biostudies-literatureWijeyesekera ANHLBI NIH HHS65-72https://www.ebi.ac.uk/biostudies/studies/S-EPMC3740387biostudies-literatureUnknown4(1)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 (lower limit of quantification) to 1000ng/mL (phenylacetylglutamine and 4-cresyl sulfate) and 3ng/mL to 3000ng/mL (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) ?mol/24-hr (men), 1145.9 (635.5) ?mol/24-hr (women); for 4-cresyl sulphate, 1002.5 (737.1) ?mol/24-hr (men), 1031.8 (687.9) ?mol/24-hr (women); for hippurate, 6284.6 (4008.1) ?mol/24-hr (men), 4793.0 (3293.3) ?mol/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.Analytical methods : advancing methods and applicationsQuantitative UPLC-MS/MS analysis of the gut microbial co-metabolites phenylacetylglutamine, 4-cresyl sulphate and hippurate in human urine: INTERMAP Study.PMC3740387R01 HL084228R01 HL050490Stamler JWijeyesekera AClarke PAYap IKRyckmans TBrown IJChan QHolmes EElliott PBictash MFidock MNicholson JKfalseQuantitative UPLC-MS/MS analysis of the gut microbial co-metabolites phenylacetylglutamine, 4-cresyl sulphate and hippurate in human urine: INTERMAP Study.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 (lower limit of quantification) to 1000ng/mL (phenylacetylglutamine and 4-cresyl sulfate) and 3ng/mL to 3000ng/mL (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) ?mol/24-hr (men), 1145.9 (635.5) ?mol/24-hr (women); for 4-cresyl sulphate, 1002.5 (737.1) ?mol/24-hr (men), 1031.8 (687.9) ?mol/24-hr (women); for hippurate, 6284.6 (4008.1) ?mol/24-hr (men), 4793.0 (3293.3) ?mol/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.2012-01-01T00:00:00Z2012 Jan2021-02-21T01:06:42Z2019-03-27T01:14:25ZS-EPMC37403872394676710.1039/C1AY05427A10.1039/c1ay05427a