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Journal of Integrative Agriculture  2016, Vol. 15 Issue (3): 694-701    DOI: 10.1016/S2095-3119(15)61098-X
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Separation and purification of deoxynivalenol (DON) mycotoxin from wheat culture using a simple two-step silica gel column chromatography
 ZHAO Xiu-mei, LI Rong-jia, ZHOU Chuang, ZHANG Jie, HE Cheng-hua, ZHENG Ya-ting, WU Wen-da
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, P.R.China
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摘要  Deoxynivalenol (DON) is a type B trichothecenes mycotoxin produced by several Fusarium species, often found in foodstuffs for humans and animals. DON is in great demand for the toxicological researches both in vivo and in vitro. In this work, wheat culture was inoculated with a Fusarium graminearum PH-1 strain for DON production. The solvent system for crude extraction was acetonitrile-water (84:16, v/v). A simple two-step silica gel column chromatography was employed to separate the DON mycotoxin from wheat culture, combined with preparative high performance liquid chromatography (preparative HPLC) to purify the compound. The solvent system for the second silica gel column chromatography was methylene chloride-methanol (17:1, v/v), which provided a good elution effect selected on thin layer chromatography (TLC). The target compound was identified by HPLC, and the chemical structure was confirmed by mass spectrometry (MS) and 1H and 13C nuclear magnetic resonance (NMR) spectroscopy. A total of 433 mg of purified DON was obtained from 1 kg of wheat culture, with a purity of 99.01%. The study had provided an easy-operating and cost-effective method to isolate an expensive compound in a simple way.

Abstract  Deoxynivalenol (DON) is a type B trichothecenes mycotoxin produced by several Fusarium species, often found in foodstuffs for humans and animals. DON is in great demand for the toxicological researches both in vivo and in vitro. In this work, wheat culture was inoculated with a Fusarium graminearum PH-1 strain for DON production. The solvent system for crude extraction was acetonitrile-water (84:16, v/v). A simple two-step silica gel column chromatography was employed to separate the DON mycotoxin from wheat culture, combined with preparative high performance liquid chromatography (preparative HPLC) to purify the compound. The solvent system for the second silica gel column chromatography was methylene chloride-methanol (17:1, v/v), which provided a good elution effect selected on thin layer chromatography (TLC). The target compound was identified by HPLC, and the chemical structure was confirmed by mass spectrometry (MS) and 1H and 13C nuclear magnetic resonance (NMR) spectroscopy. A total of 433 mg of purified DON was obtained from 1 kg of wheat culture, with a purity of 99.01%. The study had provided an easy-operating and cost-effective method to isolate an expensive compound in a simple way.
Keywords:  deoxynivalenol       silica gel column chromatography       separation       purification  
Received: 19 January 2015   Accepted:
Fund: 

This study was supported by the National Natural Science Foundation of China (31402268), the Natural Science Foundation of Jiangsu Province of China (BK20140691). A Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD), China. The Introduction of International Advanced Agricultural Science and Technology Project from the Ministry of Agriculture of China (2012-Z22).

Corresponding Authors:  ZHANG Hai-bin, Tel: +86-25-84396478,Fax: +86-25-84396586, E-mail: haibinzh@126.com     E-mail:  haibinzh@126.com
About author:  ZHAO Xiu-mei, E-mail: 2012207033@njau.edu.cn;

Cite this article: 

ZHAO Xiu-mei, LI Rong-jia, ZHOU Chuang, ZHANG Jie, HE Cheng-hua, ZHENG Ya-ting, WU Wen-da. 2016. Separation and purification of deoxynivalenol (DON) mycotoxin from wheat culture using a simple two-step silica gel column chromatography. Journal of Integrative Agriculture, 15(3): 694-701.

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