraction mixture (i.e. iso
propanol: petroleum ether: 4 N sulphuric acid; 40:10:1), 6 ml of
petroleum ether and 4 ml distilled water. The stoppered test tube was
then shaken vigorously for 15 s. After allowing sufficient time (i.e. 5-10
min) for the two layers to separate, 7.5 ml aliquot of the supernatant was
withdrawn and transferred to a 50 ml flask. After addition of 6 drops of 1
% methanolic phenolphthalein, the solution was titrated against 0.02 N
methanolic potassium hydroxide solutions. The FFA content of Shrikhand
expressed in terms of micro equivalent(µeq) per ml was calculated using
the following formula:
FFA (as % oleic acid) = V × N × 103
3P
Where, V = Net titration volume
N = Normality of KOH
P = Part of the upper layer titrated out of total volume
Water activity
The water activity of Shrikhand samples, tempered at 25°C

temperature, was measured using RotronicHygroskop Model: Hygrolab – 3
Page | 60
(M/s. Rotronicag, Switzerland) connected to sensing element (AW – DIO)

with a measuring range of 0-100 % relative humidity (RH).
The sample of Shrikhand was filled in dish/cup up to the brim to

the maximum possible extent and the dish was kept in an airtight
chamber taking care that the sample does not touch the diaphragm of the
sensor. The measuring chamber was conditioned to the temperature of
the sample. The chamber was then closed in a manner to ensure metallic
contact between the two parts. The values of water activity and
temperature were noted from the display panel when it reached the
equilibrium and a constant reading is observed.
Measurement of consistency
Compression testing of Shrikhand was done using Food Texture
Analyzer (M/s. Lloyd Instruments, LRX Plus, England; Sr. No. 160374)
using 50 Newton (N) load cell. The samples of Shrikhand, with edges of 13
mm height and 78 mm width were placed in the compression support
plate. Tempering the samples at 23±1ºC for half an hour in an air-
conditioned room (23±1ºC, 55±1 per cent RH). The Shrikhand samples
were drawn using a cork borer at 23±1ºC and were compressed at a speed
of 50 mm/min. The trigger was set at 10 gf (1gf = N/0.00980665). The
samples were compressed up to 50.0 per cent of their initial size.
Four samples were used for each experimental Shrikhand under
study and the average of these readings was reported. The textural
characteristics of Shrikhand samples were directly displayed on the
monitor of the computer as graph Load (N) vs. time (s) as well as derived
values in tabular form. A typical two-bite force-time compression curve for
fiber containing banana, oat and inulin powder and control Shrikhand is
shown in Figures 3.2A, 3.2B, 3.2C and 3.2D respectively.The hardness
result was directly taken from the displayed readings.
Page | 61
Load (N)
15.0
Texture Profile Analysis Setup
Fiber Fortified Shrikhand
Hardness1
10.0
Hardness2
5.0
Greatest Slope
Fracture
0.0
Adhesive Force
Trigger
0 5 10 15 20 25 30 35
Time (Seconds)
Figure 3.2 A representative Two-bite forceDistance Texture profile curve of

Banana powder Shrikhand sample.
Page | 62
Load (N)
Hardness1
15.0
10.0
Hardness2
5.0
Greatest Slope
Fracture
0.0
Adhesive Force
Trigger
0 5 10 15 20 25 30 35
Time (Seconds)
Figure 3.3 Representative Two-bite forceDistance Texture profile curve of

Oat powder Shrikhand sample.
Page | 63
Load (N)
Hardness1
10.0
Hardness2
5.0
Greatest Slope
Trigger
0.0
Fracture
-5.0
Adhesive Force
-10.0
0 5 10 15 20 25 30
Time (Seconds)
Figure 3.4 Representative Two-bite force Distance Texture profile curve of Inulin
powder Shrikhand sample.
Page | 64
Load (N)
Hardness1
10.0
Hardness2
5.0
Greatest Slope
Fracture
0.0
Trigger
-5.0
Adhesive Force
0 5 10 15 20 25 30
Time (Seconds)
Figure 3.5 Representative Two-bite forces Distance Texture profile curve of

control Shrikhand sample.
Page | 65
3.3.3 Microbiological analysis

All the samples were drawn aseptically and analyzed for the
probiotic count, standard plate count (SPC), yeast and mould and
coliform count. All the samples were transported, stored and handled as
described in BIS (IS: 5403 – 1969).
3.3.3.1 Preparation of samples for microbiological analysis of

Shrikhand
After sufficient cooling of Shrikhand on the 0 d, cups of sample were
removed for conducting probiotic count, coliform count, yeast and mould
count and standard plate count (SPC). Similarly thereafter at an interval
of 7 d, up to 35 d of stored Shrikhand samples were removed for microbial
analysis.
After homogenous mixing of Shrikhand in the cup with sterile

spatula, 11 g of sample was weighed aseptically in a sterile phosphate
buffer flask to prepare 1:10 dilution. After thoroughly mixing, 1 ml of
diluents from 1:10 was transferred in to 9 ml sterile phosphate buffer
tube to make 1:100 dilutions. It was further appropriately diluted using 9
ml sterile phosphate buffer to make required dilutions. One millilitre of
diluents, from suitable dilution was poured aseptically in sterile
petriplates in duplicate. The time elapsed between preparations and
pouring of appropriate dilution in sterilised petri plates normally did not
exceed 15 min.
Coliform count
The freshly prepared, unsterilized Violet Red Bile Agar (VRBA) was
meltedand sterilized at 115ºC for 15 min. then cooled at around 45°C.
Around 12-15 ml was added in plates containing 1 ml of appropriate
dilution in duplicate, mixed well and allowed to solidify. A second layer of
same agar (4 – 5ml) was overlaid in each plate. Typical colonies of coliform
bacteria were counted and expressed as cfu/g after 24 h of incubation at
37°C. Plates showing no colony growth, were incubated for a further
period of 24 h. Plates were then examined for typical colonies of coliform
Page | 66
bacteria (dark red colonies having a diameter of > 0.5 mm) and counted
(Indian Standards, IS: 5401, 1969).
Probiotic count
Probiotic count of inoculated Shrikhand samples was determined as

per the method described by Mann De et al, (1960). Petri plates
containing the appropriate dilutions were poured with melted and cooled
MRS and Bifidobacterium Selective Count Agar medium at around 45°C,
mixed properly and allowed to solidify. A second layer of same agar
(4 – 5ml) was overlaid in each plate. The petriplates were incubated at
temperature of 37°C and colonies formed on the medium were counted
after 48 h.
Standard Plate Count (SPC)
The freshly prepared, unsterilized Standard Plate Count Agar (SPC)

was melted and cooled at around 45°C. Around 15-20 ml was added in
plates containing 1 ml of appropriate dilution in duplicate, mixed well and
allowed to solidify. The plates were incubated at 37±0.5°C for 48 h and
the number of colony forming units (c.f.u.) per g of product was noted.
Yeast and Mould count
Procedure followed was according to Indian Standard, IS 5403,

(1969). The petriplates containing the diluted samples were poured with
melted and cooled potato dextrose agar medium at around 45°C (adjusted
to 3.5 pH using 10% sterile tartaric acid solution, within 30 min of
addition to the sample) mixed properly, and allowed to solidify. The plates
were incubated at temperature 22-25°C for 3-5 d before count were
recorded. Marshall (1992) also gives comparable procedure for
enumeration of yeasts and moulds from food sources.
3.3.4 Sensory evaluation
Controls as well as experimental samples of Shrikhand were

subjected to sensory evaluation by an expert trained panel of seven
judges. Judges evaluated the products using the scorecard (Appendix I)
prescribed by Bureau of Indian Standards (IS: 15348, 2003). According to
Page | 67
this scorecard, maximum score allotted to flavor is 50, body and texture
is 35, colour and appearance is 10 and package is five. Full marks i.e.
5/5 was allotted to package for all the samples, since the samples were
served in polystyrene cups with lids. Fresh product at 0 d and the stored
products (7, 14, 21, 28 and 35 d storage at 7±1°C) were given for judging.
3.4 Statistical Analysis
Completely randomized design (CRD) was used for data collected for
fresh Fiber Fortified Probiotic Shrikhand. The mean values generated from
the analysis of duplicate samples of Shrikhand obtained in four
replication were subjected to statistical analysis using completely
randomized design (CRD) as per Steel and Torrie (1980), which is
illustrated as given below:
Yij= µ + Ti + Eij
Where,
Yij= Response due to jth observation in the ith treatment
µ = General mean
Ti = Effect of ithtreatment
Eij= Error due to jth observation in the ith treatment
Page | 68
Chapter 4
RESULTS AND DISCUSSION
Milk and most dairy products are devoid of dietary fiber. With the
growing interest in dietary fiber and its health benefits, dairy industry is
gearing up for fortifying the dairy products with fiber. Moreover, the oil
content present in the structure of some food materials is a problematic
situation for consumers. In order to solve this problem, dietary fiber can be
used, which can improve the textural and sensory properties of products in
addition to being functional. Milk and milk products considered as a vehicle
for dietary fiber would not only take care of their own role in human health
but could also enhance the heath fullness of the diet as a whole. It is also
important for food materials to be delicious as well as natural.
The present investigation was planned, with the objective of developing

a technology for manufacture of a cost effective, consumer acceptable fiber
fortified probiotic Shrikhand by using probiotic culture and selection of fiber
containing powder.
This section delineates the results obtained from various experiments

during this investigation as well as discussion on the results obtained. The
data generated during the investigations was statistically analyzed by using
Completely Randomized Design and results are interpreted.
The results obtained during investigation on different aspects of

Shrikhand are covered under following phases:
4.1 PHASE-I: Preparation of Shrikhand using different fiber containing

powder
Result and Discussion
4.2 PHASE-II: Optimization of the rate of addition of fiber containing powder

from each fiber containing powder
4.3 PHASE-III: Comparison of Shrikhand containing optimized level of each

fiber containing powder to select the best fiber containing powder in fiber
fortified Shrikhand
4.4 PHASE-IV: Selection of flavouring ingredients for the fiber fortified

probiotic Shrikhand
4.5 PHASE-V: Study of the shelf life of developed fiber fortified Shrikhand at
refrigeration temperature(7 ±2 0C)
4.1 PHASE-I: PREPARATION OF SHRIKHAND USING DIFFERENT FIBER

CONTAINING POWDERS
Three fiber containing powder were incorporated in Shrikhand at three rates

viz., banana powder @ (1.0,3.0,5.0) per cent, oat powder @ (1.0,2.0,3.0) per
cent, inulin powder @ (1.0,2.0,3.0) per cent on w/w basis of expected yield of
Shrikhand. These levels were selected based on preliminary trials. The fiber
content of banana powder, oat powder and inulin powder was2.5,12.8 and
92 per cent respectively. Shrikhand was prepared using the method
described by Patel (2013) using probiotic culture as described in Section
3.1.3. After obtaining chakka from skim milk dahi, fiber-containing powders
were blended thoroughly with chakka followed by addition of sugar and
cream to prepare Shrikhand. The average yield of chakka was 310g/ lit of
skim milk and the average yield of Shrikhand 472g / lit of skim milk. The
sample of Shrikhand without addition of fiber containing powder was used as
a control. Each sample of product (approximately 50 gm) was evaluated for
the composition, physico-chemical and sensory properties.
The experimental Shrikhand prepared using these fiber-containing

ingredients at different rates are coded below in Table 4.1.
Page | 70
Table 4.1: Particulars’ of treatments and codes name of experimental

Shrikhand
Particulars’ of treatments (level of Code

addition, % on w/w basis of expected
yield of Shrikhand)
Banana powder (1.0%) B1

Oat powder (1.0%) O1
Inulin powder (1.0%) I1
Control (0%) C
4.2 PHASE-II: OPTIMIZATION OF THE RATE OF ADDITION OF

This phase was conducted to optimize the rate of addition of three

fiber-containing powders for manufacture of acceptable quality fiber fortified
probiotic Shrikhand. The fiber containing powders were incorporated in
Shrikhand at three rates each viz., banana powder @ (1.0,3.0,5.0) per cent,
oat powder @ (1.0,2.0,3.0) per cent, inulin powder @ (1.0,2.0,3.0) per cent on
w/w basis of expected yield of Shrikhand. The relevant results are described
and discussed in this section.
Page | 71
4.2.1 Optimization of rate of addition of banana powder
The experimentation involved making four batches of Shrikhand viz., C, B1,

B2 and B3. Eachtreatment was replicated four times. In all 16 batches of
Shrikhand were prepared.
4.2.1.1 Effect of varying levels of banana powder on the composition of

Shrikhand
The average composition of fiber fortified probiotic Shrikhand and

control as affected by addition of different level of the fiber containing
banana powder is presented in Table4.2.
Total solids
It can be seen from Table 4.2 that Shrikhand without fiber content of
banana powder (control) had average TS content of 58.28 per cent. This
value was significantly (P<0.05) lower than Shrikhand containing B1, B2 and
B3. These values were 58.65, 59.35 and 60.03 for B1, B2 and B3 respectively,
and found significantly (P<0.05) different from each other.
Fat
The per cent fat content of experimental Shrikhand given in Table
4.2revealed that the average fat content of control was 6.23 per cent, while
banana powder containing Shrikhand samples B1, B2, and B3 were 6.20, 5.78
and 5.72 per cent respectively. There was significant (P<0.05) difference in
fat content of control and experimental Shrikhand sample of B1. It can be
seen from the tabulated value that although there was a decrease in fat
content with increase in rate of addition of banana powder. However,
samples B2 and B3 did not show any significant difference (P>0.05) in their
fat content.
Page | 72
Protein
The protein content of banana fiber fortified probiotic Shrikhand

samples were B1 (5.79), B2 (5.78) and B3 (5.75) per cent. While it was 5.82
per cent for control, Shrikhand sample. There was significant (P<0.05)
difference in protein content. In a, control Shrikhand had significantly
(P<0.05) higher protein content (5.80 per cent) as compared to all the other
samples. It might be due to low protein content in banana powder (Appendix
IV). The protein content of Shrikhand sample B3wassignificantly lower
compared to B1 and B2. However B1 and B2did not show any significant
difference (P>0.05) in their protein content.
Table 4.2: Effect of varying levels of banana powder on the composition

of Shrikhand
Chemical composition (%)
Carbohy
Treatments Total
Fat Protein Sucrose Ash drate
solids
(By diff.)
Control – C 58.28±0.03 6.23±0.03 5.82±0.01 41.43±0.10 0.57±0.02 44.63±0.02
B1 58.65±0.01 6.20±0.03 5.79±0.01 41.40±0.08 0.65±0.01 44.98±0.02
B2 59.35±0.02 5.78±0.05 5.78±0.01 41.41±0.06 0.68±0.01 45.72±0.01
B3 60.03±0.05 5.72±0.04 5.75±0.02 41.43±0.10 0.71±0.02 46.43±0.03
SEm 0.016 0.019 0.01 0.043 0.005 0.026
CD (0.05) 0.048 0.057 0.02 NS 0.016 NS
CV (%) 0.053 0.624 0.002 0.205 1.538 1.079
Figure placed after ± indicates standard deviation
Sucrose
It can be depicted from the Table 4.2 that Shrikhand without fiber
content of banana powder (control) had average sucrose content of 41.43 per
cent. Whereas in experimental Shrikhand samples of B1, B2 and B3were
Page | 73
41.40, 41.41 and 41.43 per cent respectively. The variation in sucrose
content of all experimental samples was found to be non-significant (P>0.05).
Ash
As seen in Table 4.2, the average ash content of B1, B2, and B3 was
0.65, 0.68 and 0.71 per cent respectively. The ash content of control
(i.e. 0.57) per cent was significantly (P<0.05) lower than all the fiber fortified
samples viz., B1 B2 and B3. The ash content of B3 (i.e. 0.71 per cent) was
significantly (P<0.05) higher than all the other experimental samples.
Banana powder contains good amount of minerals (Morton, 1987). Therefore,
addition of bananas powder at higher rate in B3 resulted in significantly
higher ash content. There was significant (P<0.05) difference in ash content
of all experimental samples.
Carbohydrate
The carbohydrate content of Shrikhand samples is depicted in Table

4.2. The tabulated value reveal that carbohydrate content of sample B1, B2
and B3 were 44.98, 45.72 and 46.43 per cent respectively. Whereas control
Shrikhand had a value of 44.63 per cent. All experimental samples were
statistically not (P<0.05) different in their carbohydrate content.
Published data on proximate composition of Shrikhand as affected by

addition of banana are not available for comparison, however the values
observed in the present study are very close to those reported by Aneja et al.
(2002) and Patel, (2013).
4.2.1.2 Effect of varying levels of banana powder on the physico-

chemical properties and consistency of Shrikhand
The data on physico-chemical properties and consistency of

experimental Shrikhand samples are depicted in Table 4.3.
Page | 74
Table 4.3: Effect of varying levels of banana powder on the physico-

