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

Schriewer A., Cadenas C., Hayen H.

Forschungsartikel (Zeitschrift) | Peer reviewed

Zusammenfassung

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.

Details zur Publikation

FachzeitschriftJournal of Separation Science
Jahrgang / Bandnr. / Volume40
Ausgabe / Heftnr. / Issue22
Seitenbereich4303-4310
StatusVeröffentlicht
Veröffentlichungsjahr2017 (04.10.2017)
Sprache, in der die Publikation verfasst istEnglisch
DOI10.1002/jssc.201700617
Link zum Volltexthttps://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85034976229&origin=inward
Stichwörtermetabolites; online sample preparation; quantification

Autor*innen der Universität Münster

Hayen, Heiko
Professur für Analytische Chemie (Prof. Hayen)