SEPARATION OF STEVIOL GLYCOSIDES FROM Stevia rebaudiana USING DIFFERENT AQUEOUS EXTRACTION TECHNIQUES Chong Saw Peng * , Mustapha bin Akil and Norellia binti Bahari Agrotechnology and Bioscience Division, Malaysian Nuclear Agency (Nuclear Malaysia), Ministry of Energy, Science, Technology, Environment and Climate Change (MESTECC), Bangi, Selangor, Malaysia *Corresponding author’s email: sawpeng@nm.gov.my Received date: 14 March 2018 | Accepted date: 16 December 2019 ABSTRACT Stevia rebaudiana has recently gained the attention of the food industry as one of the natural sweeteners. The sweet favour is contributed by the glycoside compounds, especially the rebaudioside A and stevioside, which are the stevia main chemical markers. The aim of the work reported here was to compare the diferent extraction techniques of stevia leaves using diferent technologies such as the high pressure and ultrasonic on the extraction of steviol glycosides. In this paper, the extraction techniques yielding the highest glycosides from the leaves of Stevia rebaudiana were determined using hot water extraction (HWE), pressurised liquid extraction (PLE) and ultrasound-assisted extraction (UAE). The steviol glycoside yields were quantifed by two chemical markers, rebaudioside A and stevioside of Stevia rebaudiana using high- performance liquid chromatography (HPLC) analysis. The result showed that the HWE managed to obtain 1,110 mg of steviol glycosides. The PLE obtained 294 mg steviol glycosides and the UAE obtained 427.5 mg steviol glycosides. As a conclusion, the results suggested the most efcient technique for stevia extraction in this study was the HWE. Keywords: hot water extraction, pressurised liquid extraction, ultrasound-assisted extraction, steviol glycosides, rebaudioside A, stevioside Borneo International Journal of Biotechnology (BIJB) Vol. 1 (December 2020), 77 – 87 e-ISSN 2716-697X Research Article