Physico-chemical properties
Treatments
Acidity FFA Soluble Water pH Consistency
(%LA) µ.eq/g nitrogen activity (N)
(%)
C-Control 1.00±0.01 0.58±0.03 0.28±0.06 0.9389±0.01 4.33±0.05 8.15±0.06
B1 0.99±0.02 0.59±0.04 0.26±0.04 0.9388±0.01 4.35±0.06 8.35±0.06
B2 0.97±0.03 0.60±0.01 0.29±0.04 0.9379±0.01 4.36±0.07 8.53±0.05
B3 0.95±0.02 0.63±0.08 0.29±0.04 0.9371±0.01 4.38±0.05 8.73±0.05
SEm 0.012 0.023 0.023 0.002 0.028 0.027
CD (0.05) NS NS NS NS NS 0.082
C.V.% 2.356 7.600 16.172 0.530 1.305 0.632
Titratable Acidity
It can be seen from Table 4.3 that the average lactic acid content of
sample without fiber containing banana powder i.e. control Shrikhand
sample was 1.0 per cent LA, while for banana powder containing Shrikhand
samples viz., B1, B2 and B3 the values were 0.99, 0.97 and 0.95 per cent LA
respectively. The average values of acidity are well within the range of
reported values in literature (Patel, 2013). The acidity values are well below
the maximum value specified for Shrikhand by FSSA (2011). It can be seen
from Table that these was a slight decrease in acidity at the highest level of
addition viz., B3. However,the variation in titratable acidity (per cent LA) of
all experimental samples were found statistically non significant (P>0.05).
Page | 75
Free fatty acid
It can be seen from the Table 4.3 that Shrikhand without fiber content
of banana powder (control) had average free fatty acid content of 0.58 µ.eq/g.
Whereas, for banana powder containing Shrikhand samples, viz. B1 (1 per
cent), B2 (3 per cent) and B3 (5 per cent) these values were 0.59, 0.60 and
0.63 µ.eq/g respectively. The statistical values arrived at on the basis of this
data, depicted in Table 4.3 indicated non significant(P>0.05) effect of
addition of banana powder on free fatty acid content of Shrikhand.
Soluble nitrogen
It can be depicted from the Table 4.3 that soluble nitrogen content of
Shrikhand containing banana powder viz. B1 (1 per cent), B2 (3 per cent) and
B3 (5 per cent) were 0.26, 0.29 and 0.29 per cent respectively. The average
soluble nitrogen content for control was 0.28 per cent. The variation in
soluble nitrogen content of all experimental samples were statistically non-
significant (P>0.05).
Water activity
It can be depicted from the Table 4.3 that water activity of Shrikhand
containing banana powder viz.B1 (1 per cent), B2 (3 per cent) and B3 (5 per
cent) were 0.9388, 0.9379, 0.9371 respectively. Whereas Shrikhand without
fiber content of banana powder (control) had average water activity 0.9389.
The difference in water activity among all experimental Shrikhand samples
were non-significant (P>0.05).
pH
The pH of fiber containing banana powder Shrikhand samples were B1

(4.35), B2 (4.36) and B3 (4.38). While control Shrikhand C sample had, 4.33
that were higher value as compared to all other samples. However, the
variation in pH content of all experimental samples were statistically non-
Page | 76
Consistency
The consistency of the product is important criteria to measure the

body and texture characteristics such as firmness/weakness of the product.
Consistency was measured in term of N by food texture analyzer Section
3.3.1.
The value for consistency of Shrikhand without fiber content of banana

powder (control) as shown in Table 4.4 had average 8.15 N. Whereas
consistency of fiber containing banana powder Shrikhand samples B1 (1per
cent), B2 (3 per cent) and B3 (5 per cent) were 8.35, 8.53 and 8.73 N
respectively. Increase in amount of banana powder resulted in progressive
improvement in consistency. The variation in consistency of all experimental
samples were significant (P<0.05). Addition of banana powder Shrikhand was
found to exercises significant influence (P<0.05) on consistency at all the
levels studied.
Published data on proximate physico chemical properties and

consistency of Shrikhand as affected by addition of banana are not available
for comparison.
4.2.1.3 Effect of varying levels of banana powder on the sensory

characteristics of Shrikhand
In product development process, sensory quality plays a vital role in

deciding the acceptability of the product. The sensory parameter chosen to
assess the quality of Shrikhand was flavour, body and texture, colour and
appearance and total score. The result of sensory data is shown in Table 4.4.
The sensory quality of the experimental Shrikhand sample at 10±2ºC were
evaluated using a score-card shown in Appendix-I.
Page | 77
Table 4.4: Effect of varying levels of banana powder on the sensory

characteristics of Shrikhand
Sensory Characteristics
Flavor Body &texture Color & appearance Total score*
Treatments (50) (35) (10) (100)
Control 43.32±0.65 31.48±0.21 9.14±0.21 88.94±0.78
B1 45.00±0.37 32.55±0.16 8.00±0.19 90.55±0.24
B2 44.52±0.32 32.23±0.19 7.91±0.11 89.66±0.34
B3 45.25±0.66 32.71±0.11 7.02±0.16 89.98±0.63
SEm 0.263 0.087 0.086 0.264
CD (0.05) 0.811 0.268 0.265 0.814

C.V.% 1.182 0.539 2.114 2.210
*a full mark was given for package i.e. 5 out of 5.
Flavor
The flavor scores of experimental Shrikhand are depicted in Table 4.4.

The flavor scores of experimental Shrikhand prepared using supplementation
of fiber containing banana powder were 45.00, 44.52 and45.25 for B1, B2 and
B3 respectively. The average flavor score was concerned, control was
significantly (P<0.05) lower (43.32) than all three banana powder containing
Shrikhand samples. Sample B3 and B1did not show any significant (P<0.05)
difference in their flavor score. The difference in flavor score of experimental
Shrikhand samples may be attributed to increasing rate of addition of
powder that alter the flavoring score of experimental Shrikhand.
Page | 78
Body and texture
The average body and texture scores are given in Table 4.4. The body
and texture score of fiber containing banana powder Shrikhand samples
were B1, B2 and B3 for 32.55, 32.23 and 32.71 respectively. Control
Shrikhand sample had body and texture score of 31.48, which was
significantly (P<0.05) lower than all three banana fiber containing Shrikhand
samples. It might be due to increasing rate of addition of fiber in Shrikhand.
The variation in body and texture score of all experimental samples were
significant (P<0.05). While, sample B1 and B3were statistically not different
(P>0.05) in their body and texture score.
Color and appearance
of banana powder (control) had average color and appearance score 9.14.
This value was significantly higher than Shrikhand containing banana
powder viz. B1, B2 and B3. These values were 8.00, 7.91 and 7.02 for fiber
fortified B1, B2 and B3 samples respectively. The lower scores obtained to
banana powder containing Shrikhand samples were due to because of
presence of dull brownish color in such samples. The dull brownish was with
increased in level of addition of banana powder. The variation in color and
appearance score of all experimental samples were found significant
(P<0.05).
Total score
Total score is the indicative parameter of sensory quality of product in

totality; consist of flavor, body and texture, color and appearance
characteristics. The total score of experimental Shrikhand depicted in Table
4.4 was given out of 100. Shrikhand without fiber content of banana powder
(control) had average total score was 88.94. The total score of fiber
containing banana powder Shrikhand samples B1, B2 and B3 were 90.55,
Page | 79
89.66 and 89.98 respectively. However,B1, and B3 Shrikhand were found

statistically same in their total scores.
Published data on proximate sensory attributes of Shrikhand as

affected by addition of banana are not available for comparison.
Thus, from the result discussed above, it is observed that use of fiber
containing banana powder (B3) Shrikhand had superior quality in terms of
chemical, physico chemical and sensory attributes. So, B3 sample was
selected for further study. As per total score, experimental Shrikhand sample
is given preference score of B3>B1>B2>C.
4.2.2 Optimization of rate of addition of oat powder
The experimentation involved making four batches of Shrikhand viz.,

C, O1 (1 per cent), O2 (2 per cent) and O3 (3 per cent) based on expected yield
of Shrikhand. Each treatment was replicated four times. In all 16 batches of
4.2.2.1 Effect of varying levels of oat powder on the composition of

Shrikhand
The data on average composition of fresh experimental Shrikhand

sample are presented in Table 4.5.
Total solids
of oat powder (control) had average TS content of 58.28 per cent. This value
was significantly lower than Shrikhand containing O1, O2 and O3 oat powder.
These values were 58.65, 59.01 and 59.36 per cent for fiber fortified O1, O2
and O3 respectively. The difference in total solids content of all experimental
samples were significant (P<0.05). It might be due to increasing rate of
addition of powder increasing total solids content.
Page | 80
Fat
The percent fat content of oat powder containing Shrikhand is given in

Table 4.5. The average fat content of control Shrikhand sample was 6.23 per
cent, The fat contents of experimental Shrikhand prepared using
supplementation of fiber containing oat powder wereO1 (6.31 per cent), O2
(6.37 per cent) and O3 (6.46 per cent). It can be seen from the tabulated
value that there was slightly but significantly increased in fat content of
samples with increased in addition of oat fiber.
Table 4.5: Effect of varying levels of oat powder on the composition of

Shrikhand

Treatments
Total Fat Protein Sucrose Ash Carbohydrate
solids
Control – C 58.28±0.05 6.23±0.02 5.80±0.02 41.43±0.10 0.57±0.04 44.63±0.01
O1 58.65±0.03 6.31±0.01 5.90±0.02 41.40±0.08 0.65±0.04 44.70±0.04
O2 59.01±0.03 6.37±0.01 5.96±0.01 41.41±0.06 0.68±0.01 44.76±0.02
O3 59.36±0.02 6.46±0.01 6.03±0.01 41.43±0.10 0.71±0.01 44.82±0.03
SEm 0.018 0.007 0.007 0.043 0.014 0.016
CD (0.05) 0.055 0.022 0.022 NS 0.044 0.049
CV% 0.061 0.224 0.241 0.205 4.348 0.071
Protein
As shown in Table 4.5, the average protein content of control

Shrikhand C sample was 5.80 per cent, Whereas protein content of fiber
Page | 81
containing oat powder Shrikhand samples O1 (1 per cent), O2 (2 per cent) and
O3 (3 per cent) was 5.90, 5.96 and 6.03 per cent respectively. There was
significant (P<0.05) difference in the protein content of all experimental
samples. Control Shrikhand samples had significantly (P<0.05) lower protein
content compared to all the other samples. Whereas sample O3 had
significantly (P<0.05) higher protein content. This might be due to
constitution of protein content of oat fiber.
Sucrose
It can be seen from the Table 4.5 that the average sucrose content of
control Shrikhand sample was found 41.43 per cent, Whereas sucrose
content of fiber containing oat powder Shrikhand samples O1 (1 per cent), O2
(2 per cent) and O3 (3 per cent) were 41.40, 41.41 and 41.43 per cent
respectively. The variation in the total sucrose content of all experimental
samples were noticed non-significant (P>0.05).
Ash
As shown in Table 4.5 the average ash content of control Shrikhand C

sample was 0.57 per cent, Whereas ash content of fiber containing oat
powder Shrikhand samples O1 (1 per cent), O2 (2 per cent) and O3 (3 per cent)
were 0.65, 0.68 and 0.71 per cent respectively. Sample O3 had significantly
(P<0.05) higher 0.71 per cent value. There was significant (P<0.05) difference
in ash content of all experimental Shrikhand samples. However, O1 and O2
and O2 and O3 had statistically no difference (P>0.05) in their ash content.
The increased in ash content might due to the oat powder containing good
amounts of minerals contents (Appendix IV).
Carbohydrate

4.5. Carbohydrate content was determined by difference. The tabulated value
of fiber containing oat powder Shrikhand sample O1, O2 and O3 were 44.70,
Page | 82
44.76 and 44.82 per cent respectively. The value of control Shrikhand was
44.63 per cent which showed significantly (P<0.05) lower value than other
sample. All experimental samples were significantly (P<0.05) different.

addition of oat powder are not available for comparison, however the values
observed in the present study are very close to those reported by (Aneja et
al., 2002, Patel, 2013).
4.2.2.2 Effect of varying levels of oat powder on the physico-chemical


experimental Shrikhand samples are tabulated in Table 4.6.
Titratable acidity
control Shrikhand sample was 1 per cent LA, while for oat powder containing
Shrikhand samples viz., O1 (1 per cent), O2 (2 per cent) and O3 (3 per cent)
these values were0.99, 0.98 and 0.97 per cent LA respectively. Control
Shrikhand had significantly (P<0.05) higher acidity value i.e. 1.0 per cent LA
while sample O3 had lower acidity value 0.97 per cent LA. However, sample
O1, O2 and O3 did not show any statistical significant (P>0.05) difference in
their titratable acidity content.
Free fatty acids
The free fatty acids are expressed as per cent oleic acid (µ.eq/g). It can
be seen from the Table 4.6 that Shrikhand without fiber content of oat
powder (control) had average free fatty acids content of 0.60 µ.eq/g. Whereas
for oat powder containing Shrikhand samples, free fatty acids contents were
O1 (1 per cent), O2 (2 per cent) and O3 (3 per cent) had 0.62, 0.63 and 0.66
µ.eq/g respectively. Control C Shrikhand had lower FFA value i.e.
Page | 83
0.60µ.eq/g. However, not all samples showed any statistical significant

difference in their free fatty acids content.
Table 4.6: Effect of varying levels of oat powder on the physico-


Treatments (%LA) µ.eq/g nitrogen activity (N)
(%)
Control – C 1.00±0.01 0.60±0.03 0.27±0.08 0.9391±0.01 4.33±0.05 8.48±0.04
O1 0.99±0.01 0.62±0.02 0.29±0.09 0.9390±0.01 4.35±0.06 8.54±0.01
O2 0.98±0.01 0.63±0.02 0.26±0.10 0.9380±0.01 4.36±0.07 8.63±0.01
O3 0.97±0.01 0.66±0.04 0.27±0.06 0.9367±0.01 4.37±0.05 8.65±0.01
SEm 0.004 0.014 0.042 0.003 0.028 0.010
CD (0.05) 0.014 NS NS NS NS 0.032
C.V.% 0.906 4.514 30.857 0.607 1.305 0.242
Soluble nitrogen
The average soluble nitrogen content is depicted in Table 4.6. The

soluble nitrogen content of fiber containing oat powder Shrikhand samples
were O1 (0.29 per cent), O2 (0.26 per cent) and O3 (0.27 per cent) respectively.
While control Shrikhand sample had 0.27 per centsoluble nitrogen content.
The variation in soluble nitrogen content of all experimental samples were
found non-significant (P>0.05).
Page | 84
Water activity
It can be seen from the Table 4.6 that the water activity of fiber
containing oat powder Shrikhand samples O1 (1 per cent), O2 (2 per cent) and
O3 (3 per cent) were 0.9390, 0.9380, 0.9367 respectively. Whereas control
Shrikhand had average water activity 0.9391. The difference in water activity
among all experimental Shrikhand samples and control sample were not
Consistency
The value for the consistency of experimental Shrikhand samples was

measured by texture analyzer and expressed as N. The fiber containing
oat powder Shrikhand sample O3 had highest value 8.65 N followed by O2
(8.63 N) and O1 (8.54 N). Whereas sample C had significantly (P<0.05) lowest
value i.e.8.48 N. The difference of consistency among all experimental
Shrikhand samples was found significant (P<0.05). It might be due to
increasing rate of addition of powder in Shrikhand. However O2 and O3were
found statistically similar in their consistency .
pH
The pH in fiber containing oat powder Shrikhand samples were O1

(4.35), O2 (4.36) and O3 (4.37). While control Shrikhand sample had pH value
4.33. However, the variation in pH of all experimental samples were noticed
statistically non-significant (P>0.05).
Published data on proximate physico chemical and consistency of

Shrikhand as affected by addition of oat are not available for comparison.
4.2.2.3 Effect of varying levels of fiber containing oat powder on the

sensory attributes of Shrikhand
The sensory quality of the experimental Shrikhand samples was

evaluated using score card shown in Appendix-I. The sensory qualities of the
Page | 85
experimental Shrikhand samples were evaluated by expert judges. Average

value of flavour, body and texture, colour and appearance and total score
given by judges are portrayed in Table. 4.7.
Flavor
The flavor of a dairy product is the most important organoleptic

attribute upon which rests its acceptability among the consumers. It is
evident from the tabulated values that flavour score was significantly
(P<0.05) influenced by addition of fiber containing oat powder.
The flavor scores of experimental Shrikhand depicted in Table 4.7. The

flavor scores of Shrikhand prepared using supplementation of fiber
containing oat powder were O1 (43.98), O2 (43.85) and O3 (42.39). It was
observed that for control sample, the flavor score 44.05 was significantly
(P<0.05) higher as compared to all experimental samples. While O1 and O2
did not have any statistical significant difference in flavor scores. The
difference in flavor score of experimental Shrikhand samples may be due to
increasing rate of addition of oat powder, which may reduce the flavor score
of experimental Shrikhand.
Body and texture
It can be seen from Table 4.7 that the average body and texture score
of fiber containing oat powder Shrikhand samples were O1 (32.46), O2 (32.91)
and O3 (31.94). While control Shrikhand sample had body and texture score
31.64, which was significantly (P<0.05) lower from all three Shrikhand
samples. It might be due to increasing rate of addition of fiber in Shrikhand.
The variation in body and texture score of O1 and O3 experimental samples
were significant (P<0.05). The experimental sample C was criticized as weak
body and O3 was criticized as grainy texture by judges. O1 and O2 had
statistically no difference in their body and texture score.
Page | 86
Table4.7: Effect of varying levels of fiber containing oat powder on the

sensory attributes of Shrikhand
Sensory characteristics
Treatments Flavor (50) Body &texture Color& Total score*

(35) appearance (100)
(10)
Control - C 44.05±0.44 31.64±0.35 9.06±0.31 89.75±0.88
O1 43.98±0.06 32.46±0.34 8.64±0.32 90.08±0.45
O2 43.85±0.17 32.91±0.44 8.55±0.29 90.31±0.66
O3 42.39±0.22 31.94±0.25 7.96±0.17 87.29±0.63
SEm 0.130 0.176 0.139 0.326
CD (0.05) 0.401 0.541 0.430 1.005
C.V.% 0.597 1.089 3.260 2.679

Colour and appearance
It can be seen from the Table 4.7 that control Shrikhand had average
color and appearance scores of 9.06; this value is significantly higher than
Shrikhand containing O1, O2 and O3 oat powder. These values were 8.64,
8.55 and 7.96 for fiber fortified O1, O2 and O3samples respectively. The
variation in color and appearance score of all experimental samples were
found to be significant (P<0.05).
Total score

characteristics. The total score of experimental Shrikhand is depicted in
Page | 87
Table 4.7. Control Shrikhand had average total score was 89.75. The total
score of fiber containing oat powder Shrikhand samples O1 (90.08), O2
(90.31) and O3 (87.29). However, O1, O2 and control samples had statistically
no difference in their total scores. This was because of supplementation of
fiber containing oat powder in Shrikhand.

affected by addition of oat powder are not available for comparison.
containing oat powder (O2) Shrikhand had superior quality in term of
chemical, physicochemical and sensory attributes. Therefore, addition of 2
per cent oat powder in Shrikhand was selected for further study. As per total
score, experimental Shrikhand sample is given preference score of O2 > O1>
C > O3.
4.2.3 Optimization of rate of addition of inulin powder
The experimentation involved making four batches of Shrikhand viz.

