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Artefactual formation of pyruvate from in‐source conversion of lactate
Author(s) -
Trefely Sophie,
Mesaros Clementina,
Xu Peining,
Doan Mary T.,
Jiang Helen,
Altinok Oya,
Orynbayeva Zulfiya,
Snyder Nathaniel W.
Publication year - 2018
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.8159
Subject(s) - metabolomics , chemistry , metabolite , fragmentation (computing) , chromatography , biochemistry , pyruvic acid , mass spectrometry , metabolome , metabolism , electrospray ionization , biology , ecology
Rationale Lactate and pyruvate are high‐abundance products of glucose metabolism. Analysis of both molecules as part of metabolomics studies in cellular metabolism and physiology have been aided by advances in liquid chromatography/mass spectrometry (LC/MS). Methods We used ion‐pairing chromatography and negative ion mode electrospray ionization (ESI) on a QExactive HF to perform stable isotope assisted metabolomics profiling of lactate and pyruvate metabolism. Results Using an LC/MS method for polar metabolite analysis we discovered an artefactual formation of pyruvate from in‐source fragmentation of lactate. Surprisingly, this in‐source fragmentation has not been previously described, thus we report this identification to warn other investigators. This artefact was detected by baseline chromatographic resolution of lactate and pyruvate by LC with confirmation of this artefact by stable isotope labeling of lactate and pyruvate. Conclusions These findings have immediate implications for metabolomics studies by LC/MS and direct infusion MS, especially in negative ion mode, whereby users should resolve lactate from pyruvate or robustly quantify the potential formation of pyruvate from higher abundance lactate in their assays.