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

doi:10.1039/c1ay05427a

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

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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 language	English (US)
Pages (from-to)	65-72
Number of pages	8
Journal	Analytical Methods
Volume	4
Issue number	1
DOIs	https://doi.org/10.1039/c1ay05427a
State	Published - Jan 2012

ASJC Scopus subject areas

Analytical Chemistry
General Chemical Engineering
General Engineering

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10.1039/c1ay05427a

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Wijeyesekera, A., Clarke, P. A., Bictash, M., Brown, I. J., Fidock, M., Ryckmans, T., Yap, I. K. S., Chan, Q., Stamler, J., Elliott, P., Holmes, E., & Nicholson, J. K. (2012). Quantitative UPLC-MS/MS analysis of the gut microbial co-metabolites phenylacetylglutamine, 4-cresyl sulphate and hippurate in human urine: INTERMAP Study. Analytical Methods, 4(1), 65-72. https://doi.org/10.1039/c1ay05427a

@article{a248d46a3d624440a1f5e3d9af219705,

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

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.",

author = "Anisha Wijeyesekera and Clarke, \{Philip A.\} and Magda Bictash and Brown, \{Ian J.\} and Mark Fidock and Thomas Ryckmans and Yap, \{Ivan K.S.\} and Queenie Chan and Jeremiah Stamler and Paul Elliott and Elaine Holmes and Nicholson, \{Jeremy K.\}",

year = "2012",

month = jan,

doi = "10.1039/c1ay05427a",

language = "English (US)",

volume = "4",

pages = "65--72",

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publisher = "Royal Society of Chemistry",

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Wijeyesekera, A, Clarke, PA, Bictash, M, Brown, IJ, Fidock, M, Ryckmans, T, Yap, IKS, Chan, Q, Stamler, J, Elliott, P, Holmes, E & Nicholson, JK 2012, 'Quantitative UPLC-MS/MS analysis of the gut microbial co-metabolites phenylacetylglutamine, 4-cresyl sulphate and hippurate in human urine: INTERMAP Study', Analytical Methods, vol. 4, no. 1, pp. 65-72. https://doi.org/10.1039/c1ay05427a

TY - JOUR

T1 - Quantitative UPLC-MS/MS analysis of the gut microbial co-metabolites phenylacetylglutamine, 4-cresyl sulphate and hippurate in human urine

T2 - INTERMAP Study

AU - Wijeyesekera, Anisha

AU - Clarke, Philip A.

AU - Bictash, Magda

AU - Brown, Ian J.

AU - Fidock, Mark

AU - Ryckmans, Thomas

AU - Yap, Ivan K.S.

AU - Chan, Queenie

AU - Stamler, Jeremiah

AU - Elliott, Paul

AU - Holmes, Elaine

AU - Nicholson, Jeremy K.

PY - 2012/1

Y1 - 2012/1

N2 - 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.

AB - 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.

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

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