MATERIALS AND METHODS

The present research entitled “Enrichment of filled milk paneer by the use of
medicinal and aromatic herbs” was conducted in the Students Training Dairy and
Research Laboratory of Warner School of Dairy and Food Technology and Nutrition
Research Laboratory of Ethelind School of Home Science, Sam Higginbottom
Institute of Agriculture Technology & Sciences. Materials used in the experiment and
techniques employed in the treatment of milk for preparation of filled milk paneer,
different tests and evaluation of the product involved are dealt within this chapter.

3.1 Materials Required:

Skim milk

Skim milk powder

Citric acid

Amyl alcohol

Gerber acid (H2SO4)

Vegetable oil (Saffola)

Sodium hydroxide

Hydrochloric acid (Conc.)

Sulphuric acid (Conc.)

Phenolphthalein indicator

Zinc sulphate

Lactose

Mercuric oxide

Sodium sulphate

Boric acid

Methyl red indicator

Methyl blue indicator

Petroleum eather

Herbs- Basil, Mint and Fenugreek Leaves

Material and Methods Page 31

Nutrient Agar

Macconkey Agar

Potato Dextrose Agar and salt

3.2 METHODS:

3.2.1- Collection of Milk:

Whole milk and skim milk were collected from students training dairy, SHIATS,
Allahabad for each replication. The skim milk was standardised to desired level of fat
and SNF percentage by using skim milk and saffola oil.

3.2- Testing of whole milk:

(a)Sampling of milk

Sampling of milk was carried out as per the procedure laid down in I.S. 1479 (Part-I),
1960.

(b) Determination of fat percentage of whole milk

The milk was tested for fat percentage as per the procedure laid down in IS:1224
(Part- I), 1977 (Gerbar method).

(c) Determination of SNF percentage of milk

The SNF percent of milk was determined as per the procedure given in exercise 30 of
Manual in Dairy Chemistry ICAR, 1972(Appendix-A).

3.3- Testing of skim milk:

(a) Sampling of skim milk:

Sampling of skim milk was carried as per the procedure laid down in IS:1979 (Part-
I), 1960.

(b) Determination of fat percentage of skim milk: The skim milk was tested for fat
percent as per the procedure given in IS: 1224 (Part-I), 1977 (Gerbar method).

(c) Determination of SNF percentage of skim milk:

The procedure given in exercise 30 of Manual in Dairy Chemistry, ICAR (1974) was
followed to calculate the SNF percentage of milk (Appendix-A).

Material and Methods Page 32

3.4. Standardization of whole milk for preparing paneer:

The whole milk was standardised to 5.5 percent fat and 8.5 percent SNF and was used
for preparing paneer. Paneer thus prepared was represented as T0, and served as
control for filled milk paneer. The SNF percentage of milk was maintained by
addition of skim milk powder (Appendix-B).

3.5. Preparation of herbs extract:

The herbs that are basil, mint, and fenugreek represented as H1, H2, and H3
respectively, procured from local market and change in to paste by using mixer and
grinder separately with the addition of 50 ml of water in to 100 g of leaves and
pressed through muslin cloth for extraction of liquid. Extract was used in the
proportion of 1%, 2% and 3% in filled milk paneer and represented as L1, L2 and L3
respectively.

3.6. Preparation of experimental paneer: For the preparation of experimental

paneer, milk was standardized at two levels 5% fat and 8.5% SNF and 5.5% fat and
8.5% SNF and then heated to 85oC. At the same time basil leaves, mint leaves and
fenugreek leaves were cleaned and extracted. The heated milk was cooled to 70oC.
Extract of basil leaves, mint leaves and fenugreek leaves was added into the milk.
The level of herbs was 1%, 2%, and 3% separately, these levels of extract were
indicated as L1, L2, L3, respectively (see Fig. 1).

Milk

Heating to 60-65oC

Pre-calculated amount of vegetable oil (saffola) is heated 65oC

Blending of vegetable oil (saffola) with milk

Mixing/Stabilizing the vegetable oil at 65oC by using mixture and grinder (Mixy)

Pasteurization (72 oC for 15 seconds)

Filled milk

(Fig. no. 1 Preparation of filled milk)

Material and Methods Page 33

3.7. Preparation of filled milk paneer: For the preparation of filled milk pre
calculated amount of saffola and skim milk were heated to 65oc and mixed by
blending it in to mixer and grinder. Milk fat was standardized at two different level
5% fat and 8.5 % SNF (F1) and 5.5 % fat and 8.5% SNF (F2).The extract was added in
to milk at 85oc and holds it for 5 minutes at same temp. In filled milk same procedure
was adopted as used for preparing control paneer (see fig. 2).

