Chromatographic separation of 24(R),25-dihydroxyvitamin D3 and 25-hydroxyvitamin D3-26,23-lactone using a cyano-bonded phase packing.
A high-performance liquid chromatographic system is described for the baseline resolution of 25-hydroxyvitamin D3, 24(R),25-dihydroxyvitamin D3 and 25-hydroxyvitamin D3-26,23-lactone, the three principal circulating metabolites of vitamin D3 in the vitamin D-replete animal. The system is based upon a cyano-bonded phase packing and the solvent hexane--isopropanol--methanol (94:5:1). Of particular interest is the strong retention of carbonyl-containing vitamin D metabolites. The new system can be used for unequivocal analysis of vitamin D metabolites in plasma samples from clinical and animal studies and in the separation and identification of renal metabolites generated in vitro.
D015650:24,25-Dihydroxyvitamin D 3; D000818:Animals; D001667:Binding, Competitive; D002112:Calcifediol / Q000031:analogs & derivatives* / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004032:Diet; D004100:Dihydroxycholecalciferols / Q000302:isolation & purification*; D007668:Kidney / Q000378:metabolism; D008297:Male; D011485:Protein Binding; D051381:Rats; D011919:Rats, Inbred Strains
D004100:Dihydroxycholecalciferols; D015650:24,25-Dihydroxyvitamin D 3; C026396:calcifediol lactone; D002112:Calcifediol