Chemical and Pharmaceutical Bulletin
Online ISSN : 1347-5223
Print ISSN : 0009-2363
ISSN-L : 0009-2363
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Development of a Rapid and Detailed Structural Identification System with an On-Line Immobilized Enzyme Reactor Integrated into LC-NMR
Yousuke KashimaYoshito Okabayashi
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2010 Volume 58 Issue 3 Pages 423-425

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Abstract

Immobilized enzyme reactors (IMERs) integrated into an LC-NMR system were developed for rapid and detailed structural identification of enzymatic reaction products. An on-column enzymatic reaction was achieved for immobilized cytochrome-c and dog microsomes. After the reaction, these products were analyzed by LC-NMR without any work-up processes. The immobilized cytochrome-c column integrated into LC-NMR was used to characterize the reaction product formed on N-demethylation by measurement of 1H-NMR. In the case of reaction taking place on the microsome column, a glucuronidation product was identified by 1H-NMR and 1H–1H correlated spectroscopy. The chemical structures of the enzymatic reaction products could be elucidated by IMER-LC-NMR without the need for authentic samples or isolation processes.

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© 2010 The Pharmaceutical Society of Japan
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