On-line coupling of high-performance liquid chromatography to a continuous-flow enzyme assay based on electrospray ionization mass spectrometry

A.R. de Boer; T. Letzel; D.A. van Elswijk; H. Lingeman; W.M.A. Niessen; H. Irth

doi:10.1021/ac035380w

On-line coupling of high-performance liquid chromatography to a continuous-flow enzyme assay based on electrospray ionization mass spectrometry

A.R. de Boer, T. Letzel, D.A. van Elswijk, H. Lingeman, W.M.A. Niessen, H. Irth

BioAnalytical Chemistry

Research output: Contribution to Journal › Article › Academic › peer-review

Abstract

Liquid chromatography (LC) was coupled on-line to a continuous-flow enzymatic assay using electrospray ionization mass spectrometry (ESI-MS) as readout for the screening of enzyme inhibitors in complex samples. Inhibitors were detected by changes in the concentration of the enzymatic reaction products, indicating the inhibition of enzymatic activity. The molecular masses of the inhibitors were determined with high certainty by using retention time matching and peak shape comparison. Due to the high matching accuracy, baseline separation of coeluting analytes was not necessary in order to identify the correct masses of the bioactive compounds. The continuous-flow system was successfully applied for the screening of complex samples, such as natural extracts. For a red clover extract, detection limits of 0.3-0.8 μmol/L were obtained. System validation was performed by determining the IC

Original language	English
Pages (from-to)	3155-61
Journal	Analytical Chemistry
Volume	76
Issue number	11
DOIs	https://doi.org/10.1021/ac035380w
Publication status	Published - 2004

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title = "On-line coupling of high-performance liquid chromatography to a continuous-flow enzyme assay based on electrospray ionization mass spectrometry",

abstract = "Liquid chromatography (LC) was coupled on-line to a continuous-flow enzymatic assay using electrospray ionization mass spectrometry (ESI-MS) as readout for the screening of enzyme inhibitors in complex samples. Inhibitors were detected by changes in the concentration of the enzymatic reaction products, indicating the inhibition of enzymatic activity. The molecular masses of the inhibitors were determined with high certainty by using retention time matching and peak shape comparison. Due to the high matching accuracy, baseline separation of coeluting analytes was not necessary in order to identify the correct masses of the bioactive compounds. The continuous-flow system was successfully applied for the screening of complex samples, such as natural extracts. For a red clover extract, detection limits of 0.3-0.8 μmol/L were obtained. System validation was performed by determining the IC",

author = "{de Boer}, A.R. and T. Letzel and {van Elswijk}, D.A. and H. Lingeman and W.M.A. Niessen and H. Irth",

year = "2004",

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T1 - On-line coupling of high-performance liquid chromatography to a continuous-flow enzyme assay based on electrospray ionization mass spectrometry

AU - de Boer, A.R.

AU - Letzel, T.

AU - van Elswijk, D.A.

AU - Lingeman, H.

AU - Niessen, W.M.A.

AU - Irth, H.

PY - 2004

Y1 - 2004

N2 - Liquid chromatography (LC) was coupled on-line to a continuous-flow enzymatic assay using electrospray ionization mass spectrometry (ESI-MS) as readout for the screening of enzyme inhibitors in complex samples. Inhibitors were detected by changes in the concentration of the enzymatic reaction products, indicating the inhibition of enzymatic activity. The molecular masses of the inhibitors were determined with high certainty by using retention time matching and peak shape comparison. Due to the high matching accuracy, baseline separation of coeluting analytes was not necessary in order to identify the correct masses of the bioactive compounds. The continuous-flow system was successfully applied for the screening of complex samples, such as natural extracts. For a red clover extract, detection limits of 0.3-0.8 μmol/L were obtained. System validation was performed by determining the IC

AB - Liquid chromatography (LC) was coupled on-line to a continuous-flow enzymatic assay using electrospray ionization mass spectrometry (ESI-MS) as readout for the screening of enzyme inhibitors in complex samples. Inhibitors were detected by changes in the concentration of the enzymatic reaction products, indicating the inhibition of enzymatic activity. The molecular masses of the inhibitors were determined with high certainty by using retention time matching and peak shape comparison. Due to the high matching accuracy, baseline separation of coeluting analytes was not necessary in order to identify the correct masses of the bioactive compounds. The continuous-flow system was successfully applied for the screening of complex samples, such as natural extracts. For a red clover extract, detection limits of 0.3-0.8 μmol/L were obtained. System validation was performed by determining the IC

U2 - 10.1021/ac035380w

DO - 10.1021/ac035380w

M3 - Article

SN - 0003-2700

VL - 76

SP - 3155

EP - 3161

JO - Analytical Chemistry

JF - Analytical Chemistry

IS - 11

ER -

On-line coupling of high-performance liquid chromatography to a continuous-flow enzyme assay based on electrospray ionization mass spectrometry

Abstract

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