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Application of automated matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry for the measurement of enzyme activities
Author(s) -
Kang MinJung,
Tholey Andreas,
Heinzle Elmar
Publication year - 2001
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/rcm.376
Subject(s) - chemistry , mass spectrometry , analyte , chromatography , analytical chemistry (journal) , desorption , time of flight mass spectrometry , surface enhanced laser desorption/ionization , matrix assisted laser desorption/ionization , ionization , matrix (chemical analysis) , sample preparation in mass spectrometry , electrospray ionization , organic chemistry , adsorption , ion
Sample preparation methods and data acquisition protocols were optimized for the application of matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOFMS) to high‐throughput quantitative analysis of low molecular mass substrates and products of an enzyme‐catalyzed reaction. Using a deuterlum‐labeled internal standard, precise standard curves were obtained (r 2 = 0.9998) over two orders of magnitude of concentration of rac ‐1‐phenylethylamine (PEA), which is converted to 2‐methoxy‐ N ‐[(1 R )‐1‐phenylethyl]acetamide (MET) by a lipase‐catalyzed reaction with ethylmethoxyacetate (EMA) as second substrate. Reliable relative standard deviations were achieved (≤5%) using automated analysis with peak intensity ratios between 0.2 and 5 of analyte to internal standard. This method permitted quantitative analysis of the lipase reaction, producing results comparable to those from gas chromatographic (GC) analysis in the dynamic range of GC. This work shows that MALDI‐TOFMS can be applied for the high‐throughput screening of enzymes. Copyright © 2001 John Wiley & Sons, Ltd.