Indian Journal of Natural Products and Resources

Vol. 3 (4), December 2012, pp. 501-505

Formulation and evaluation of herbal gel

Ganesh Misal1*, Gouri Dixit2 and Vijay Gulkari1
Department of Pharmacognosy, 2 Department of Pharmaceutics,
J. L. Chaturvedi College of Pharmacy, Nagpur-440 010, Maharashtra, India
1

Received 30 June 2011; Accepted 17 July 2012

The present study has been undertaken with the aim to formulate and evaluate the polyherbal gel containing
Cassia alata Linn., Cassia tora Linn. and Cynodon dactylon Pers. extract. The gel formulation was designed by using
methanolic extract of leaves of C alata and C tora and aerial part of C. dactylon in varied concentrations (1, 2 and 4%).
Topical anti-inflammatory activity of gel was also evaluated. The gel was prepared by using Carbopol 940 (1% w/v),
C. alata, C. tora and C. dactylon extract, ethanol, propylene glycol 400, methyl paraban, propyl paraben, EDTA,
tri-ethanolamine and required amount of distilled water. The prepared gels were evaluated for physical appearance, pH,
spread ability, skin irritation to observe toxicity or side effects and also for anti inflammatory activity. It was inferred from
the results that gel formulations were good in appearance and homogeneity. The values of spread ability indicated that these
polyherbal gels were easily spread able by small amount of shear. Viscosity of polyherbal gels were determined by using
Brookfield viscometer and were ranging between 4500 to 4900 centipoise. The gels showed significant inhibition in
carrageenan induced paw oedema and formalin induced paw oedema in Wistar rat models.
Keywords: Anti-inflammatory, Carbopol 940, Cassia alata, Cassia tora, Cynodon dactylon, Herbal gel, Topical.
IPC code; Int. cl. (2011.01) A61K 36/00, A61K 36/482, A61K 36/899, A61K 127/00, A61K 135/00, A61P 29/00

Introduction
Topical application of gels at pathological sites offer
great advantage in a faster release of drug directly to site
of action, independent of water solubility of the drug as
compared to creams and ointments1,2. Cassia alata Linn.
and C. tora Linn. (Family: Caeasalpiniaceae) is
phytochemically rich in steroids, alkaloids, tannins,
triterpenes, flavanoids and anthraquinone glycoside.
Both C. tora and C. alata have been found to be
used traditionally for their various therapeutic
properties
like,
anticancer3
and
oral
4,5
anti-inflammatory , antibacterial6, antioxidant7,
skin disorder and wound-healing activities8. The
plant Cynodon dactylon Pers. (Family: Poaceae) is
phytochemically rich in steroids, tannins, flavonoids,
carbohydrates and alkaloids and has been found to
be used traditionally for various therapeutic
properties like, antiviral9, antidiabetic10, antifungal11,
antibacterial12, antioxidant13, antiulcer14 skin disorder
and wound-healing activities15,16. The present study
was
undertaken to evaluate the
topical
anti-inflammatory potential of C. alata, C. tora
andC. dactylon.
_______________
*Correspondent author:
E-mail: ganesh15misal@gmail.com, dixit.gouri@yahoo.com

The present investigation involves the preparation

of gel formulations of C. alata, C. tora and
C. dactylon (methanolic extract in varied
concentration) followed by the evaluation for physical
appearance, pH, viscosity, spread ability and for
anti-inflammatory activity.
Materials and Methods
Plant collection and authentication

The leaves of Cassia tora and C. alata were

collected in August 2010 at Hingna, MIDC area of
Nagpur. The aerial part of Cynodon dactylon was
collected from medicinal plant garden of
J. L. Chaturvedi college of Pharmacy, Nagpur. The
collected plants were authenticated at Department of
Botany, R.T. M. Nagpur University, Nagpur. Voucher
specimens (9549, 9550 and 9551) are maintained in
the Department.
Chemicals

Carbopol 934 (Merck Ltd), Diclofenac sodium

(Zim lab India), Ethanol (Loba Chem India), Methyl
Paraban (Loba Chem India), Propyl Paraben
(Loba Chem India), Propylene glycol-400 (Loba
Chem India), Tri-ethanolamine (Loba Chem India),
Carragennan (Sigma Aldrich), Formalin (Loba Chem
India) were purchased from the market.

