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Published on: December 15, 2015
Use of chlorite to improve HPLC detection of pyridoxal 5'-phosphate
Karen L Ericson1, J Dennis Mahuren, Yvonne M Zubovic
1Indiana University-Purdue University, Fort Wayne, 2101 East Coliseum Blvd., Fort Wayne, IN 46805-1499, USA. ericsonk@ipfw.edu
Abstract:
The sensitivity of fluorescent detection of the biologically active form of Vitamin B-6, pyridoxal 5'-phosphate (PLP), in biological samples has been improved approximately four-fold by adopting chlorite as a post-column derivatization reagent (instead of bisulfite) in high-performance liquid chromatography (HPLC) separation. Chlorite oxidizes PLP to the more fluorescent 4-pyridoxic acid 5'-phosphate, and avoids the toxicity and heating of the cyanide procedure. Detection of another major metabolite, 4-pyridoxic acid (4-PA), is not effected. Detection of pyridoxal (PL) is slightly lowered due to eluting at a lower pH.
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