Determination of 25-Hydroxyvitamin D in Human Plasma Using High-Performance Liquid Chromatography-Tandem Mass Spectrometry

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• 作者：Naoko Tsugawa ; Yoshitomo Suhara ; Maya Kamao ; and Toshio Okano
• 刊名：Analytical Chemistry
• 出版年：2005
• 出版时间：May 1, 2005
• 年：2005
• 卷：77
• 期：9
• 页码：3001 - 3007
• 全文大小：165K
• 年卷期：v.77,no.9(May 1, 2005)
• ISSN：1520-6882

文摘

We report here the development of a precise and sensitivemethod to determine 25-hydroxyvitamin D (25-OH-D₂/-D₃) in human plasma using high-perfomance liquidchromatography-tandem mass-mass spectrometry withatomospheric pressure chemical ionization (LC-APCI-MS/MS). The method involves the use of deuterated 25-OH-D₃ as an internal standard compound for 25-OH-D₂/-D₃,which was synthesized in our laboratory, and the selectionof a precursor and product ion with a MS/MS multiplereaction monitoring method. The average intraassay andinterassay variation values (relative standard deviation)were 5.7 and 2.5%, respectively, for 25-OH-D₃ and 4.5and 5.1%, respectively, for 25-OH-D₂. The average spikedrecoveries from authentic compounds added to normalhuman plasma samples for 25-OH-D₃ and 25-OH-D₂were 103.8 and 98.8%, respectively. Mean plasma concentrations of 25-OH-D₃ and 25-OH-D₂ in healthy subjects were 20.5 and 0.4 ng/mL, respectively. We concludethat this novel LC-APCI-MS/MS method would be usefulfor the evaluation of the vitamin D status in postmenopausal women and elderly subjects and provide usefulinformation in the diagnosis of vitamin D insufficiency/deficiency, as well as for the treatment and prevention ofosteoporosis with vitamin D.