Non-degradative extraction and simultaneous quantitation of riboflavin, flavin mononucleotide, and flavin adenine dinucleotide in foods by HPLC
1992
Russell, L.F. | Vanderslice, J.T.
The published tables of nutrient composition only list values for the total riboflavin (TRF) content of foods and much of this data was generated by methods of questionable accuracy. There are no published methods for simultaneously determining the three principal forms of vitamin B2, i.e. riboflavin (RF), flavin mononucleotide (FMN), and flavin adenine dinucleotide (FAD), in foods. A simultaneous high performance liquid chromatographic (HPLC) separation of RF, FMN and FAD has been developed on polymer-based columns, using a mobile phase gradient of acetonitrile in citrate-phosphate buffer, pH 5.50. The natural fluorescence of the flavins was used for detection, and a RF analog, 7-ethyl-8-methyl-riboflavin, was used as the internal standard. A two-step extraction procedure, using methylene chloride, methanol, and the citrate-phosphate buffer, was also developed. The recoveries from spiked food samples demonstrated that the flavins were stable under these extraction conditions. The commercial FMN and FAD standards were found to contain significant quantities of impurities, and the appropriate correction factors were developed and applied to the HPLC results. This method was successfully applied to a variety of foods that represented significant dietary sources of vitamin B2, including raw and cooked meats, dairy products, eggs, and cereal products. The results of the HPLC analyses generally agreed with published values for TRF.
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