Quantitative determination of pyruvate, the standard substance for Reitman-Frankel method.

DOI Web Site Web Site Open Access

Bibliographic Information

Other Title
  • Reitman-Frankel法の標準物質としてのピルビン酸の含量測定法
  • Reitman-Frankelホウ ノ ヒョウジュン ブッシツ ト シテ ノ ビルビンサン ノ ガンリョウ ソクテイホウ

Search this article

Description

Following the mechanism of color reaction (Fig.4) in Reitman-Frankel assay method of transaminase activity, 2, 4-dinitrophenylhydrazone of pyruvic acid. (2, 4-DZ) was purified and used as a standard substance, and the quantity of pyruvate was determined colorimetrically (TableI). <BR>One milliliter of 0.8mM 2, 4-dinitrophenylhydrazine (2, 4-D) in 0.4N HCl was added to 2ml of 0.2mM 2, 4-DZ in 0.08N NaOH (standard).Simultaneously, 1ml of1.2 mM 2, 4-D (see Fig.3) in 0.25N HCl was added to 2ml of 0.2mM pyruvate (sample).After equilibration for 30minutes at 25°C, the both solutions resulted in similar concentrations of 2, 4-D, 2, 4-DZ, pyruvate and HCl (pH 1.20, see Fig. 2). To each of the solutions, 10 ml of 1 N NaOH was added to make the pH 13, and the absorbance at 520mμ (Fig.1) was measured after standing for 15 minutes. By comparing the absorbance of sample solution to that of standard, the concentration of pyruvate was determined.<BR>Content of purified sample of lithium pyruvate was assayed as 99.8% (σ=0.34%) by this method while as 96.8% (σ=0.53%) by iodometric titration.

Journal

  • BUNSEKI KAGAKU

    BUNSEKI KAGAKU 19 (5), 694-698, 1970

    The Japan Society for Analytical Chemistry

Keywords

Details 詳細情報について

Report a problem

Back to top