Sunteți pe pagina 1din 6

Santhosh Aruna, et al.

/ International Journal of Advances in Pharmaceutical Research

IJAPR
Available Online through
www.ijapronline.org

Research Paper
ISSN: 2230 7583

FORMULATION AND EVALUATION OF A WOUND HEALING GEL


CONTAINING SYZYGIUM CUMINI LEAF EXTRACT
Santhosh Aruna Mamidi*, N.Santhi Priya, Sravani Avula, Bhavani B,
Gopi chand U, Fathima SK, Rama Rao.Nadendla.
Department of Pharmaceutics, Chalapathi Institute of Pharmaceutical Science, Lam, Guntur
Received on 09 01 - 2015

Revised on 14 01- 2015

Accepted on 25 02 2015

ABSTRACT
Wounds are defined as the disruption of anatomical and functional integrity of living tissue. Wound healing is an
intricate and continual cascade of events, with various cellular and biochemical processes, ultimately resulting in the
reconstruction and regeneration of damaged tissue. Plants and their extracts have immense potential for the
management and treatment of wounds. The phyto-medicines for wound healing are not only cheap and affordable
but are also safe as hyper sensitive reactions are rarely encountered with the use of these agents. These natural
agents induce healing and regeneration of the lost tissue by multiple mechanisms. Various plant products have been
used in treatment of wounds over the years. Wound healing herbal extracts promote blood clotting, fight infection,
and accelerate the healing of wounds. The aim of present study was to prepare and evaluate the wound healing
activity of herbal gel. Our present study reveals that the new polyherbal formulations posses potent wound healing
activity, which could be a good choice of remedy for wound healing.
KEY WORDS: Wound healing, phyto-medicines, regeneration, polyherbal formulations
INTRODUCTION
Plants have bear the basis of many traditional
medicines throughout the world for thousands of
years and continue to provide new remedies to
mankind1.Correct knowledge of such crude drugs is
very important aspect in preparation, safety and
efficacy of the herbal products. Pharmacognosy is a
simple and reliable tool, by which complete
informations of the crude drugs can be obtained2.
Syzygium cumini Linn (family Myrtaceae),
commonly known as Jamun (Hindi), is a medicinal
plant and utilizable species. Common names are Java
plum, Black plum, Jambul and Indian Blackberry 3.
Corresponding author
M.Santhosh Aruna
Assistant Professor
Department of pharmaceutics
Chalapathi Institute of Pharmaceutical Sciences,
Lam, Guntur
Andhra Pradesh, India- 522034.
E mail: santhosharuna.kathi@gmail.com
Mobile no: 91-8143275848

IJAPR /April. 2015/ Vol. 6/Issue.04/ 82 87

The original home of jamun is India, distributed


throughout India, in forest up to 1800m usually along
the bank and moist localities .The sprouts are
refrigerant, carminative &astringent to bowels.
Powdered seeds are used as remedy in diabetes and
in metrorrhagia 4. As per Unani system of medicine,
they act as liver tonic, enrich blood, strengthen teeth
and gums and form good lotion for removing
ringworm infection of the head. Leaves have been
used in traditional medicine as a remedy for diabetes
mellitus in many countries. The leaves are also used
to strengthen the teeth and gums, to treat leucorrhoea,
stomachalgia,
fever,
gastropathy
strangury,
Dermopathy, constipation and to inhibit
in the
faeces5. The major phyto constituents are reported to
contain vitamin C, gallicacid, tannins, anthocyanins,
includes cyanidin, petunidin, malvidin, glycoside and
other components 6, 7.
Wound is defined as the disruption of the
cellular and anatomic continuity of a tissue. Wound
may be produced by physical, chemical, thermal,
microbial or immunological insult to the tissues

82

Santhosh Aruna, et al. / International Journal of Advances in Pharmaceutical Research


