| 固定化β-葡萄糖醛酸酶反应器的制备及其在4-甲基亚硝胺基-1-(3-吡啶)-1-丁醇的O-糖苷化合物分析中的应用 |
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| 引用本文： | 王婷,姚二民,邓楠,边阳阳,刘萍萍,张柯,陈千思,李斌.固定化β-葡萄糖醛酸酶反应器的制备及其在4-甲基亚硝胺基-1-(3-吡啶)-1-丁醇的O-糖苷化合物分析中的应用[J].色谱,2018,36(3):245-252. |
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| 作者姓名： | 王婷 姚二民 邓楠 边阳阳 刘萍萍 张柯 陈千思 李斌 |
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| 作者单位： | 1. 中国烟草总公司郑州烟草研究院, 河南 郑州 450001;
2. 郑州大学第一附属医院医学研究中心, 河南 郑州 450052;
3. 郑州轻工业学院, 河南 郑州 450001 |
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| 基金项目： | 国家自然科学基金项目（21505158，81600046）. |
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| 摘 要： | 采用溶胶-凝胶法,以丙烯酰胺作为单体,制备了基于有机-硅胶杂化整体柱的β-葡萄糖醛酸酶反应器。优化了硅酸甲酯和γ-(甲基丙烯酰氧)丙基三甲氧基硅的物质的量的比、丙烯酰胺和聚乙二醇的用量,以及水浴温度等制备条件,获得了孔隙均匀、通透性良好、机械强度高的有机-硅胶杂化整体柱。采用光学显微镜和扫描电镜表征杂化整体柱。进一步将β-葡萄糖醛酸酶共价键合在整体柱上,以4-甲基亚硝胺基-1-(3-吡啶)-1-丁醇(NNAL)的O-糖苷化合物(NNAL-O-Gluc)为底物研究酶反应器的水解效果,实验结果证明酶反应器在室温条件下的水解效率大大提高,实现了NNAL-O-Gluc高效水解与分析,解决了目前NNAL-O-Gluc分析中前处理水解效率低的问题。
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| 关 键 词： | 有机-硅胶杂化整体柱 β-葡萄糖醛酸酶 固定化酶反应器 4-甲基亚硝胺基-1-(3-吡啶)-1-丁醇的O-糖苷化合物 |
| 收稿时间： | 2017-12-11 |
Preparation of immobilized β-glucuronidase enzyme reactor for the analysis of O-glucuronides of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol |
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| WANG Ting,YAO Ermin,DENG Nan,BIAN Yangyang,LIU Pingping,ZHANG Ke,CHEN Qiansi,LI Bin.Preparation of immobilized β-glucuronidase enzyme reactor for the analysis of O-glucuronides of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol[J].Chinese Journal of Chromatography,2018,36(3):245-252. |
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| Authors: | WANG Ting YAO Ermin DENG Nan BIAN Yangyang LIU Pingping ZHANG Ke CHEN Qiansi LI Bin |
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| Institution: | 1. Zhengzhou Tobacco Research Institute of China National Tobacco Corporation, Zhengzhou 450001, China;
2. Medical Research Centre, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China;
3. Zhengzhou University of Light Industry, Zhengzhou 450001, China |
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| Abstract: | An immobilized β-glucuronidase enzyme reactor (IMER) based on organic-silica gel hybridized capillary column was prepared by the sol-gel method with acrylamide (AAM) as the organic functional monomer. The effects of the amounts of tetramethyl orthosilicate (TMOS), 3-glycidoxypropyltrimethoxysilane (γ-MAPS), acrylamide (AAM), polyethylene glycol (PEG), and the water bath temperature on the quality of the microscopic structure were studied. After optimization,the monolithic column with good permeability and high mechanical strength was obtained. Then β-glucuronidase was covalently immobilized, and the hydrolysis efficiency of the β-glucuronidase IMER was investigated. As the substrate, O-glucuronides of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL-O-Gluc) were hydrolyzed at room temperature, indicating the high hydrolysis efficiency of the β-glucuronidase IMER, which demonstrated its high potential for the high throughput analysis of tobacco metabolites in the future. |
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| Keywords: | organic silica hybrid monolithic column β-glucuronidase immobilized enzyme reactor (IMER) O-glucuronides of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL-O-Gluc) |
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