Development of HILIC-MS/MS method for acyl-CoAs covering short- to long-chain species in a single analytical run

Abstract:
      Acyl-CoAs play a significant role in numerous physiological and metabolic processes making it important to assess their concentration levels for evaluating metabolic health. Considering the important role of acyl-CoAs, it is crucial to develop an analytical method that can analyze these compounds. Due to the structural variations of acyl-CoAs, multiple analytical methods are often required for comprehensive analysis of these compounds, which increases complexity and the analysis time. In this study, we have developed a method using a zwitterionic HILIC column that enables the coverage of free CoA and short- to long-chain acyl-CoA species in one analytical run. Initially, we developed the method on a QTOF instrument for the identification of acyl-CoA species, optimizing their chromatography and retention times. Later, a targeted HILIC-MS/MS method was created in scheduled multiple reaction monitoring mode on a QTRAP instrument. The performance of the method was evaluated based on various parameters such as linearity, precision, recovery and matrix effect. This method was applied to identify the difference in acyl-CoA profiles in HepG2 cells cultured in different conditions. Our findings revealed an increase in levels of acetyl-CoA, medium- and long-chain acyl-CoA while a decrease in the profiles of free CoA in the starved state, indicating a clear alteration in the fatty acid oxidation process.

Citation: [Preprint]

Date Published: 11th Sep 2023

Registered Mode: by DOI

help Submitter
Citation
Singh, M., Kiyuna, L. A., Odendaal, C., Bakker, B. M., Harms, A. C., & Hankemeier, T. (2023). Development of HILIC-MS/MS method for acyl-CoAs covering short- to long-chain species in a single analytical run. In []. American Chemical Society (ACS). https://doi.org/10.26434/chemrxiv-2023-h0w52
Activity

Views: 420

Created: 22nd Sep 2023 at 19:39

Last updated: 22nd Sep 2023 at 19:39

help Tags

This item has not yet been tagged.

help Attributions

None

Powered by
(v.1.14.2)
Copyright © 2008 - 2023 The University of Manchester and HITS gGmbH