A validated RP‑HPLC method for quantifcation of steviol glycoside: Rebaudioside a in extracts of stevia rebaudiana leaf

Kolate, Nikhil S and Mishra, Himanshu and Kini, Suvarna G (2021) A validated RP‑HPLC method for quantifcation of steviol glycoside: Rebaudioside a in extracts of stevia rebaudiana leaf. Chromatographia, 84. pp. 21-26. ISSN 000-95893

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Abstract

RP-HPLC-PDA method has been developed and validated for the quantifcation of Steviol glycoside: Rebaudioside A in aqueous extract of Stevia rebaudiana plant leaf. Rebaudioside A was separated on a Phenomenex Luna C18 analytical column with column temperature 40 °C using an isocratic solvent system consisting (70:30 v/v) mixture of 10 mmol/L sodium phosphate bufer and acetonitrile having pH-2.6 at 1.0 mL/min fow rate with detection at 210 nm wavelength. The method has been validated for linearity, precision, accuracy, detection limit, quantitation limit and robustness. The proposed method linearity for Rebaudioside A with a regression coefcient of 0.9999 was obtained within the range of 0.0005–1.5 mg/mL. The relative standard deviation was less than 2.0% in intra-day and inter-day precision. The proposed method’s accuracy was determined by a three diferent level recovery study. The recovery average was between 95.20 and 96.77 percent. Rebaudioside A levels of LOD and LOQ were 0.01 and 0.05 mg/mL respectively. The leaves from eight diferent Indian territories were extracted by sonication extraction method. Rebaudioside A content obtained in the dried leaves was within the range of 2.39–7.53 percent w/w. The method proposed is simple, rapid and reproducible; therefore, it is appropriate for routine analysis of Rebaudioside A in plant material as well as fnished products which are used commercially as high-intensity sweeteners across the globe to replace sugar.

Item Type: Article
Uncontrolled Keywords: Stevia rebaudiana; rebaudioside A; quantitation; RP-HPLC method.
Subjects: Pharmacy > MCOPS Manipal > Pharmaceutical Chemistry
Depositing User: KMC Library
Date Deposited: 08 Sep 2021 10:35
Last Modified: 08 Sep 2021 10:35
URI: http://eprints.manipal.edu/id/eprint/157233

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