Fast and quantitative analysis of branched-chain amino acids in biological samples using a pillar array column Amino Acid Analysis

Yanting Song, Katsuya Takatsuki, Muneki Isokawa, Tetsushi Sekiguchi, Jun Mizuno, Takashi Funatsu, Shuichi Shoji, Makoto Tsunoda

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

In this study, a fast and quantitative determination method for branched-chain amino acids (BCAAs), namely leucine, isoleucine, and valine, was developed using a pillar array column. A pillar array column with low-dispersion turns was fabricated on a 20 × 20-mm2 microchip using multistep ultraviolet photolithography and deep reactive ion etching. The BCAAs were fluorescently labeled with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F), followed by reversed-phase separation on the pillar array column. The NBD derivatives of the three BCAAs and an internal standard (6-aminocaproic acid) were separated in 100 s. The calibration curves for the NBD-BCAAs had good linearity in the range of 0.4-20 μM, using an internal standard. The intra- and interday precisions were found to be in the ranges of 1.42-3.80 and 2.74-6.97 %, respectively. The accuracies for the NBD-BCAA were from 90.2 to 99.1 %. The method was used for the analysis of sports drink and human plasma samples. The concentrations of BCAAs determined by the developed method showed good agreements with those determined using a conventional high-performance liquid chromatography system. As BCAAs are important biomarkers of some diseases, these results showed that the developed method could be a potential diagnostic tool in clinical research.

Original languageEnglish
Pages (from-to)7993-7999
Number of pages7
JournalAnalytical and Bioanalytical Chemistry
Volume405
Issue number25
DOIs
Publication statusPublished - 2013 Oct 1

Keywords

  • 4-fluoro-7-nitro-2,1, 3-benzoxadiazole
  • Fluorescence
  • Human plasma
  • Internal standard
  • Microchip

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry

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