Development of a rapid and detailed structural identification system with an on-line immobilized enzyme reactor integrated into LC-NMR.
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 (1)H-NMR. In the case of reaction taking place on the microsome column, a glucuronidation product was identified by (1)H-NMR and (1)H-(1)H 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.
Kashima
Yousuke
Y
Developmental Research Laboratories, Shionogi & Co., Ltd, Osaka 553-0002, Japan. yousuke.kashima@shionogi.co.jp
Okabayashi
Yoshito
Y
D015060:1,2-Dipalmitoylphosphatidylcholine / Q000737:chemistry; D000818:Animals; D019149:Bioreactors; D002851:Chromatography, High Pressure Liquid; D045304:Cytochromes c / Q000737:chemistry* / Q000378:metabolism; D004285:Dogs; D004789:Enzyme Activation; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008861:Microsomes / Q000737:chemistry; D015394:Molecular Structure; D009862:Online Systems
D004800:Enzymes, Immobilized; D015060:1,2-Dipalmitoylphosphatidylcholine; D045304:Cytochromes c