C, I1 (1 per cent), I2 (2 per cent) and I3 (3 per cent) based on expected yield of
Shrikhand. Each treatment was replicated four times. In all 16 batches of
4.2.3.1 Effect of varying levels of fiber containing inulin powder on the

The data on chemical composition of experimental Shrikhand samples

are presented in Table 4.8.
Total solids content
It can be seen from the Table 4.8, that the average total solids content
of control Shrikhand (C) averaged 58.28 per cent. While for fiber containing
inulin powder fortified in Shrikhand samples, total solids contents were
58.67, 59.03 and 59.40 per cent for samples I1, I2 and I3 respectively. There
Page | 88
was significant difference (P<0.05) in total solids content of Shrikhand

samples. Control Shrikhand had significantly (P<0.05) lower i.e. 58.28per
cent total solids content as compared to all other fiber fortified Shrikhand
samples. Whereas sample I 3 containing 3 per cent inulin powder had a
significantly (P<0.05) highest total solid (59.40 per cent). Increasing rate of
addition of inulin resulted in increased total solids content of Shrikhand.
Table 4.8: Effect of varying levels of inulin powder on the composition

of Shrikhand
Composition (%)
Treatments
solids
Control – C 58.28±0.05 6.23±0.02 5.80±0.01 41.42±0.01 0.57±0.02 44.63±0.01
I1 58.67±0.03 6.19±0.04 5.73±0.03 41.42±0.01 0.63±0.03 44.23±0.03
I2 59.03±0.05 6.10±0.05 5.69±0.04 41.44±0.01 0.64±0.05 43.85±0.06
I3 59.40±0.02 6.05±0.04 5.61±0.07 41.43±0.01 0.64±0.04 43.49±0.03
SEm 0.021 0.019 0.022 0.005 0.018 0.021
CD (0.05) 0.063 0.059 0.066 NS NS 0.065
C.V.% 0.070 0.623 0.755 0.026 5.815 0.095
Fat
The percent fat content of inulin powder containing Shrikhand is given

in Table 4.8. The average fat content of control Shrikhand sample was 6.23
per cent, while for fiber containing inulin powder Shrikhand samples were
6.19, 6.10 and 6.05 per cent for I1, I2 and I3 samples respectively. There was
significant (P<0.05) difference in fat content of control sample and fiber
containing Inulin powder Shrikhand samples. However, samples I2 and I3
had statistically no difference (P>0.05) in their fat content. It can be seen
Page | 89
from tabulated value that there was a slight decrease in fat content with
increase in rate of addition of fiber containing powder.
Protein
It can be seen from Table 4.8 that the average protein content of
control (C) Shrikhand sample was 5.80 per cent, Whereas protein content of
fiber containing inulin powder Shrikhand samples were I1 (1 per cent), I2 (2
per cent) and I3 (3 per cent) were 5.73, 5.69 and 5.61 per cent respectively.
There was statistically significant (P<0.05) difference in the protein content
of all experimental samples.
Sucrose
It can be seen from Table 4.8 that the average sucrose content of
control Shrikhand sample was 41.42 per cent, Whereas sucrose content of
fiber containing inulin powder Shrikhand samples I1 (1 per cent), I2 (2 per
cent) and I3 (3 per cent) were 41.42, 41.44 and 41.43 respectively.The
variation in the total sucrose content of all experimental samples were found
statistically non-significant (P>0.05).
Ash
The average ash content of control Shrikhand C sample was 0.57 per
cent (Table 4.8). The ash content of fiber containing inulin powder Shrikhand
samples I1 (1 per cent), I2 (2 per cent) and I3 (3 per cent) were 0.63, 0.64 and
0.64per cent respectively. There was no significant (P>0.05) difference in ash
content of all experimental Shrikhand samples.
Carbohydrate

4.8. Carbohydrate content was determined by difference. The carbohydrate
content of fiber containing inulin powder Shrikhand sample I1, I2 and I3 were
44.23, 43.85 and 43.49 per cent respectively. The control Shrikhand had
Page | 90
44.63 per cent carbohydrate, which showed significantly (P<0.05) higher

value than other sample. All experimental samples were significantly
(P<0.05) different from each other.

addition of inulin are not available for comparison, however the values
observed in the present study are very close to those reported by (Aneja et
al., 2002, Patel, 2013).
4.2.3.2 Effect of varying levels of fiber containing inulin powder on the

physico-chemical properties and consistency of Shrikhand

experimental Shrikhand samples are depicted in Table 4.9.
Titratable acidity
without fiber containing inulin powder (control) Shrikhand sample was 1.0
per cent LA, while for inulin powder containing Shrikhand samples, these
values were I1 (1 per cent), I2 (2 per cent) and I3 (3 per cent) had 0.99, 0.98
and 0.97 per cent LA respectively. Control Shrikhand had higher acidity
value i.e. 1.00 per cent LA.The variation titratable acidity (per cent LA)
content of all experimental samples were found statistically not significant
(P<0.05).
Free fatty acids
The free fatty acids are expressed as per cent oleic acid (µ.eq/g). It can
be seen from the Table 4.9 that Shrikhand without fiber content of inulin
powder (control) had average free fatty acids content of 0.55 µ.eq/g. Whereas
for inulin powder containing Shrikhand samples viz. I1 (1 per cent), I2 (2 per
cent) and I3 (3 per cent) these were 0.59, 0.61 and 0.62 µ.eq/g respectively.
Control C Shrikhand had lower FFA value i.e. 0.55 µ.eq/g. However, sample
Page | 91
I1, I2 and I3did not show any statistical significant difference in their free fatty
acids content. The variation in FFA (µ.eq/g) content of all experimental
samples were found non-significant (P>0.05).
Table 4.9: Effect of varying levels of fiber containing inulin powder on

the physico-chemical properties and consistency of Shrikhand
Treatments Soluble
Acidity FFA Water Consistency
nitrogen pH
(%LA) µ.eq/g activity (N)
(%)
Control – C 1.00±0.02 0.55±0.02 0.21±0.04 0.9375±0.01 4.38±0.05 8.37±0.06
I1 0.99±0.02 0.59±0.05 0.22±0.02 0.9372±0.01 4.35±0.06 8.51±0.04
I2 0.98±0.02 0.61±0.03 0.24±0.01 0.9369±0.01 4.33±0.05 8.58±0.07
I3 0.97±0.03 0.62±0.04 0.25±0.02 0.9365±0.01 4.32±0.05 8.62±0.08
SEm 0.010 0.018 0.013 0.003 0.026 0.033
CD (0.05) NS NS NS NS NS 0.101
C.V.% 2.076 5.945 11.097 0.573 1.198 0.772
Soluble nitrogen
The soluble nitrogen content of fiber containing inulin powder

Shrikhand samples I1 (0.22 per cent), I2 (0.24 per cent) and I3 (0.25 per cent)
is shown in Table 4.9. While control Shrikhand sample had soluble nitrogen
content of 0.21per cent. The difference of soluble nitrogen among all
experimental Shrikhand samples were statisticallynon significant (P>0.05).
Page | 92
Water activity
It can be seen from Table 4.9 that the water activity of fiber containing
inulin powder Shrikhand samples were I1 (1 per cent), I2 (2 per cent) and I3
(3 per cent) were 0.9372, 0.9369, 0.9365 respectively. Whereas Shrikhand
without fiber content of inulin powder (control) had average water activity of
0.9375. The difference in water activity among all experimental Shrikhand
samples were statistically non-significant (P>0.05).
pH
The pH content in fiber containing inulin powder Shrikhand samples

were I1 (4.35), I2 (4.33) and I3 (4.32). While control Shrikhand sample had a
pH of 4.38. However, the variation in pH content of all experimental samples
were noticed non-significant (P>0.05).
Consistency
The value for the consistency of experimental Shrikhand samples was

measured by texture analyzer, it is expressed in N. The fiber containing
inulin powder Shrikhand sample I3 had significantly (P<0.05) highest value
8.62 N followed by I2 (8.58 N) and I1 (8.51 N). Whereas sample C had
significantly (P<0.05) lowest value i.e.8.37N. The difference of consistency
among all experimental Shrikhand samples was found significant (P<0.05). It
was might be due to increasing rate of addition of inulin powder in
Shrikhand. However there was no statistically difference (P>0.05) observed in
sample I1, I2 and I2 & I3.
Published data on proximate physico chemical properties and

consistency of Shrikhand as affected by addition of inulin powder are not
available for comparison.
Page | 93
4.2.3.3 Effect of varying levels of inulin powder on the sensory

attributes of Shrikhand
The effect of varying levels of inulin powder on the sensory attributes

of Shrikhand shown in Table 4.10. The sensory quality of the experimental
Shrikhand sample were evaluated using a score-card suggested by Indian
standards (Appendix-I).
Flavor
The flavor scores of experimental Shrikhand depicted in Table 4.10

varied from 45.50 (C) to 45.64 (I2). The flavor scores of experimental
Shrikhand prepared using supplementation of fiber containing inulin powder
Shrikhand were I1 (45.53), I2 (45.64) and I3 (45.60). While, for control
Shrikhand score was 45.50. The average flavor score for, control sample C
was lower than all three fiber containing Shrikhand samples. However, the
variation in flavor score of all experimental samples were found non-
Body and texture
The average body and texture scores of inulin powder containing

Shrikhand given in Table 4.10. The body and texture scores of sample I1, I2
and I3 were 33.14, 33.48 and 33.52 respectively. The control Shrikhand
sample scored 32.01 which was significantly (P<0.05) lower than all three
Shrikhand samples. The variation in body and texture score of all
experimental samples were found not significant (P>0.05).
Page | 94
Table 4.10: Effect of varying levels of inulin powder on the sensory

Treatments
Flavor (50) Body &texture Color& Total score*
(35) appearance (100)
(10)
Control – C 45.50±0.09 32.01±0.55 9.35±0.18 91.86±0.73
I 1% 45.53±0.19 33.14±045 9.48±0.24 93.15±0.52
I 2% 45.64±0.23 33.48±0.56 9.48±0.20 93.60±0.66
I3% 45.60±0.24 33.52±0.52 9.48±0.21 93.60±0.57
SEM 0.098 0.262 0.104 0.384
CD (0.05) NS 0.807 NS 1.182
C.V.% 0.430 1.585 2.195 2.456

The color and appearance scores of the Shrikhand samples, as

presented in Table 4.10 revealed non-significant differences amongst
samples. Fiber fortifiedsamplesI1, I2 and I3 were given the scores 9.48, 9.48,
9.48 respectively while for the Shrikhand sample without fiber fortification
was 9.35. Addition of inulin powder in Shrikhand resulted in brighter and
whiter appearance in inulin-fortified samples compared to control. The
variation in color and appearance score of all experimental samples were
observed non-significant (P>0.05).
Page | 95
Total score
The total score of experimental Shrikhand depicted in Table 4.10 was

given out of 100 and indicated that total scores ranged from I1 (93.15) to I3
(93.60). Control C sample had lower total score (91.86) than experimental
Shrikhand. Among all experimental Shrikhand samples I2 and I3 had
significantly (P<0.05) higher total score compared to I 1 and control.
Statistically samples I1, I2 and I3 were not significantly (P>0.05) different in
total score from each other. This may be due to experimental Shrikhand
supplement with fiber containing inulin powder resulting in smoother
product with glossy appearance.
It is evident that there was no significant difference amongst all levels

tried for total score. Due to higher cost of Inulin as ingredient, lower rate of
addition (1 per cent) was selected for further study.

affected by addition of inulin powder are not available for comparison.
From this part of the study, it is observed that Shrikhand sample I1,
had superior quality in terms of chemical and physico – chemical attributes.
As per total score, experimental Shrikhand sample is given preference score
of I1 > I2> I3> C.
4.2.4 FIBER CONTENTS INDIFFERENT EXPERIMENTAL SHRIKHAND
Crude fiber of different experimental Shrikhand were estimated by

official method described in Analysis of the Association of Official Analytical
Chemist (AOAC, 1990), which are depicted in Table 4.11.
Page | 96
Table4.11: Fiber content in different Shrikhand samples
Treatments (%)Crude fiber
B1 0.03%
B2 0.08%
B3 0.13%
O1 0.12%
O2 0.24%
O3 0.36%
I1 0.92%
I2 1.84%
I3 2.8%
The fiber content in Shrikhand samples, as depicted in Table 4.11 was

analyzed to be 0.03 per cent in B1(1 per cent), 0.08 per cent in B2 (3 per
cent), 0.13 per cent in B3 (5 per cent), 0.12 per cent in O1(1 per cent), 0.24
per cent in O2 (2 per cent) and 0.36 per cent in O3 (3 per cent) and 0.92 per
cent in I1 (1 per cent), 1.84 per cent in I 2 (2 per cent), 2.8 per cent in I 3 (3 per
cent). With increasing the rate of addition of fiber, an increasing fiber
content in samples Shrikhand that was observed. Samples fortified with
inulin powder had higher fiber contents because of presence of higher fiber
content in inulin powder i.e. 92 per cent (Appendix IV).
4.3 PHASE-III: COMPARISON OF SHRIKHAND CONTAINING OPTIMIZED

LEVEL OF EACH FIBER CONTAINING POWDER TO SELECT THE BEST
FIBER CONTAINING POWDER IN FIBER FORTIFIED SHRIKHAND
Selection was done with objective of determining the different rate of

different fiber level that would lead to the most acceptable product in terms
of sensory scores, compositional attributes, textural attributes and physico-
chemical attributes. Based on result obtained in Section 4.2.1, 4.2.2 and
Page | 97
4.2.3 Shrikhand prepared from using banana powder (B3 at rate of 3 per
cent) oat powder (O2 at rate of 2 per cent) and from inulin (I1 at rate of 1 per
cent) were selected and the chemical composition, physico chemical
properties, sensory attributes consistency and microbial quality were
compared with control (C) in this part of the study.
4.3.1 Chemical composition
The data on average composition of fresh experimental Shrikhand

sample are presented in Table 4.12.
Total solids
The average total solids content of control Shrikhand (C), as shown in

Table 4.12, was 58.28 percent, while for fiber fortified Shrikhand samples I1,
O2 and B3 contained 58.67, 59.01 and 60.03 per cent total solids
respectively. There was significant difference (p<0.05) in total solid content of
all samples. It may be due to fortification of powder containing fiber in
Shrikhand.
Fat
The average fat content of control Shrikhand sample was 6.23 percent,
while for experimental fiber fortified probiotic Shrikhand it was I1 (6.19), O2
(6.37) and B3 (5.72) per cent (Table 4.12). There was significant (P<0.05)
difference in the fat content of control Shrikhand and fiber fortified
Shrikhand made using oat and banana powder. However, I1 and control were
not significantly (P>0.05) different from each other.
Page | 98
Table 4.12: Average composition of fiber fortified probiotic Shrikhand

Treatments
solids
Control - C 58.28±0.03 6.23±0.03 5.80±0.01 41.45±0.04 0.57±0.01 44.63±0.01
I1 58.67±0.03 6.20±0.04 5.73±0.03 41.44±0.04 0.63±0.03 44.23±0.03
O2 59.01±0.03 6.37±0.01 5.96±0.01 41.47±0.04 0.68±0.01 44.76±0.02
B3 60.03±0.05 5.72±0.04 5.76±0.02 41.45±0.03 0.71±0.02 46.44±0.03
SEm 0.017 0.015 0.009 0.019 0.009 0.015
CD (0.05) 0.053 0.048 0.029 NS 0.028 0.047
C.V.% 0.058 0.505 0.325 0.089 2.7.09 0.067
Protein
It can be seen from the Table 4.12 that protein content of the control C
Shrikhand was 5.80 per cent. While in experimental fiber fortified Shrikhand,
it was I1 (5.73 per cent), O2 (5.96 per cent) and B3 (5.76 per cent). Sample O2
(5.96 per cent) had significantly (P<0.05) higher protein content compared to
I1 and I3 samples. This is because oat powder had good amount of protein
(Appendix IV). Tabulated value showed significant (P<0.05) difference in all
experimental Shrikhand samples, whereas sample I 1 and B3were found
statistically not significant (P>0.05).
Sucrose
The total sucrose content of control Shrikhand averaged 41.45 per

cent, whereas in fiber fortified samples it was I 1 (41.44 per cent), O2 (41.47
Page | 99
per cent), B3 (41.45 per cent). The variation in sucrose content of all samples
were noticed non-significant (P>0.05) (Table 4.12).
Ash
The average ash content of control sample was C (0.57 per cent) and
fiber fortified Shrikhand sample were I1 (0.63 per cent), O2 (0.68 per cent)
and B3 (0.71 per cent). Tabularized value showed significant (P<0.05)
difference in their ash content.
Carbohydrate
It can be seen from the Table 4.12 that the carbohydrate (by
difference) content of C Shrikhand was 44.63 percent. While fiber fortified
samples varied from 44.23 to 46.44 percent. There was significant (P<0.05)
difference in the carbohydrate content of control Shrikhand and fiber fortified
Shrikhand.
containing additives influenced the total solids, protein and ash content. It
may be because of supplementation of powder containing fiber to Shrikhand.
In fiber containing banana and oat powder having appreciable amount of
protein and carbohydrate, fat and mineral content tabulated values showed
significant (P<0.05) difference so ultimately it affects on total solids..
Whereas for sucrose it can be said that use of different fiber containing
powder supplement in Shrikhand showed non-significant (P>0.05) difference.
Comparative data on the use of fiber containing additives for Shrikhand
manufacturing are not available. However, the compositional values of all the
fiber-fortified probiotic Shrikhand prepared in the present study are
comparable to those reported by different research co-workers for Shrikhand
(Prajapati et al., 1992; Shukla et al., 2007 and Patel, 2013).
Page | 100
4.3.2 Physico -chemical properties and Consistency

experimental Shrikhand samples are depicted in Table 4.13
Acidity
The acidity was found to be 1.0 per cent LA. In control, Shrikhand
sample. While for fortified Shrikhand samples I1, O2 and B3, these were0.99,
0.98 and 0.95 per cent LA respectively (Table 4.13). There was significant
(P<0.05) difference observed in acidity of Shrikhand samples. However, the
acidity of control, I1 and O2 Shrikhand were statistically at par.
Table 4.13: Physico-chemical properties and Consistency of fiber

