Abstract
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 various temperatures from 20 °C to 79 °C. HPLC results suggested that temperatures between 40 °C and 60 °C would be ideal conditions for better separation of steviol glycosides. Additional HPLC studies at five different temperatures 40 °C, 45 °C, 50 °C, 55 °C, and 60 °C under three different pH 2.4, 2.6, and 2.8 were performed and results suggested that temperatures 50 °C and 55 °C at pH 2.4 would be ideal condition for better separation of steviol glycosides.
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