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High-performance liquid chromatographic (HPLC) analysis of thiamin pyrophosphokinase activity in rat liver
S Egoramaiphol1, A Henke-Saipt, E Klapp-Kunsemüller
1Institut für Ernährungswissenschaft, Justus Liebig Universität, Giessen, FRG.
Abstract:
A HPLC method for the determination of TPPK activity in rat liver is presented. The assay comprises an extraction of TPPK from the liver into sodium phosphate buffer, pH 7.3. TPP formed in the TPPK reaction from thiamin was converted to thiochrome and thiochrome was absorbed onto Shandon APS Hypersil. The mobile phase was acetonitrile/potassium phosphate buffer, pH 8.2, and elution was monitored spectrofluorometrically.