Abstract

ISOLATION, IDENTIFICATION AND QUANTITATIVE ANALYSIS OF PIPERINE FROM PIPER NIGRUM LINN. OF VARIOUS REGIONS OF KERALA BY RP-HPLC METHOD

Ajmal H.*

ABSTRACT

Piperine is the naturally occurring alkaloid that gives the spice; Piperine is extracted from Piper nigrum, Linn. It is medicinally used as Anti-inflammatory, Anti-malarial, Cure for Stomach Ache, Weight Loss, Anti-leukemia, Fever prevention and reducer, Treatment for Snake Venom Poisoning and Anti-epileptic, also helpful in increasing the absorption of certain vitamins such as Selenium, Vitamin B and Beta-Carotene. Extraction and Standardization of piperine is performed. In Standardization, Solubility, Total ash content, Acid insoluble ash, TLC, UV Spectroscopy, HPLC were performed. Piperine was extracted successfully, it was found to be soluble in ethanol and chloroform. Piperin extracted from Piper nigrum Linn. By refluxing 32 hrs with chloroform. Black pepper is collected from various regions of kerala at the month of august and january. The various geographical sources of black pepper are Wayanad, Idukki, Kozhikode, Alappuzha, Palakkad and Thiruvananthapuram district. Six pepper samples are collected from above six different district and extracted. A simple, rapid and precise RP-HPLC method has been developed for the quantitative determination of Piperine from Piper nigrum Linn. (Black pepper). Chromatographic analysis was carried out on Shimadzu phenomenex C18 column (250mm x 4.6mm, 5μm particle size). Quantitation was performed using a UV detector at 343 nm. RP-HPLC was performed by using ACN and aqueous acetic acid 1as mobile phase. The method was validated for linearity, precision, accuracy and can be effectively used to evaluate quality of black pepper. Quantitation, based on peak areas, achieved by reference to purified piperine as external standard.

Keywords: Back pepper, Piperine, Isolation, Chloroform extract, TLC, RP-HPLC, UV spectrum, IR spectrum.