Hummer L, Riis B J, Christiansen C, Rickers H
Scand J Clin Lab Invest. 1985 Nov;45(7):611-9. doi: 10.3109/00365518509155268.
Reliable assays to determine the vitamin D metabolites are useful aids in the study of disorders involving vitamin D metabolism, and in the evaluation of the response in patients receiving vitamin D treatment. We report here the establishment in our laboratory of a method capable of measuring 25(OH)D, (including 25(OH)D2 and 25(OH)D3), 1,25(OH)2D and 24,25(OH)2D in a single blood sample. The method involves methanol/dichloromethane extraction and Sephadex LH-20 chromatography. The monhydroxylated fraction was purified on Lipidex 5000 and separated in 25(OH)D2 and 25(OH)D3 on high pressure liquid chromatography (HPLC), followed by ultra violet absorbance (UV) detection. The dihydroxylated fraction was separated by HPLC and quantified by protein-binding assays. The method is precise and accurate. The vitamin D metabolites were measured in different groups of patients and in normal subjects.
用于测定维生素D代谢产物的可靠分析方法，有助于研究涉及维生素D代谢的疾病，并有助于评估接受维生素D治疗患者的反应。我们在此报告，我们实验室建立了一种能够在一份血样中测定25(OH)D（包括25(OH)D2和25(OH)D3）、1,25(OH)2D和24,25(OH)2D的方法。该方法包括甲醇/二氯甲烷萃取和Sephadex LH - 20色谱法。单羟基化部分在Lipidex 5000上纯化，然后通过高压液相色谱（HPLC）分离为25(OH)D2和25(OH)D3，随后进行紫外吸收（UV）检测。双羟基化部分通过HPLC分离，并通过蛋白质结合分析进行定量。该方法精确且准确。我们在不同组的患者和正常受试者中测定了维生素D代谢产物。
