Simple isocratic high-performance liquid chromatographic method for the separation of the six vitamers of vitamin B6.
A simple, sensitive and fast isocratic high-performance liquid chromatographic method has been developed for the separation of all six biologically active forms (vitamers) of vitamin B6. The separation is accomplished using a strong cation-exchange column and a mobile phase of 0.1 M ammonium dihydrogenphosphate adjusted to pH 4.0. All six vitamers are separated within 20 min at a flowrate of 1 ml/min. The concentration of the vitamers is determined with a fluorescence detector (excitation 290 nm; emission 389 nm). The within-run precision of the method expressed as the coefficient of variation is below 5% at the 25 pmol level. Pyridoxal 5'-phosphate can be determined using either pre- or post-column derivatization with sodium bisulfite. Application of the method to cell-free yeast culture media is presented.
Argoudelis
C J
CJ
University of Illinois, Department of Food Science, Urbana 61801.
D002851:Chromatography, High Pressure Liquid; D007536:Isomerism; D011736:Pyridoxine / Q000302:isolation & purification*; D012015:Reference Standards; D012440:Saccharomyces / Q000378:metabolism; D013050:Spectrometry, Fluorescence
D011736:Pyridoxine