INDIAN J NAT PROD RESOUR, DECEMBER 2012

502
Preparation of plant extracts

Evaluation of gel formulations

The collected fresh leaves of C. alata, C. tora and

aerial parts of C. dactylon were dried in hot air oven
at 40C to avoid degradation of phytoconstituents.
After drying plant materials were coarsely powdered
with Willey mill and kept in well closed container.
About 185, 100 and 125 g powder of each specimen,
respectively were defatted with Pet. ether (60-80) in
Soxhlet apparatus. After defatting, it was further
extracted with methanol. The collected extracts were
concentrated on rotary evaporator and concentrated
extracts were kept in vacuum dryer until used.

Evaluation parameters of gel formulations are

given in Table 2 and they are:
pH The pH value of polyherbal gel formulation
was determined by using a pH meter. The
measurement was performed at 1, 30, 60, 90 days
after preparation to detect any pH changes with time.
Appearance and Homogeneity All developed gels
were tested for physical appearance and homogeneity
by visual observation.
Viscosity The measurement of viscosity of the
prepared polyherbal gel was done with Brookfield
viscometer (Model RVTDV II). The reading was
taken at 100 rpm using spindle no. 6.
Spread ability The spread ability of the gel
formulations was determined by measuring the
spreading diameter of 1 g of gel between two horizontal
plates (20 cm 20 cm) after one minute. The standard
weight applied on the upper plate was 125 g.
Skin irritation studies The Wistar rats of either sex
weighing 150-200 g were used for this test. The intact
skin was used. The hairs were removed from the rat
3 days before the experiment. The gels containing extracts
were used on test animal. Gel base was applied on the
back of animal taken as control. The animals were treated
daily up to seven days and finally the treated skin was
examined visually for erythema and edema.

Animals

The Wister rats weighing between 150-200 g were

procured from Animal house of J. L. Chaturvedi College
of Pharmacy, R.T.M. Nagpur University, Nagpur and
maintained under constant conditions (temperature 25
2C, humidity 40-60%, 12 h light/12 h dark cycle).
During maintenance the animals received a diet of food
pellet supplied from animal house and water ad libitum.
These experiments were approved by the Institutional
Animal Ethics Committee, R.T.M. Nagpur University,
Nagpur (IAEC No. 648/02/6/CPCSEA).
Preparation of topical gels17- 22

The gel was prepared using the dried methanolic

extract of Cassia alata, C. tora and Cynodon dactylon.
The gel was prepared using Carbapol-940 (1%),
propylene glycol 400, ethanol, methyl paraben, propyl
paraben, EDTA, tri-ethanolamine and distilled water
in a quantity sufficient to prepare 100 g of gel in case
of blank gel. Water required for these formulations
was divided in to two parts. In one part the exact
amount of extract was dissolved and to this calculated
quantity of propylene glycol 400 and ethanol was
added and in other part, carbapol-940 was dissolved
and to this solution methyl paraben, propyl paraben
and EDTA was added. Both of these solutions were
mixed in a beaker and tri-ethanolamine was added to
the mixture dropwise to obtain the gel consistency.
The same procedure was used for preparation of
Diclofenac sodium gel as standard. Composition of
gel formulation is given in Table 1.

Assessment of anti-inflammatory activity23, 24

Carrageenan induced rat paw edema 23

Pedal inflammation in animal was produced

according to the method described by Winter et al
(1962)24. Rats were divided in 9 groups of six rats in
each. Group I was applied with gel base and served as
control. Group II standard (Diclofenac sodium Gel
0.5%) and served as reference. Group III-IX
application of 1 g of 1, 2, 4% gel of Cassia alata,
C. tora and Cynodon dactylon each, respectively.
The edema was induced by injecting 0.1 ml of
carrageenan (1% w/v) in normal saline into the sub
planter region of the left hind paw, after 1 hour of
drug application. Paw thickness was measured with
the help of Digital Vernier caliper at 0, 30, 60, 120,
180, 240 and 300 min after administration of

Table 1 Composition of polyherbal gels

Formulation
C.A., C.T., C.D. Gel 1%
C.A., C.T., C.D. Gel 2%
C.A., C.T., C.D. Gel 4%

Carbapol-940
(%)
1
1
1

Extract Propylene glycol Ethanol Methyl paraben

(%)
(%)
(%)
(%)
1
2
4

4
4
4

C.A. - Cassia alata, C.T. - Cassia tora, C.D. - Cynodon dactylon

3
3
3

0.2
0.2
0.2

Propyl paraben
(%)

EDTA
(%)

Water
(%)

