Phosphates of riboflavin and riboflavin analogs: A reinvestigation by high-performance liquid chromatography

✍ Scribed by Peter Nielsen; Peter Rauschenbach; Adelbert Bacher

• Publisher: Elsevier Science
• Year: 1983
• Tongue: English
• Weight: 743 KB
• Volume: 130
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(83)90600-0

✦ Synopsis

Phosphoric acid esters of riboflavin can be easily separated by reverse-phase high-performance liquid chromatography using eluants of 0.1 M ammonium formate in aqueous methanol. Commercial FMN preparations contained seven different flavin phosphates; the content of riboflavin S-phosphate was 70-7590 and is in agreement with previous studies. Millimole amounts of crude FMN can be processed by preparative HPLC. The method permits the preparation of >99%pure S-FUN. The following compounds were isolated in pure form and their structures determined: riboflavin 4'-phosphate, riboflavin 3'-phosphate, riboflavin 4,5'diphosphate; riboflavin 3',4'diphosphate, and riboflavin 3',5'-diphosphate. The latter compound binds tightly to apoflavodoxin from Megasphaeru elsdenii (&, = 9.7 X lo-' M). The bound flavin has high catalytic activity, thus representing a novel type of FMN analog. A wide variety of structural analogs of FMN can be obtained in pure form by preparative HPLC.