Quantification of riboflavin, riboflavin 5'-phosphate and flavin adenine dinucleotide in plasma and urine by high-performance liquid chromatography.
A high-performance liquid chromatographic method with fluorimetric detection for the quantification of riboflavin (RB), riboflavin 5'-phosphate (FMN), and flavin adenine dinucleotide (FAD) in plasma, whole blood, and urine is described. Under isocratic conditions with a reversed-phase column, the compounds are completely resolved and eluted within 9 min. Plasma proteins are precipitated with acetonitrile followed by shaking the aqueous phase with chloroform. Urine samples are diluted and injected directly. The reproducibility of this method for the quantification of RB in plasma has a between-day coefficient of variation of 6%. The application of this method is illustrated by analyzing plasma and urine samples from a human subject who received an intravenous dose of FMN equivalent to 25 mg of RB.
College of Pharmacy, University of Arizona, Tucson 85721.
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Lopez-Anaya
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Mayersohn
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002851:Chromatography, High Pressure Liquid; D005486:Flavin Mononucleotide / Q000032:analysis* / Q000097:blood / Q000652:urine; D005182:Flavin-Adenine Dinucleotide / Q000032:analysis* / Q000097:blood / Q000652:urine; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D008297:Male; D008875:Middle Aged; D012256:Riboflavin / Q000032:analysis* / Q000097:blood / Q000652:urine; D013050:Spectrometry, Fluorescence
D007202:Indicators and Reagents; D005182:Flavin-Adenine Dinucleotide; D005486:Flavin Mononucleotide; D012256:Riboflavin