Development and validation of an ultra-performance liquid chromatography quadrupole time of flight mass spectrometry method for rapid quantification of free amino acids in human urine

Amino Acids
Richard JoyceRuey Leng Loo

Abstract

An ultra-performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-qTOF-MS) method using hydrophilic interaction liquid chromatography was developed and validated for simultaneous quantification of 18 free amino acids in urine with a total acquisition time including the column re-equilibration of less than 18 min per sample. This method involves simple sample preparation steps which consisted of 15 times dilution with acetonitrile to give a final composition of 25 % aqueous and 75 % acetonitrile without the need of any derivatization. The dynamic range for our calibration curve is approximately two orders of magnitude (120-fold from the lowest calibration curve point) with good linearity (r (2) ≥ 0.995 for all amino acids). Good separation of all amino acids as well as good intra- and inter-day accuracy (<15 %) and precision (<15 %) were observed using three quality control samples at a concentration of low, medium and high range of the calibration curve. The limits of detection (LOD) and lower limit of quantification of our method were ranging from approximately 1-300 nM and 0.01-0.5 µM, respectively. The stability of amino acids in the prepared urine samples was found to be stable for 72 h at 4 °C, a...Continue Reading

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Citations

Feb 2, 2017·Journal of Separation Science·Ana Paola VilchesDan Bylund
Aug 10, 2017·The Analyst·O BegouG Theodoridis
Aug 29, 2020·Amino Acids·Weihua XuNing Zhang
May 26, 2018·Biomedical Chromatography : BMC·Jie ZhaoFrank J Gonzalez
Feb 9, 2019·Frontiers in Cellular Neuroscience·Aiping LiuMingbang Wang
Nov 5, 2019·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Juan LiHong-Min Liu
Apr 21, 2021·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Eirini StampinaParaskevas D Tzanavaras
Oct 20, 2018·Analytical Chemistry·William M Gilliland, J Michael Ramsey
Dec 4, 2021·Mass Spectrometry Reviews·Annelaure DamontJean-Claude Tabet

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Methods Mentioned

BETA
ion-exchange chromatography
electrophoresis

Software Mentioned

QuanLynx
Accq
Tag
Masslynx

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