Continuation of Reversed-Phase HPLC Analysis Studies of Steviol Glycosides Isolated From Stevia rebaudiana Bertoni

Abstract

Additional High Performance Liquid Chromatography (HPLC) studies were performed on the nine sweet steviol glycosides reported in Joint Expert Committee on Food Additives (JECFA) namely rebaudioside A, steviolbioside, stevioside, rubusoside, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside F, and dulcoside A isolated from the leaves of Stevia rebaudiana. Using Reversed-Phase (RP) HPLC method, individual retention times and area for nine naturally occurring ent -kaurane diterpene glycosides of S. rebaudiana have been determined at five different temperatures: 40, 45, 50, 55, and 60 ºC at three different pH 2.4, 2.6, and 2.8. HPLC results suggested that temperatures 50 and 55 ºC at pH 2.4 would be ideal condition for better separation of steviol glycosides.