A reliable LC-MS/MS method for the quantification of natural amino acids in mouse plasma: Method validation and application to a study on amino acid dynamics during hepatocellular carcinoma progression

Zhenzhen Liu, Mei Juan Tu, Chao Zhang, Joseph L. Jilek, Qian Yu Zhang, Aiming Yu

Research output: Contribution to journalArticle

Abstract

A simple and fast LC-MS/MS method was developed and validated for simultaneous quantification of 20 proteinogenic L-amino acids (AAs) in a small volume (5 μL) of 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 AAs 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 LC-MS/MS method as a specific, accurate, and reliable assay. This LC-MS/MS method was thus utilized to compare the dynamics of individual plasma AAs between healthy and orthotopic hepatocellular carcinoma (HCC) xenograft mice housed under identical conditions. Our results revealed that, 5 weeks after HCC tumor progression, plasma L-arginine concentrations were significantly decreased in HCC mice while L-alanine and L-threonine levels were sharply increased. These findings support the utilities of this LC-MS/MS method and the promise of specific AAs as possible biomarkers for HCC.

Original languageEnglish (US)
Pages (from-to)72-81
Number of pages10
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume1124
DOIs
StatePublished - Aug 15 2019

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Hepatocellular Carcinoma
formic acid
Plasmas
Amino Acids
Biomarkers
Threonine
Heterografts
Isotopes
Alanine
Arginine
Tumors
Assays
Positive ions
Flow rate
Ions
Recovery
Water
Monitoring
Neoplasms

Keywords

  • Amino acids
  • Biomarker
  • Hepatocellular carcinoma
  • LC-MS/MS
  • Tumor progression

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry
  • Cell Biology

Cite this

@article{472772434bf44843b374d4299fe740b4,
title = "A reliable LC-MS/MS method for the quantification of natural amino acids in mouse plasma: Method validation and application to a study on amino acid dynamics during hepatocellular carcinoma progression",
abstract = "A simple and fast LC-MS/MS method was developed and validated for simultaneous quantification of 20 proteinogenic L-amino acids (AAs) in a small volume (5 μL) of 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 AAs 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 LC-MS/MS method as a specific, accurate, and reliable assay. This LC-MS/MS method was thus utilized to compare the dynamics of individual plasma AAs between healthy and orthotopic hepatocellular carcinoma (HCC) xenograft mice housed under identical conditions. Our results revealed that, 5 weeks after HCC tumor progression, plasma L-arginine concentrations were significantly decreased in HCC mice while L-alanine and L-threonine levels were sharply increased. These findings support the utilities of this LC-MS/MS method and the promise of specific AAs as possible biomarkers for HCC.",
keywords = "Amino acids, Biomarker, Hepatocellular carcinoma, LC-MS/MS, Tumor progression",
author = "Zhenzhen Liu and Tu, {Mei Juan} and Chao Zhang and Jilek, {Joseph L.} and Zhang, {Qian Yu} and Aiming Yu",
year = "2019",
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TY - JOUR

T1 - A reliable LC-MS/MS method for the quantification of natural amino acids in mouse plasma

T2 - Method validation and application to a study on amino acid dynamics during hepatocellular carcinoma progression

AU - Liu, Zhenzhen

AU - Tu, Mei Juan

AU - Zhang, Chao

AU - Jilek, Joseph L.

AU - Zhang, Qian Yu

AU - Yu, Aiming

PY - 2019/8/15

Y1 - 2019/8/15

N2 - A simple and fast LC-MS/MS method was developed and validated for simultaneous quantification of 20 proteinogenic L-amino acids (AAs) in a small volume (5 μL) of 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 AAs 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 LC-MS/MS method as a specific, accurate, and reliable assay. This LC-MS/MS method was thus utilized to compare the dynamics of individual plasma AAs between healthy and orthotopic hepatocellular carcinoma (HCC) xenograft mice housed under identical conditions. Our results revealed that, 5 weeks after HCC tumor progression, plasma L-arginine concentrations were significantly decreased in HCC mice while L-alanine and L-threonine levels were sharply increased. These findings support the utilities of this LC-MS/MS method and the promise of specific AAs as possible biomarkers for HCC.

AB - A simple and fast LC-MS/MS method was developed and validated for simultaneous quantification of 20 proteinogenic L-amino acids (AAs) in a small volume (5 μL) of 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 AAs 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 LC-MS/MS method as a specific, accurate, and reliable assay. This LC-MS/MS method was thus utilized to compare the dynamics of individual plasma AAs between healthy and orthotopic hepatocellular carcinoma (HCC) xenograft mice housed under identical conditions. Our results revealed that, 5 weeks after HCC tumor progression, plasma L-arginine concentrations were significantly decreased in HCC mice while L-alanine and L-threonine levels were sharply increased. These findings support the utilities of this LC-MS/MS method and the promise of specific AAs as possible biomarkers for HCC.

KW - Amino acids

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KW - Hepatocellular carcinoma

KW - LC-MS/MS

KW - Tumor progression

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