Treatments
(%LA) (µ.eq/g) nitrogen activity (N)
(%)
Control - C 1.0±0.02 0.53±0.01 0.29±0.06 0.939±0.01 4.33±0.03 8.45±0.12
I1 0.99±0.02 0.56±0.03 0.28±0.05 0.939±0.01 4.34±0.03 8.53±0.12
O2 0.98±0.02 0.58±0.05 0.27±0.06 0.938±0.01 4.36±0.01 8.65±0.10
B3 00.95±0.02 0.58±0.04 0.29±0.04 0.937±0.01 4.38±0.02 8.73±0.12
SEm 0.010 0.017 0.026 0.003 0.012 0.060
CD (0.05) 0.032 NS NS NS NS 0.186
C.V.% 2.104 6.21 18.27 0.56 0.550 1.406
Page | 101
Free fatty acids
It can be seen from the Table 4.13 that the FFA values of experimental
samples are expressed in µ.eq/g, which were C (0.53), I1 (0.56), O2 (0.58) and
B3 (0.58) µ.eq/g. There is no significant (P>0.05) difference observed in the
free fatty acids content of control Shrikhand and fiber fortified Shrikhand.
Soluble nitrogen
The soluble nitrogen content in probiotic Shrikhand samples were C

(0.29 per cent) I1 (0.28 per cent), O2 (0.27 per cent) and B3 (0.29 per cent) as
shown in Table 4.13. The difference of soluble nitrogen among all
experimental Shrikhand samples were found statistically non-significant
(P>0.05).
Water activity
It is revealed from Table 4.13 that the water activity of experimental

Shrikhand samples varied from B3 (0.937) to C (0.939). Sample B 3 had lower
water activity as compared to all experimental samples. However the
difference of water activity among all experimental Shrikhand samples were
found non-significant (P>0.05).
pH
The pH was found to be 4.33 in control Shrikhand sample. While for

fortified Shrikhand samples I1, O2 and B3, it was 4.34, 4.36 and 4.38
respectively, (Table 4.13). There was no significant (P>0.05) difference
observed in pH values of all Shrikhand samples.
Consistency
Hardness is the peak force that results from being compressed to a

given distance, time or percent of deformation. The consistency of the
Shrikhand is an important criterion to measure the body and texture
Page | 102
characteristics such as firmness/weakness of the Shrikhand. In this study,

consistency was measured by food texture analyzer and is expressed in N.
It can be seen from Table 4.13 that the consistency of experimental

Shrikhand ranged from 8.45 to 8.73 N. B3 sample had the highest (8.73 N)
value but based on sensory evaluation the body and texture were not
satisfactory and thus, given a lower score due to excessively firm body (Table
4.14). Sample C was less consistent and thus resulted in a weak body, which
fetched a lower score on the sensory scale. Sample I 1 and O2 were
statistically at par and were optimally consistent. The variation in
consistency was found to be significant (P<0.05).
Supporting data on this aspect of study is not available for

comparison. However, the values of physico-chemical properties of
Shrikhand obtained in present study are comparable to those reported by
Patel (2013). There are no available data of consistency (N) of Shrikhand as
analyzed by texture analyzer for comparison.
4.3.3 Sensory attributes
Fiber fortified probiotic Shrikhand samples were subjected to judging

for various sensory attributes viz. (i) flavor (ii) body and texture (iii) color and
appearance and (iv) total score. The data obtained while analyzing the fiber-
fortified probiotic Shrikhand samples are represented in Table 4.14. The
sensory quality of experimental Shrikhand samples was evaluated using a
scorecard shown in Appendix-1.
Flavor
The flavor of any food product including Shrikhand is most important

sensory attributes, as far as consumer preference is concerned because it
determines its acceptance and rejection. The flavor scores of experimental
Shrikhand (Table 4.14) varied from 44.07 to 46.14. The flavor scores of
experimental Shrikhand prepared using supplementation of powder
Page | 103
containing fiber ingredients along with sucrose and cream, it was observed
that for freshly made product, the flavor score was significantly (P<0.05)
higher for sample I 1 (46.14) following O2 (46.11), B3 (44.39) and C (44.07). As
far as the fresh product samples average flavor score was concerned, control
sample C was significantly (P<0.05) lower from all three Shrikhand samples,
while sample I1 and O2 were statistically at par (P>0.05).
The difference in flavor score of experimental Shrikhand samples may

be due to different rate of addition of powder containing fibers, which alter
the flavor score of experimental Shrikhand.
Table 4.14: Sensory characteristics of fresh fiber fortified probiotic

Shrikhand
Body Color & Total score*

Treatments Flavor (50) &texture appearance (10) (100)
(35)
Control - C 44.07±0.64 31.50±0.32 9.17±0.24 89.74±0.98
I1 46.14±0.82 32.64±0.63 9.48±0.44 93.27±1.74
O2 46.11±0.91 33.25±0.80 9.46±0.44 93.82±2.05
B3 44.39±0.27 30.93±0.25 7.17±0.52 87.67±0.13
SEm 0.353 0.278 0.213 0.717
CD (0.05) 1.08 0.86 0.66 2.21
CV (%) 1.56 1.73 4.79 1.67

Page | 104
Body and texture
It can be revealed from Table 4.14 that the score for body and texture of
experimental Shrikhand samples ranged from C (31.50) to O 2 (33.25). Body
and texture score of I1 and O2 samples were significantly (P<0.05) higher
than control. Among experimental Shrikhand sample scores of body and
texture B3 was significantly (P<0.05) lower than all other samples. This
clearly indicates the supplementation of fiber containing banana powder in
the current investigation had positive impact on body and texture of samples
I1 and O2 were statistically (P<0.05) at par. While B3 sample was criticized as
excessively firm body and sample C was criticized as weak body as compared
to other samples by judges.
It can be seen from Table 4.14 that the color and appearance score of
experimental Shrikhand samples ranged from 7.17 to 9.48. The experimental
sample I1and O2had significantly (P<0.05) high score compared to B3 sample.
However, I1 and O2 were yielded statistically similar to color and appearance
of the Shrikhand. It clearly indicates the supplementation of fiber containing
powder in the current investigation resulted in improved color and
appearance scores. The tabulated value show color and appearance of
samples I1 and O2 were statistically at par (P>0.05).
Total score

characteristics. The total score of experimental Shrikhand depicted in Table
4.14 indicated that the values ranged from B3 (87.67) to O2 (93.82). B3
sample had significantly lower acceptability score. Among all experimental
Shrikhand O2 had significantly (P<0.05) higher acceptability score
indicating superior acceptability among experimental samples. The
Page | 105
Shrikhand samples I1 and O2 were found statistically at par (P>0.05). As per

total score, experimental Shrikhand sample is given preference score of O2 >I1
>B3> C
Thus, addition of fiber containing oat powder at the rate of 2 per cent
was found most suitable to get good quality of probiotic Shrikhand.
4.3.4 Microbiological analysis
The microbiological quality of experimental Shrikhand samples are

measure as log cfu/g in terms of probiotic count, standard plate count, yeast
and mold and coliform count are depicted in Table 4.15.
Probiotic count
All the four freshly made Shrikhand samples had probiotic count
varying from 24 to 29 × 107cfu / ml (log value 8.397 to 8.475), which were
statistically significant (P<0.05). Probiotic culture used in experiment had
higher count than minimum count (6.00 log cfu/g) to state that the product
as probiotic. Tabularized value showed that sample C had significantly
(P<0.05) lower counts, while sample I1 had significantly (P<0.05) higher
counts, it may be due to supplementation of inulin and oat beta-glucan as
prebiotic. A prebiotic is a selectively fermented ingredient that promotes
specific changes in the composition and/or activity of the gastrointestinal
micro biota, which, in turn, confers benefits on host well-being, and health
(Pasquale et al., 2012). Sample I1 and O2 were statistically at par and were
optimally consistent. The variation in probiotic count was found to be
significant (P<0.05).
Standard plate count
The standard plate count (SPC) of experimental fresh Shrikhand

samples was expressed in log cfu/g. The SPC count of experimental fresh
Shrikhand samples varied from 1.354 to 1.402, which showed in Table 4.15.
Page | 106
The SPC count of all experimental Shrikhand samples had showed non-
significant (P>0.05) difference.
Yeasts and molds count
Yeasts and molds perhaps, are one of the most important groups of
microbes present among several other groups of spoilage micro flora in
fermented milk products, including Shrikhand. Yeasts and molds are mainly
contaminated to the product sample through air (Tamime and Robinson,
1999).
Shrikhand samples were found free from yeast and mould in fresh
product samples (Table 4.15). Patel, (2013) studied the manufacture of
probiotic Shrikhand and found that the yeast and mold count ranged from
1.549 to 1.621 log cfu/g, which was within specified limit of FSSA (2011) i.e.
max.50/g.
Table 4.15 Microbiological quality of fiber fortified probiotic Shrikhand
Microbiological analysis (log cfu/g)
Treatments Probiotic Count Standard Plate Yeast & Mold Coli form Count
Count Count
C 8.397±0.02 5.233±0.04 Nil Nil
I1 8.475±0.03 5.274±0.05 Nil Nil
O2 8.460±0.02 5.272±0.06 Nil Nil
B3 8.411±0.01 5.243±0.02 Nil Nil
SEm 0.010 0.022 - -
CD (0.05) 0.03 NS - -
C.V.% 0.24 0.84 - -
Page | 107
Coliform count
The presences of coliform bacteria in dairy products are suggestive of

insanitary conditions or practice followed during production, processing and
storage (Speck, 1984). The confirmation of presence of fecal coliforms in the
products, further, indicates probable and alarming presence of potent
human pathogens too. In the present work to adjudge mainly, the extent of
sanitary practice followed during manufacturing, or otherwise, the coliform
count of control along with experimental Shrikhand samples were carried
out.
It was found that during entire course of study, the coliforms in all the
four fresh experimental Shrikhand samples were absent in 1gm Shrikhand
sample. Absence of coliform count indicates that the hygienic condition
adapted during manufacturing as well as the sanitation of the cups carried
out using hypochlorite solution was adequate. The results were in
confirmation with the coliforms count carried out by some workers for
Shrikhand samples. When Shrikhand was prepared using mixed cultures of
Streptococcus lactis and Streptococcus lactissubsp. diacetylactis. Upadhyay
et al. (1984) found that coliform count was absence in 0.1 g of Shrikhand
sample, which was within specified limit of FSSA (2011) i.e. max. 50/g.
Optimization was done with objective of determining the final fiber

containing powder with rate of addition that would lead to the most
acceptable product in terms of sensory scores, compositional attributes,
textural i.e. consistency and physico-chemical as well as microbiological
attributes. As per total score, experimental Shrikhand sample is given
preference score of O2 >I1 >B3> C.
Page | 108
4.4 PHASE - IV: SELECTION OF FLAVOURINGINGRDIENTS FOR THE

FINAL PREPARED SHRIKHAND
This part of the study was conducted to select the most preferred
flavor for finalized fiber fortified probiotic Shrikhand from amongst two
flavouring ingredients viz. cardamom and rajbhog. Shrikhand was prepared
according to the procedure described in Figure 3.1. Oat powder was added to
chakka at the rate of 2 per cent by weight. The two flavours viz., cardamom
and rajbhog were selected based on preliminary screening. Rate of addition
of cardamom as flavouring substance was 0.2 percent by weight of
Shrikhand. The rate of addition was decided based on preliminary trials.
Swapna and Suvarna (2013) added cardamom at much higher rate @1.6
percent by weight of chakka during preparation of Shrikhand. Rajbhog
flavour was added @ 0.05 percent by weight of Shrikhand. The flavouring
ingredients were added to Shrikhand after mixing cream with chakka and
passing it through mesh. In this Section the sensory profile of three
treatments viz., control (no flavour addition), cardamom and rajbhog
flavoured probiotic fiber fortified Shrikhand were compared and data
presented in Table 4.16.
Flavour
Flavor is most important sensory attributes, as far as consumer like is

concerned. Flavor is only the things that are enhancing affinity of consumers
towards food products. The flavor scores of experimental Shrikhand prepared
using supplementation of finalized powder containing fiber ingredient along
with flavor substance i.e. cardamom had significantly (P<0.05) highest score
(46.76), whereas control sample without flavor added had lowest score
(43.68). All three samples were significantly (P<0.05) different in their flavor
score.
Page | 109
Table 4.16 Selection of flavor substance for the final prepared

Shrikhand
Effect of varying flavor substances on sensory characteristics
Color&
Treatment Body & Total score*
Flavor (50) Appearance
Texture (35) (100)
(10)
Control– C 43.68±0.87 32.54±0.63 8.67±0.12 89.85±1.25
Cardamom 46.76±0.60 33.30±0.43 8.82±0.31 93.88±1.17
Rajbhog 44.62±0.71 33.30±0.49 8.53±0.33 91.45±0.88
SEm 0.329 0.234 0.122 0.497
CD(0.05) 1.01 NS NS 1.53
CV% 1.59 1.58 3.15 1.26

*A full mark was given for package i.e. 5 out of 5.
Body and Texture
It can be seen from tabulated values (Table 4.16) that the scores for
body and texture of Shrikhand samples varied from 32.54 to 33.30. The
highest body and texture score was obtained in cardamom containing
samples (33.30).While lowest body and texture score was noticed in control
samples (32.54). The variation in body and texture was found to be non-
Color and appearance is most appealing attribute of any food product.

Color and appearance of the product is the visual perception, which in turn,
defines product quality. The color and appearance of the samples ranged
Page | 110
from 8.53 to 8.82. The difference of color and appearance among all
experimental flavor substances samples were non-significant (P>0.05).
However, the cardamom used Shrikhand had the highest color and
appearance score (8.82).
Total score
It can be seen from table 4.16 that the total score of samples were
varied from 89.85 to 93.88. Shrikhand containing cardamom flavor had
significantly the highest total score i.e. 93.88, while control had the lowest
total score i.e.89.85. The difference of total score among all experimental
Shrikhand containing flavor substances were found significantly (P<0.05)
different.
Thus, it can be concluded that use of cardamom as flavoring

ingredient yields the oat fiber containing Shrikhand was superior with
respect to sensory properties.
4.4.1 PREPARATION OF OPTIMIZED FIBER FORTIFIED PROBIOTIC

SHRIKHAND
For manufacture of good quality of fiber-fortified probiotic Shrikhand,
the method of manufacture is developed from above study. For this purpose,
skim milk (0.1 per cent fat), cream (60 per cent fat), probiotic culture, sugar,
fiber containing oat powder and cardamom as flavoring substance were
used. Method of manufacture of fiber fortified probiotic Shrikhand is given in
Figure.4.1.
Page | 111
Skim milk (0.1% fat)
Heat treatment (90˚C/ 10 min)
Cooling to 39˚C
Inoculation with probiotic starter culture ABY-3 (@0.1g/lit)
Incubation (39°C/7 hr) (curd-0.7-0.8% LA)
Whey drainage (8-9 h)
Chakka
Addition of sugar (72.5% on DM of chakka) + 2% of oat powder
On basis of expected yield of Shrikhand
Addition of cream (6.2% fat in Shrikhand)
Mixing
Passing through mesh (no.30)
Shrikhand
Addition of flavoring ingredients (0.2 per cent cardamom powder)
Packing and Storage (7±2ºC)
Figure. 4.1 Method for fiber fortified probiotic Shrikhand
Page | 112
The developed product was analyzed for its gross composition, physico
chemical properties, consistency, sensory quality and microbiological profile.
The results are presented in Table 4.17.
It can be seen from Table 4.17 that the compositional parameters and
acidity are well within the range of reported literature (Patel, 2013). All the
compositional, parameters except ash on dry matter basis meet the legal
requirement specified for Shrikhand by FSSA (2011). The ash content of
developed fiber fortified probiotic Shrikhand was found to be 0.68 per cent
and total Ash (on dry matter basis), per cent by weight was 1.1. This value is
higher than the legal limit of 0.9 totals Ash (on dry matter basis). This could
be attributed to the high mineral content of oat fiber. (Appendix IV).
As seen in Table 4.17 the total score obtained by the product was
93.28. According grading system suggested by Indian standards (IS 15348,
2003) a product can be rated as excellent if the total scored exceed 90.00.
Hence the developed product can be considered as excellent product as per
Indian standards (IS: 15348, 2003) grading system.
According to IS 5402 (2002) and ISO:4833 (1991) a product can be

labeled as probiotic if it meets the minimum standard requirements of
probiotic organisms of one million cfu/g. The probiotic count of the
developed product is much higher than these requirements i.e. 8.46 log
cfu/g. This effect could be attributed to prebiotic effect of oat fiber. This
result is in corroboration with those obtained by Vasiljevic et al., (2007) who
also reported that of 0.5 per cent oat β-glucan to yoghurt resulted in
enhanced survival rate of probiotic bacteria.
Page | 113
Table 4.17 Gross composition of oat fiber fortified probiotic Shrikhand
Constraints Values
Composition
Total solids(%) 59.01
Fat(%) 6.37
Milk Fat (on dry matter basis), % by wt 10.79
Protein 5.72
Milk protein (on dry matter basis), % by wt 9.69
Carbohydrate 44.76
Sucrose 41.47
Sucrose (on dry matter basis), % by wt 70.27
Ash 0.68
Total Ash (on dry matter basis), % by wt 1.1
Fiber content 0.24
Physico-chemical properties and Consistency
Acidity (%LA) 0.98
Free Fatty Acids (µ.eq/g) 0.58
Soluble nitrogen (%) 0.27
Water activity (aw) 0.938
pH 4.36
Consistency (N) 8.65
Sensory Quality
Flavor (50) 46.11
Body & Texture (35) 33.25
Color& Appearance (10) 9.46
Package (5) 5.00
Total score (100) 93.82
Microbiological Profile
Probiotic Count(log cfu/g) 8.460
Standard Plate Count (log cfu/g) 5.272
Yeast &Mould Count (log cfu/g) Nil
Coliform count Nil
Page | 114
4.4.2: Estimation of cost of production of fiber fortified probiotic

