Hydrophilic interaction liquid chromatography tandem mass spectrometry analysis of malonyl‐coenzyme A in breast cancer cell cultures applying online solid‐phase extraction

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EarlyView Article

  • Published: Oct 4, 2017
  • Author: Alexander Schriewer, Cristina Cadenas, Heiko Hayen
  • Journal: Journal of Separation Science

Abstract

Cofactors such as coenzyme A and its derivatives acetyl‐coenzyme A and malonyl‐coenzyme A are involved in many metabolic pathways. Due to trace level concentrations in biological samples and the high reactivity of cofactors, a fast, sensitive, and selective method for quantification is mandatory. In this study, online solid‐phase extraction was coupled successfully to hydrophilic interaction liquid chromatography with tandem mass spectrometry for isolation of analytes in complex matrix and quantification by external calibration. Online solid‐phase extraction was carried out by application of a weak anion‐exchange column, whereas hydrophilic interaction liquid chromatography separation was performed on an amide modified stationary phase. Sample preparation of the extracts before the analysis was reduced to a centrifugation and dilution step. Moreover, the applied online solid‐phase extraction significantly reduced matrix effects and increased the signal‐to‐noise ratio. The limit of detection and the limit of quantification were in the lower nanomolar range. Finally, the applicability of this method was demonstrated on MCF‐7 breast cancer cell cultures, a commonly used model system, where acetyl‐coenzyme A and malonyl‐coenzyme A were determined using standard addition procedure in concentrations of 1.98 μM and 41 nM, respectively.

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