COMPARATIVE ESTIMATION OF PIPERINE IN AYURVEDIC LOZENGES BY UV SPECTROSCOPIC METHOD.

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 (λ_max 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 λ_max 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.

References

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