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A targeted ultra performance liquid chromatography - tandem mass spectrometric assay for tyrosine and metabolites in urine and plasma: application to the effects of antibiotics on mice

A targeted ultra performance liquid chromatography - tandem mass spectrometric assay for tyrosine and metabolites in urine and plasma: application to the effects of antibiotics on mice
A targeted ultra performance liquid chromatography - tandem mass spectrometric assay for tyrosine and metabolites in urine and plasma: application to the effects of antibiotics on mice

Tyrosine plays a key role in mammalian biochemistry and defects in its metabolism (e.g., tyrosinemia, alkaptonuria etc.) have significant adverse consequences for those affected if left untreated. In addition, gut bacterially-derived p-cresol and its metabolites are of interest as a result of various effects on host xenobiotic metabolism. A fit-for-purpose quantitative ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) assay was developed to target and quantify tyrosine and eleven metabolites in urine and plasma. Dansylation, using dansyl chloride, was used to improve chromatographic and mass spectral properties for tyrosine and nine phenolic metabolites, with detection using positive electrospray ionisation (ESI). The sulfate and glucuronide conjugates of p-cresol, where the phenol group was blocked, were quantified intact, using negative ESI via polarity switching during the same run. Sample preparation for urine and plasma involved deproteinization by solvent precipitation (of acetonitrile:isopropyl alcohol (1:1 v/v)) followed by in situ dansylation in 96 well plates. To minimize sample and solvent usage, and maximize sensitivity, analysis was performed using microbore reversed-phase gradient UPLC on a C8 phase with a 7.5 min. cycle time. The coefficients of variation obtained were <15%, with lower limits of quantification ranging from 5 to 250 nM depending upon the analyte. The method was applied to plasma and urine samples obtained from mice placed on a high tyrosine diet with one subgroup of animals subsequently receiving antibiotics to suppress the gut microbiota. Whilst plasma profiles were largely unaffected by antibiotic treatment clear reductions in the amount of p-cresol sulfate and p-cresol glucuronide excreted in the urine were observed for these mice.

Biofluids, Dansyl chloride, Metabolites, P-cresol conjugates, Quantification, Tyrosine
1570-0232
1-9
Letertre, Marine P M
dac01b82-106d-4480-a123-7747c51dcf4e
Myridakis, Antonis
e44d6247-d3ea-4b0e-992e-431cb4d95302
Whiley, Luke
af3461fa-e393-40ae-8dba-d09cb1a22d02
Camuzeaux, Stéphane
6b867d8a-f607-4c73-af41-0ae23fe0d7d3
Lewis, Matthew R
ee27221b-8582-41d4-94d1-ea36b74c5378
Chappell, Katie E
7b9a74a7-7f1c-45f0-9a8b-cec23d47e39b
Thaikkatil, Annie
aada82dc-a74c-4730-b18b-abede4a67491
Dumas, Marc-Emmanuel
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Nicholson, Jeremy K
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Swann, Jonathan R
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Wilson, Ian D
d7da811b-6cc9-4166-82d0-e780c95122d2
Letertre, Marine P M
dac01b82-106d-4480-a123-7747c51dcf4e
Myridakis, Antonis
e44d6247-d3ea-4b0e-992e-431cb4d95302
Whiley, Luke
af3461fa-e393-40ae-8dba-d09cb1a22d02
Camuzeaux, Stéphane
6b867d8a-f607-4c73-af41-0ae23fe0d7d3
Lewis, Matthew R
ee27221b-8582-41d4-94d1-ea36b74c5378
Chappell, Katie E
7b9a74a7-7f1c-45f0-9a8b-cec23d47e39b
Thaikkatil, Annie
aada82dc-a74c-4730-b18b-abede4a67491
Dumas, Marc-Emmanuel
15b9f5b3-0452-465e-8f0a-d9ac2e94da30
Nicholson, Jeremy K
72991774-7e08-4592-ae57-e7dcc2ec158e
Swann, Jonathan R
7c11a66b-f4b8-4dbf-aa17-ad8b0561b85c
Wilson, Ian D
d7da811b-6cc9-4166-82d0-e780c95122d2

Letertre, Marine P M, Myridakis, Antonis, Whiley, Luke, Camuzeaux, Stéphane, Lewis, Matthew R, Chappell, Katie E, Thaikkatil, Annie, Dumas, Marc-Emmanuel, Nicholson, Jeremy K, Swann, Jonathan R and Wilson, Ian D (2021) A targeted ultra performance liquid chromatography - tandem mass spectrometric assay for tyrosine and metabolites in urine and plasma: application to the effects of antibiotics on mice. Journal of Chromatography B, 1164, 1-9, [122511]. (doi:10.1016/j.jchromb.2020.122511).

Record type: Article

Abstract

Tyrosine plays a key role in mammalian biochemistry and defects in its metabolism (e.g., tyrosinemia, alkaptonuria etc.) have significant adverse consequences for those affected if left untreated. In addition, gut bacterially-derived p-cresol and its metabolites are of interest as a result of various effects on host xenobiotic metabolism. A fit-for-purpose quantitative ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) assay was developed to target and quantify tyrosine and eleven metabolites in urine and plasma. Dansylation, using dansyl chloride, was used to improve chromatographic and mass spectral properties for tyrosine and nine phenolic metabolites, with detection using positive electrospray ionisation (ESI). The sulfate and glucuronide conjugates of p-cresol, where the phenol group was blocked, were quantified intact, using negative ESI via polarity switching during the same run. Sample preparation for urine and plasma involved deproteinization by solvent precipitation (of acetonitrile:isopropyl alcohol (1:1 v/v)) followed by in situ dansylation in 96 well plates. To minimize sample and solvent usage, and maximize sensitivity, analysis was performed using microbore reversed-phase gradient UPLC on a C8 phase with a 7.5 min. cycle time. The coefficients of variation obtained were <15%, with lower limits of quantification ranging from 5 to 250 nM depending upon the analyte. The method was applied to plasma and urine samples obtained from mice placed on a high tyrosine diet with one subgroup of animals subsequently receiving antibiotics to suppress the gut microbiota. Whilst plasma profiles were largely unaffected by antibiotic treatment clear reductions in the amount of p-cresol sulfate and p-cresol glucuronide excreted in the urine were observed for these mice.

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More information

Accepted/In Press date: 14 December 2020
e-pub ahead of print date: 22 December 2020
Published date: 1 February 2021
Keywords: Biofluids, Dansyl chloride, Metabolites, P-cresol conjugates, Quantification, Tyrosine

Identifiers

Local EPrints ID: 448390
URI: http://eprints.soton.ac.uk/id/eprint/448390
ISSN: 1570-0232
PURE UUID: 8c5d383f-c150-44e1-ae54-806fadf905bd
ORCID for Jonathan R Swann: ORCID iD orcid.org/0000-0002-6485-4529

Catalogue record

Date deposited: 21 Apr 2021 16:32
Last modified: 17 Mar 2024 04:00

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Contributors

Author: Marine P M Letertre
Author: Antonis Myridakis
Author: Luke Whiley
Author: Stéphane Camuzeaux
Author: Matthew R Lewis
Author: Katie E Chappell
Author: Annie Thaikkatil
Author: Marc-Emmanuel Dumas
Author: Jeremy K Nicholson
Author: Ian D Wilson

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