SUPRIYADI, 051217097306 (2017) PENGEMBANGAN METODE ANALISIS GLIKOSIDA STEVIOL SECARA HPLC-ELSD DALAM UPAYA MENUNJANG PENJAMINAN MUTU PRODUK STEVIA. Disertasi thesis, Universitas Airlangga.

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Abstract

Stevia rebaudiana Bertoni is now being considered as a possible sucrose substitute due to its pleasant organoleptic properties and associated health benefits. The Evaporative Light Scattering Detector (ELSD) is a valuable HPLC detector for detection of non-UV absorbing compounds, such as stevioside, rebaudioside A, rebaudioside C and dulcoside A. The objective of the presen study was to develop and validate a selective HPLC-ELSD method for determination of steviol glycosides contained in Stevia rebaudiana, mainly stevioside, rebauside A, rebaudioside C, and dulcoside A. The chromatographic separation of stevioside, rebaudioside A, rebaudioside C, and dulcoside A was achieved using Phenomenex Luna column 250 mm x 4.6 mm i.d. in isocratic system mode with amobile phase of acetonitrile-water (35 : 65). The temperature of nebulization and evaporization of the ELS detector was set at 40oC and 50oC, respectively. The good separation of stevioside, rebaudioside A, rebaudioside C, and dulcoside A was obtained, yielding the resolution of all the analytes more than 1.5. All the validation parameters like specificity, linearity, range, accuracy and precision met the acceptance criteria according to ICH guidelines. The proposed HPLC-ELSD method is simple and sensitive for the simultaneously detection and determination of of stevioside, rebaudioside A, rebaudioside C and dulcoside A contained in Stevia rebaudiana. The method was successfully applied for the determination of the samples product of Stevia rebaudiana.

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