Premium
A reliable LC‐MS/MS method for targeted analysis of amino acid metabolome: Method validation and application to studies on amino acid dynamics during tumor progression
Author(s) -
Liu Zhenzhen,
Tu Mei-Juan,
Zhang Chao,
Jilek Joseph Lionel,
Zhang Qian-Yu,
Yu Ai-Ming
Publication year - 2020
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2020.34.s1.03701
Subject(s) - formic acid , chemistry , ammonium formate , chromatography , selected reaction monitoring , metabolome , amino acid , analyte , hepatocellular carcinoma , metabolomics , mass spectrometry , tandem mass spectrometry , biochemistry , cancer research , medicine
A simple and fast LC‐MS/MS method was developed and validated for targeted quantification of 20 proteinogenic L‐amino acids (AAs) in mouse plasma. Chromatographic separation was achieved on an Intrada Amino Acid column within 13 min via gradient elution with an aqueous solution containing 100 mM ammonium formate and an organic mobile phase containing acetonitrile, water and formic acid ( v : v : v = 95: 5: 0.3), at the flow rate of 0.6 mL/min. Individual AAs and corresponding stable‐isotope‐labeled AA internal standards were analyzed by multiple reaction monitoring (MRM) in positive ion mode under optimized conditions. Method validation consisted of linearity, sensitivity, accuracy and precision, recovery, matrix effect, and stability, and the results demonstrated this method as a specific, accurate, and reliable assay. This LC‐MS/MS method was thus utilized to examine the dynamics of plasma AA metabolome during tumor progression in an orthotropic hepatocellular carcinoma (HCC) xenograft mouse model. The results reveal a significantly lower Arg concentration as well as higher Ala and Thr levels during HCC progression. These findings support the utilities of this LC‐MS/MS method and the promise of specific AAs as possible biomarkers for HCC. Support or Funding Information National Cancer Institute (grant No. R01CA225958) National Institute of General Medical Sciences (R01GM113888)