Shrikhand
The cost of product is an important consideration for its
commercialization and booming marketing. The cost of any products
depends upon many variable factors like cost of raw materials; cost of
processing etc. the economic feasibility of manufacturing of Shrikhand was
also evaluated. An attempt is made here to determine the approximate cost
of Shrikhand as outlined in Table 4.18.
Table 4.18 Estimated cost of production of fiber fortified probiotic

Shrikhand (for 10 liters milk)
Raw materials required Quantity (kg) Price ( ) /kg Unit price ( )
(A)
Skim milk 10 32 320
Sugar 1.3 35 45.5
Probiotic starter culture 0.001 30550 30.55
Cream (60%fat) 0.318 300 95.4
Oat powder 0.0944 180 16.99
Cardamom 0.00944 1074 10.14
Total (A) 518.58

Processing cost (B)
+ 103.72
Packaging cost (C)
622.30
Total (A+B+C)
Quantity of Shrikhand 4.72kg / 10 liters of milk
Cost of Shrikhand per kg Rs131.84
Page | 115
Note:
 Processing cost and packaging cost: approx. 20 per cent of the cost of
Ingredients.
 Current market price of cardamom flavored Shrikhand – Rs 160 / kg
4.5 PHASE – V: STUDY OF THE SHELF LIFE OF PREPARED SHRIKHAND

AT REFRIGERATION TEMPERATURE (7 ±2 0C)
The nature of Shrikhand as like other dairy/food products during

storage is crucial for its commercial success. Shelf life of the product is most
important from manufacturing and consumer point of view. To make
Shrikhand as viable product, it should have sufficient shelf life. Being highly
acidic and intermediate moisture product and preserved by sugar in nature,
Shrikhand resists spoilage organisms for quite some time yet moulds and
other acid tolerant, organisms start proliferation after 4 to 5 d at room
temperature; Hence the product should preferably be stored at refrigeration
temperature. The growth of microorganisms brings about various changes in
product and spoils the taste of the product during storage. For the present
study the developed fiber fortified probiotic Shrikhand prepared according to
procedure described in Figure 4.1 was packed in polystyrene cups (sanitized
cups with hypochlorite solution having 150 ppm available chlorine) and
storage related changes were monitored at 7 d interval i.e. 0, 7,14, 21, 28
and 35 d of storage.
The fiber fortified Shrikhand packed in PS cups was stored at 7±2ºC

up to 35d. The packed product was analyzed for its titratable acidity, free
fatty acids, soluble nitrogen, water activity, consistency, pH, sensory
attributes and microbial quality (Standard Plate count, yeast and Mold
count, Probiotic count and Coliform Count) of stored samples of Shrikhand
were monitored at predetermined time intervals after every 7 d of refrigerated
storage or until the product was rejected on sensory basis. The results of
Page | 116
three replicates on storage related changes in Shrikhand are presented and

discussed in this section.
4.5.1 EFFECT OF STORAGE ON PHYSICO CHEMICAL AND

CONSISTENCY OF SHRIKHAND
Changes in titratable acidity
Shrikhand is fermented dairy product, specially known for its sweet –

sour taste. Titratable acidity of Shrikhand plays a very important role in
producing sweet – sour flavor of Shrikhand. The influence of storage period
on titratable acidity of Shrikhand sample during storage is collated in Table
4.20 and Figure 4.2.
Table 4.19 Effect of storage on physico- chemical properties and

consistency of Shrikhand
Physico - chemical properties

Storage Soluble
Acidity FFA Water Consistency
period (D) nitrogen pH
(%LA) (µ.eq/g) activity (N)
(%)
0 0.98±0.01 0.64±0.02 0.24±0.01 0.9391 4.37±0.05 7.81±0.01
7 1.02±0.01 0.80±0.01 0.27±0.03 0.9378 4.33±0.01 8.00±0.02
14 1.08±0.02 0.97±0.02 0.34±0.01 0.9369 4.23±0.01 8.21±0.02
21 1.17±0.02 1.05±0.02 0.37±0.01 0.9355 4.17±0.03 8.45±0.01
28 1.23±0.02 1.09±0.01 0.41±0.03 0.9334 4.12±0.02 8.88±0.01
35 1.33±0.02 1.13±0.03 0.44±0.01 0.9330 4.06±0.01 9.23±0.03
SEm 0.007 0.009 0.009 0.001 0.016 0.009
CD(0.05) 0.022 0.026 0.029 0.003 0.05 0.027
CV% 1.512 1.815 5.385 0.183 0.95 0.207
Page | 117
1.4
1.2
1
Acidity (%LA)
0.8
0.6 Acidity
0.4
0.2
0
0 7 14 21 28 35
Storage period (Days)
Figure 4.2: Influence of Storage period on the Titratable Acidity (% LA)

of Shrikhand at refrigerated temperature (7±2ºC)
It can be seen from tabulated values that storage period had a

significant (P<0.05) effect on lactic acid content of Shrikhand stored at
refrigeration temperature.
Fresh Shrikhand had the acidity value of 0.98 per cent LA which
increased significantly (P<0.05) to 1.33 per cent LA on 35 d of storage. The
observed increased acidity value might be due to lactose fermentation or
microbial changes in product during storage. A marked linear increased in
acidity of experimental Shrikhand samples during storage at refrigerated
temperature could also be attributed to presence of viable starter culture as
well as other adventitious organisms, giving greater biochemical activity in
the product.
No data on changes in acidity during storage periods are available for

comparison for fiber-fortified probiotic Shrikhand. However, Patel (2013)
reported the acidity of probiotic Shrikhand ranged from 1.068 to 1.123 per
cent LA on storage at refrigerated temperature (7±2ºC) for 28 d. Sharma and
Page | 118
Zariwal (1980) reported that acidity of Shrikhand was 1.38 to 1.50 per cent
LA on storage at refrigerated (10±3ºC) temperature for 45 d. While Prajapati
(1989) reported an increased acidity of Shrikhand from 1.22 to 1.46 per cent
LA on storage at refrigerated (8 to 10ºC) temperature for 45 d. Thus, it can
be concluded that refrigerated storage temperature (7±2ºC) restrict the
changes in acidity and gives an acceptable product even at 35 d of storage.
Even after 35 d of refrigerated storage (7±2 ºC), the acidity of sample was
well below 1.4 per cent LA, the maximum permissible limit allowed by FSSA
(2011) for plain Shrikhand.
Changes in free fatty acids
Lipolytic changes in most of dairy products are related with

extracellular thermo stable lipases produced by microorganisms. The off
flavors that accompany with breakdown of milk fat by lipases are due to fatty
acids (FFA). In some of the products, such change under controlled
conditions within limit is desired, whereas in others it has deleterious effect
on their shelf life. In view of importance of release of FFA content in relation
to the organoleptic quality of Shrikhand, the change in FFA content of
Shrikhand during storage are depicted in Table 4.20 and Figure 4.3 at
refrigeration temperature. The FFA (in terms of oleic acid) contents is
expressed as µ.eq/g of product.
It can be seen from tabulated values that storage period had shown
significant effect (P<0.05) on FFA content of Shrikhand stored at refrigeration
temperature. The FFA content of fiber-fortified probiotic Shrikhand ranged
from 0.64 (0 d) to 1.13 (35 d) during storage. The FFA content of fresh
Shrikhand on initial was significantly (P<0.05) lower (0.64 µ.eq/g) which
increased significantly (P<0.05) up to 1.13 (µ.eq/g) on 35 d of storage period.
However, the FFA content of Shrikhand sample at 0 d and 7 d were found
statistically similar to each other (P>0.05). All of the samples were still
acceptable even on 35th d of storage on basis of sensory analysis.
Page | 119
1.2
0.8
FFA (µ.eq/g)
0.6
FFA
0.4
0.2
0
0 7 14 21 28 35
Figure 4.3: Influence of Storage period on the free fatty acids (µ.eq/g) of
Shrikhand at refrigerated temperature (7±2ºC)
Nakae and Elliott (1965) reported that volatile fatty acids in fermented
milk may not be the result of lipid metabolism only, but may also arise from
breakdown of other milk constituents, which may show considerable
variation in volatile acidity at different temperatures. Certain extra-cellular
lipases, which are synthesized by some organisms that survive high heat
treatment. The increase in FFA during storage of Shrikhand is suggestive of
the probable residual enzymatic activity. However, the increasing trend of
FFA content during storage of probiotic Shrikhand was in agreement with the
findings of Desai (1983) who reported increase in FFA content from 2.29 to
3.73 (µ.eq/g) during storage for 40 d at 10ºC, while Patel (2013) reported
increased FFA content from 0.980 to 1.250(µ.eq/g) for 28 d at 7±2ºC and
Prajapati (1989) also noticed increased FFA content from 2.19 to 4.41
(µ.eq/g) during storage at 8 to 10ºC for 45 d.
The observed rise in free fatty acids during storage of product might be
a result of degradation of lipids due to lipolytic activity of intrinsic and/or
bacterial lipase or due to lipolytic activity of surviving. The data pertaining
Page | 120
to the effect of fiber-fortified probiotic Shrikhand on FFA profile during

storage of Shrikhand is lacking, hence it is not possible to compare the
findings of the present study. The possible reason for increased in FFA
content might be due to enzymes that survived and released during the
growth of microorganisms.
Changes in soluble nitrogen contents
The soluble nitrogen content can be considered as a major for

monitoring proteolysis changes in Shrikhand during storage, which is the
consequence of degradation of protein content of Shrikhand by the heat
resistant proteolysis enzymes. Dave (1991) and Kalita (1990) studied the
NPN (non protein nitrogen) content of dahi at ambient and refrigerated
storage temperatures and found that NPN content of dahi was significantly
influenced by levels of total solids and types of cultures uses at both storage
temperatures. Higher the total solids resulted in higher the NPN content.
The values of change in soluble nitrogen content of Shrikhand stored

at refrigeration temperature are depicted in Table 4.20 and Figure 4.4. It can
be revealed that storage period had significant effect (P<0.05) on soluble
nitrogen content of Shrikhand at storage temperature. Fresh Shrikhand had
soluble nitrogen content 0.24 per cent, which was increased significantly to
0.44 per cent upon storage of 35 d.
Page | 121
0.5
0.45
0.4
0.35
Soluble nitrogen (%)
0.3 Soluble
0.25 nitrogen
0.2
0.15
0.1
0.05
0
0 7 14 21 28 35
Figure 4.4: Influence of storage period on the soluble nitrogen (per cent)
of Shrikhand at refrigerated temperature (7±2ºC)
Published data on soluble nitrogen content of fiber-fortified probiotic

Shrikhand are not available however, the values of soluble nitrogen content
of probiotic Shrikhand in the range of 0.153 to 0.515 per cent was obtained
by Patel (2013). The data of soluble nitrogen content obtained in the present
study can be comparable to them. Sharma and Zariwala (1980) and Salunke
(2006) reported the values of soluble nitrogen content of Shrikhand in the
range of 0.3262 to 0.510 and 0.177 to 0.239 percent respectively.
Thus, from the discussion it can be seen that there was significant
(P<0.05) variation in the soluble nitrogen content of all fiber fortified
probiotic Shrikhand, and during storage there was increase in soluble
nitrogen content. The same trend is also for non-probiotic Shrikhand.
Survival of heat resistant bacteria groups and heat stable enzymes capable
of protein breakdown. This can be considered for proportionately higher
soluble nitrogen content during storage.
Page | 122
Changes in water activity
Water activity (aw) is defined as the vapour pressure of water in the

product divided by that of the pure water at the same temperature. Higher
water activity substance tends to support more microorganisms. Bacteria
usually require atleast 0.91 and fungi atleast 0.7 a w for their growth. Thus,
water activity is helpful in deciding the shelf life of the product.
0.94
0.939
0.938
0.937
Water activity
0.936
0.935
0.934
0.933 Water
0.932 activity
0.931
0.93
0.929
0 7 14 21 28 35
Figure 4.5 Influence of Storage period on the Water activity of

The values pertaining to change in a w of Shrikhand are shown in Table

4.20 and Figure.4.5 at refrigeration temperature. The mean value revealed
that the water activity of Shrikhand was influenced significantly (P<0.05) by
the storage period at refrigeration temperature (7±2ºC). During storage, the
water activity of Shrikhand decreased significantly from 0.9391(0 d) to
0.9330 (35 d storage). There was significantly (P<0.05) decreased in water
activity of experimental Shrikhand sample from 0 d to 35 d of storage.
Page | 123
Patel (2013) reported that the value of water activity of probiotic

Shrikhand ranged from 0.9368 to 0.9329 for 0 d to 28 d of storage at
refrigeration temperature (7±2ºC). From the above discussion, it can be seen
that, during storage period of 35 d water activity has been gradually
decreased. If the water activity is low then, the microbiological deterioration
of the product will be comparatively less.
Changes in pH
The acceptability of milk products like Shrikhand is determined by the

pH. These changes, therefore, are critical from product quality point of view,
especially in context to long-term storage. Owning to this fact, the changes
in pH during storage of Shrikhand was studied.
It can be seen from tabulated values (Table 4.20 and Figure.4.6) that
storage period had a significant (P<0.05) effect on pH of Shrikhand stored at
refrigeration temperature (7±2ºC). It can also be observed that there was
significant progressive decreased in pH was found during storage.
Fresh Shrikhand had the pH value 4.81 which decreased significantly

(P<0.05) to 4.37 on 35 d of storage at refrigeration temperature (7±2ºC). No
data on changes in pH during storage are available for comparison for fiber-
fortified probiotic Shrikhand.
Page | 124
4.4
4.35
4.3
4.25
4.2
4.15
pH
4.1 pH
4.05
4
3.95
3.9
0 7 14 21 28 35
Figure 4.6 Influence of Storage period on the pH of Shrikhand at

Change in consistency
Consistency of Shrikhand is one of the attributes, which play an

important role in acceptance of the product by the consumers. To ascertain
response of storage period to change in consistency of Shrikhand during
refrigerated storage (7±2ºC), texture analyzer readings were recorded at
regular interval. The same are set out in Table 4.20 and Figure 4.7
The data collated in Table 4.20 revealed that the consistency of

Shrikhand, measured in terms of texture analyzer readings in N, ranged from
7.81 (at 0 d) to 9.23 (at 35 d). The consistency of Shrikhand is significantly
(P<0.05) affected by storage period. The consistency value for Shrikhand
increased from 7.89 (0 d) to 9.23 at 35th d of storage. From the Figure 4.7, it
can be clearly observed that the trend of increase in the consistency reading.
Page | 125
9.5
9
Consistency
8.5
(N)
Consistency
8
7.5
7
0 7 14 21 28 35
Figure 4.7 Influence of Storage period on the consistency of Shrikhand

at refrigerated temperature (7±2ºC)
The storage period was found to influence the consistency of

Shrikhand significantly (P<0.05) when the product was stored at refrigeration
temperature. With advancement in the period of storage, Shrikhand became
progressively firmer, as indicated by the higher values. At 35 d of storage,
Shrikhand samples had the highest value (i.e. greatest firmness) which
differed significantly (P<0.05) from fresh samples. The progressive firming of
Shrikhand during storage period could be attributed to possible decline in
moisture content and change in body and texture of structural matrix of
Shrikhand. Patel (1982) has reported loss of moisture, causing slight drying
of surface of Shrikhand on storage.
The present findings could not be supported and compared in view of

non-availability of any published reports on such aspect of fiber-fortified
probiotic Shrikhand. Nevertheless, results of decrease in penetrometer
reading of probiotic Shrikhand could be comparable with data presented by
Patel (2013), which decreased from 28.52 to 26.09.
Page | 126
4.5.2 EFFECT OF STORAGE ON SENSORY PROPERTIES OF SHRIKHAND
The fate of any food product has always rested on the acceptance of
the product by consumers. All of the dairy products have tendency to
develop certain specific and non-specific organoleptic shortcomings during
storage and Shrikhand is no exception to eat. The data on the sensory
quality of Shrikhand referring to flavor, body and texture, color and
appearance and total score after a storage period of 35 d at refrigeration
temperature (7±2ºC) are depicted in Table 4.21. The score card used for
judging was as shown in appendix 1.
Table 4.20: Effect of storage period on sensory properties
Influence of period of storage at refrigerated temperature (7±2°C) on