Wounds are physical injuries that results in an
opening and break of the skin that cause disturbance
in the normal skin anatomy and function. They result
in the loss of continuity of epithelium with or without
the loss of underlying connective tissue8. Wound may
be produced by physical, chemical, thermal,
microbial or immunological insult to the tissues.
Topical gel drug administration is a
localized drug delivery system anywhere in the body
through ophthalmic, rectal, vaginal and skin as
topical routes. Skin is one of the most extensive and
readily accessible organs on human body for topical
administration and is main route of topical drug
delivery system. Topical application of drugs offers
potential advantages of delivering the drug directly to
the site of action and acting for an extended period of
time. It can penetrate deeper into skin and hence give
better absorption1. They are deemed more effective
less toxic than conventional formulations due to the
bilayer composition and structure.
The U.S.P. defines gels as a semisolid
system consisting of dispersion made up of either
small inorganic particle or large organic molecule
enclosing and interpenetrated by liquid. Gels consist
of two phase system in which inorganic particles are
not dissolved but merely dispersed throughout the
continuous phase and large organic particles are
dissolved in the continuous phase, randomly coiled in
the flexible chains 5.
Research on wound healing agents is one of
the developing areas in modern biomedical sciences.
Many of the synthetic drugs currently used for the
treatment of wounds are not only expensive but also
pose problems such as allergy, drug resistance etc
and this situation has forced scientists to seek
alternative drugs . In spite of tremendous
development in the field of synthetic drugs during
recent era, they are found to have some or other side
effects, whereas plants still hold their own unique
place, by the way of having no side effects.
MATERIALS AND METHODS
Plant Collection
The leaves of the plant syzygium cumini were
collected from the medicinal plant garden of
Chalapathi institute of pharmaceutical sciences.
Collected plant material was subjected for
identification based on herbarium specimens at
department of Pharmacognosy, Chalapathi institute
of pharmaceutical sciences, Guntur. The voucher
specimen no ANU/CIPS/Identi /Tech/2014/24.
Macroscopic Study
The leaves measuring about 10 to 15 cm long and 4
to 6 cm wide. These are entire, ovateoblong,
sometimes lanceolate and also acuminate, coraceous,

IJAPR /April. 2015/ Vol. 6/Issue.04/ 82 87

tough and smooth with shine above. The fragrant


flowers of Jamun are small, nearly 5 mm in diameter.
These are arranged in terminal trichotomous panicles
greenish white in color.
Microscopy: Transverse section of S. cumini leaves
showed following featuresEpidermis: Two to three layered epidermis.
Mesophyll: It is composed of isodiametric thin
walled parenchymatous ground cells which are
packed with simple starch grains. In the mid-rib
region, the vascular bundles show xylem, towards
upper epidermis and phloem on the lower side. Starch
grains, oil globules, tannin cells and stone cells are
also visible.
Phytochemical Investigation: The leaves were
washed properly & cut into small pieces before being
subjected to cold maceration for seven days. The
solvent used was ethanol (95%), after 7 days; the
macerates were filtered through muslin cloth &
concentrated using rotary evaporator. The ethanolic
extracts were tested for the presence of various phyto
constituents like tannins, alkaloids, carbohydrates,
flavonoids, sterols, & glycosides etc9.
Extraction of syzygium cumini:
The powdered leaves were used for extraction. The
powder is extracted in soxhalet apparatus with
ethanol. The extract obtained was dried in a vacuum
evaporator under reduced pressure and below50C to
give a dried residue. The product is stored in
desiccator for further studies.
Animals collection:
Wister Males rats (150-200gm) were procured from
Animal House of Chalapathi institute of
pharmaceutical sciences, lam, Guntur & were fed a
standard diet, water was provided & they were
acclimated 7 days before entry into subsequent study.
The protocol was approved by Institutional Animal
Ethics Committee (IAEC).
Formulations of Gels:
Carbopol 934P NF 0.1gm was measured and was
dispersed in 10ml of distilled water and mixed by
stirring continuously in a magnetic stirrer at 800rpm
for 1 h. Glycerol 5ml was added to the mixture under
continuous stirring. The mixture was neutralized by
drop-wise addition of 50% triethanolamine (w/w).
Mixing was continued until a transparent gel was
formed. The extract of syzygium cumini was
incorporated into the gel base and mixed
continuously for uniformity. Three types of gels are
formulated of which F I containing syzygium cumini
leaf extract and F II is a poly herbal gel where a
mixture of sesame oil and leaf extract and F III
contains sesame oil F IV is control.
Evaluation of Gel

83

Santhosh Aruna, et al. / International Journal of Advances in Pharmaceutical Research


