Skip to main content
SpringerLink
Log in

Gas chromatographic–mass spectrometric analysis of the tripeptide glutathione in the electron-capture negative-ion chemical ionization mode

  • Short Communication
  • Published:
Amino Acids Aims and scope Submit manuscript

Abstract

The dicarboxylic tripeptide glutathione (GSH) is the most abundant intracellular thiol. GSH analysis by liquid chromatography is routine. Yet, GSH analysis by gas chromatography is challenged due to thermal instability and lacking volatility. We report a high-yield laboratory method for the preparation of 2H-labeled GSH dimethyl ester ((d3Me)2-GSH) for use as internal standard (IS) which was characterized by LC–MS/MS. For GC–MS analysis, the dimethyl esters of GSH and the IS were derivatized with pentafluoropropionic (PFP) anhydride. Electron-capture negative-ion chemical ionization of the (Me)2-(PFP)3-GSH provided high sensitivity. We encourage increasing use of GC–MS in the analysis of amino acids as their Me-PFP derivatives in the ECNICI mode.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Abbreviations

CID:

Collision-induced dissociation

ECNICI:

Electron-capture negative-ion chemical ionization

EI:

Electron ionization

ESI:

Electrospray ionization

GC–MS:

Gas chromatography–mass spectrometry

GSH:

Glutathione, reduced

LC–MS/MS:

Liquid chromatography–tandem mass spectrometry

Me:

Methyl

PAR:

Peak area ratio

PFP:

Pentafluoropropionyl

PFPA:

Pentafluoropropionic anhydride

References

  • Capitan P, Malmezat T, Breuillé D, Obled C (1999) Gas chromatographic–mass spectrometric analysis of stable isotopes of cysteine and in biological samples. J Chromatogr B 732:127–135

    Article  CAS  Google Scholar 

  • Giustarini D, Dalle-Donne I, Tsikas D, Rossi R (2009) Oxidative stress and human diseases: origin, link, measurement, mechanisms, and biomarkers. Crit Rev Clin Lab Sci 46:241–281

    Article  PubMed  CAS  Google Scholar 

  • Hanff E, Böhmer A, Jordan J, Tsikas D (2014) Stable-isotope dilution LC–MS/MS measurement of nitrite in human plasma after its conversion to S-nitrosoglutathione. J Chromatogr B 970:44–52

    Article  CAS  Google Scholar 

  • Iwasaki Y, Saito Y, Nakano Y, Mochizuki K, Sakata O, Ito R, Saito K, Nakazawa H (2009) Chromatographic and mass spectrometric analysis of glutathione in biological samples. J Chromatogr B 877:3309–3317

    Article  CAS  Google Scholar 

  • Kayacelebi AA, Knöffel AK, Beckmann B, Hanff E, Warnecke G, Tsikas D (2015) Measurement of unlabeled and stable isotope-labeled homoarginine, arginine and their metabolites in biological samples by GC–MS and GC–MS/MS. Amino Acids 47:2023–2034

    Article  PubMed  CAS  Google Scholar 

  • Küster A, Tea I, Sweeten S, Rozé JC, Robins RJ, Darmaun D (2008) Simultaneous determination of glutathione and cysteine concentrations and 2H enrichments in microvolumes of neonatal blood using gas chromatography–mass spectrometry. Anal Bioanal Chem 390:1403–1412

    Article  PubMed  CAS  Google Scholar 

  • Michaelsen JT, Dehnert S, Giustarini D, Beckmann B, Tsikas D (2009) HPLC analysis of human erythrocytic glutathione forms using OPA and N-acetyl-cysteine ethyl ester: evidence for nitrite-induced GSH oxidation to GSSG. J Chromatogr B 877:3405–3417

    Article  CAS  Google Scholar 

  • Monostori P, Wittmann D, Karg E, Túri S (2009) Determination of glutathione and glutathione disulfide in biological samples: an in-depth review. J Chromatogr B 877:3331–3346

    Article  CAS  Google Scholar 

  • Tsikas D, Raida M, Sandmann J, Rossa S, Forssmann WG, Frölich JC (2000) Electrospray ionization mass spectrometry of low-molecular-mass S-nitroso compounds and their thiols. J Chromatogr B 742:99–108

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors thank the DFG (Deutsche Forschungsgemeinschaft) for financial support (Grant TS 60/4-1).

Author information

Authors and Affiliations

  1. Bioanalytical Research Laboratory for NO, Oxidative Stress and Eicosanoids, Centre of Pharmacology and Toxicology, Hannover Medical School, Carl-Neuberg-Str. 1, 30625, Hannover, Germany

    Dimitrios Tsikas, Erik Hanff, Arslan Arinc Kayacelebi & Anke Böhmer

Authors
  1. Dimitrios Tsikas
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Erik Hanff
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Arslan Arinc Kayacelebi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Anke Böhmer
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Dimitrios Tsikas.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical standard

The work does not report studies in animals and humans.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 60 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Tsikas, D., Hanff, E., Kayacelebi, A.A. et al. Gas chromatographic–mass spectrometric analysis of the tripeptide glutathione in the electron-capture negative-ion chemical ionization mode. Amino Acids 48, 593–598 (2016). https://doi.org/10.1007/s00726-015-2133-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00726-015-2133-8

Keywords

Search

Navigation