Storage the sensory score of Shrikhand
period
Body & Color&
(d) Flavor (50) Total score (100)
texture (35) appearance (10)
0 45.66 ± 0.58 33.26 ± 0.12 9.61 ± 0.22 93.53 ± 0.78
7 42.75±0.08 32.37±0.24 9.08±0.16 89.2±0.32
14 39.91±0.09 31.68±0.24 8.41±0.20 85.00±0.09
21 38.72±0.03 30.04±0.62 7.79±0.13 81.55±0.51
28 35.34±0.19 28.94±0.59 7.15±0.13 76.43±0.70
35 33.79±0.07 28.36±0.80 7.08±0.05 74.23±0.70
SEM 0.13 0.25 0.08 0.28
CD (0.05) 0.39 0.77 0.24 0.88
C.V.% 0.64 1.63 1.94 0.73
Page | 127
Flavor
Fermented milk products have a pleasing flavor from a blend of a clean

acid taste and a delicate aromatic flavor. It should be free from undesirable
off flavors like flat, metallic, yeasty or bitter. During storage, Shrikhand
undergoes various physico – chemical and microbiological changes which
tend to affect the flavor of the product. Flavor is the most important
parameter of sensory profile as it along with other attributes decides the
acceptability of Shrikhand.
Shrikhand samples were subjected to judging and grading for various

sensory attributes viz. (i) flavor, (ii) Body and texture, (iii) color and
appearance, and (iv) Total score. The data obtained during storage for the
Shrikhand samples are represented in Table 4.21 and Figure 4.8.
50
45
40
35
Flavour score
30
25
20 Flavour
15
10
5
0
0 7 14 21 28 35
Figure 4.8 Influence of Storage period on the Flavor of Shrikhand at

Page | 128
Desai and Gupta (1986) suggested that decline in score of any sensory
attributes of Shrikhand up to 60 per cent of its maximum score makes the
product organoleptically unacceptable.
It can be seen from tabulated values that storage period had

significant (P<0.05) effect on flavor of Shrikhand at refrigeration temperature.
It was observed that for the freshly made product, the flavor score decreased
from 45.66 to 33.79 at 35 d of storage at refrigerated temperature
significantly.
The fiber-fortified probiotic Shrikhand sample showed the low flavor

score may be the result of high acidity developed in the sample and the
sample criticized too sour by judges during the storage from 0 d to 35 d.
A marked linear decrease in flavor score of Shrikhand was noticed

during storage at refrigerated temperature that might be due to
appearance of flavor defects such as sour, fermented, yeasty and unnatural
at different stages of assessment. Development of such flavor defects can be
attributed to biochemical changes induced by bacteria and their enzymes.
As information in flavor changes of the fiber-fortified probiotic

Shrikhand is lacking, the observation of this study could not be compared.
However, results obtained for probiotic Shrikhand reported by Patel (2013)
ranged from 42.34 to 34.50 on 28 d of refrigeration temperature (7±2ºC).
Results obtained from market Shrikhand reported by Salunke et al. (2006)
and Jain et al. (2003) ranged from 42.11 to 38.04 and 44.84 to 36.11
respectively are in similar pattern during storage.
Body and texture
Various sensory attributes collectively determines their acceptability of

Shrikhand and one such attribute is body and texture score. During storage,
Shrikhand undergoes various physico chemical changes that affect the
sensory profile of the product as like body and texture. Sensory perception of
Page | 129
body and textural attributes are liable to change in accordance with the
overall caused to consistency and water retention ability of the product via
physical and biological changes during storage. The body and textural
characteristics stand next to flavor in the overall
34
33
Body &texture score
32
31
30
Body
29
&texture
28
27
26
25
0 7 14 21 28 35
Figure 4.9 Influence of Storage period on the Body and texture of

Acceptability of the Shrikhand. The results obtained are delineated in Table

4.21 and Figure.4.9 at refrigerated temperature (7±2º C) storage.
It is evident from tabulated value that the body and texture score of
Shrikhand during storage declined significant (P<0.05) with the advancement
of storage period regardless at refrigeration temperature. At zero d the
highest score (33.26) was observed, it was significantly (P<0.05) higher than
all the storage intervals while at 35 d of storage the lowest score (28.39) was
observed which was found significantly (P<0.05) lower than all the storage
intervals.
The declined in scores at refrigeration temperature could be mainly

attributed to the gradual increase in hardness and decrease in moisture
Page | 130
content, as criticized by the judges. The observed decrease in body and

texture score of Shrikhand during storage of product at refrigeration
temperature might be results of various rheological, chemical and microbial
changes.
Research work carried out on probiotic Shrikhand prepared using

different probiotic cultures had body and texture score ranged from 24.93 to
22.60 (Patel, 2013). Salunke et al. (2006) studied on market samples and
reported that body and texture score ranged from 26.65 to 22.88 for market
samples of Maharashtra and Jain et al. (2003) reported body and texture
score ranged from 26.89 to 24.11 for market samples of Gujarat during
storage.
Color and appearance of Shrikhand is another contributor that plays a

vital role in its acceptance by consumer. As is the case with all of the dairy
products during storage, Shrikhand undergoes various physiochemical,
microbial changes, which affect the color and appearance of the product. The
consumer judges to accept or reject the product for further sensory
evaluation based on its color and appearance. The results obtained are
delineated in Table 4.21 and Figure. 4.10 for storage at refrigeration
temperature.
Page | 131
12
Colour & appearance score

10
6
Colour &
4
appearance
2
0
0 7 14 21 28 35
Figure 4.10 Influence of Storage period on the Color and appearance of

It can be seen from the tabulate values that color and appearance
score of Shrikhand is significantly (P<0.05) affected by the storage period at
refrigeration temperature.
The color score of Shrikhand during storage was found to decrease

with the increase in storage period. Fresh Shrikhand had a score of 9.61 on
0 d which decreased significantly (P<0.05) to 7.08 at 35 d of storage at
refrigeration temperature. At 35 d score for color and appearance was
significantly (P<0.05) lower than all other storage intervals.
The observed decrease in color and appearance score might be due to

the interaction of lactose and protein that changes the color to some extent.
In absence of published data on effect of storage period on color and

appearance of fiber fortified probiotic Shrikhand curing storage of Shrikhand,
it is not possible to compare color and appearance profile of Shrikhand
samples. However, Patel (2013) prepared probiotic Shrikhand and reported
Page | 132
decrease in trend of color and appearance score of Shrikhand ranging from

12.98 to 11.59 during storage at refrigeration temperature. Salunke (2006)
have reported decrease in trend of color and appearance score of Shrikhand
ranging from 13.38 to 10.96 during storage at refrigeration temperature.
Total score
As a whole, the acceptability of Shrikhand exclusively depends on its

organoleptic characteristics, which are based on the compositional,
physicochemical as well as microbiological characteristics. These attributes
of Shrikhand that governs organoleptic characteristics of the product, in
terms are dependent on the flavor, body and texture, color and appearance,
type of package and temperature of storage. In view of these facts, the
changes in the total score of Shrikhand were studied and results are
presented in Table 4.21 and Figure. 4.11 for storage at refrigeration
temperature.
The total score, which describes the overall quality of the product, is
the collective score of body and texture, flavor and color and appearance.
Consequently, it follows the trend noticed with the individual organoleptic
parameter studied.
The total score of Shrikhand stored at refrigeration temperature was

found to decrease significantly (P<0.05) with the increase in storage period.
Fresh Shrikhand has a total score of 93.53, which was significantly (P<0.05)
highest score but decreased significantly to 74.23 at 35 d of storage.
Shrikhand score at 0, 7, 14, 21, 28 and 35 d of storage were statistically
different from each other however; the product was still acceptable by judges
on sensory basis.
A marked linear decrease in total score of Shrikhand during storage at

refrigeration temperature could be attributed to lower color and appearance,
body and texture and flavor score of the Shrikhand.
Page | 133
100
90
80
70
Total score
60
50
40 Toatl score
30
20
10
0
0 7 14 21 28 35
Figure 4.11 Influence of Storage period on the total score of Shrikhand

at refrigerated temperature (7±2ºC)
In absence of published information on effect of storage period of the

total score of fiber-fortified probiotic Shrikhand during storage, it is not
possible to compare data obtained in the present study. However, the result
obtained here in this study are very close to the results obtained for the
sensory profile of probiotic Shrikhand by Patel (2013) ranging from 80.25 to
68.68 for storage at refrigeration temperature. Salunke et al. (2006) and Jain
et al. (2003) reported sensory profile of market sample ranging from 86.75 to
75.7 and 89.22 to 77.45 respectively during storage.
Therefore, it can be concluded that total score or in turn the

organoleptic attribute of the product follows the trend that was evident in
individual attributes of Shrikhand. The overall sensory scores gave a shelf life
of more than 35 d at refrigeration temperature and found to acceptable on
sensory basis.
Page | 134
4.5.3 EFFECT OF STORAGE ON MICROBIOLOGICAL QUALITY OF

SHRIKHAND
Most of the milk products are highly perishable in nature; this

perishablility of dairy products is mostly ruled by microbiological quality of
dairy product. The microbiological quality of dairy products like Shrikhand
becomes more important owing to the sole dependence of product shelf life
on the growth of microorganisms in product during storage. Most of the
physico – chemical changes like FFA content, soluble nitrogen content,
change in acidity/pH etc. that in turn decides the fate of Shrikhand during
storage are profoundly affected by the presence and growth of various
microorganisms. Taking these facts in to consideration, Shrikhand packed in
PS cups was judged for its microbiological quality during storage at
refrigeration temperature (7±2ºC).
The microbiological status viz., Standard Plate Count, Yeast and

Mould Count, Probiotic Count and coli form count is presented in Table 4.22
for storage at refrigeration temperature (7±2ºC). All the samples of fresh and
stored Shrikhand had zero count of coli form on zero d as well as all through
the storage indicating hygienic manufacturing.
Probiotic Count
Probiotic are defined as “a preparation of or a product containing

viable, defined microorganisms in sufficient numbers, which alter the micro
flora (by implantation or colonization) in a compartment of the host and by
that exert beneficial health effects in this host” (Schrezeneir and de Vrese
2001). The growing interest of consumers towards therapeutic products has
led to incorporation of probiotic cultures in different milk products. Probiotic
have been therapeutically to modulate immunity, improve digestive process,
prevent cancer, improve lactose intolerance, etc. (Devshete et al., 2012).
Lactic acid bacteria decreases serum cholesterol levels, increases vitamin B
content in the product (Grill et al., 2000). The probiotic count should be
Page | 135
more than 6.000 log cfu/g or 106cfu/g in order to prescribed /designate the
product as probiotic product.
Table 4.21 Effect of storage on microbiological quality
Effect of various period on microbiology quality (log cfu /g)

Storage period
(d) Probiotic count Standard plate Yeast and mould
count count
0 8.73±0.02 5.29±0.01 Nil
7 8.50±0.05 5.46±0.01 1.53±0.22
14 8.40±0.05 5.62±0.06 1.59±0.20
21 8.15±0.04 5.70±0.01 1.69±0.27
28 7.75±0.03 5.76±0.05 1.85±0.27
35 7.50±0.06 5.85±0.05 2.06±0.16
SEM 0.021 0.019 0.113
CD (0.05) 0.066 0.059 0.348
C.V.% 0.521 0.679 12.199
The average probiotic count of all experimental Shrikhand samples

during storage at refrigeration temperature is tabulated in Table 4.21and
graphically presented in Figure 4.12.
Page | 136
9
8.8
8.6
Probiotic count
8.4
(log cfu/g)
8.2
8
Probiotic
7.8
7.6 count
7.4
7.2
7
6.8
0 7 14 21 28 35
Figure 4.12 Influence of Storage period on the Probiotic count of

The statistical analysis for storage period of Shrikhand up to 35 d, at

interval of 7 d indicated that the probiotic count for all d of storage were
differ significantly (P<0.05). Results revealed that highest probiotic count
was recorded in Shrikhand prepared (8.73 log cfu/g) on zero d and their after
it were gradually decreased with increasing of storage. The significantly
(P<0.05) lowest score was recorded in Shrikhand prepared (7.50 log cfu/g) on
35 d of storage. There was decline in probiotic count that can be observed
clearly from the graph. Nevertheless, in the Shrikhand sample probiotic
count remained well above 107 even after the end of refrigerated storage for
35 d. This indicates that there was not much difference in probiotic count
during storage period.
As suggested by different workers (Speck 1976; Gilliland et al., 1978;

Goldin and Gorbach, 1980; Kurmann and Rasic, 1991; Shah, 2000;
Lourens-Hattingh and Viljoen, 2001; Shah et al., 2001), the viable count of
probiotic culture that should be available to the consumers’ for therapeutic
Page | 137
benefits should be in the range of 106 to 109 in product, which is present in

Shrikhand sample, even after refrigerated storage for 35 d. This could be an
added an advantages as some times during storage the probiotic count drops
below the required level (Shah et al., 2001). Researchers found that addition
of 0.5 per cent inulin, oat beta-glucan to yogurt resulted in prolonged
survival of B. lactis (Vasiljevic et al., 2007).
However, Patel, (2013) prepared probiotic Shrikhand using probiotic

cultures. He reported that the probiotic count of Shrikhand at zero d was
7.44, 8.72, 9.42, 8.24, and 9.28 log cfu/g that decreased to 6.83, 8.03, 8.61,
6.70 and 8.963 log cfu/g for PC 1, PC 2, PC 3, PC 4 and PC 5 respectively on
28 d of storage at refrigerated (7±2ºC) temperature. The probiotic count of
Shrikhand decreased significantly at each interval of storage refrigerated
(7±2ºC) temperature.
This decline in probiotic count may be due to the result of higher

acidity produced by the accompanying lactic acid producing in the
combination with the probiotic culture, in probiotic Shrikhand. However,
decline was more prominent due to presence of sugar in product.
Standard Plate Count (SPC)
Standard plate count is the collective enumeration of the overall

microbiological quality of the product, after production and during its
storage period. It gives an overall idea about the status of the Shrikhand in
terms of its overall microbiological quality during storage. It was determined
with a view to know the number of total bio-load of the product. The mean
Standard plate count (SPC), expressed as log cfu/g. Therefore, the standard
plate count of Shrikhand packed in PS cup was analyzed for its standard
plate count during storage at refrigerated (7±2ºC) temperature and result is
presented in Table 4.22 and Figure 4.13.
Page | 138
The SPC values of Shrikhand during storage, at refrigeration

temperature were increased with the increase in storage period. It can be
seen from tabulated values that the storage period had significant effect
(P<0.05) on SPC of Shrikhand stored at 7±2ºC as predetermined time
interval.
Fresh Shrikhand sample had a SPC count of 5.29 log cfu/g but
increased significantly (P<0.05) to 5.85 log cfu/g after 35 d of storage. From
sensory point of view Shrikhand sample was still acceptable after 35 d of
refrigerated storage.
5.9
5.8
Standard plate count
5.7
5.6
(log cfu/g)
5.5
standard
5.4
plate count
5.3
5.2
5.1
5
0 7 14 21 28 35
Figure 4.13 Influence of Storage period on the Standard plate count of

Mechanized method of Shrikhand manufacturing showed significantly

(P<0.05) lower values of SPC than the product from traditional
manufacturers, whereas Shrikhand manufactured by traditional method in
commercial sector was at par with Shrikhand manufactured by traders. This
is due to the fact that Shrikhand manufactured on mechanized line is
Page | 139
thermized and then packed in sanitized polystyrene cups, both of which

helps in keeping the lower counts in the product.
Such a wide variation in the SPC may be ascribed to variation in the

pre and post contamination of the product, initial load, varying amount and
type of culture used, level and type of contaminants, acidity/pH of the
product, sugar concentration, storage temperature and time and quality of
packaging materials, hygienic conditions maintained during and post
manufacture etc.
In absence of published data on effect of storage period on fiber

fortified Shrikhand on the SPC of Shrikhand during storage, comparison was
not possible. However, market study done by Salunke (2006) showed similar
trend of SPC ranging from 5.59 to 8.36 log cfu/g.
Prajapati et al. (1993) reported that as the intensity of heat treatment

to the product during thermization increased, there was decreased in total
viable count. This is true with the Shrikhand made by mechanized method
that showed lower SPC owing to heat treatment.
From above foregoing discussion, it can be concluded that the sample

is acceptable and have shelf life of 35 d of storage under refrigeration from
microbiological point of view.
Yeast and mold count
The values represented in Table 4.22 and Figure 4.14 are the yeast and
mold count of Shrikhand expressed in terms of log cfu/g, when stored at
refrigerated temperature (7±2ºC) for the period of 35 d.
The average values of YMC for stored Shrikhand ranged from (7 d)1.53
to (35 d) 2.06 log cfu/g. There was significant effect (P<0.05) of storage
interval of storage at refrigerated temperature. Shrikhand sample was
Page | 140
found1.53-logcfu/g on seven of storage. One of 35 it was significantly higher