1. Physical evaluation: The color, appearance and
the feel on application of the prepared herbal gel
formulations were noted and the results are shown in
Table 4
2. Subjective Properties: Subjective properties such
as consistency, texture and Irritation are observed and
shown in Table 5
3. pH measurement: The pH of the gel was
determined by using a digital dissolved in 50 ml
water and pH was determined by dipping the glass
electrode completely into gel solution system so as to
cover the electrode. Then instrument reading in terms
of pH are tabulated in the Table 6. The pH was
studied for 30 days.
4. Viscosity: The measurement of viscosity of the
prepared gel was done with a Brookfield Viscometer.
The gels were rotated at 0.3, 0.6 and 1.5 rotations per
minute. At each speed, the corresponding dial reading
was noted. The viscosity of the gel was obtained by
multiplication of the dial reading with factor given in
the Brookfield Viscometer catalogues 10.
5. Spreadability: It indicates the extent of area to
which gel readily spreads on application to skin or
affected part. The therapeutic potency of a
formulation also depends upon its spreading value.
Spreadability is expressed in terms of time in seconds
taken by two slides to slip off from gel which is
placed in between the slides under the direction of
certain load. Lesser the time taken for the separation
of two slides, better the spreadibility. It is calculated
by using the formula
S = M. L / T
where,
M = wt. tied to upper slide
L= length of glass slides
T = time taken to separate the slides
6. Stability testing: Since the period of stability
testing can be as long as two year, it is time
consuming and expensive. Therefore it is essential to
device a method that will help rapid prediction of
long term stability of drug. The accelerated stability
testing is defined as the validated method by which
the product stability maybe predicted by storage of
the product under condition that accelerated the
change in defined and predictable manner. The
stability studies of formulated gels were carried out at
40C, 250C, 450C and at a room temperature for the
period of one month. The effect of temperature,
humidity and time on the physical characterization of
the gels was evaluated for assessing the stability of
prepared formulation. The result was shown in Table
7
7. Antimicrobial activity: The anti-microbial
activity of each formulation was assessed by
measuring the zone of inhibition in nutrient agar

IJAPR /April. 2015/ Vol. 6/Issue.04/ 82 87

medium, employing pseudomonas aeruginosa and


Staphylococcus aureus as test organisms 11,
8. Wound healing studies: An excision wound
model was used for studying wound healing activity.
Albino rats (Wistar strain) of both sexes weighing
between 150200 g were randomly divided into 8
groups of six animals each. The back of the each
animal was shaved and prepared after washing with
spirit. An area of about 2 sq.cm was defined with a
marker on the shaven back of the animals. The
circular marked area was excised with its full
thickness using a surgical sterile blade and scissors
under phenobarbitone anesthesia.
RESULT AND DISCUSSION
Physical Evaluation of Gels:
The results of various physical parameters for
evaluation of the prepared herbal gel Formulation are
reported below. The physical parameter such as
color, appearance, and feel on application are
observed and shown in Table No 4. From the
physical evaluation the color of prepared herbal gels
was dark green, faint yellow and light yellow as the
color of extracts was green and yellow. Appearance
of gel was translucent and it was smooth on
application. So it shows significant physical
evaluation parameters. The subjective properties
such as consistency, texture and irritation are
observed. and are shown in Table No.5. The
subjective properties such as consistency were good
and texture of prepared herbal gel was found to be
smooth. No skin irritation was thereon application of
gel to the skin surface. So it can be used safely. The
pH value of gel formulation were studied at room
temperature are change in pH is observed and shown
in Table No.6. pH value of prepared herbal gel
incorporating the medicinal plant extract was studied
by using digital pH meter Systronics. (pH meter type
335). The pH was studied for 30 days at room
temperature. All four formulations were in range of
6.35 - 6.52 pH at initial phase. As we go from
epidermis to dermis, pH of the skin increases and
attained the neutral value i.e. 7. So gel formulation
having pH range 6.2 to 7.0 are desirable to skin since
they do not interfere with the physiology of skin.
The Stability testing of the different formulation was
shown in Table No.8. The prepared herbal gel
formulations were subjected to accelerated stability
testing. The prepared herbal gel were store at 4oC,
250C,450C in refrigeration, room temperature and
oven for a period of 30 days to study effect of
temperature and at different humidity condition. The
physical parameter were evaluated during study
period the result of study indicates that preparation
are physically stable at 450C.
Anti-microbial activity of the formulations

