Volume 35, Issue 14 p. 1821-1827
Research Article

Comprehensive characterization of Stevia Rebaudiana using two-dimensional reversed-phase liquid chromatography/hydrophilic interaction liquid chromatography

Qing Fu

Qing Fu

School of Pharmacy, East China University of Science and Technology, Shanghai, P. R. China

Key Lab of Separation Science for Analytical Chemistry,, Key Lab of Natural Medicine, Liaoning Province,, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, P. R. China

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Zhimou Guo

Corresponding Author

Zhimou Guo

Key Lab of Separation Science for Analytical Chemistry,, Key Lab of Natural Medicine, Liaoning Province,, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, P. R. China

Additional correspondence: Dr. Zhimou Guo at Dalian Institute of Chemical Physics, Dalian, China. E-mail: [email protected] Fax: +86-411-84379539

Correspondence: Prof. Dr. Xinmiao Liang, School of Pharmacy, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, P. R. China

E-mail:[email protected]

Fax: +86-411-84379539

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Xiuli Zhang

Xiuli Zhang

Key Lab of Separation Science for Analytical Chemistry,, Key Lab of Natural Medicine, Liaoning Province,, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, P. R. China

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Yanfang Liu

Yanfang Liu

Key Lab of Separation Science for Analytical Chemistry,, Key Lab of Natural Medicine, Liaoning Province,, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, P. R. China

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Xinmiao Liang

Xinmiao Liang

School of Pharmacy, East China University of Science and Technology, Shanghai, P. R. China

Key Lab of Separation Science for Analytical Chemistry,, Key Lab of Natural Medicine, Liaoning Province,, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, P. R. China

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First published: 17 July 2012
Citations: 60

Abstract

Two-dimensional reversed-phase liquid chromatography/hydrophilic interaction liquid chromatography (2D-RPLC/HILIC) system was successfully applied for comprehensive characterization of steviol glycosides from Stevia rebaudiana. The experiments were performed in offline mode using an XCharge C18 column in first dimension and an XAmide column in second dimension. In first dimension, preliminary separation of Stevia aqueous extract was accomplished and 30 fractions were collected. Then fractions 1–20 were selected for further purification and 13 compounds with high purity were obtained in second dimension. Comprehensive characterization of these compounds was completed by determination of their retention time, accurate molecular weight, diagnostic fragmentation ions, and nuclear magnetic resonance spectroscopy. As a result, all nine known steviol glycosides, as well as other four steviol glycosides were fully purified. The result demonstrated that this procedure is an effective approach for the preparative separation and comprehensive characterization of steviol glycosides in Stevia. This 2D-RPLC/HILIC method will be a promising tool for the purification of low-abundance compounds from natural products.