Preparation of immobilized β-glucuronidase enzyme reactor for the analysis of O-glucuronides of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol

doi:10.3724/SP.J.1123.2017.11050

Abstract

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.

Key words: β-glucuronidase, O-glucuronides of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL-O-Gluc), immobilized enzyme reactor (IMER), organic silica hybrid monolithic column

CLC Number:

O658

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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