CIRCULAR PAPER CHROMATOGRAPHY Part X. Separation, Identification and Quantitative Estimation of Riboflavin and Flavin Compounds

Abstract

A method for lhe separation, identification and quantitative determination of riboflavin and its derivatives (flavin mononucleotide, flavinadenine dinucleotide, lumiflavin and lumichrome) by circular paper chromatography is described. A number of solvenl combinations have been tried in an attempt to obtain the best separation of the flavins, and IT-butanol-acetic acid·water in the proportion 40: 10: 50 was found to give the best resolution both for qualitative charactedsation as well as quantitative evaluation. Quantitative determination of the individual flavins was made by cutting out the bands from the chromatograms (after identification under ultra-violet light), eluting them with water and measuring the fluorescence in a fluorimeter. Spectrophotometric analysis was also carrJed out as a check-up and the recovery was fotmd to be satisfactory. Some applications of the method for the study of riboflavin metabolism in a mutant yeast BY2, arc briefly described. The advantages and possible applications of the technique are discussed.