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

17 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

Fingerprint

Branched Chain Amino Acids
Amino Acids
Chemical analysis
Plasma (human)
Aminocaproic Acid
Isoleucine
Reactive ion etching
Valine
High performance liquid chromatography
Biomarkers
Photolithography
Sports
Leucine
Phase separation
Calibration
High Pressure Liquid Chromatography
Ions
Derivatives
Research

Keywords

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

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry

Cite this

Fast and quantitative analysis of branched-chain amino acids in biological samples using a pillar array column Amino Acid Analysis. / Song, Yanting; Takatsuki, Katsuya; Isokawa, Muneki; Sekiguchi, Tetsushi; Mizuno, Jun; Funatsu, Takashi; Shoji, Shuichi; Tsunoda, Makoto.

In: Analytical and Bioanalytical Chemistry, Vol. 405, No. 25, 10.2013, p. 7993-7999.

Research output: Contribution to journalArticle

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