Abstract

DEVELOPMENT AND VALIDATION OF HPTLC METHOD FOR DETERMINATION OF CAFFEINE IN FOOD, BEVERAGE AND MEDICINAL PREPARATIONS

Ankit Raghuwanshi, Jinu John*, C.T. Aravindakumar

ABSTRACT

A simple, precise and accurate HPTLC method was developed for the determination of caffeine in different coffee products, beverage and pharmaceutical products. Caffeine from different samples such as natural coffee bean, locally marketed coffee powder, instant coffee mix, Cola drink and tablets was extracted with dichloromethane. Analysis was performed on silica gel G 60 F254 HPTLC plates with chloroform and methanol in the proportion of 25:1 (v/v) as mobile phase. Samples were applied with Linomat V under nitrogen gas flow. Caffeine gave a clear band with an Rf value 0.24±0.02. The densitometric analysis was performed CAMAG TLC scanner 4 at 274 nm. The linear regression analysis of data for the calibration curve showed good linearity over a concentration range of 50 ng to 200 ng/spot with a regression value of 0.99953 and standard deviation of 1.59%. The regression equation of the calibration curve was Y=687.946 + 24.936 * X. The limit of detection and limit of quantification of caffeine was 0.198 ng spot-1 and 0.599 ng spot-1. Using the developed method, the concentration of caffeine in complex matrix containing samples such as marketed coffee powder, soft drink and tablet was determined.

Keywords: Caffeine, HPTLC method, beverage and pharmaceutical preparation