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Quantitation of circulating hydroxyvitamin D3 in human plasma by a continuous cleanup/concentration procedure prior
F Ortiz-Boyer1, J M Fernández-Romero, M D Luque de Castro
1Department of Analytical Chemistry, Faculty of Sciences, University of Cordoba, Spain.
Abstract:
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).