Quantitation of branched-chain amino acids in ascites by capillary electrophoresis with light-emitting diode-induced fluorescence detection

Po Ling Chang*, Tai Chia Chiu, Tsang En Wang, Kuang Chun Hu, Yu Hsien Tsai, Cho Chun Hu, Ming Jong Bair, Huan Tsung Chang

*Corresponding author for this work

Research output: Contribution to journalJournal Article peer-review

15 Scopus citations

Abstract

Branched-chain amino acids (BCAAs) are one of the important biomarkers for monitoring liver disease such as hepatitis or hepatoma. In this communication, we present the determination of the concentrations of BCAA in ascites by CE light-emitted diode-induced fluorescence (LEDIF) using 1.5% m/v poly(ethylene oxide) (average Mv: ∼8000000g/mol) that was prepared in 10mM sodium tetraborate solution (pH 9.3). Naphthalene-2,3-dicarboxaldehyde was used to derivatize 15 amino acids (AAs) to form naphthalene-2,3-dicarboxaldehyde (NDA)-AA derivatives prior to CE analysis. The separation of 15 NDA-AA derivatives was accomplished within 15min, with RSD values of <5.8% (within-day) and 7.4% (between-days) with respect to their migration times. The limits of detection for the tested BCAAs ranged from 10.6 to 10.9nM. We determined the concentrations of three BCAAs - leucine, isoleucine and valine - in ascites by applying a standard addition method, with recovery percentages ranging from 93.9 to 111%. The results obtained from this CE-LEDIF method is in good agreement with those by a gold standard method using an AA analyzer. We have found that the concentrations of the three BCAAs in ascites obtained from patients suffering from liver diseases were lower than those from healthy individuals. Our approach is highly efficient, sensitive, and cost-effective, which holds great potential for the diagnosis of liver diseases.

Original languageEnglish
Pages (from-to)1080-1083
Number of pages4
JournalElectrophoresis
Volume32
Issue number9
DOIs
StatePublished - 04 2011
Externally publishedYes

Keywords

  • Branched-chain amino acids
  • CE
  • Light-emitting diode-induced fluorescence
  • Naphthalene-2,3-dicarboxaldehyde
  • Poly(ethylene oxide)

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