5 gm of gel sample was
taken in a 250 mL flask and 30 mL of acetic acid and chloroform solution were
added and swirled gently.6 0.5 mL of potassium iodide solution was added with
continuous shaking and 30 mL of water was added thereafter. The solution was
then titrated with 0.1 M sodium thiosulfate solution with vigorous shaking
until yellow color almost disappears. Then 0.5 mL of 1% starch was added and
titration was continued with vigorous shaking to release all iodine from
chloroform layer, until blue colour disappeared.6, 40
Peroxide value = S × M × 1000/gm
sample
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Where S = mL of sodium thiosulfate and M
= Molarity of sodium thiosulfate solution.
Total fatty
matter determination
2 gm of gel sample was
taken and 20-25 mL of 1:1 dilute HCl was added to it. It was then heated on a water bath till the
solution becomes clear.6 The sample (aqueous phase) was drawn in a 250 mL
separating funnel and then allowed to cool at room temperature. 50 mL of
petroleum ether (organic phase) was then added in the funnel and shaked and
left for separation to occur.6 The organic phase was collected. The above
aqueous layer partitioned twice with same quantity of petroleum ether. The
organic layers were collectively evaporated to obtain residue which was
consequently washed with water. The residue was filtered and sodium sulfate was
added to it. The mixture was again filtered, the extract was dried and the
content was determined.6, 41
Total
fatty matter (%) by mass = 100×M1/M2;
M1
= mass of residue; M2 = mass of sample in gram.
Product evaluation on skin (Patch test)
Twenty
volunteers were selected whose ages were in between (20-35) years. Prior to the
study consent form was filled by each of them.6 Volunteers having serious
skin diseases, asthma were excluded from the study. Patch test was performed on
the forearms of each volunteer to determine any possible reaction(s) to the
formulation.6, 42 The prepared formulations A, B and C along with base alone
were applied on the forearms of the volunteers separately. Adhesive tape was
used to fix them in place and the test sites were marked. The patches were left
in place for 48 h, during which care was taken not to wash the applied area.6
After 48 hour, the patches were removed and reading was taken one hour later. Skin
was examined for any redness, itching, or blemishes. These visible signs along
with any itchy or irritable sensations indicated that there is something wrong
to the product. Clear skin devoid of aforesaid visible signs indicated that the
product is safe to use.6, 40, 43
Stability
of polyherbal gel
Stability of base and formulations (A, B and C) were
studied at applied different storage conditions and checked for their physical characteristics
like color, appearance, odor and centrifugation test (for 30 days).6
Antibacterial
activity of polyherbal gel
The prepared formulations were
screened for their antibacterial activity by Disc plate method.6, 44-47 It
was tested on nutrient medium against S.
aureus, B. subtilis, A. niger and E.
coli which are representative types of Gram positive and Gram negative
organisms. The activity was determined by measuring the
diameter of zone of inhibition recorded.6
Results
Evaluation
of poly herbal gel
Physical analysis of the prepared polyherbal gel
It was observed that the
freshly prepared formulations were off white to yellow in color (Table 2). Regarding the base and the
formulation A, B and C, there was no change in color, odor and appearance up to
the observation period of 30 days at 80C and 40°C using different
storage conditions, however Formulations A, B and C were stable 6.
pH
of the prepared formulations
It was found to be in
the range of 6.62 to 7.08, kept at different storage conditions for 30 days. pH
of the formulations and base kept at 8°C for one month did not show much change
and data were significant over control (base) during one month (p < 0.05).
Interestingly at 40°C, formulation A exhibited elevated change in pH (7.08),
while the others remained slightly stable during one month study. Data of
formulations A, B and C at 40°C were found to be significant Table 3.
Viscosity
test
Viscosity of the
formulations, kept at storage conditions for 30 days was found to be within the
range. The data of viscosity in formulations A, B, C and Control at 8°C and at
40°C were significant Table 3.
Centrifugation
Centrifugation test for
base and formulation kept at different storage conditions were performed for 30
days. No phase separation after centrifugation was found in formulations A, B,
C and base at 8 and 40°C during one month study Table 2.
Spreadability
Spreadability of base
and formulations (A, B and C) were studied. All the formulations and base were
found to possess good spreadability.
Acid
value, peroxide value and total fatty matter determination
Acid value, peroxide
value and total fatty matter for base and formulations kept at different
storage conditions were observed for 30 days and values for base and formulations
A, B, C were found within the range Table
3. Acid value was found to be in the range of 2.30 to 2.91, peroxide value
was found to be in the range of 1.64 to 1.88 and total fatty matters were found
to be in the range of 15.01 to 15.5 for the formulations and base kept at
different storage conditions for 30 days. Data of acid values of the
formulations and base were found to be significant (p < 0.05) during one month of stability study. Peroxide value
data in formulations A, B, C and control at 8 and 40°C were found to be
significant (p <0.05). Total fatty
matter data were found to be significant in all except formulation A, B, C and control
at 8 and 40°C Table 3.
