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- Title
Development and validation of a rapid, selective, and sensitive LC-MS/MS method for simultaneous determination of d- and l-amino acids in human serum: application to the study of hepatocellular carcinoma.
- Authors
Han, Minlu; Xie, Mengyu; Han, Jun; Yuan, Daoyi; Yang, Tian; Xie, Ying
- Abstract
A validated liquid chromatography-tandem mass spectrometry method was developed for the simultaneous determination of d- and l-amino acids in human serum. Under the optimum conditions, except for dl-proline, l-glutamine, and d-lysine, the enantioseparation of the other 19 enantiomeric pairs of proteinogenic amino acids and nonchiral glycine was achieved with a CROWNPAK CR-I(+) chiral column within 13 min. The lower limits of quantitation for l-amino acids (including glycine) and d-amino acids were 5-56.25 μM and 0.625-500 nM, respectively, in human serum. The intraday precision and interday precision for all the analytes were less than 15%, and the accuracy ranged from −12.84% to 12.37% at three quality control levels. The proposed method, exhibiting high rapidity, enantioresolution, and sensitivity, was successfully applied to the quantification of d- and l-amino acid levels in serum from hepatocellular carcinoma patients and healthy individuals. The serum concentrations of l-arginine, l-isoleucine, l-aspartate, l-tryptophan, l-alanine, l-methionine, l-serine, glycine, l-valine, l-leucine, l-phenylalanine, l-threonine, d-isoleucine, d-alanine, d-glutamate, d-glutamine, d-methionine, and d-threonine were significantly reduced in the hepatocellular carcinoma patients compared with the healthy individuals (<italic>P</italic> < 0.01). d-Glutamate and d-glutamine were identified as the most downregulated serum markers (fold change greater than 1.5), which deserves further attention in hepatocellular carcinoma research.Simultaneous determination of d- and l-amino acids in human serum from hepatocellular carcinoma patients and healthy individuals. AA amino acid, HCC hepatocellular carcinoma, LC liquid chromatography, MS/MS tandem mass spectrometry, NC normal control, TIC total ion chromatogram<graphic></graphic>
- Subjects
HIGH performance liquid chromatography; AMINO acids; BLOOD plasma; HEPATOTOXICOLOGY; METABOLITES
- Publication
Analytical & Bioanalytical Chemistry, 2018, Vol 410, Issue 10, p2517
- ISSN
1618-2642
- Publication type
Academic Journal
- DOI
10.1007/s00216-018-0883-3