0.02
0.02
0.02

0.03
0.03
0.03

Up to 100
Up to 100
Up to 100

MISAL et al: FORMULATION AND EVALUATION OF HERBAL GEL

carrageenan. The dosage details are presented in

Table 3 and 4.
Formalin-induced rat paw edema25

The formalin-induced rat paw edema model was

used for acute as well as chronic inflammation on the
basis of formalin concentration. For chronic model
2% of formalin in saline was used. Formalin-induced
edema is biphasic, an early neurogenic component is
mediated by substance P and bradykinin followed by
a tissue mediated response where histamine, 5-HT,
prostaglandin are known to be involved. The dosage
details are given in Table 5.
The % inhibition of edema was calculated by formula:
% Inhibition= 1- (-/b-y) 100
where,
= paw thickness of test animal after treatment
= initial paw thickness of test animal
Table 2 Evaluation parameters of prepared polyherbal gels
Formulation pH Appearance Homo- Spreading Viscosity
geneity diameter
(cp)
after 1 min
(mm)
Diclo gel
C.A. 1%,
C.T. 1%
C.D 1%
C.A. 2%
C.T. 2%
C.D. 2%
C.A. 4%
C.T. 4%
C.D. 4%

6.15
6.35
6.30
6.31
6.45
6.50
6.49
6.45
6.46
6.48

White
Light green
Light green
Light green
Light green
Light green
Light green
Light green
Light green
Light green

Good
Good
Good
Good
Good
Good
Good
Good
Good
Good

55
48
42
44
42
40
42
40
39
41

4500
4700
4700
4800
4700
4700
4800
4700
4800
4800

503

b= paw thickness of control animal after treatment

y= initial paw thickness of control animal.
Statistics

All values are expressed as mean SEM. P value

calculated by comparing with control by ANOVA
followed by Bonferroni post test. P values lower than
0.05 were considered significant.
Results and Discussion
The dried powder of leaves and aerial part were
defatted with petroleum ether (60-80C) and extracted
with methanol. The average % practical yield of
methanolic extract of C. alata, C. dactylon and
C. tora were found to be 18.41, 12.30 and 8.152%,
respectively. Polyherbal gels of methanolic extract of
C. alata (1%), C. tora (2%) and C. dactylon (4%) are
prepared by using Carbapol-940. Composition of
herbal gels is shown in Table 1. The physicochemical
properties of the gel formulation were shown in Table 2.
From the results it is concluded that all the gel
formulations showed good appearance and
homogeneity. The physical appearance of the gel
formulations was green in nature. The pH of the gel
formulations was in the range of 6.150.04 to
6.980.03, which lies in the normal pH range of the
skin and with time no skin irritation was observed.
There was no significant change in pH values with
time (varied from 0.05 to 0.20). Viscosity of
polyherbal gels were determined by using Brookfield
viscometer and were ranging between 4700 to 4800

Table 3 Anti-inflammatory activity of different gel formulations

Formulation
Increase in paw edema (mm)
Initial/ 0 h
h
1h
2h
3h
4h
5h
Control
(Gel base)
8.36 0.012
8.89 0.029
9.21 0.022
8.94 0.016
7.92 0.026
5.11 0.027 8.10 0.021
Standard
5.20 0.026 5.88 0.023*** 6.14 0.023*** 6.65 0.027** 6.97 0.021*** 6.70 0.012*** 5.80 0.026***
Cassia alata Gel
1%
5.11 0.020 6.18 0.026*** 6.24 0.027*** 6.96 0.029** 7.95 0.027*** 7.11 0.024*** 6.25 0.013***
2%
5.05 0.017 6.18 0.026*** 6.21 0.033*** 6.94 0.024** 7.84 0.027*** 7.08 0.013*** 6.04 0.026***
4%
5.05 0.024 6.17 0.026*** 6.21 0.022*** 6.90 0.021** 7.25 0.027*** 7.02 0.022*** 5.81 0.016***
Cynodon dactylon Gel
1%
5.02 0.045 6.64 0.033*** 6.89 0.055*** 7.45 0.061** 7.77 0.055*** 7.16 0.038*** 6.27 0.044***
2%
5.11 0.031 6.18 0.045*** 6.51 0.045*** 7.15 0.072** 7.69 0.059*** 6.98 0.040*** 6.23 0.044***
4%
5.04 0.031 6.16 0.051*** 6.46 0.051*** 6.91 0.053** 7.15 0.057*** 7.00 0.039*** 5.82 0.044***
Cassia tora Gel
1%
5.04 0.026 6.75 0.152*** 7.02 0.025** 7.55 0.026*** 7.85 0.026*** 7.43 0.027*** 6.58 0.030***
2%
5.05 0.040 6.70 0.044*** 7.00 0.078** 7.50 0.060*** 7.82 0.044*** 7.40 0.032*** 6.48 0.172***
4%
5.05 0.027 6.23 0.044*** 6.68 0.062** 7.15 0.052*** 7.21 0.060*** 6.63 0.026*** 6.05 0.049***
n: Number of animals. (n= 6)
(i) 1 g of formulation was applied to the planter surface of the right hand paw by gently rubbing 50 times with index finger.
(ii) Values are mean S.E.M. *P <0.05, **P <0.01, ***P <0.001. P value calculated by comparing with control by ANOVA followed by
Bonferroni post test.

