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

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