2.06 (P<0.05) log cfu/g. fresh Shrikhand samples found to free from yeast
and mold at refrigeration temperature.
2.5
2
Yeast and mold count
(log cfu/g)
1.5
Yeast and
mold
1 count
0.5
0
0 7 14 21 28 35
Figure 4.14 Influence of Storage period on the Yeast and mold count of
However, Patel (2013) reported yeast and mold count (log cfu/g) for
probiotic Shrikhand was 1.563, 1.616, 1.552, 1.55, 1.621 and 1.596 log
cfu/g. at zero d, which increased to 2.226, 2.263, 2.252, 2.203, 2.257 and
2.243 log cfu/g. at the 28thd of refrigerated temperature (7±2ºC) storage in
NPC, PC 1, PC 2, PC 3, PC 4 and PC 5 respectively.
Thus, it can be concluded on the basis of this part of the study that
the shelf life of fiber-fortified probiotic Shrikhand when stored at refrigerated
temperature was 35 d.
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Chapter 5
SUMMARY AND CONCLUSION
Milk and milk products considered as a vehicle for dietary fiber would
not only take care of their own role in human health but could also enhance
the heath fullness of the diet as a whole. Therefore, the present study was
undertaken in the following phases.
PHASE-I: Preparation of Shrikhand using different fiber containing powder
PHASE-II: Optimization of the rate of addition of fiber containing powder from

each fiber containing powder
PHASE-III: Comparison of Shrikhand containing optimized level of each fiber

containing powder to select the best fiber containing powder in fiber fortified
Shrikhand
PHASE-IV: Selection of flavouring ingredients for the fiber fortified probiotic

Shrikhand
PHASE-V: Study of the shelf life of developed fiber fortified Shrikhand at

refrigeration temperature (7 ±2 0C)
PHASE-1 PREPARATION OF SHRIKHAND USING DIFFERENT FIBER

CONTAINING POWDER AT DIFFERENT RATE OF ADDITION

rates viz., banana powder @ (1.0, 3.0, 5.0) per cent, oat powder @ (1.0,2.0,3.0)
per cent, inulin powder @ (1.0,2.0,3.0) per cent on w/w basis of expected yield
of Shrikhand. These levels were selected based on preliminary trials. The fiber
content of banana powder, oat powder and inulin powder was 2.5, 12.8 and 94
per cent respectively. Shrikhand was prepared using the method described by
Summary and Conclusion
Patel (2013) using probiotic culture i.e. thermophilic lactic acid freeze-dried
DVS (Direct Vat Set) culture (ABY-3). After obtaining chakka from skim milk
dahi, fiber-containing powders were blended thoroughly with chakka followed
by addition of sugar and cream to prepare Shrikhand. The average yield of
chakka was 310 g/ l of skim milk; and the average yield of Shrikhand – 472 g /
l of skim milk. The sample of Shrikhand without addition of fiber containing
powder was used as a control.
PHASE-II: OPTIMIZATION OF THE RATE OF ADDITION OF FIBER

CONTAINING POWDER FROM EACH FIBER CONTAINING POWDER
This phase was conducted to optimize the rate of addition of three fiber-
containing powders for manufacture of acceptable quality of fiber-fortified
probiotic Shrikhand. The fiber containing powders were incorporated in
Shrikhand at three rates each viz., banana powder @ (1.0,3.0,5.0) per cent, oat
powder @ (1.0,2.0,3.0) per cent, inulin powder @ (1.0,2.0,3.0) per cent on w/w
basis of expected yield of Shrikhand. Each treatment was replicated four times.
The samples were analyzed for their chemical composition, physico chemical
and sensory attributes.
Optimization of rate of addition of banana powder
Effect of varying levels of banana powder on the composition of

Shrikhand
Total solids content of control was 58.28 per cent. This value was
significantly (P<0.05) lower than Shrikhand containing B1, B2 and B3. These
values were 58.65, 59.35 and 60.03 for B1, B2 and B3 respectively. The average
fat content of control was 6.23 per cent, while banana powder containing
Shrikhand samples B1, B2,B3 were 6.20, 5.78 and 5.72 per cent respectively.
There was significant (P<0.05) difference in fat content of control and
experimental Shrikhand samples. The protein content of banana fiber fortified
probiotic Shrikhand samples were B1 (5.79), B2 (5.78) and B3 (5.75) per cent.
While it was 5.82 per cent for control, Shrikhand sample. There was significant
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(P<0.05) difference in protein content. Control had average sucrose content of

41.43 per cent. Whereas in experimental Shrikhand samples, it were 41.40,
41.41 and 41.43 per cent for B1, B2 and B3 respectively. The variation in
sucrose content of all experimental samples was found to be non-significant
(P>0.05). The average ash content of B1, B2,and B3 was 0.65, 0.68and 0.71
percent respectively. The ash content of control (i.e. 0.57 per cent) was
significantly (P<0.05) lower than B1, B2 and B3. The ash content of B3 (i.e. 0.71
per cent)was significantly (P<0.05) higher than all the other experimental
samples. The carbohydrate content of B1, B2 and B3 were 44.98, 45.72 and
46.43 per cent respectively whereas control had a value of 44.63 per cent. All
experimental samples were significantly (P<0.05) different in carbohydrate
content.
Effect of varying levels of banana powder on the physico-chemical

The average lactic acid content of samples without fiber containing

banana powder i.e. control Shrikhand sample was 1.0 per cent LA, while for
banana powder containing Shrikhand samples viz., B1, B2 and B3 the values
were 0.99, 0.97 and 0.95 per cent LA respectively. The variation in titratable
acidity (per cent LA) of all experimental samples were non-significant
(P>0.05).Control had average free fatty acid content of 0.58 µ.eq/g. whereas, for
banana powder containing Shrikhand samples, viz., B1 (1 per cent), B2 (3per
cent) and B3 (5per cent) these were 0.59, 0.60 and 0.63 µ.eq/g respectively.
There was no significant (P>0.05) effect of addition of banana powder on free
fatty acid content of Shrikhand.
Soluble nitrogen content of C, B1, B2 and B3 were 0.28, 0.26, 0.29 and
0.29 per cent respectively. The variation in soluble nitrogen content of all
experimental samples were non-significant (P>0.05).The water activity of C, B1,
B2and B3were0.9389, 0.9388, 0.9379, 0.9371 respectively. The difference in
water activity among all experimental Shrikhand samples were non-significant
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(P>0.05).The pH of C, B1, B2 and B3 were 4.33, 4.35, 4.36, and 4.38

respectively. The variation in pH content of all experimental samples were non-
significant (P>0.05). The consistency of Shrikhand samples C, B1, B2 and B3
was 8.15, 8.35, 8.53 and 8.73 N respectively. The variation in consistency of all
experimental samples were found significant (P<0.05).
Effect of varying levels of banana powder on the sensory characteristics of

Shrikhand
The flavour scores of C, B1, B2 and B3 were 43.32, 45.00, 44.52, and 45.25
respectively. The difference in flavour score of experimental Shrikhand samples
may be attributed to increasing rate of addition of powder, which alters the
flavouring score of experimental Shrikhand. The average body and texture
scores of C, B1, B2 and B3 were 31.48, 32.55, 32.23, and 32.71 respectively.
The variation in body and texture score of all experimental samples were
significant (P<0.05). The average colour and appearance scores of C, B1, B2 and
B3 were 9.14, 8.00, 7.91 and 7.02 respectively. The variation in colour and
appearance score of all experimental samples were significant (P<0.05).The
total score ofC, B1, B2 and B3 were 88.94, 90.55, 89.66 and 89.98 respectively.
containing banana powder (B3) Shrikhand had superior quality in terms of
chemical, physicochemical and sensory attributes. So, B 3 sample was selected
for use in phase III. Thus, among three levels of banana powder, addition of 2
per cent banana powder was selected for further study.
Optimization of rate of addition of oat powder
Effect of varying levels of oat powder on the composition of Shrikhand
TS content of C, O1, O2 and O3were 58.28, 58.65, 59.01 and 59.36 per
cent respectively. The difference in total solids content of all experimental
samples were significant (P<0.05). The average fat content of C, O1, O2, and O3
were 6.23, 6.31, 6.37 and 6.46 respectively. There was significant (P<0.05)
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difference in fat content of all samples. The average protein content of C, O1, O2
and O3 were 5.80, 5.90, 5.96 and 6.03 per cent respectively. There was
significant (P<0.05) difference in the protein content of all experimental
samples. The average sucrose content of C, O1, O2 and O3 were 41.43, 41.40,
41.41 and 41.43 per cent respectively. The variation in the total sugar content
of all experimental samples were non-significant (P>0.05). The average ash
content of control C, O1, O2 and O3 were 0.57, 0.65, 0.68 and 0.71per cent
respectively. There was significant (P<0.05) difference in ash content of all
experimental Shrikhand samples. The carbohydrate content of C, O1, O2 and O3
were 44.63, 44.70, 44.76 and 44.82 per cent respectively. All experimental
samples were significantly (P<0.05) different.
Effect of varying levels of oat powder on the physico-chemical properties

of Shrikhand
The average lactic acid content of C, O1, O2 and O3 were 1, 0.99, 0.98 and 0.97
per cent LA respectively. The variation titratable acidity (per cent LA) content of
all experimental samples were found significant (P<0.05).
The average free fatty acids content of C, O1, O2 and O3 were 0.60, 0.62, 0.63
and 0.66 µ.eq/g respectively. The variation in FFA (µ.eq/g) content of all
experimental samples were non-significant (P>0.05). The soluble nitrogen
content of C, O1, O2 and O3 were 0.27, 0.29, 0.26 and 0.27 per cent. The
variation in soluble nitrogen content of all experimental samples were non-
significant (P>0.05). The water activity of C, O1, O2 and O3 were 0.9391,
0.9390, 0.9380 and 0.9367 respectively. The difference in water activity among
all experimental Shrikhand samples were non-significant (P>0.05).The value for
the consistency of O 3 had highest value 8.65 N followed by O2 (8.63 N) and O1
(8.54 N). Whereas sample C had significantly (P<0.05) lowest value i.e.8.48 N.
The difference of consistency among all experimental Shrikhand samples was
significant (P<0.05). The pH in fiber containing oat powder Shrikhand samples
were O1 (4.35), O2 (4.36) and O3 (4.37). While control Shrikhand sample had
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4.33. However, the variation in pH of all experimental samples were non-

Effect of varying levels of fiber containing oat powder on the sensory

The flavour scores of Shrikhand prepared using supplementation of fiber

containing oat powder were O1 (43.98), O2 (43.85) and O3 (42.39) and C (44.05).
The variation in flavour score of all experimental samples were significant
(P<0.05). The body and texture score of fiber containing oat powder Shrikhand
samples were O1 (32.46), O2 (32.91) and O3 (31.94). While control Shrikhand
sample had body and texture score 31.64. The variation in body and texture
score of all experimental samples were significant (P<0.05). The experimental
sample C was criticized as weak body and O 3 was criticized as grainy texture
by judges. The average colour and appearance scores of C, O1, O2 and O3were
9.06, 8.64, 8.55 and 7.96 respectively. The variation in colour and appearance
score of all experimental samples were found to be significant (P<0.05). The
average total score of C, O1, O2 and O3 were 89.75, 90.08, 90.31 and 87.29
respectively. Thus, from the result discussed above, it is observed that use of
fiber containing oat powder (O2) Shrikhand had superior quality in term of
chemical, physico chemical and sensory attributes. So, O2 sample was selected
for use in phase III. Thus, addition of 2 per cent oat powder is yielding
Shrikhand better quality compared to other rate of addition of oat powder.
Optimization of rate of addition of inulin powder
Effect of varying levels of fiber containing inulin powder on the

The average total solids content of C, I1, I2 and I3 were 58.28, 58.67,
59.03 and 59.40 per cent respectively. There was significant difference (P<0.05)
in total solids content of Shrikhand samples. Control Shrikhand had
significantly (P<0.05) lower i.e. 58.28 per cent total solids content as compared
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to all other fiber fortified Shrikhand samples. The average fat content of C, I1, I2
and I3 were 6.23, 6.19, 6.10 and 6.05 per cent respectively. There was
significant (P<0.05) difference in fat content of control sample and fiber
containing Inulin powder Shrikhand samples. The average protein content of C,
I1, I2 and I3 were 5.80, 5.73, 5.69 and 5.61 per cent respectively. There was
statistically significant (P<0.05) difference in the protein content of all
experimental samples. The average sucrose content of C, I1, I2 and I3 were
41.42, 41.42, 41.44 and 41.43 respectively. The variation in the total sucrose
content of all experimental samples were non-significant (P>0.05).The average
ash content of C, I1, I2 and I3 were 0.570.63, 0.64 and 0.64 per cent
respectively. There was no significant (P>0.05) difference in ash content of all
experimental Shrikhand samples. The carbohydrate content of fiber containing
inulin powder Shrikhand sample I1, I2 and I3 were 44.23, 43.85 and 43.49 per
cent respectively. The value of control Shrikhand had 44.63 per cent. All
experimental samples were found significantly (P<0.05) different from each
other.
Effect of varying levels of fiber containing inulin powder on the physico-

chemical properties of Shrikhand
The average lactic acid content of control Shrikhand sample was 1.0 per
cent LA, while for Shrikhand with inulin i.e. I1, I2 and I3 had 0.99, 0.98 and 0.97
per cent LA respectively. The variation in titratable acidity (per cent LA) content
of all experimental samples were significant (P<0.05). The average free fatty
acids content of C, I1, I2 and I3 were0.55 µ 0.59, 0.61 and 0.62 µ.eq/g
respectively. The variation in FFA (µ.eq/g) content of all experimental samples
were non-significant (P>0.05). The soluble nitrogen content of C,I1, I2 and I3
were0.21, 0.22, 0.24 and 0.25 per cent. The difference of soluble nitrogen
among all experimental Shrikhand samples were statistically non-significant
(P>0.05). The water activity of C, I1, I2 and I3 were 0.9375, 0.9372, 0.9369, and
0.9365 respectively. The difference in water activity among all experimental
Shrikhand samples were statistically non-significant (P>0.05).The pH content in
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fiber containing inulin powder Shrikhand samples were I1 (4.35), I2 (4.33) and I3
(4.32). While control Shrikhand sample had a pH of 4.38. However, the
variation in pH content of all experimental samples were non-significant
(P>0.05). The consistency of C, I1, I2 and I3 were 8.37, 8.51, 8.58 and 8.62
respectively. There is no statistically difference (P>0.05) was found in sample I1,
I2 and I3.
Effect of varying levels of inulin powder on the sensory attributes of

Shrikhand
The flavour scores of C, I1, I2 and I3 were 45.50, 45.53, 45.64 and 45.60.
However, the variation in flavour score of all experimental samples were non-
significant (P>0.05). The average body and texture scores of C, I1, I2 and I3 were
32.01, 33.14, 33.48 33.52.The control Shrikhand sample scored 32.01 which
was significantly (P<0.05) lower than all three Shrikhand samples. The
variation in body and texture score of all experimental samples were found not
significant (P>0.05). The colour and appearance scores of C, I1, I2 and I3 were
9.35, 9.48, 9.48, and 9.48 respectively. The variation in colour and appearance
score of all experimental samples were non-significant (P>0.05).The total score
of C, I1, I2 and I3 were 91.86, 93.15 93.60 and 93.60 respectively. Statistically
samples I1, I2 and I3 were not significantly different (P>0.05). Use of fiber
containing inulin powder at each level of addition in yielded (I1) Shrikhand with
superior quality in term of chemical, physico – chemical attributes also.
Nevertheless, looking to the high cost of inulin I 1 (1 per cent) was selected for
further study.
FIBER CONTENTS IN DIFFERENT EXPERIMENTAL SHRIKHAND
The fiber content in Shrikhand samples was analyzed to be 0.03 per cent
in B1 (1 per cent), 0.08 per cent in B2 (3 per cent), 0.13 per cent in B3 (5 per
cent), 0.12 per cent in O1 (1 per cent), 0.24 per cent in O2 (2 per cent) and 0.36
per cent in O3 (3 per cent) and 0.92 per cent in I1 (1 per cent), 1.84 per cent in
I2 (2 per cent), 2.8 per cent in I3 (3 per cent), With increasing the rate of
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addition of fiber an increasing fiber content in samples Shrikhand that was

observed.
PHASE-III: COMPARISON OF SHRIKHAND CONTAINING OPTIMIZED LEVEL

OF EACH FIBER CONTAINING POWDER TO SELECT THE BEST FIBER
CONTAINING POWDER IN FIBER FORTIFIED SHRIKHAND
This phase was conducted to select the best fiber containing powder in
fiber fortified Shrikhand. Shrikhand was prepared from using banana powder
(B3), oat powder (O2) and inulin (I1) were selected and the chemical composition,
physico chemical properties, consistency, sensory attributes and microbial
quality were compared with control (C).
The average total solids content of C, I1, O2 and B3 were 58.28, 58.67,
59.01 and 60.03 percent respectively. There was significant difference (p<0.05)
in total solid content of all samples. The average fat content of control C, I1, O2
and B3 were 6.23, 6.19, 6.37 and 5.72 per cent. There was significant (P<0.05)
difference in the fat content of control Shrikhand and fiber fortified Shrikhand,
while I1 and control were not significantly (P>0.05) different. Protein content of
the C was 5.80. While in experimental fiber fortified Shrikhand, these were I1
(5.73), O2 (5.96) and B3 (5.76). These value showed significant (P<0.05)
difference in all experimental Shrikhand samples, whereas sample I1 and O2
were not significantly different (P>0.05). The total sucrose content of control
Shrikhand averaged 41.45 per cent, whereas in fiber fortified samples these
were I1 (41.44), O2 (41.47), B3 (41.45). The variation in sugar content of all
samples were non-significant (P>0.05).The average ash content of control
sample was C (0.57 per cent) and fiber fortified Shrikhand sample were I1
(0.63 per cent), O2 (0.68 per cent) and B3 (0.71 per cent). These values showed
significant (P<0.05) difference. The carbohydrate (by difference) content of C
sample 44.63 per cent. While fiber fortified samples varied from 44.23 to 46.44
percent. There was significant (P<0.05) difference in the carbohydrate content
of control Shrikhand and fiber fortified Shrikhand.
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Physico -chemical properties and Consistency of Shrikhand
The acidity was found to be 1.0 per cent LA. In control, Shrikhand
sample. While for fortified Shrikhand samples I1, O2 and B3, these were 0.99,
0.98 and 0.95 per cent LA respectively. There was significant (P<0.05)
difference observed in lactic acid contents of Shrikhand. The FFA value of
experimental samples are expressed in µ.eq/g, which were C (0.53), I 1 (0.56),
O2 (0.58) and B3 (0.58) µ.eq/g. There is no significant (P>0.05) difference in the
free fatty acids content of control Shrikhand and fiber fortified Shrikhand. The
soluble nitrogen content of C, I1, O2 and B3 were 0.29, 0.28, 0.27 and 0.29 per
cent respectively. The difference of soluble nitrogen among all experimental
Shrikhand samples was non-significant (P>0.05). The water activity of
experimental Shrikhand samples varied from B3 (0.937) to C (0.939). However
the difference of water activity among all experimental Shrikhand samples were
non-significant (P>0.05). The pH of C, I1, O2 and B3 were 4.33 4.34, 4.36 and
4.38respectively. There was no significant (P>0.05) difference observed in Table
value. The consistency of C, I1, O2 and B3 were 8.45, 8.53, 8.65 and 8.73 N. B3
sample had highest value but based on sensory evaluation the body and
texture were not satisfactory and thus, given a lower score due to excessively
firm body. Sample C was less consistent and thus resulted in a weak body,
which fetched a lower score on the sensory scale. Sample I 1 and O2 were
statistically at par and were optimally consistent. The variation in consistency
was found to be significant (P>0.05).
Sensory attributes
The flavours scores of C, I1, O2 and B3 were 44.07, 46.14, 46.11and

