We report here the development of a precise and sensitive method to determine 25-hydroxyvitamin D (25-OH-D2/ -D3) in human plasma using high-perfomance liquid chromatography−tandem mass−mass spectrometry with atomospheric 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 selection of a precursor and product ion with a MS/MS multiple reaction monitoring method. The average intraassay and interassay variation values (relative standard deviation) were 5.7 and 2.5%, respectively, for 25-OH-D3 and 4.5 and 5.1%, respectively, for 25-OH-D2. The average spiked recoveries from authentic compounds added to normal human plasma samples for 25-OH-D3 and 25-OH-D2 were 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 conclude that this novel LC-APCI-MS/MS method would be useful for the evaluation of the vitamin D status in postmenopausal women and elderly subjects and provide useful information in the diagnosis of vitamin D insufficiency/deficiency, as well as for the treatment and prevention of osteoporosis with vitamin D.
我们在此报告了一种精确灵敏的方法,该方法使用高效
液相色谱-串联质谱-质谱法,结合常压
化学电离(LC-APCI-MS/MS),测定人血浆中的25-羟基
维生素D(25-OH-D2/-D3)。该方法使用
氘代25-OH-D3作为25-OH-D2/-D3的内部标准化合物,该化合物由我们的实验室合成,并使用MS/MS多重反应监测法选择母离子和子离子。25-OH-D3的平均内标和间标变异值(相对标准偏差)分别为5.7%和2.5%,25-OH-D2的平均内标和间标变异值分别为4.5%和5.1%。在正常人血浆样品中添加真实化合物后,25-OH-D3和25-OH-D2的平均加标回收率分别为103.8%和98.8%。健康受试者的25-OH-D3和25-OH-D2平均血浆浓度分别为20.5和0.4 ng/mL。我们的结论是,这种新型LC-APCI-MS/MS方法可用于评估绝经后妇女和老年受试者的
维生素D状态,为诊断
维生素D不足/缺乏症提供有用的信息,以及用于治疗