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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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。
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 theβ-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.
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 theβ-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.
引文
[1]Hoffmann D,Hoffmann I,El-Bayoumy K.Chem Res Toxicol,2001,14(7):767
[2]Khariwala S S,Scheuermann T S,Berg C J,et al.Nicotine Tob Res,2013,16(5):600
[3]Pleasance E D,Stephens P J,O’Meara S,et al.Nature,2010,463(7278):184
[4]Kassem N O F,Kassem N O,Liles S,et al.Am J Health Behav,2015,39(5),680
[5]Zhang H,Liu M.Journal of Baoji University of Arts and Sciences(Natural Science Edition),2007,27(4):287张红,刘铭.宝鸡文理学院学报(自然科学版),2007,27(4):287
[6]Schrader E,Hirsch-Ernst K I,Scholz E,et al.Drug Metab Dispos,2000,28(2):180
[7]Lu W Y,Ferguson S G,Nichols D S,et al.J Pharmaceut Biomed,2016,131:327
[8]Chung C J,Lee H L,Yang H Y,et al.Sci Total Environ,2011,409(9):1638
[9]Kotandeniya D,Carmella S G,Ming X,et al.Anal Chem,2015,87(3):1514
[10]Carmella S G,Le K,Upadhyaya P,et al.Chem Res Toxicol,2002,15(4):545
[11]Homaei A A,Sariri R,Vianello F,et al.J Chem Biol,2013,6(4):185
[12]Ma J F,Zhang L H,Liang Z,et al.TrAC-Trends Anal Chem,2011,30(5):691
[13]Yuan H M,Zhang L H,Zhang Y K.J Chromatogr A,2014,1371:48
[14]Valikhani D,Bolivar J M,Viefhues M,et al.ACS Appl Mater Inter,2017,9(40):34641
[15]Naldi M,Cernigoj U.Talanta,2017,167:143
[16]Song J Y,Su P,Yang Y,et al.Chinese Journal of Chromatography,2017,35(3):260宋佳一,苏萍,杨烨,等.色谱,2017,35(3):260
[17]Lin H,Ou J J,Zhang Z B,et al.Anal Chem,2012,84:2721
[18]Lin Z,Tan X Q,Yu R F,et al.J Chromatogr A,2014,1355:228
[19]Liu Z,Lin Y B,Lv H D.Int Mater Rev,2006,20(12):137
[20]Hu J W,Shen Y H,Qin W J,et al.Chemical Journal of Chinese University,2012,33(2):268胡佳薇,申烨华,秦伟捷,等.高等学校化学学报,2012,33(2):268
[21]Kotandeniya D,Carmella S G,Ming X,et al.Anal Chem,2015,87(3):1514
[22]Zhou L Q,Zhang J,Tian F,et al.Chinese Journal of Chromatography,2012,31(4):355周廉淇,张姣,田芳,等.色谱,2012,31(4):355
[23]Wang Z Q,Wang Z W,Zhang Y G.Food Research and Development,2011,32(7):148王振强,王振伟,张耀光,食品研究与开发,2011,32(7):148
[2]Khariwala S S,Scheuermann T S,Berg C J,et al.Nicotine Tob Res,2013,16(5):600
[3]Pleasance E D,Stephens P J,O’Meara S,et al.Nature,2010,463(7278):184
[4]Kassem N O F,Kassem N O,Liles S,et al.Am J Health Behav,2015,39(5),680
[5]Zhang H,Liu M.Journal of Baoji University of Arts and Sciences(Natural Science Edition),2007,27(4):287张红,刘铭.宝鸡文理学院学报(自然科学版),2007,27(4):287
[6]Schrader E,Hirsch-Ernst K I,Scholz E,et al.Drug Metab Dispos,2000,28(2):180
[7]Lu W Y,Ferguson S G,Nichols D S,et al.J Pharmaceut Biomed,2016,131:327
[8]Chung C J,Lee H L,Yang H Y,et al.Sci Total Environ,2011,409(9):1638
[9]Kotandeniya D,Carmella S G,Ming X,et al.Anal Chem,2015,87(3):1514
[10]Carmella S G,Le K,Upadhyaya P,et al.Chem Res Toxicol,2002,15(4):545
[11]Homaei A A,Sariri R,Vianello F,et al.J Chem Biol,2013,6(4):185
[12]Ma J F,Zhang L H,Liang Z,et al.TrAC-Trends Anal Chem,2011,30(5):691
[13]Yuan H M,Zhang L H,Zhang Y K.J Chromatogr A,2014,1371:48
[14]Valikhani D,Bolivar J M,Viefhues M,et al.ACS Appl Mater Inter,2017,9(40):34641
[15]Naldi M,Cernigoj U.Talanta,2017,167:143
[16]Song J Y,Su P,Yang Y,et al.Chinese Journal of Chromatography,2017,35(3):260宋佳一,苏萍,杨烨,等.色谱,2017,35(3):260
[17]Lin H,Ou J J,Zhang Z B,et al.Anal Chem,2012,84:2721
[18]Lin Z,Tan X Q,Yu R F,et al.J Chromatogr A,2014,1355:228
[19]Liu Z,Lin Y B,Lv H D.Int Mater Rev,2006,20(12):137
[20]Hu J W,Shen Y H,Qin W J,et al.Chemical Journal of Chinese University,2012,33(2):268胡佳薇,申烨华,秦伟捷,等.高等学校化学学报,2012,33(2):268
[21]Kotandeniya D,Carmella S G,Ming X,et al.Anal Chem,2015,87(3):1514
[22]Zhou L Q,Zhang J,Tian F,et al.Chinese Journal of Chromatography,2012,31(4):355周廉淇,张姣,田芳,等.色谱,2012,31(4):355
[23]Wang Z Q,Wang Z W,Zhang Y G.Food Research and Development,2011,32(7):148王振强,王振伟,张耀光,食品研究与开发,2011,32(7):148