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Clinical Chemistry 27: 1757-1760, 1981;
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Clinical Chemistry, Vol 27, 1757-1760, Copyright © 1981 by American Association for Clinical Chemistry

Simultaneous determination of 25-hydroxyvitamin D, 24,25- dihydroxyvitamin D, and 1,25-dihydroxyvitamin D in plasma or serum

MJ Jongen, WJ van der Vijgh, HJ Willems, JC Netelenbos and P Lips

We describe a simultaneous assay for the principal vitamin D metabolites: 25-hydroxyvitamin D, 24-25-dihydroxyvitamin D, and 1,25- dihydroxyvitamin D. Special attention has been paid to simplification of the extensive extraction and purification procedures used in previously described simultaneous assays. All three metabolites were isolated with a single extraction step, followed by only one gradient liquid-chromatographic procedure. For final quantitation we used competitive protein binding assays, involving readily available binding proteins and commercially purchased tritiated vitamin D metabolites. Concentrations in the plasma of healthy subjects (mean age, 27 years), sampled during December were 51 (SD 17) nmol/L, 4.1 (SD 1.3) nmol/L, and 124 (SD 26) pmol/L for 25-hydroxyvitamin D, 24,25-dihydroxyvitamin D and 1,25-dihydroxyvitamin D, respectively. Intra- and interassay CVs for the three metabolites were 4.4 and 3.9%, 6.7 and 8.0%, and 7.0 and 4.8%, respectively.


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 Endocr. Rev.Home page
P. Lips
Vitamin D Deficiency and Secondary Hyperparathyroidism in the Elderly: Consequences for Bone Loss and Fractures and Therapeutic Implications
Endocr. Rev., August 1, 2001; 22(4): 477 - 501.
[Abstract] [Full Text] [PDF]


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Copyright © 1981 by the American Association for Clinical Chemistry.