84

Santhosh Aruna, et al. / International Journal of Advances in Pharmaceutical Research


The gel base without the herbal extracts did not
shown any zone of inhibition. The zone of inhibition
was found to increase on increasing the herbal drug
concentration. Hence the results of this study confirm
that the herbs possess anti-bacterial activity and this
will help keep the wound area sterile, thus promoting
wound healing. This fact supports a faster wound
healing in the treated groups compared with the
control group.

Wound contraction studies


The results of wound contraction studies indicate that
all the formulations enhance wound healing in open
wounds. The rate of wound contraction was found to
reach a maximum on the 12th day in the treated
groups. The gel formulations produced better wound
contraction compared with the marketed formulations

Table 1: Physicochemical Parameters


Parameter
Value (%)
Total Ash
12
Acid Insoluble Ash
1.5
Water soluble ash
3
Loss on drying
32
Methanol soluble extractive
4
Ethanol soluble extractive
4.2
Petroleum ether soluble extractive
3
Benzene soluble extractive
1.8

Chemical constituents
Carbohydrates
Flavanoids
Phytosterols
Glycosides
Alkaloids
Tannin & phenolics

Table 2: Phytochemical Screening


Test
Molischs Reagent
Benedicts Reagent
Shinoda Test
Salkowskis
Legal test
Dragonodroffs reagent
Ferric chloride solution
Lead acetate solution

Table 3:
INGREDIENTS
Plant extract
Sesame oil
Carbopol 934 (0.1%)

Parameter
Color
Appearance
Feel on application

Parameters
Consistency
Texture
Irritation

Preparation of medicated formulations


FI
F II
F III
1g
0.2ml
0.2 ml
0.2ml
0.1 g
0.1 g
0.1g

Table 4: Physical Evaluation of Gels


FI
F II
Dark green
Yellow
Translucent
Translucent
Smooth
Smooth

Table 5: Subjective properties of Gels


FI
F II
Good
Good
Smooth
Smooth
_
_

IJAPR /April. 2015/ Vol. 6/Issue.04/ 82 87

Result
(+)ve
(+)ve
(-)ve
(-)ve
(+)ve
(-)ve
(+)ve
(+)ve

CONTROL
0.1 g

F III
White
Transcluent
Smooth

F III
Good
Smooth
_

85

Santhosh Aruna, et al. / International Journal of Advances in Pharmaceutical Research


Table 6: pH of the Gels:
FI
F II
6.37
6.50
6.38
6.49
6.40
6.47
6.41
6.48
6.39
6.47
6.39
6.48

Time in days
0
2
7
14
22
30

Formulation
Formulation I
Formulation II
Formulation III

F III
6.39
6.38
6.39
6.40
6.39
6.38

Table 7: spread ability and viscosity


Spreadability (mm)
Viscosity (cp )
55
4500
48
4700
45
4500

Formulation

Initial colour

Formulation I
Formulation II
Formulation III

Dark green
Yellow
White

Table 8: Stability testing


I month
II month
+
+
+

III month

+
+
+

IV month

+
+
+

+
+
+

Table 9: Anti microbial studies (Zone of inhibition shown by the formulations).


Formulation
P. aeruginosa (mm*)
S.aureus (mm*)
Formulation I
14
6
Formulation II
24
10
Formulation III
26
13
Table 10: wound contraction studies
Area of wound during different days of observation (%)
Treatment in days

12

16

21

Control

06.60
0.7160

15.60
0.7065

36.94
0.9410

53.44
0.7819

66.38
0.5671

Formulation I

1.88**
0.5347

25.38**
0.5043

47.71**
0.5809

67.60**
0.5994

79.05**
0.5802

Formulation II

07.72*
0.7276

18.16*
0.5369

37.10*
0.7287

54.27*
0.5468

67.94*
0.5477

Formulation III

8.27**
0.7821

19.38**
0.6963

38.88**
0.5567

56.22**
1.060

72.27**
0.5802

Marketed

1 3.88
29.38
53.94
73.27
88.33
0.7487
0.7272
0.6522
0.6408
0.5671
Values are expressed as mean SEM, N= 6, *p<0.05, **p<0.01 Vs Group 6 One way ANOVA followed by
Dunkeertannets test
CONCLUSION
Various topical application dosage forms like creams,
ointments, liniments, lotions, gels and jellies have
been in use for many decades. Gels and jellies are
although age old formulations, they have now gained
more and more importance and the extensive studies

