Ishige Takayuki, Satoh Mamoru, Ogawa Shoujiro, Nishimura Motoi, Higashi Tatsuya, Nomura Fumio
Rinsho Byori. 2015 Apr;63(4):457-64.
Vitamin D is pro-hormone that has important roles in calcium metabolism in the intestine, kidneys, and bone. Many studies have indicated that vitamin D deficiency causes not only rickets and osteomalacia, but also increases the risk of various diseases such as cancer and autoimmune disease. Of the many vitamin D metabolites, 25-hydroxyvitamin D[25OH-D], 1α,25-dihydroxyvitamin D[1,25(OH)2-D], and 24,25-dihydroxyvitamin D [24,25 (OH) 2-D] are important for assessing vitamin D metabolism. They are circulating, biologically active, and major inactive forms of vitamin D metabolite, respectively. Immunoassays are widely used for the measurement of serum/plasma vitamin D metabolites. With such an assay, however, the accurate quantification of these metabolites is difficult because of cross-reactivity. Liquid chromatography-tandem mass spectrometry [LC-MS/MS] is the gold standard method for analyzing these metabolites due to its high sensitivity and selectivity. Derivatization with a Cookson-type reagent is a key technique for robust and sensitive analysis by LC-MS/MS. A Cookson-type reagent rapidly and quantitatively reacts with the s-cis-diene structure of vitamin D metabolites, and markedly enhances the ionization efficiency. Using a recently developed Cookson-type reagent (DAPTAD), we successfully established an LC-MS/MS-based method for analyzing serum vitamin D metabolites including 250H-D, 1,25 (OH) 2-D, and 24,25 (OH) 2-D. Thus, LC-MS/MS can be a powerful tool for the accurate determination of vitamin D metabolism. In this review, we describe the advantages of LC-MS/MS for the determination of vitamin D metabolites.
维生素D是一种前体激素，在肠道、肾脏和骨骼的钙代谢中发挥着重要作用。许多研究表明，维生素D缺乏不仅会导致佝偻病和骨软化症，还会增加患癌症和自身免疫性疾病等各种疾病的风险。在众多维生素D代谢产物中，25-羟基维生素D[25OH-D]、1α,25-二羟基维生素D[1,25(OH)2-D]和24,25-二羟基维生素D[24,25(OH)2-D]对于评估维生素D代谢至关重要。它们分别是维生素D代谢产物的循环形式、生物活性形式和主要无活性形式。免疫测定法广泛用于血清/血浆维生素D代谢产物的测量。然而，使用这种测定法时，由于交叉反应性，难以对这些代谢产物进行准确的定量。液相色谱-串联质谱法[LC-MS/MS]因其高灵敏度和选择性，是分析这些代谢产物的金标准方法。用库克森型试剂衍生化是通过LC-MS/MS进行稳健且灵敏分析的关键技术。库克森型试剂能与维生素D代谢产物的s-顺式二烯结构快速定量反应，并显著提高电离效率。使用最近开发的库克森型试剂（DAPTAD），我们成功建立了一种基于LC-MS/MS的方法来分析血清维生素D代谢产物，包括250H-D、1,25(OH)2-D和24,25(OH)2-D。因此，LC-MS/MS可以成为准确测定维生素D代谢的有力工具。在这篇综述中，我们描述了LC-MS/MS在测定维生素D代谢产物方面的优势。
