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Analytical and Bioanalytical Chemistry

, Volume 407, Issue 22, pp 6681–6688 | Cite as

LC-MS/MS-based analysis of coenzyme A and short-chain acyl-coenzyme A thioesters

  • Stefan Neubauer
  • Dinh Binh Chu
  • Hans Marx
  • Michael Sauer
  • Stephan Hann
  • Gunda KoellenspergerEmail author
Research Paper

Abstract

Absolute quantification of intracellular coenzyme A (CoA), coenzyme A disulfide, and short-chain acyl-coenzyme A thioesters was addressed by developing a tailored metabolite profiling method based on liquid chromatography in combination with tandem mass spectrometric detection (LC-MS/MS). A reversed phase chromatographic separation was established which is capable of separating a broad spectrum of CoA, its corresponding derivatives, and their isomers despite the fact that no ion-pairing reagent was used (which was considered as a key advantage of the method). Excellent analytical figures of merit such as high sensitivity (LODs in the nM to sub-nM range) and high repeatability (routinely 4 %; N = 15) were obtained. Method validation comprised a study on standard purity, stability, and recoveries during sample preparation. Uniformly labeled U13C yeast cell extracts offered ideal internal standards for validation purposes and for a quantification exercise in the rumen bacterium Megasphaera elsdenii.

Keywords

Coenzyme A Short-chain acyl-coenzyme A LC-MS/MS Metabolite profiling Megasphaera elsdenii Stability 

Supplementary material

216_2015_8825_MOESM1_ESM.pdf (258 kb)
ESM 1 (PDF 257 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Stefan Neubauer
    • 1
    • 2
  • Dinh Binh Chu
    • 1
    • 3
  • Hans Marx
    • 4
    • 5
  • Michael Sauer
    • 2
    • 4
    • 5
  • Stephan Hann
    • 1
    • 2
  • Gunda Koellensperger
    • 2
    • 6
    Email author
  1. 1.Department of Chemistry, Division of Analytical ChemistryUniversity of Natural Resources and Life Sciences-BOKUViennaAustria
  2. 2.Austrian Center of Industrial Biotechnology (ACIB)ViennaAustria
  3. 3.Department of Analytical Chemistry, School of Chemical EngineeringHanoi University of Science and TechnologyHanoiVietnam
  4. 4.Department of BiotechnologyUniversity of Natural Resources and Life Sciences-BOKUViennaAustria
  5. 5.CD Laboratory for Biotechnology of GlycerolViennaAustria
  6. 6.Department of Analytical Chemistry, Faculty of ChemistryUniversity of ViennaViennaAustria

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