Clemens T L, Adams J S, Nolan J M, Holick M F
Clin Chim Acta. 1982 Jun 3;121(3):301-8. doi: 10.1016/0009-8981(82)90239-x.
An assay for vitamin D consisting of high-pressure liquid chromatography (HPLC) and ultraviolet (UV) absorbance detection has been developed and used to measure circulating vitamin D concentrations in human subjects during summer and winter and after deliberate exposure to ultraviolet radiation. Extracts of 2-4 ml of serum were initially fractionated on silica Sep-Pak cartridges followed by reverse-phase HPLC and finally quantitated by UV-absorbance during straight-phase HPLC. Using these methods, we determined the normal range for circulating vitamin D in Boston subjects to be less than 0.5 ng/ml to 25 ng/ml (n = 30); subjects sampled during summer months had higher concentrations of vitamin D than those sampled during winter months. In subjects exposed to a single quantitative dose of ultraviolet radiation (UVR), large transient increases in circulating vitamin D3 were observed. Concentrations rose 30-50 fold over the first days after exposure before returning to basal levels by one week.
一种由高压液相色谱（HPLC）和紫外（UV）吸光度检测组成的维生素D检测方法已被开发出来，并用于测量人类受试者在夏季、冬季以及故意暴露于紫外线辐射后的循环维生素D浓度。最初将2 - 4毫升血清提取物在硅胶Sep - Pak柱上进行分离，然后进行反相HPLC，最后在正相HPLC期间通过UV吸光度进行定量。使用这些方法，我们确定波士顿受试者循环维生素D的正常范围为低于0.5纳克/毫升至25纳克/毫升（n = 30）；夏季采样的受试者维生素D浓度高于冬季采样的受试者。在暴露于单一定量剂量紫外线辐射（UVR）的受试者中，观察到循环维生素D3有大幅短暂升高。暴露后的头几天浓度升高了30 - 50倍，一周后恢复到基础水平。
