Liquid chromatography-tandem mass spectrometric method for the determination of salivary 25-hydroxyvitamin D3:: a noninvasive tool for the assessment of vitamin D status
Liquid chromatography-tandem mass spectrometric method for the determination of salivary 25-hydroxyvitamin D3:: a noninvasive tool for the assessment of vitamin D status
复制标题
DOI：
10.1007/s00216-007-1780-3
复制
发表时间：
2008-05-01
影响因子：
4.3
通讯作者：
Shimada, Kazutake
中科院分区：
文献类型：
--
作者：
Higashi, Tatsuya;Shibayama, Yujin;Shimada, Kazutake
建立了人唾液中25-羟基维生素D-3[25(OH)D-3]的高效液相色谱-电喷雾电离-串联质谱仪(LC-ESI-MS/MS)测定方法。唾液用乙腈去蛋白，用Strata-X色谱柱纯化，用库克森型试剂4-苯基-1，2，4-三唑啉-3，5-二酮(PTAD)衍生化，然后进行LC-MS/MS。与完整的25(OH)D-3相比，PTAD衍生物更容易在正ESI-MS中电离，并在MS/MS过程中有效地产生特征产物离子。甲胺作为流动相添加剂，也有效地提高了测定的灵敏度。定量采用选择性反应监测，以25-羟基维生素D-4为内标。该方法可以用1.0ml的样品重复性好、准确地定量唾液中的25(OH)D-3，其定量限为2.0pg/ml，并考察了该方法在临床研究中的适用性。血清25(OH)D-3水平与唾液25(OH)D-3水平呈直线正相关(r(2)=0.830)。该方法还能够检测到补充维生素D-3后唾液25(OH)D-3水平的增加。
A sensitive liquid chromatography-electrospray ionization-tandem mass spectrometric (LC-ESI-MS/MS) method for the determination of 25-hydroxyvitamin D-3 [25(OH)D-3] in human saliva has been developed and validated. The saliva was deproteinized with acetonitrile, purified using a Strata-X cartridge, derivatized with a Cookson-type reagent, 4-phenyl-1,2,4-triazoline-3,5-dione (PTAD), and subjected to LC-MS/MS. The PTAD derivative was much more easily ionized in positive-ESI-MS and efficiently produced a characteristic product ion during MS/MS, compared to the intact 25(OH)D-3. Methylamine was used as the mobile phase additive, and also effectively enhanced the assay sensitivity. Quantification was based on selected reaction monitoring, and 25-hydroxyvitamin D-4 was used as the internal standard. This method allowed the reproducible and accurate quantification of salivary 25(OH)D-3 using a 1.0-ml sample, and the limit of quantitation for 25(OH)D-3 was 2.0 pg/ml. The applicability of the developed method for clinical studies was then examined. There was a positive linear relationship (r(2) = 0.830) between the serum 25(OH)D-3 level, which is conventionally used as a means of assessing the vitamin D status, and the salivary 25(OH)D-3 level measured using the proposed method. The method also enabled the detection of the increase in the salivary 25(OH)D-3 level after the supplementation of vitamin D-3.