Quantitative determination of leucine concentration and the abundance of C-leucine in human plasma by LC-MS/MS
Objective: To evaluate the catabolism level of amino acid and investigate the amino acid requirement of the body,a specific,sensitive and efficient LC-MS/MS method will be developed for the simultaneous determination of leucine concentration and 13C-leucine enrichment
in human plasma. Method: The plasma samples were extracted with acetonitrile.The HPLC separation was performed on an Atlantics(4.6 mm× 100 mm,3.5 μm)column using acetonitrile-water(0.1% formic acid)5:95 as mobile phase with the isocratic flow rate at 1 mL·min-1.Detection
was performed on a tandem quadrapole mass spectrometer using positive electrospray ionization;multiple reactions monitoring(MRM)scan mode was applied to detect mass transitions of leucine(m/z:132.1→86.0)and 13C-leucine(m/z:133.1→86.0).Each analytical run
was 3 min. Results: The calibration curves were linear over the concentration range of 50-1000 nmol·mL-1 and the abundance ratio range of 0.01-0.5(r > 0.99).The intra and inter batch precision(RSD,%)and accuracy(RE,%)met the acceptance criteria. Conclusion:
A sensitive and specific LC-MS/MS method for the determination of leucine concentration and 13C-leucine enrichment in human plasma is developed.The method is validated with respect to specificity,accuracy,precision and stability.Taken together,the results demonstrate the established
method is suitable to evaluate the catabolism level of protein and amino acid in clinical research.
PLEASE NOTE: The full text of this publication is in Chinese.
Keywords: 13C-leucine; MRM; isotope abundance; leucine; oxidative decomposition of amino acid; plasma sample
Document Type: Research Article
Publication date: 01 January 2013
- Journal of Pharmaceutical Analysis is a professional journal administrated by China Association for Science and Technology, sponsored by Chinese Pharmaceutical Association, held by National Institutes for Food and Drug Control, edited and published by the Editorial Board of Journal of Pharmaceutical Analysis, and publicly distributed at home and abroad. It is one of the first group of journals that created by academic leaders in the field of drug control. It originated from Drug Control Communication which magazine was founded in 1951 and later been renamed in 1981. In 2005, it was changed from bi-monthly journal to monthly journal. Journal of Pharmaceutical Analysis is among core journals in nature sciences and even other Chinese core journals, whose main columns are research reports, communications, review, forum, information and etc. It mainly reports the latest research results, explore new theories, introduce new development, spread new techniques, and promote new methods of pharmaceutical analysis. Its contents include peer-reviewed research reports, research presentation and summary reviews on all aspects of pharmaceutical analysis including drug development, pharmaceutical manufacturing, clinical research, drug safety, quality assessment, and market surveillance. Main authors and target reader are professional and technical personnel engaged in research, application as well as higher education in pharmaceutical analysis.
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