High throughput LC-MS/MS method for the simultaneous analysis of multiple vitamin D analytes in serum.
High throughput LC-MS/MS method for the simultaneous analysis of multiple vitamin D analytes in serum.
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DOI：
10.1016/j.jchromb.2016.01.049
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发表时间：
2016-03-01
期刊：
影响因子：
--
通讯作者：
Keevil BG
中科院分区：
文献类型：
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作者：
Jenkinson C;Taylor AE;Hassan-Smith ZK;Adams JS;Stewart PM;Hewison M;Keevil BG
最近的研究表明，维生素D缺乏与常见的人类健康问题的风险增加有关。为了确定维生素D的“状态”，大多数常规分析方法定量一种特定的维生素D代谢物，25-羟基维生素D3（25 OHD 3）。然而，维生素D的特点是复杂的代谢途径，同时测量多种维生素D代谢物可能会提供更准确的维生素D状态的解释。为了解决这个问题，我们开发了一种高通量液相色谱-串联质谱（LC-MS/MS）方法来分析多种维生素D分析物，特别强调差向异构体代谢物的分离。建立了一种支持性液-液萃取（SLE）和LC-MS/MS方法，用于定量10种维生素D代谢物以及分离干扰7α-羟基-4-异戊烯-3-酮（7αC4）同量异位素（胆汁酸前体），并通过分析人血清样品进行了验证。在116名健康受试者的队列中，25-羟基维生素D3（25 OHD 3）、3-表-25-羟基维生素D3（3-epi-25-hydroxyvitamin D3，25 OHD 3）和25-羟基维生素D3（25-hydroxyvitamin D3，（3-epi-25 OHD 3），24，25-二羟维生素D3（24 R，25（OH）2D 3），1，25-二羟维生素D3（1α，25（OH）2D 3）和25-羟基维生素D2（25 OHD 2）可使用220 μl血清定量，25 OHD 3和24 R，25（OH）2D 3显示显著的季节性变化。这种高通量LC-MS/MS方法为评估维生素D对人类健康和疾病的影响提供了一种新的策略。
Recent studies suggest that vitamin D-deficiency is linked to increased risk of common human health problems. To define vitamin D ‘status’ most routine analytical methods quantify one particular vitamin D metabolite, 25-hydroxyvitamin D3 (25OHD3). However, vitamin D is characterized by complex metabolic pathways, and simultaneous measurement of multiple vitamin D metabolites may provide a more accurate interpretation of vitamin D status. To address this we developed a high-throughput liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to analyse multiple vitamin D analytes, with particular emphasis on the separation of epimer metabolites. A supportive liquid-liquid extraction (SLE) and LC-MS/MS method was developed to quantify 10 vitamin D metabolites as well as separation of an interfering 7α-hydroxy-4-cholesten-3-one (7αC4) isobar (precursor of bile acid), and validated by analysis of human serum samples. In a cohort of 116 healthy subjects, circulating concentrations of 25-hydroxyvitamin D3 (25OHD3), 3-epi-25-hydroxyvitamin D3 (3-epi-25OHD3), 24,25-dihydroxyvitamin D3 (24R,25(OH)2D3), 1,25-dihydroxyvitamin D3 (1α,25(OH)2D3), and 25-hydroxyvitamin D2 (25OHD2) were quantifiable using 220 μl of serum, with 25OHD3 and 24R,25(OH)2D3 showing significant seasonal variations. This high-throughput LC-MS/MS method provides a novel strategy for assessing the impact of vitamin D on human health and disease.