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Quantitation of low molecular mass substrates and products of enzyme catalyzed reactions using matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry
Author(s) -
Kang MinJung,
Tholey Andreas,
Heinzle Elmar
Publication year - 2000
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/1097-0231(20001115)14:21<1972::aid-rcm119>3.0.co;2-5
Subject(s) - chemistry , derivatization , substrate (aquarium) , acetamide , mass spectrometry , hydroxylamine , benzaldehyde , catalysis , chromatography , reagent , organic chemistry , oceanography , geology
Relative peak‐height ratios of products to substrates determined by MALDI‐TOFMS allow the quantitative analysis of enzyme catalyzed reactions for screening purposes. Two examples were investigated: the first one was a lipase catalyzed reaction which produces 2‐methoxy‐ N ‐[(1 R )‐1‐phenylethyl]acetamide (MET) using rac ‐α‐phenylethylamine (PEA) as substrate. The second one was the pyruvate decarboxylase catalyzed formation of (1 R )‐1‐hydroxy‐1‐phenyl‐2‐propanone (PAC) with benzaldehyde (BzA) as substrate. Here the corresponding oximes were analyzed after derivatization using hydroxylamine. The standard curves (r 2 = 0.985 for MET, r 2 = 0.991 for PAC) were linear over two orders of magnitude for MET and PAC concentrations. After optimization of the sample preparation an average relative standard deviation of 12.5% was obtained in both cases. Copyright © 2000 John Wiley & Sons, Ltd.