Institute of Clinical Chemistry, Hospital of the University of Munich, Marchioninistr. 15, D-81377 Munich, Germany.
J Steroid Biochem Mol Biol. 2010 Aug;121(3-5):565-73. doi: 10.1016/j.jsbmb.2010.02.025. Epub 2010 Mar 4.
Hypovitaminosis D is a highly prevalent condition and quantification of serum 25-hydroxyvitamin D3 is accepted to be the most useful marker for the assessment of the individual vitamin D status. Due to the increasing awareness of the prevalence and potential health consequences of hypovitaminosis D, the request numbers for 25-hydroxyvitamin D quantification are growing rapidly in many countries. Automated protein binding assays (based on the use of vitamin D-binding protein or antibodies) for the quantification of 25-hydroxyvitamin D3 are available which enable convenient high-throughput analyses in a routine setting; there is, however, substantial concern about accuracy and analytical reliability of these assays. Several LC-MS/MS methods for the quantification of 25-hydroxyvitamin D3 in serum have been described and in a growing number of clinical laboratories this technology is used routinely for vitamin D monitoring. It is justified to assume that LC-MS/MS enables more reliable analyses of 25-hydroxyvitamin D concentrations compared to protein binding assays. In particular the ability to co-quantify the naturally occurring 25-hydroxyvitamin D3 and 25-hydroxyvitamin D2 which is derived primarily from food fortification is a relevant advantage of LC-MS/MS over protein binding assays. This review describes the background of 25-hydroxyvitamin D measurement, compares published LC-MS/MS methods, discusses problems, strengths and limitations of these assays and compares the application characteristics of LC-MS/MS with those of protein binding assays and HPLC-UV.
维生素 D 缺乏症是一种高发疾病，血清 25-羟维生素 D3 的定量检测被认为是评估个体维生素 D 状态最有用的标志物。由于人们对维生素 D 缺乏症的流行程度及其潜在健康后果的认识不断提高，许多国家对 25-羟维生素 D 定量检测的需求数量正在迅速增长。目前已有基于维生素 D 结合蛋白或抗体的自动化蛋白结合测定法（protein binding assays）可用于 25-羟维生素 D3 的定量检测，这些方法能够在常规设置中进行方便的高通量分析；然而，人们对这些检测方法的准确性和分析可靠性存在很大的担忧。目前已经有几种用于血清 25-羟维生素 D3 定量检测的 LC-MS/MS 方法，并且在越来越多的临床实验室中，这项技术已被常规用于维生素 D 监测。可以合理地假设 LC-MS/MS 能够比蛋白结合测定法更可靠地分析 25-羟维生素 D 浓度。特别是能够同时定量检测天然存在的 25-羟维生素 D3 和主要来自食物强化的 25-羟维生素 D2，这是 LC-MS/MS 相对于蛋白结合测定法的一个相关优势。本文描述了 25-羟维生素 D 测量的背景，比较了已发表的 LC-MS/MS 方法，讨论了这些检测方法的问题、优势和局限性，并比较了 LC-MS/MS 与蛋白结合测定法和 HPLC-UV 的应用特点。
