Clinical Chemistry
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Clinical Chemistry 23: 855-862, 1977;
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Clinical Chemistry, Vol 23, 855-862, Copyright © 1977 by American Association for Clinical Chemistry

Sequential-saturation-type assay for serum 25-hydroxyvitamin D

G Ellis and K Dixon

We describe a method for measuring 25-hydroxyvitamin D in serum. The serum is denatured with ethanol, and the extract purified by chromatography on silicic acid to prevent potential interference from vitamin D and by other substances. 25-Hydoxyvitamin D is assayed by sequential saturation analysis in Tris-HCI buffer, pH 8.6, containing the surfactant Triton X-405 (2 ml/liter) to solubilize the steroid. Diluter, unpurifed normal human serum is used as the binder. The method is sensitive and precise. The dose-response curve is linear. The assay normally gives high count rates (12 000 to 20 000 cpm for the zero standard) and consequently 25-hydroxyvitamin D of low specific activity can be used if necessary. Serum 25-hydroxyvitamin D was determined in normal and osteomalacic subjects.


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Copyright © 1977 by the American Association for Clinical Chemistry.