IJAPR /April. 2015/ Vol. 6/Issue.04/ 82 87

on their release properties have revealed that the


active ingredients in gel based formulations are better
percutaneous absorbed than from creams and
ointment bases. Thus the present research work
suggests that herbal gel formulation holds a
tremendous potential against wound healing and can

86

Santhosh Aruna, et al. / International Journal of Advances in Pharmaceutical Research


prove to be a safe and efficacious remedy for treating
wounds. However an elaborate protocol for the
clinical trials is needed to be designed and
implemented to check the anti-acne activity on
human volunteers.
ACKNOWLEDGEMENTS
We would like to sincerely thank the management
and Principal of Chalapathi institute of
pharmaceutical sciences, Guntur, for letting us avail
the facilities of the College. We are also thanking to
department of biotechnology and department of
pharmacology, their co-operation for the Invivo
studies.
REFERENCES
1. Patel SS. Morphology and Pharmacology of
Passiflora edulis. Journal of Herbal
medicine and Toxicology. 2009; 3(1):1-6.
2. KR and Basu BD. Indian Medicinal Plants.
Vol. II. 1975:1052-53.
3. Nadkarni KM. Indian Materia medica. Vol.
I, book depot, Bombay, India, 1954; 51618.
4. Sharma
P
and
Mehta
PM.
In
Dravyagunavignyan. (The Chowkhamba
Vidyabhawan), Part II & III, Varanasi.
1969; 586.
5. Gowri Shyamala S and Vasantha K.
Phytochemical Screening and Antibacterial
Activity of Syzygium cumini (L.)
(Myrtaceae)Leaves Extracts. International
Journal of Pharm Tech Research. 2010 ;(
2):1569-1573.

6. Martinez SB and Del Valle MJ. Storage


stability and sensory quality of duhat
Syzygium Cumini Linn. Anthocyanins as
food colorant. UP Home Economic Journal.
1981 ;( 9):1.
7. Wealth of India. Raw materials, New Delhi:
CSIR, Vol X. 1976; 100104.
8. Charde M.S., Fulzele S.V., Satturwar P.:
Wound Healing and Anti-inflammatory
potential of Madhu Ghrita, Indian Journal
of pharmaceutical science, 2006; 68(1); 2631.
9. Khandwal
KR.
Practical
Pharmacognosy.10th
edition,
Nirali
Prakashan, Pune. 2003:158.
10. Harding K.G., Morris H.L.; healing chronic
wounds. Science, medicine, and the future
.2002; 324,160-163.
11. Stadelmann WK, Digenis AG, Tobin GR.
Physiology and healing dynamics of
chronic cutaneous wounds. Am. J. Surg.
1998; 176: 26-38.
12. Madden JW, Peacock J. Studies on the
biology of collagen during wound healing.
I. Rate of collagen synthesis and deposition
in cutaneous wounds of the rat. Surgery
1968; 64:288-294.

INTERNATIONAL CONGRESS IN PHARMACY AND HEALTH SCIENCES


Pharma Science Tech Association, Foundation No: AP/PSTA/56/2012.
Please visit for Details: www.icphsmembership.com
Totally three types
FICPHS (Fellowship in International Congress in Pharmacy and Health Sciences), MICPHS (Member in International Congress in
Pharmacy and Health Sciences), AMICPHS (Associate Member in International Congress in Pharmacy And Health Sciences)
Eligibility
FICPHS: Ph.D in Chemistry/ Pharmacy / M.Sc / M.Pharm with 2 years experience, MICPHS: M.Sc / M.Pharm
2 years experience, AMICPHS: B.Sc (or) B.Pharmacy

IJAPR /April. 2015/ Vol. 6/Issue.04/ 82 87

(or) B.Sc / B.Pharm with

87