Avoiding Misannotation of In-Source Fragmentation Products as Cellular Metabolites in Liquid Chromatography–Mass Spectrometry-Based Metabolomics

Xu, Yi-Fan; Lu, Wenyun; Rabinowitz, Joshua D.

doi:10.1021/ac504118y

Title: Avoiding Misannotation of In-Source Fragmentation Products as Cellular Metabolites in Liquid Chromatography–Mass Spectrometry-Based Metabolomics

Journal Article · Wed Jan 14 23:00:00 EST 2015 · Analytical Chemistry

DOI: https://doi.org/10.1021/ac504118y · OSTI ID:1344897

Xu, Yi-Fan ^[1]; Lu, Wenyun ^[2]; Rabinowitz, Joshua D. ^[3]

Princeton Univ., NJ (United States). Lewis Sigler Inst. for Integrative Genomics and Dept. of Chemistry; Office of Scientific and Technical Information (OSTI)
Princeton Univ., NJ (United States). Lewis Sigler Inst. for Integrative Genomics
Princeton Univ., NJ (United States). Lewis Sigler Inst. for Integrative Genomics and Dept. of Chemistry

Liquid chromatography–mass spectrometry (LC-MS) technology allows for rapid quantitation of cellular metabolites, with metabolites identified by mass spectrometry and chromatographic retention time. Recently, with the development of rapid scanning high-resolution high accuracy mass spectrometers and the desire for high throughput screening, minimal or no chromatographic separation has become increasingly popular. Furthermore, when analyzing complex cellular extracts, however, the lack of chromatographic separation could potentially result in misannotation of structurally related metabolites. Here, we show that, even using electrospray ionization, a soft ionization method, in-source fragmentation generates unwanted byproducts of identical mass to common metabolites. For example, nucleotide-triphosphates generate nucleotide-diphosphates, and hexose-phosphates generate triose-phosphates. We also evaluated yeast intracellular metabolite extracts and found more than 20 cases of in-source fragments that mimic common metabolites. Finally and accordingly, chromatographic separation is required for accurate quantitation of many common cellular metabolites.

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Research Organization:: Princeton Univ., NJ (United States)

Sponsoring Organization:: USDOE Office of Science (SC)

Grant/Contract Number:: SC0006839; SC0012461

OSTI ID:: 1344897

Journal Information:: Analytical Chemistry, Journal Name: Analytical Chemistry Journal Issue: 4 Vol. 87; ISSN 0003-2700

Publisher:: American Chemical Society (ACS)Copyright Statement

Country of Publication:: United States

Language:: English

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