Filled milk

Standardized to 5% fat Standardized to 5.5% fat Medicinal and aromatic herbs

and 8.5% SNF and 8.5% SNF (Basil, Mint, Fenugreek leaves)

Heating 85oC Cleaning of herbs

Holding for 5 minutes Extraction of leaves

Cooling up to 70oC (in water bath)

Add extract of leaves into the milk at

1% 2% 3%
L1 L2 L3

Coagulation at 70oC
(with 1% citric acid solution)

Straining through muslin cloth

Hoopping and pressing

Immersion of paneer in chilled water (4oC) and

rubbing of 1% salt by weight

(Fig no. 2 Preparation of experimental paneer)

Material and Methods Page 34

3.8 Preparation of coagulant: For each of the replication, 1 litre of 1% citric acid
solution was prepared by dissolving 10 g of citric acid in 1 litre of distilled water.

3.9 Plan of Work:-

3.9.1 Type of milk:
I. 5% fat and 8.5% SNF II. 5.5% fat and 8.5% SNF - 2

3.9.2Type of medicinal and aromatic herbs:

I. Basil II. Mint III. Fenugreek - 3

3.9.3 Levels of medicinal and aromatic herbs

I. 1% II. 2% III. 3% - 3

3.9.4 Temperature of coagulation: 70oC - 1

3.9.5 Coagulant: Citric acid - 1
3.9.6 Packaging: Alluminium foil - 1
3.9.7 Storage: 0, 5, 10.15,20,25 days - 6

No of replications - 3
Total no of replications 2x3x3x5x3+1x3 - 273

Treatment combinations: Two different level of milk (5% and 5.5%), three types of
herbs (basil, mint and fenugreek) and their three levels of herbs (1%, 2%, 3%) were
used for preparation of filled milk paneer and control filled milk paneer was made on
5% level of fat. Prepared filled milk paneer were compared with control paneer in
which treatments was absent.

Material and Methods Page 35

 The different treatment combinations used in the experiment were represented as
follows:

T0 Control paneer at level of 5% fat and 8.5% S.N.F

2 F1H1L1 paneer made by using 1% basil extract at level of 5% fat and 8.5% S.N.F
3 F1H1L2 paneer made by using 2% basil extract at level of 5% fat and 8.5% S.N.F
4 F1H1L3 paneer made by using 3% basil extract at level of 5% fat and 8.5% S.N.F
5 F1H2L1 paneer made by using 1% mint extract at level of 5% fat and 8.5% S.N.F
6 F1H2L2 paneer made by using 2% mint extract at level of 5% fat and 8.5% S.N.F
7 F1H2L3 paneer made by using 3% mint extract at level of 5% fat and 8.5% S.N.F
8 F1H3L1 paneer made by using 1% fenugreek extract at level of 5% fat and 8.5% S.N.F
9 F1H3L2 paneer made by using 2% fenugreek extract at level of 5% fat and 8.5% S.N.F
10 F1H3L3 paneer made by using 3% fenugreek extract at level of 5% fat and 8.5% S.N.F
11 F2H1L1 paneer made by using 1% basil extract at level of 5.5% fat and 8.5% S.N.F
12 F2H1L2 paneer made by using 2% basil extract at level of 5.5% fat and 8.5% S.N.F
13 F2H1L3 paneer made by using 3% basil extract at level of 5.5% fat and 8.5% S.N.F
14 F2H2L1 paneer made by using 1% mint extract at level of 5.5% fat and 8.5% S.N.F
15 F2H2L2 paneer made by using 2% mint extract at level of 5.5% fat and 8.5% S.N.F
16 F2H2L3 paneer made by using 3% mint extract at level of 5.5% fat and 8.5% S.N.F
17 F2H3L1 paneer made by using 1% fenugreek extract at level of 5.5% fat and 8.5% S.N.F
18 F2H3L2 paneer made by using 2% fenugreek extract at level of 5.5% fat and 8.5% S.N.F
19 F2H3L3 paneer made by using 3% fenugreek extract at level of 5.5% fat and 8.5% S.N.F

3.10 Assessment of quality –The quality of paneer was analyzed on the basis of
sensory, chemical and microbial parameters throughout the investigation.

3.10.1 Sensory attributes: The sensory attributes were assess for its Flavour and
taste, Body and texture, Colour and appearance and overall acceptability. Sensory
evaluation of paneer was done by a panel of five judges by using “9 point hedonic
score card”. Scores were allocated for various parameters. The samples were placed
before the judges with sample code by random number table to avoid positional or
order biasness. The panel of judges graded the coded samples of the product
(Appendix-C).

