Determination of Riboflavin and Flavocoenzymes in Human Blood Plasma by High-Performance Liquid Chromatography

A high-performance liquid chromatographic method for determining riboflavin, flavin adenine dinucleotide, and flavin mononucleotide in human blood plasma is presented. Flavocoenzymes are determined as flavin mononucleotide after acid hydrolysis of flavin adenine dinucleotide. Metabolites are separated by reversed-phase column chromatography and quantified by their native fluorescence. Criteria of quality are (riboflavin/flavocoenzymes): coefficients of variation 2.8/4.6% (intra-assay) and 2.8/4.4% (inter-assay); recovery 82.4/94.4%; detection limit < 3.0/9.0 nmol/l. Because sample preparation requires only few steps, and the retention times are short ( < 5 min), this method is recommended for use in routine analysis.