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A validated UPLC-MS/MS assay for the quantification of amino acids and biogenic amines in rat urine

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Title: A validated UPLC-MS/MS assay for the quantification of amino acids and biogenic amines in rat urine
Authors: Gray, N
Plumb, RS
Wilson, ID
Nicholson, JK
Item Type: Journal Article
Abstract: A UPLC-MS/MS assay, employing a reversed-phase separation, has been applied to the analysis of a number of common amino acids and biogenic amines in rat urine. Analytes were derivatised, using 6‑aminoquinolyl‑N‑hydroxysuccinimidyl carbamate (AccQTag Ultra™). Derivatisation with this reagent, by increasing the hydrophobicity of the analytes, enables better retention by improving reversed-phase chromatographic properties and also improves ionisation efficiency to enhance MS-detection. The method allows for the determination of 38 amino compounds in 7.5 min, including baseline resolution of critical isomers. The assay has been validated for the absolute quantification of 29 amino compounds in rat urine, over a concentration range of 0.6–200 μM, for the purpose of exploratory metabolite phenotyping. Acceptable linearity (R2 > 0.995) and intra- and inter-day accuracy (<20.7%) and precision (<20.1%) for these analytes was achieved. The limits of detection ranged from 1.2–12 fmol on column with 20 μL of sample. The remaining nine amines examined were not accurately quantified by this method but can be monitored for relative/fold change in biological samples. The use of the method is exemplified by the monitoring of changes in healthy male Sprague-Dawley rat urinary amino acid concentrations over a 7-day period.
Issue Date: 1-Feb-2019
Date of Acceptance: 27-Dec-2018
URI: http://hdl.handle.net/10044/1/77642
DOI: 10.1016/j.jchromb.2018.12.028
ISSN: 1570-0232
Publisher: Elsevier
Start Page: 50
End Page: 57
Journal / Book Title: Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume: 1106
Copyright Statement: © 2018 Elsevier Ltd. All rights reserved. This manuscript is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International Licence http://creativecommons.org/licenses/by-nc-nd/4.0/
Keywords: Science & Technology
Life Sciences & Biomedicine
Physical Sciences
Biochemical Research Methods
Chemistry, Analytical
Biochemistry & Molecular Biology
Chemistry
Amino compounds
Metabolic phenotyping
Quantification
Validation
6-aminoquinolyl-N-hydrotrysuccinimidyl carbamate
PERFORMANCE LIQUID-CHROMATOGRAPHY
PRECOLUMN DERIVATIZATION
HUMAN PLASMA
CAPILLARY-ELECTROPHORESIS
HIGH-THROUGHPUT
CARBAMATE
QUANTITATION
NORMALIZATION
6‑aminoquinolyl‑N‑hydroxysuccinimidyl carbamate
Amino compounds
Metabolic phenotyping
Quantification
Validation
Amino Acids
Aminoquinolines
Animals
Biogenic Amines
Biological Assay
Carbamates
Chromatography, High Pressure Liquid
Male
Rats
Rats, Sprague-Dawley
Sensitivity and Specificity
Tandem Mass Spectrometry
Animals
Rats
Rats, Sprague-Dawley
Biogenic Amines
Carbamates
Aminoquinolines
Amino Acids
Biological Assay
Chromatography, High Pressure Liquid
Sensitivity and Specificity
Male
Tandem Mass Spectrometry
Science & Technology
Life Sciences & Biomedicine
Physical Sciences
Biochemical Research Methods
Chemistry, Analytical
Biochemistry & Molecular Biology
Chemistry
Amino compounds
Metabolic phenotyping
Quantification
Validation
6-aminoquinolyl-N-hydrotrysuccinimidyl carbamate
PERFORMANCE LIQUID-CHROMATOGRAPHY
PRECOLUMN DERIVATIZATION
HUMAN PLASMA
CAPILLARY-ELECTROPHORESIS
HIGH-THROUGHPUT
CARBAMATE
QUANTITATION
NORMALIZATION
0301 Analytical Chemistry
0601 Biochemistry and Cell Biology
1115 Pharmacology and Pharmaceutical Sciences
Publication Status: Published
Online Publication Date: 2018-12-29
Appears in Collections:Department of Surgery and Cancer