Quantitation of circulating hydroxyvitamin D3 in human plasma by a continuous cleanup/concentration procedure prior to HPLC-UV detection.
A method for the determination of hydroxyvitamin D3 metabolites (25-hydroxyvitamin D3, 24,25-dihydroxyvitamin D3 and 1,25-didydroxyvitamin D3) based on a continuous cleanup/ preconcentration procedure coupled with HPLC and UV-detection is reported here. The method exhibits a linear range between 0.05 and 100 ng/ml (r2 = 0.9917) with CV values lower than 6.5%, and has been checked by applying it to plasma samples from a hospital with acceptable recoveries. The results compare well with those obtained by routine radioimmunoassay (y = 2.784+/-1.37 + 0.333+/-0.05 sigma(yx), r = 0.8233, n = 19 for 25-hydroxyvitamin D3). The sampling frequency was 4 h(-1); 12 analytes h(-1).
Department of Analytical Chemistry, Faculty of Sciences, University of Cordoba, Spain.
F
F
Ortiz-Boyer
J M
JM
Fernández-Romero
M D
MD
Luque de Castro
J M
JM
Quesada
D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation* / Q000379:methods*; D006801:Humans; D006887:Hydroxycholecalciferols / Q000097:blood*; D011863:Radioimmunoassay; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet
D006887:Hydroxycholecalciferols