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1 2014

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Preparative separation of steviol glycosides from Stevia rebaudiana bertoni by macroporous resin and preparative HPLC

Lv X. H., Kuang P. Q., Yuan Q. P., Liang H., Zheng G. Q.

Acta Chromatographica

2014

Vol. 26, no. 1

123--135

In this article, a rapid, novel, and effective method was presented to separate pure steviol glycosides from leaves of Stevia rebaudiana Bertoni, including stevioside, rebaudioside A, and rebaudioside C, using resin-based column chromatography followed by preparative high-performance liquid chromatography under hydrophilic interaction liquid chromatography (HILIC) mode. The separation procedure was first optimized on an analytical HILIC column and then scaled up on a preparative HILIC column using a mobile phase consisting of 83% acetonitrile in water with flow rate of 10 mL•min -1 at 25°C. 200 mg of the crude extract containing 68.1% steviol glycosides yielded 79.2 mg stevioside, 9.4 mg rebaudioside C, and 33.7 mg rebaudioside A, with the purities of 97.5%, 96.8%, and 97.2%, respectively. Those products were identified by MS, 1H NMR, and 13C NMR.