ISBN:
978-81-85694-67-2
Abstract
Ayurvedic
lozenges are immensely popular in today’s world, because due to its highly
palatable taste, cheap, soothing effect, counter-irritant effect, decreasing abdominal
flatulence, and treating Anorexia. Herbs of Piper species are one of the
most important ingredients of Ayurvedic lozenges and Piperine is the
main chemical constituent of herbs of Piper species. In the present
study an attempt was made for the estimation of piperine in ayurvedic lozenges
sample by using UV-spectrophotometer. Three samples of lozenges from different
reputed brand have been selected for this purpose. The amount of piperine in
three samples (S-1, S-2, and S-3) was found to be 1.55, 1.36 and 1.40 µg/ml
respectively. Recovery studies were carried out by standard addition method and
the average % recovery of the three samples S-1, S-2, S-3 were found to be 98.62%,
99.32%, and 98.35% respectively.
Introduction
Lozenges
are small, medicated candy intended to be dissolved slowly in the mouth to
lubricate and soothe irritated tissues of the throat and also have carminative
effect. Several ayurvedic companies come forward in today’s FMCG field to
market throat lozenges, and carminative digestive lozenges. Trikatu[1-3]
is an important ayurvedic formulation, which have synergistic effect, and is
used in several classical anti-tussive, expectorant[2] preparation
and also to fight, acidity[3] and anorexia. Trikatu consists
of Piper longum, Piper nigrum and Zingiber officinalis[4]. Piperine
is an important heterocyclic alkaloid of pyridine and piperidine skeletal structures
[4] found in the herbs of Piper species. These herbs enhanced the
activities of antioxidant enzymes--superoxide dismutase, catalase, glutathione reductase
and glutathione-S-transferase in both gastric and intestinal mucosa, suggesting
a gastrointestinal protective role which is the
main responsible factor for the therapeutic effect of these ayurvedic lozenges.
Apart from the classical use, recent study has proved that Piperine have
antidepressant activity [5-6], anti-inflammatory activity[7],
gastro protective effect[8-9], immune-modulator effect[10],
bioavailability enhancer[11] etc. Hence the amount of piperine in
this type of lozenges may be a parameter, to determine the quality of ayurvedic
lozenges. In the present study a comparative evaluation of the three most
popular branded ayurvedic lozenges on the basis of Piperine content.
|
MOLECULAR Wt
|
285.342
|
|
MOLECULAR FORMULA-
|
C17H19NO3
|
Materials and methods
Gift
sample of Piperine standard was taken from Merck chemicals, India. Ayurvedic
lozenges from three different brands were collected from local vendor, and named
them as S-1, S-2 and S-3. UV absorbance was recorded using “Varian UV-
Spectrophotometer” with Carry-100 software.
Preparation of standard solution
for calibration curve of ciprofloxacin
Stock
solution of Piperine was prepared by dissolving 0.01gram in 10 ml of methanol.
Standard solutions were prepared from stock solution in the range of 2 to 20
µg/ml in methanol. The absorbance of piperine was measured at 342 nm (λmax
for piperine) against water as blank. Mean of three readings were taken for
each standard solutions. Calibration curve was plotted between absorbance and
concentration.
Table1:
Standard curve of Piperine
|
Standard
|
Conc.
(µg/ml)
|
Mean*
|
SD
|
%RSD
|
|
Std.
1
|
0.0
|
0.0005
|
0.0007
|
>100
|
|
Std.
2
|
2.0
|
0.1150
|
0.0003
|
0.26
|
|
Std.
3
|
4.0
|
0.2375
|
0.0006
|
0.24
|
|
Std.
4
|
8.0
|
0.5258
|
0.0007
|
0.13
|
|
Std.
5
|
12.0
|
0.8425
|
0.0000
|
0.00
|
|
Std.
6
|
16.0
|
1.1795
|
0.0004
|
0.03
|
|
Std.
7
|
20.0
|
1.5454
|
0.0006
|
0.04
|
*Value expressed as mean
of three readings
Preparation
of sample solution
Five
lozenges of S-1 were taken in a mortar and pestle, and powdered. The powdered sample
was dissolved in the water q.s, and then chloroform is added to the water containing
the dissolved sample. The bi-phasic mixture is then taken in a separating
funnel and slowly shaken. The chloroform part which contains piperine is
separated out, the chloroform is evaporated in water bath, and TLC is carried
out with the piperine extract, for its conformation, with the application of standard
Piperine spot, on the same plate. Then methanol is added to the piperine extract,
and the solution is diluted. The mobile phase is made up of chloroform and
methanol in the ratio of (95:5), the absorbance of sample solutions was measured
at 342 nm against methanol as blank. Mean of three readings were taken for each
sample solutions, the process was repeated with the sample, S-2 and S-3.
|
Sample
|
Conc.
(µg/ml)*
|
SD
|
%RSD
|
|
S1
|
1.55
|
0.0003
|
0.24
|
|
S2
|
1.36
|
0.0002
|
0.19
|
|
S3
|
1.40
|
0.0002
|
0.21
|
Results
and discussion.
This
method involves the measurement of UV absorbance at 342 nm for piperine corresponding
to the absorption maxima. The absorbance characteristics showed that piperine obeys
Beer’s Lambert law within the concentration range from 2-20 µg/ml at the λmax
342 nm. Calibration equation for this method was found to be Abs = 0.07733*Concentration, with Correlation Coefficient
is of 0.99569.
Recovery
Studies were carried out by standard addition method & the average %
recovery of the four samples S1, S2 and S3 were found to be 98.62%, 99.32% and 98.35
% respectively. Results obtain from the recovery study indicating the accuracy
& the precision of the method.
Conclusion
The
developed method was found to be accurate & simple. We came to know that
all the three samples of lozenges from different manufacturers contains considerably
amount of piperine, and thus they bear the necessary quality, for therapeutic
efficacy.
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