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 language | English (US) |
|---|---|
| Pages (from-to) | 72-81 |
| Number of pages | 10 |
| Journal | Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences |
| Volume | 1124 |
| DOIs | |
| State | Published - Aug 15 2019 |
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Keywords
- Amino acids
- Biomarker
- Hepatocellular carcinoma
- LC-MS/MS
- Tumor progression
ASJC Scopus subject areas
- Analytical Chemistry
- Biochemistry
- Clinical Biochemistry
- Cell Biology
Cite this
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. / Liu, Zhenzhen; Tu, Mei Juan; Zhang, Chao; Jilek, Joseph L.; Zhang, Qian Yu; Yu, Aiming.
In: Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, Vol. 1124, 15.08.2019, p. 72-81.Research output: Contribution to journal › Article
}
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
KW - Biomarker
KW - Hepatocellular carcinoma
KW - LC-MS/MS
KW - Tumor progression
UR - http://www.scopus.com/inward/record.url?scp=85066800759&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85066800759&partnerID=8YFLogxK
U2 - 10.1016/j.jchromb.2019.05.039
DO - 10.1016/j.jchromb.2019.05.039
M3 - Article
C2 - 31177050
AN - SCOPUS:85066800759
VL - 1124
SP - 72
EP - 81
JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
JF - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
SN - 1570-0232
ER -