Quantitative UPLC-MS/MS analysis of the gut microbial co-metabolites phenylacetylglutamine, 4-cresyl sulphate and hippurate in human urine: INTERMAP Study

Anisha Wijeyesekera, Philip A. Clarke, Magda Bictash, Ian J. Brown, Mark Fidock, Thomas Ryckmans, Ivan K.S. Yap, Queenie Chan, Jeremiah Stamler, Paul Elliott, Elaine Holmes, Jeremy K. Nicholson*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

26 Scopus citations

Abstract

The role of the gut microbiome in human health, and non-invasive measurement of gut dysbiosis are of increasing clinical interest. New high-throughput methods are required for the rapid measurement of gut microbial metabolites and to establish reference ranges in human populations. We used ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) - positive and negative electrospray ionization modes, multiple reaction monitoring transitions - to simultaneously measure three urinary metabolites (phenylacetylglutamine, 4-cresyl sulphate and hippurate) that are potential biomarkers of gut function, among multi-ethnic US men and women aged 40-59 from the INTERMAP epidemiologic study (n = 2000, two timed 24-hr urine collections/person). Metabolite concentrations were quantified via stable isotope labeled internal standards. The assay was linear in the ranges 1ng mL -1 (lower limit of quantification) to 1000ng mL -1 (phenylacetylglutamine and 4-cresyl sulfate) and 3ng mL -1 to 3000ng mL -1 (hippurate). These quantitative data provide new urinary reference ranges for population-based human samples: mean (standard deviation) 24-hr urinary excretion for phenylacetylglutamine was: 1283.0 (751.7) μmol/24-hr (men), 1145.9 (635.5) μmol/24-hr (women); for 4-cresyl sulphate, 1002.5 (737.1) μmol/24-hr (men), 1031.8 (687.9) μmol/24-hr (women); for hippurate, 6284.6 (4008.1) μmol/24-hr (men), 4793.0 (3293.3) μmol/24-hr (women). Metabolic profiling by UPLC-MS/MS in a large sample of free-living individuals has provided new data on urinary reference ranges for three urinary microbial co-metabolites, and demonstrates the applicability of this approach to epidemiological investigations.

Original languageEnglish (US)
Pages (from-to)65-72
Number of pages8
JournalAnalytical Methods
Volume4
Issue number1
DOIs
StatePublished - Jan 2012

ASJC Scopus subject areas

  • Analytical Chemistry
  • Chemical Engineering(all)
  • Engineering(all)

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