High performance liquid chromatography assay for alanine amino transferase

S. Canepari, V. Carunchio, A. M. Girelli, A. Messina

Research output: Contribution to journalArticle

Abstract

An improved method of alanine-amino transferase determination is proposed. The reaction is carried out with alanine and 2-oxoglutarate as substrates and analysis is by HPLC on a reversed-phase chromatographic system using a C18 column and tetrabutylammonium phosphate in phosphate buffer (pH 7.0)-acetonitrile as mobile phase. The enzyme activity was determined by directly following the formation of pyruvic acid without employing any secondary reaction, which is necessary in the spectrophotometric method. The detection limit of pyruvic acid is 10 pmole μl-1 and the standard deviation for the enzymatic activity of standard solutions is 5.4%. Furthermore under the chromatographic conditions selected it is possible to detect the presence of some intermediate species.

Original languageEnglish
Pages (from-to)23-26
Number of pages4
JournalChromatographia
Volume32
Issue number1-2
DOIs
Publication statusPublished - Jul 1991

Keywords

  • Alanine amino transferase
  • Column liquid chromatography
  • Enzyme assay

ASJC Scopus subject areas

  • Analytical Chemistry
  • Clinical Biochemistry

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