Simplified method for the determination of 25-hydroxy and 1alpha,25-dihydroxy metabolites of vitamins D2 and D3 in human plasma. Application to nutritional studies.
A simplified method for the determination of 25-hydroxy and 1alpha,25-dihydroxy metabolites of vitamins D2 and D3 in human plasma was developed. Plasma samples were deproteinizated and applied to a Bond Elut C18OH cartridge to separate 25-hydroxyvitamin D (25-OH-D) and 1alpha,25-dihydroxyvitamin D [1,25(OH)2D] fractions. The 25-OH-D fraction was purified by a Bond Elut C18 cartridge and 25-OH-D2 and 25-OH-D3 were assayed by HPLC using a Zorbax SIL column. The 1,25(OH)2D fraction obtained above was subsequently applied to HPLC using a Zorbax SIL column to separate 1,25(OH)2D2 and 1,25(OH)2D3 fractions which were determined by a radioreceptor assay (RRA) using calf thymus receptor. The method was applied to nutritional studies.
Takeuchi
A
A
Department of Hygienic Sciences, Kobe Pharmaceutical University, Higashinada-ku, Japan.
Ishida
Y
Y
Sekimoto
H
H
Masuda
S
S
Okano
T
T
Nishiyama
S
S
Matsuda
I
I
Kobayashi
T
T
D015652:25-Hydroxyvitamin D 2 / Q000097:blood*; D000328:Adult; D000368:Aged; D002112:Calcifediol / Q000097:blood*; D002117:Calcitriol / Q000097:blood*; D002851:Chromatography, High Pressure Liquid; D004872:Ergocalciferols / Q000097:blood* / Q000172:deficiency / Q000378:metabolism / Q000627:therapeutic use; D005527:Food, Fortified; D006801:Humans; D006887:Hydroxycholecalciferols / Q000627:therapeutic use; D007223:Infant; D008875:Middle Aged; D014983:Xeroderma Pigmentosum / Q000097:blood / Q000188:drug therapy
D004872:Ergocalciferols; D006887:Hydroxycholecalciferols; D015652:25-Hydroxyvitamin D 2; C003564:1,25-dihydroxyergocalciferol; D002117:Calcitriol; D002112:Calcifediol; C008088:alfacalcidol