Optimization of rebaudioside a extraction from Stevia Rebaudiana (Bertoni) and quantification by high perfomance liquid chromatography analysis

摘要

A solid-liquid extraction and an HPLC method for determination of rebaudioside Audfrom the leave parts of Stevia rebaudiana were developed. Separation method consisted ofudsolvent extraction of leaf powder using various solvents like petroleum ether, methanol, diethyludether and butanol followed by its purification using high performance liquid chromatography inudorder to obtain bioactive compound rebaudioside A. This solvent selection is very importantudprior to alternative extraction methods since it can be used as a pre-extraction solvents, mainudsolvents, or co-solvents. The problem of hydrolysable components and solvent removaluddifficulties in the conventional extraction led us to study further the effects of solvent propertiesudon the conventional extraction using Soxhlet method in order to determine the best solvent orudsolvent mixture for high extraction yield of S. rebaudiana. The chromatographic separation wasudrealized using a C18 column, mobile phase consisting of methanol: water with UV detection atud210 nm. Based on the yield of extraction and glycosides content, methanol was found to be bestudsolvent.