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Turbulent flow chromatography for the reduction of matrix effects in electrospray ionization mass spectrometry‐based enzyme assays
Author(s) -
Liesener André,
Karst Uwe
Publication year - 2005
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200500090
Subject(s) - chemistry , chromatography , analyte , mass spectrometry , ion suppression in liquid chromatography–mass spectrometry , electrospray ionization , matrix (chemical analysis) , trypsin , electrospray , analytical chemistry (journal) , liquid chromatography–mass spectrometry , enzyme , biochemistry
Abstract Turbulent flow chromatography (TFC) is presented as a means to reduce ion suppression in simultaneous multianalyte mass spectrometric bioassays. In this study, the effects of enzymes present in the sample on the signal response of five analytes were simultaneously investigated over a protein content range from 0 to 38 μg/mL by means of direct flow injection MS. As model enzymes, trypsin, thrombin, and chymotrypsin were selected. Without employment of TFC, both signal suppression and signal enhancement, depending on the nature of the analyte and the amount of matrix in the sample, were observed. Generally, these matrix effects were found to be intolerably large. The deviation from the mean signal response as a measure of deterioration was found to be between 14 and 112%. The addition of an excess of methanol as means of sample clean‐up was investigated and found not to be sufficient. By employing TFC for online sample preparation, it was possible to reduce the matrix effects to a minimum for all model systems investigated. In case of trypsin the distortion could be lowered from 41.9 to 2.6%. Thus, TFC is considered to be a highly valuable tool for improving the sensitivity and reliability in the monitoring of enzymatic conversions by means of MS.