超高效液相色谱-串联质谱法测定干血点中25-羟基维生素D

发布时间：2018-12-28 13:26

【摘要】：目的建立干血点中25-羟基维生素D2[25(OH)D2]和D3[25(OH)D3]的超高效液相色谱-串联质谱(UHPLC-MS/MS)同时测定方法。方法干血点经过定量取样,加水超声提取后加乙腈沉淀蛋白,再使用乙酸乙酯:正己烷(2:1,V/V)萃取,氮气吹干后加PTAD试剂衍生,氮吹干后用流动相复溶,使用Waters BEH C18柱分离、电喷雾正离子(ESI+)模式和多反应监测(MRM)检测,同位素内标法定量。结果 25(OH)D2和25(OH)D3分别在全血浓度为5~167ng/m L和1.7~167ng/m L的范围内呈良好的线性关系,相关系数r0.999;检出限(LOD)分别为1.5ng/m L和0.25ng/m L,定量限(LOQ)分别为5ng/m L和1.7ng/m L;样品高低两水平的加标回收率均80%,日内精密度均9%,日间精密度均10%。结论该方法需血量少、灵敏、快速、准确,适用于人群维生素D营养状况的监测。
[Abstract]:Objective to establish a method for simultaneous determination of 25-hydroxyvitamin D _ 2 [25 (OH) D _ 2] and D _ 3 [25 (OH) D _ 3] in dry blood spots by ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Methods after quantitative sampling, ultrasonic extraction with water, acetonitrile precipitated protein was added. Ethyl acetate: n-hexane (2: 1 V / V) was extracted, nitrogen was dried and PTAD reagent was added. After nitrogen was dried, mobile phase was used to dissolve the protein. Waters BEH C18 column separation, electrospray positive ion (ESI) model and multi-reaction monitoring (MRM) were used. Results there was a good linear relationship between 25 (OH) D2 and 25 (OH) D 3 within the range of 5~167ng/m L and 1.7~167ng/m L, respectively. The correlation coefficient was r 0.999. The detection limit (LOD) was 1.5ng/m L and 0.25ng/m L, and the quantitative limit (LOQ) was 5ng/m L and 1.7ng/m L. The recoveries of the samples at both levels were 80, 9 and 10, respectively. Conclusion this method needs less blood volume, is sensitive, rapid and accurate, and is suitable for the monitoring of vitamin D nutritional status.
【作者单位】：上海市普陀区疾病预防控制中心;上海市疾病预防控制中心;
【分类号】：R151

【相似文献】