Thiol profiling in cancer cell lines by HPLC-mass spectrometry

STAR Protoc. 2021 Dec 8;2(4):100977. doi: 10.1016/j.xpro.2021.100977. eCollection 2021 Dec 17.

Abstract

We describe a protocol for identifying cellular thiol metabolites such as cysteine and cystine in adherent cells using high performance liquid chromatography (HPLC) tandem mass spectrometry-based metabolomics. We applied a modified extraction and sample derivatization protocol to accurately quantify the intracellular levels of labile thiol species and to inhibit oxidation prior to analysis. For complete details on the use and execution of this protocol, please refer to Liu et al. (2020) and Koppula et al. (2021).

Keywords: Cancer; Mass Spectrometry; Metabolism.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Culture Techniques
  • Cell Line, Tumor
  • Chromatography, High Pressure Liquid / methods*
  • Humans
  • Metabolomics / methods*
  • Neoplasms* / chemistry
  • Neoplasms* / metabolism
  • Sulfhydryl Compounds* / analysis
  • Sulfhydryl Compounds* / chemistry
  • Sulfhydryl Compounds* / metabolism
  • Tandem Mass Spectrometry / methods*

Substances

  • Sulfhydryl Compounds