44.39 respectively. C was significantly (P<0.05) lower from all three Shrikhand
samples, while sample I1 and O2 were at par (P>0.05). The score for body and
texture of C, I1, O2 and B3 were 31.50, 32.64, 33.25 and 30.93 respectively.
Among experimental Shrikhand samples scores of body and texture of B3 was
significantly (P<0.05) lower than all other samples. This clearly indicates the
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supplementation of fiber containing powder in the current investigation had

positive impact on body and texture of samples I 1 and O2 were statistically
(P<0.05) at par. While B3 sample was criticized as excessively firm body and
sample C was criticized as weak body as compared to other samples by judges.
The colour and appearance score of C, I1, O2 and B3 were 9.17, 9.48, 9.46 and
7.17. These value show colour and appearance of samples I 1 and O2 were
statistically at par (P>0.05).The total score was ranged from B3 (87.67) to O2
(93.82). B3 sample had significantly lower acceptability score. Among all
experimental Shrikhand O2 had significantly (P<0.05) higher acceptability
score indicating superior acceptability among experimental samples,
statistically samples I1 and O2 were at par (P>0.05).
Microbiological analysis
Probiotic count of control was significantly (P>0.05) lowest 8.397 log

cfu/g whereas count of fiber fortified samples varied from 8.41 (B 3) to 8.475
(I1). The SPC count of experimental fresh Shrikhand samples varied from 1.354
to 1.402 log cfu/g. The SPC count of experimental Shrikhand samples varied
non significantly (P>0.05) different among all the experimental samples.
Shrikhand samples were found free from yeast& mould, and coliform in fresh
product samples.
PHASE - IV: SELECTION OF FLAVOURINGINGREDIENTS FOR THE FINAL

PREPARED SHRIKHAND
Comparison of cardamom and rajbhog as the flavouring ingredients for

the fiber fortified probiotic Shrikhand with control without flavour ingredients
revealed that at Shrikhand containing cardamom flavour had significantly the
highest total score i.e. 93.88, while control had the lowest total score i.e.89.85.
The gross composition of the developed cardamom flavour probiotic fiber

fortified Shrikhand was total solids 59.01 per cent, milk fat 6.37 per cent,
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consistency of the developed cardamom flavour probiotic fiber fortified
to state that product is probiotic. The estimated cost of the developed fiber
current market rate of Rs. 160/kg for cardamom flavour Shrikhand.
PHASE – V: STUDY OF THE SHELF LIFE OF PREPARED SHRIKHAND AT

REFRIGERATION TEMPERATURE (7 ±2 0C)
The fiber fortified Shrikhand was packed in PS (Polystyrene) cups and

stored at 7±2ºC to study its shelf life. The fiber fortified Shrikhand packed in PS
cups was stored at 7±2ºC up to 35 d. The packed product was monitored for its
rejected on sensory basis. Fresh Shrikhand had the acidity value of 0.98 per
cent LA. Which increased significantly (P<0.05) to 1.33 per cent LA on 35 d of
storage. The FFA ranged from 0.64(0 d) to 1.13 (35 d) during storage. Fresh
Shrikhand had soluble nitrogen content 0.24 per cent, which increased
significantly (P<0.05) to 0.44 per cent upon storage of 35 d. During storage, the
water activity of Shrikhand decreased significantly from 0.9391 (0 d) to 0.9330
(35 d). Fresh Shrikhand had the pH value 4.81 with decreased significantly
(P<0.05) to 4.37 on 35 d of storage. The consistency of Shrikhand ranged from
7.81 (at 0 d) to 9.23 (at 35 d).
Storage period had significant (P<0.05) effect on flavour of Shrikhand at

refrigeration temperature. The flavour score of the product was 45.66 on 0 day.
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There was significant (P<0.05) decreased in flavour score of Shrikhand at each

interval of storage. On 35 d storage there was significantly (P<0.05) lower
flavour score i.e. 33.79. Body and texture score of Shrikhand during storage
the highest score (33.26) was observed, which was significantly (P<0.05) higher
than all the storage intervals. At 35 d of storage, the lowest score (28.39) was
observed which was found significantly (P<0.05) lower than all the storage
intervals. The colour score of Shrikhand during storage was found to decrease
with the increase in storage period. Fresh Shrikhand had a score of 9.61 on 0
day which was found to decrease significantly (P<0.05) to 7.08 at 35 d of
storage under refrigeration temperature. At 35 d, score of colour was
significantly (P<0.05) lower than all other storage intervals. The total score of
Shrikhand stored at refrigeration temperature was found to decrease
significantly (P<0.05) with the increased in storage period. Fresh Shrikhand has
a total score of 93.53, which was significantly (P<0.05) highest score but
decreased significantly at each interval of storage. The overall sensory scores
gave a shelf life of more than 35 d at refrigeration temperature and found
acceptable on sensory basis.

(7.50 log cfu/g) on 35 d of storage. There was decline in probiotic count that
can be observed clearly from the graph. Nevertheless, in the Shrikhand sample
probiotic count remained well above 10 7 even after the end of refrigerated
storage for 35 d. Fresh Shrikhand sample had a SPC count of 5.29 log cfu/g
but increased significantly (P<0.05) to 5.85 log cfu/g after 35 d of storage. The
average values of YMC for stored Shrikhand ranged from (7 d) 1.53 to (35 d)
2.37 log cfu/g. There was significant effect (P<0.05) of storage period
independently on yeast and mould count of product sample on each interval of
storage at refrigerated temperature. Shrikhand sample was found 1.53-logcfu/g
Page | 154
on 7 d of storage. On 35 d it was significantly higher 2.06 (P<0.05) log cfu/g.

fresh Shrikhand samples found to free from yeast and mould at refrigeration
temperature.
The following conclusion can be drawn from this study:
 An acceptable quality cardamom flavoured fiber fortified probiotic

Shrikhand by addition oat powder at the rate of 2 per cent by weight
expected yield of Shrikhand
 The probiotic count of the developed product was 8.46-log cfu/g, which
was much higher than minimum count (6.00 log cfu/g) to state that
product is probiotic.
 The estimated cost of product was found to be Rs. 131.84/kg compared
to current market rate of Rs. 160/kg for cardamom flavour Shrikhand.
 The product met the all the compositional and microbiological
requirements as specified by FSSAI (2010) for Shrikhand except for
higher ash content. The fiber content of the product was found to be 0.24
per cent.
 The product had a shelf life of 35 d when stored at refrigeration
temperature (7±2ºC).
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xx
Appendix I
Evaluation Card for Shrikhand (IS:15348, 2003)
Name: Date: Time:
Sr. Characteristics Maximum Sample Code

No. Score
1 Flavour 50
2 Body and 35
texture
3 Colour and 10
appearance
4 Package 5
5 Total score 100
Guidelines for deducting perceived defects
Sr. Characteristics Defects Degree of Defects

No.
1 Flavour Bitter Slight Definite Pronoun
Cooked
Fermented
Foreign
Fruity
High acid
Lacks
sweetness 8 12 18
Oxidized,
rancid
Salty,
too strong
Unnatural,
yeasty
2 Body and Coarse, gummy
texture texture hard,
leaky, weak 5 9 17
3 Colour and Dull, mouldy,
appearance unclean, wet 1 2 5
4 Package Improperly 1 2 4
sealed
Soiled, torn
Comments:
Preference:
Signature:
i
Appendix II
Buffer and Media for Microbiological Analysis
1. Sterilized Skim Milk

Skim milk powder was reconstituted and then distributed in test
tubes and flasks in required quanties. It was then autoclaved to temperature
of 121ºC (15 psi) and holding at this temperature for 20 min.
2. Phosphate buffer for preparing dilutions (IS: 1479, part-III, 1962):
Stock solution of phosphate buffer was prepared by dissolving 34.0 g
of KH2PO4 in 500 ml distilled water pH was adjusted to 7.2 with 0.1 N
NaOH solution and the total volume was made up to 1 litre with distilled
water. For use as dilution water, 1.25 ml of the stock phosphate buffer
solution was diluted to 1 litre with distilled water. It was then filled in 9 ml
aliquots in tubes and sterilized by autoclaving by at 121 ºC for 20 min.
3. Standard Plate Count Agar (SPCA)
Tryptone 5.0 g
Yeast 2.5 g
extract
Glucose 1.0 g
Agar 15.0 g
Distilled 1.0 L
water
Dissolve ingredients in distilled water by boiling, and adjust to pH 7.1

±0.1. Dispense into flask and autoclave for 15min. at 1210C.
4. Potato Dextrose Agar (PDA)
Infusion from white Potatoes 200 ml

Dextrose 20 g
Agar 15 g
Suspend ingredients in 1000 ml distilled water and heat the mixture

to boiling to dissolve; distribute into flask and autoclave for 15min. at
1210C. When used as plating medium, melt it and acidify to pH 3.5 with
sterile 10 per cent tartaric acid.
ii
5. Violet Red Bile Agar (VRBA)
The quantity suggested 41.53 g of dehydrated powder of VRBA was
suspended in 1000 ml of distilled water and allow them to stand for few
min. The contents were heated with stirring to boiling temperature and held
for 2 min, to dissolve the completely, the pH of the medium was adjusted to
7.4±0.1. it was aseptically distributed in the 100 ml quantities in sterilized
flasks of 250 ml capacity. The medium was allowed to cool at 45 to 46ºC
and used for plating.
Composition of Violet Red Bile Agar:
Ingredients g/litre
Peptic digest of animal tissue 7.00
Yeast extract 3.00
Lactose 10.00
Bile salt mixture 1.50
Sodium Chloride 5.00
Neutral red 0.03
Crystal violet 0.002
Agar 15.00
6. MRS Agar Medium

The suggested quantity 67.15 g of dehydrated powder of was
suspended in 1000 ml of distilled water and allow them to stand for few
min. The contents were heated to boiling, to dissolve the completely. The
medium was cooled and the pH of the medium was adjusted to 6.5±0.2. The
medium was aseptically distributed in the 100 ml quantities in sterilized
flasks of 250 ml capacity. The medium was allowed to cool at 45 to 46ºC
and used for plating.
iii
Composition of MRS Agar:
Ingredients g/litre
Protease peptone 10.00
Beef extract 10.00
Yeast extract 5.00
Dextrose 20.00
Polysorbate 80 1.00
Ammonium citrate 2.00
Sodium citrate 5.00
Magnesium sulphate 0.10
Magnesium sulphate 0.05
Dipotassium phosphate 2.00
Agar 12.00
iv
Appendix III
Sharpe extraction Solution:
(a) Stock solution:
Glacial acetic acid 57.5ml
Sodium acetate, 3H2O 136.1g
Sodium chloride 47.0g
Calcium chloride (anhydrous) 8.9g
Make volume up to 1 lit. by distilled water
(b) Working Solution:
Stock solution 250 ml

Distilled water 750ml
Working solution 1000 ml
Thisis the sharp’s extraction solution and pH was adjusted to 5.5 by

0.1 sodium hydroxide just before use.
v
Appendix IV
Composition of Banana powder
Constituents Amounts (%)

Protein 5.12
Fat 3.2
Ash 2.46
Moisture 4.30
Fiber 2.5
Composition of Oat powder

Protein 14.2
Fat 14.2
Ash 3.4
Moisture 4.2
Fiber 12.2
Composition of Inulin powder

Fiber 92
Fructose, glucose 6
vi
Table 2.2: Composition of Shrikhand
Total
Protein Fat(per Sucrose Titrable Ash (per
Author Source solids
(per cent) cent) (per cent) acidity/pH cent)
(per cent)
Aneja et al.
Experimental 60 ≥7 5 42 - -
(1978)
Patel and Abd-
EL-Salman Experimental 57-60 6.5-7 5-6 40-43 1.05-1.10 % LA
(1986)
Sharma and
Prepared by UF 60 - 6 41 - -
Reuter (1992)
Mital (1977) Experimental 64.34-65.52 5.33-6.13 1.53-5.56 55.55-53.76 - -
Desai et al. Unhomogenize
61.93 6.73 6.07 41.32 1.14%LA -
(1985) d
Desai et al.
Homogenized 61.84 6.69 6.39 42.37 1.13%LA -
(1985)
Upadhyay and Mechanized
58.6 7.4 5.5 41 1.05-1.10%LA 0.41
Pandya (1997) process
Upadhyay and Organized
59.6 6.4 6.0 45.1 - 0.46
Pandya (1997) dairies
Upadhyay and Small sector
63.6 6.2 5.4 49.4 - 0.34
Pandya (1997) manufacturer
Kulkarni et al.
Experimental 61.0 11.5 10.0 - 4.2-4.4 0.5
(2000)
Nilofar et al.
Experimental 57.26 8.73 7.14 40 - 0.61
(2012)
Shambharkar
Experimental 65.65 6.15 8.05 41.11 1.86% LA -
et al. (2011)
Mehta (2013) Market samples 45-52 5.4-9.2 5.34-7.61 - 0.89% LA 4.7-5.9
10 | P a g e

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DEVELOPMENT OF FIBER FORTIFIED

DEPARTMENT OF DAIRY TECHNOLOGY

REGISTRATION NO: 04-2024-2012

DEPARTMENT OF DAIRY TECHNOLOGY

REGISTRATION NO: 04-2024-2012

Name of Student M ajor Advisor

DEPARTMENT OF DAIRY TECHNOLOGY

The present investigation was planned with the objective of developing

Three fiber containing powder were incorporated in Shrikhand at three

In the next part of the study comparison of Shrikhand, containing

Comparison of cardamom and rajbhog as the flavoring substances for

The gross composition of the developed cardamom flavor probiotic fiber

developed product can be considered excellent as per Indian standards (IS

The fiber fortified Shrikhand was packaged in PS cups and stored at

Storage period had significant (P<0.05) effect on flavor of Shrikhand at

Highest probiotic count was recorded in Shrikhand prepared (8.73 log

This is to certify that the thesis entitled “DEVELOPMENT OF

FIBER FORTIFIED PROBIOTIC SHRIKHAND” submitted by

Ms. Priyanka Suvera (Registration No. : 04-2024-2012) in partial

fulfillment of the requirements for the degree of Master of

Technology in Dairy Technology of Anand Agricultural University

is a record of bonafide research work carried out by him under my

Place: Anand P. S. Prajapati

I am also thankful to my wonderful Dairy Technology Department where I share and

Place : Anand (PriyankaSuvera)

No. Title Page

2.2.2 Manufacture of Shrikhand using membrane Technology 31

4.2.3.2 Effect of varying levels of fiber containing

Table No. Title Page No.

2.1 FSSA standards for Shrikhand 09

Fig. No. Title Page No.

DEPARTMENT OF DAIRY TECHNOLOGY

REGISTRATION NO: 04-2024-2012

DEPARTMENT OF DAIRY TECHNOLOGY

REGISTRATION NO: 04-2024-2012

Name of Student Major Advisor

DEPARTMENT OF DAIRY TECHNOLOGY

The present investigation was planned with the objective of developing

Three fiber containing powder were incorporated in Shrikhand at three

In the next part of the study comparison of Shrikhand, containing

Comparison of cardamom and rajbhog as the flavoring substances for

The gross composition of the developed cardamom flavor probiotic fiber

developed product can be considered excellent as per Indian standards (IS

The fiber fortified Shrikhand was packaged in PS cups and stored at

Storage period had significant (P<0.05) effect on flavor of Shrikhand at

Highest probiotic count was recorded in Shrikhand prepared (8.73 log

This is to certify that the thesis entitled “DEVELOPMENT OF

FIBER FORTIFIED PROBIOTIC SHRIKHAND” submitted by

Ms. Priyanka Suvera (Registration No. : 04-2024-2012) in partial

fulfillment of the requirements for the degree of Master of

Technology in Dairy Technology of Anand Agricultural University

is a record of bonafide research work carried out by him under my

Place: Anand P. S. Prajapati

I am also thankful to my wonderful Dairy Technology Department where I share and

Place : Anand (PriyankaSuvera)

No. Title Page

2.2.2 Manufacture of Shrikhand using membrane Technology 31

4.2.3.2 Effect of varying levels of fiber containing

Table No. Title Page No.

2.1 FSSA standards for Shrikhand 09

Fig. No. Title Page No.

Fermented milk and milk products occupy a place in satisfying

Shrikhand in Sanskrit means “Sandal”. Shrikhand, the product might