Material and Methods Page 36

3.10.2 Chemical content: The chemical content of the value added filled milk paneer
and its nutrient availability of experimental paneer as well as control was determined
through standard procedure as given below-

3.10.2.1 Sampling of paneer:

The procedure given in Manual in Dairy Chemistry, ICAR (1972), for sampling of
cheese was followed.

3.10.2.2 Determination of moisture percentage:

The moisture percentage of paneer was determined as per the procedure given in
exercise 31 of Manual in Dairy Chemistry, ICAR (1972) (Appendix-D).

3.10.2.3 Determination of fat percentage:

The fat percentage of paneer was determined as per procedure given in I.S.I hand
book of food analysis (Appendix-E).

3.10.2.4 Determination of protein percentage in paneer:

The total protein content of paneer was determined by the method of Menefee and
Overman (1940) (Appendix-F).

3.10.2.5 Determination of Lactose percentage in paneer:

The lactose percentage in paneer sample was determined by calorimetric method as

described by Action (1977).

3.10.2.6 Determination of Ash percentage in paneer:

Ash percentage was determined according to the methods described in AOAC (1980)
(Appendix-G).

3.10.2.7 Determination of Iron in paneer:

Iron was determined according to the methods described in AOAC (1980) (Appendix-
H).

Material and Methods Page 37

3.10.2.8 Determination of Calcium in paneer:

Calcium was determined according to the methods described in AOAC (1980)

(Appendix-I).

3.10.2.9 Determination of total carotene in paneer:

Total carotene was determined according to the methods described in AOAC (1980)
(Appendix-J).

3.10.2.10 Determination of total energy:

Physiological energy value

Kcal/100 gm= (4 x protein %) + (4 x CHO %) + (9 x fat %)

3.10.3 Microbial attributes: In microbial evaluation of the filled milk paneer

standard plate count, yeast and moulds and coliform were determined as per the
standard given below.

3.10.3.1 Preparation of dilutions:

Dilutions of paneer were prepared as per the procedure laid down in the Manual in
Dairy Bacteriology, ICAR (1972) (Appendix-K).

3.10.3.2 Testing of paneer for Standard Plate Count:

SPC were determined as per procedure laid down in the manual in Dairy
Bacteriology, ICAR (1972) (Appendix-L).

3.10.3.3 Testing of paneer for Yeast and Mould Count:

YMC were determined as per procedure laid down in the manual in Dairy
Bacteriology, ICAR (1972) (Appendix-M).

3.10.3.4 Testing of paneer for Coliform Count:

Coliform Count was determined as per procedure laid down in the manual in Dairy
Bacteriology, ICAR (1972) (Appendix-N).

Material and Methods Page 38

3.11 CONSUMER ACCEPTABILITY:

The consumer acceptability of the product was judged by subjecting all combinations
of paneer to a group of 30 consumers (Appendix-C).

3.12 COST ANALYSIS OF PANEER:

Cost analysis of filled milk paneer was estimated on the basis of yield of paneer and
raw ingredients purchased from the local market of Allahabad, used during
preparation.

 Milk
 Skimmed milk
 Vegetable oil (Saffola)
 Herbs (basil, mint and fenugreek)
 Skimmed milk powder
 Coagulant (citric acid)
3.13 STATISTIAL ANALYSIS:

In order to study the effects of various fat level, type of herbs and level of herbs at
different storage periods of filled milk paneer, a laboratory experiment was conducted
and required data were collected and tabulated. The data obtained were statistically
analysed for its validity on the basis of factorial completely randomized design. SAS
method was used to analyse the observed data. Tables of analysis of variance are
given below-

3.16.1: Structure of analysis of variance for chemical attributes-

Source d.f S.S M.S.S F.cal F table value

5% 1% 0.1
%
F 1
H 2
L 2
FXH 2
FXL 2
HXL 4
FXHXL 4
Treat vs control 1
Error 36
Total 54

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3.16.2: Structure of analysis of variance for sensory and microbial attributes-

Source d.f S.S M.S.S F.cal F table value

5% 1% 0.1%
F 1
H 2
L 2
FXH 2
FXL 2
HXL 4
FXHXL 4
Treat vs cont. 1
S 5
FXS 5
HXS 10
LXS 10
FXHXS 10
FXLXS 10
HXLXS 20
FXHXLXS 20
Treat vs cont. x S 5
Error 228
Total 341

Material and Methods Page 40