Use of chlorite to improve HPLC detection of pyridoxal 5'-phosphate.
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.
Ericson
Karen L
KL
Indiana University-Purdue University, Fort Wayne, 2101 East Coliseum Blvd., Fort Wayne, IN 46805-1499, USA. ericsonk@ipfw.edu
Mahuren
J Dennis
JD
Zubovic
Yvonne M
YM
Coburn
Stephen P
SP
D002712:Chlorides / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000592:standards; D006801:Humans; D010084:Oxidation-Reduction; D011732:Pyridoxal Phosphate / Q000097:blood* / Q000737:chemistry; D011735:Pyridoxic Acid / Q000031:analogs & derivatives / Q000097:blood; D015203:Reproducibility of Results
D002712:Chlorides; D011735:Pyridoxic Acid; D011732:Pyridoxal Phosphate; C062598:4-pyridoxic acid 5'-phosphate; C001599:chlorite