INDIAN J NAT PROD RESOUR, DECEMBER 2012

504

centipoise. The values of spread ability indicated that

the gels were easily spreadable by small amount of
shear. Spreading diameter after 1 min (mm) of
prepared gel was between 38-55 mm which indicates
good spread ability of polyherbal gels. The results of
skin irritancy studies indicated that the prepared gels
were free from dermatological reaction. The gels were
evaluated for anti-inflammatory activity by using
carrageenan-induced rat paw edema and formalininduced rat paw edema topically. Statistical analysis
showed that the edema inhibition by formulation
containing extracts were significantly differing from
Table 4 Percent inhibition for anti-inflammatory activity
Treatment
After 3 h (%)

Percent inhibition
After 5 h (%)

Standard 56.83331.012057
78.65167 3.077156
Cassia alata Gel
1%
30.73667 0.866685*** 59.435 0.9219***
2%
31.95833 0.632861*** 64.77167 1.46761***
4%
46.34667 0.5072***
72.96 0.735314
Cynodon dactylon Gel
1%
32.93333 2.041139*** 55.52167 1.168918***
2%
37.07833 1.607943*** 60.14833 4.570212***
4%
48.54167 0.895198*** 72.245 0.875166*
Cassia tora Gel
1%
31.47 0.237241***
45.2 1.457767***
2%
33.175 0.740543***
50.18 0.855625***
4%
47.32333 0.975***
64.42 0.489518***
n: Number of animals. (n = 6)
(i) 1 g of formulation was applied to the planter surface of the
right hand paw by gently rubbing 50 times with index finger.
(ii) Values are mean S.E.M. *P <0.05, **P <0.01, ***P <0.001.
P value calculated by comparing with control by ANOVA
followed by Bonferroni post test.
Table 5 Anti-inflammatory activity of different gel
formulations
Treatment

Initial/0 day
Control
5.11 0.131
Standard
5.10 0.132
Cassia alata Gel
1%
5.11 0.131
2%
5.05 0.126
4%
5.05 0.131
Cynodon dactylon Gel
1%
5.11 0.130
2%
5.05 0.151
4%
5.10 0.092
Cassia tora Gel
1%
5.10 0.062
2%
5.04 0.088
4%
5.05 0.108

Increase in rat paw oedema

After 9 day
Percent inhibition
7.05 0.083
6.15 0.103
45.88 4.744677
6.82 0.096 11.86 4.585198***
6.61 0.096*** 19.59 1.692329***
6.41 0.084
29.30 3.856368
6.82 0.104 11.86 4.585198***
6.63 0.126*** 18.56 2.349822***
6.55 0.103 25.26 3.325367***
6.85 0.090
9.8 2.765303***
6.69 0.187*** 14.95 1.009449***
6.53 0.108 23.72 0.198224***

control group at all the concentration tested. The

results showed that the anti-inflammatory effect of the
formulation containing 4% of C. alata gel was better
than the effect of other gel formulation (Tables 3 & 4).
The highest inhibition was found at 300 min. post
carrageenan injection, which is supposed to be due to
inhibition of late phase mediators, arachidonic acid
product and prostaglandins, of acute inflammation
induced by carrageenan. Formalin-induced rat paw
edema model was used for acute as well as chronic
inflammation on the basis of formalin concentration.
For chronic model, 2% of formalin in saline is used.
Statistical analysis showed that the edema inhibition by
formulation containing extracts were significantly
differing from control group at all the concentration
tested. As compared to acute inflammation, in chronic
model, it showed no significant result as compared to
standard.
The
results
showed
that
the
anti-inflammatory effect of the formulation containing
4% of C. alata gel was better than the effect of other
gel formulation (Table 5).
Conclusion
It is inferred from results that the gel formulations
are good in appearance, homogeneity and easily
spread able and showed significant inhibition in
carrageenan induced and formalin induced paw
oedema in Wistar rat models. The results also showed
that the anti-inflammatory effect of the formulation
containing 4% of C. alata gel was better than the
effect of other gel formulation.
Acknowledgement
The authors express their sincere thanks to the
management of the J. L. Chaturvedi College of
Pharmacy, Nagpur, for encouraging and providing
research facilities and we are thankful to Principal
Dr. D. R. Chaple for the generous help and
coordination during project work.
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