利用大孔吸附树脂与高速逆流色谱纯化脱氧雪腐镰刀菌烯醇-3-葡萄糖苷  

Purification of Deoxynivalenol-3-Glucoside by Using Macroporous Adsorption Resin Combined with High-Speed Counter-Current Chromatography

作　　者：陈龙云 胡俊强 何灿 史建荣[2] 徐剑宏 王刚[1,2] CHEN LongYun;HU JunQiang;HE Can;SHI JianRong;XU JianHong;WANG Gang(School of Food and Biological Engineering,Jiangsu University,Zhenjiang 212013,Jiangsu;Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base,Ministry of Science and Technology/Key Laboratory for Agro-Product Safety Risk Evaluation(Nanjing),Ministry of Agriculture and Rural Affairs/Key Laboratory for Control Technology and Standard for Agro-Product Safety and Quality,Ministry of Agriculture and Rural Affairs/Collaborative Innovation Center for Modern Grain Circulation and Safety/Institute of Food Safety and Nutrition,Jiangsu Academy of Agricultural Sciences,Nanjing 210014)

机构地区：[1]江苏大学食品与生物工程学院,江苏镇江212013 [2]江苏省农业科学院农产品质量安全与营养研究所/江苏省食品质量安全重点实验室-省部共建国家重点实验室培育基地/农业农村部农产品质量安全控制技术与标准重点实验室/农业农村部农产品质量安全风险评估实验室(南京)/农业农村部农产品质量安全控制技术与标准重点实验室/江苏省现代粮食流通与安全协同创新中心,南京210014

出　　处：《中国农业科学》2025年第8期1627-1637,共11页Scientia Agricultura Sinica

基　　金：国家重点研发计划(2023YFD1301003);国家自然科学基金(32161143034);江苏省科技计划专项资金(BE2022786);江苏省农业科技自主创新资金(CX(23)3003)。

摘　　要：【目的】脱氧雪腐镰刀菌烯醇-3-葡萄糖苷(DON-3G)是粮食产品中最常见的隐蔽型真菌毒素,但高昂的底物价格限制了对其深入研究。开发联用大孔吸附树脂与高速逆流色谱(HSCCC)方法,以实现酶促转化产物中DON-3G的大量纯化。【方法】采用静态吸附试验筛选不同类型吸附树脂或离子交换树脂;利用单因素水平试验优化最佳吸附条件与洗脱条件并拟合吸附热力学模型;筛选合适的双相体系并采用HSCCC分离DON-3G纯物质;最终通过紫外吸收光谱与核磁共振波谱确证产物结构,通过高效液相色谱分析产物纯度。【结果】采用XAD-4大孔吸附树脂吸附反应体系中DON-3G,吸附容量达269.23μg·g^(-1);洗脱溶剂采用40%甲醇水溶液,可在5倍柱体积内洗脱DON-3G。采用正丁醇﹕三氟乙酸﹕水为1﹕0.01﹕1作为HSCCC双相体系,在流速1 mL·min^(-1)、转速1 000 r/min、柱温35℃、双向洗脱模式下可以实现底物DON与产物DON-3G的分离,DON-3G回收率79.7%,液相色谱分析表明产物纯度为97.46%。未反应DON可以通过反向洗脱回收。【结论】本方法可实现单批次100mg以上的DON-3G分离纯化,为针对DON-3G的后续研究奠定了基础。【Objective】Deoxynivalenol-3-glucoside (DON-3G) is the most commonly found masked mycotoxin in cereals,however,the high cost of substrates hinders further studies on it.This study aimed to develop an efficient and affordable method for the large-scale purification of DON-3G from the enzymatic transformation product.【Method】Static adsorption experiments were utilized to screen various types of adsorption and ion-exchange resins,and a single-factor experimental design was employed to determine the optimal adsorption and desorption conditions.The adsorption thermodynamic model was established to fit the adsorption behavior.The pure material of DON-3G was then prepared using high-speed counter-current chromatography (HSCCC),and the biphasic systems were screened.The UV and NMR spectroscopy were used to confirm the product's structure,and high-performance liquid chromatography (HPLC) was used to assess its purity.【Result】With a maximal adsorption capacity of269.23μg·g^(-1) resin,the XAD-4 resin was determined to be the most effective macroporous resin for adsorbing DON-3G from the reaction system.The 40%methanol solution was used as the desorption solvent,because it could elute DON-3G within 5 bed volume DON-3G and unreacted DON were separated using a biphasic system with n-butanol/trifluoroacetic acid/water (1:0.01:1) at a flow rate of 1 mL·min^(-1),a rotation speed of 1 000 r/min,and a column temperature of 35℃under the bidirectional elution mode.With optimal conditions,the HSCCC process produced DON-3G with a purity of 97.46%and the recovery rate was 79.7%.Unreacted DON could be recovered by reverse elution.【Conclusion】Over 100 mg of DON-3G might be separated and purified using this approach in a single run,paving the way for the subsequent research targeting DON-3G.

关 键 词：脱氧雪腐镰刀菌烯醇-3-葡萄糖苷 隐蔽型毒素 分离纯化 大孔吸附树脂 高速逆流色谱 

分 类 号：S482.2[农业科学—农药学]