Studies on the analysis of 25-hydroxyvitamin D(3) by isotope-dilution liquid chromatography-tandem mass spectrometry using enzyme-assisted derivatisation.
Studies on the analysis of 25-hydroxyvitamin D(3) by isotope-dilution liquid chromatography-tandem mass spectrometry using enzyme-assisted derivatisation.
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DOI：
10.1016/j.bbrc.2014.01.088
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发表时间：
2014-04-11
影响因子：
3.1
通讯作者：
Griffiths, William J.
中科院分区：
文献类型：
--
作者：
Abdel-Khalik, Jonas;Crick, Peter J.;Carter, Graham D.;Makin, Hugh L.;Wang, Yugin;Griffiths, William J.
吉拉德P衍生化分析25-羟基维生素D3的新方法该方法也适用于维生素D3、1-α、25-和24，25-二羟基维生素D3。通过对方法的改进，还可以分析3-表-25-羟基维生素D3。25-羟基维生素D(25-羟基维生素D3和25-羟基维生素D2)的血清总浓度目前被用作维生素D状态的指标。维生素D缺乏被认为与多种疾病有关，因此需要准确和精确的参考方法来定量测定25-羟基维生素D。本文以25-[26，26，26，27，27，27-2H6]羟基维生素D3为内标，建立了一种分析成人血清中维生素D代谢物的酶辅助衍生化新方法。从血清中提取25-羟基维生素D是用乙腈进行的，这被证明比乙醇更有效。胆固醇氧化酶被用来氧化维生素D代谢物中的3-β-羟基，然后用Girard P试剂对新形成的3-oxo基团进行衍生化。17-β-羟基类固醇脱氢酶10选择性氧化25-羟基维生素D3的3-α-羟基同分异构体中的3-α-羟基。采用同位素-稀释液相色谱-串联质谱法进行定量。对成人血清进行了25-羟基维生素D3的回收实验，回收率为102-106%。此外，除25-羟基维生素D3外，在成人血清中还检测到24，25-二羟基维生素D3和其他未知的二羟基代谢物。
New method for the analysis of 25-hydroxyvitamin D3 exploiting Girard P derivatisation. Method also applicable to vitamin D3, 1α,25- and 24,25-dihydroxyvitamin D3. By modification of the method 3-epi-25-hydroxyvitamin D3 can also be analysed. The total serum concentration of 25-hydroxyvitamins D (25-hydroxyvitamin D3 and 25-hydroxyvitamin D2) is currently used as an indicator of vitamins D status. Vitamins D insufficiency is claimed to be associated with multiple diseases, thus accurate and precise reference methods for the quantification of 25-hydroxyvitamins D are needed. Here we present a novel enzyme-assisted derivatisation method for the analysis of vitamins D metabolites in adult serum utilising 25-[26,26,26,27,27,27-2H6]hydroxyvitamin D3 as the internal standard. Extraction of 25-hydroxyvitamins D from serum is performed with acetonitrile, which is shown to be more efficient than ethanol. Cholesterol oxidase is used to oxidize the 3β-hydroxy group in the vitamins D metabolites followed by derivatisation of the newly formed 3-oxo group with Girard P reagent. 17β-Hydroxysteroid dehydrogenase type 10 is shown to oxidize selectively the 3α-hydroxy group in the 3α-hydroxy epimer of 25-hydroxyvitamin D3. Quantification is achieved by isotope-dilution liquid chromatography–tandem mass spectrometry. Recovery experiments for 25-hydroxyvitamin D3 performed on adult human serum give recovery of 102–106%. Furthermore in addition to 25-hydroxyvitamin D3, 24,25-dihydroxyvitamin D3 and other uncharacterised dihydroxy metabolites, were detected in adult human serum.
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影响因子：
5.8
作者：
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通讯作者：
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影响因子：
4.1
作者：
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通讯作者：
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影响因子：
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作者：
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作者：
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DOI：
10.1016/j.jsbmb.2003.07.007
发表时间：
2003-11-01
影响因子：
4.1
作者：
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通讯作者：
Yang, SY