Measurement of 25-hydroxyvitamin D3 and 25-hydroxyvitamin D2 in clinical settings

A quick and simple method for the selective measurement of 25-hydroxyvitamin D3 (25OHD3) and 25-hydroxyvitamin D2 (25OHD2) is described. It includes a rapid sample preparation technique and a combination of a selective radioimmunoassay for 25OHD3 and a competitive protein-binding assay using vitamin D-binding protein for the determination of total 25OHD, including 25OHD3 and 25OHD2. The method was compared with a procedure which include methanol/methylene chloride extraction and chromatography on Sephadex LH 20, and a procedure which includes HPLC and final quantification by u. v. detection. The methods were applied to three groups of patients in order to obtain information on how far assay procedures could be simplified for use in the clinical settings. It is concluded that the method described is applicable for following patients on vitamin D2 therapy. When groups of patients have to be compared, the mean values of the estimates are comparable, whether a simple method or a laborious method is used. Hence, the selection of assay method should take into account the clinical problem and the cost of the analysis.