Development of an LC-MS Method for Measuring TNF in Human Vaginal Tissue
Development of an LC-MS Method for Measuring TNF in Human Vaginal Tissue
复制标题
DOI：
10.1093/chromsci/48.3.219
复制
发表时间：
2010-03-01
影响因子：
1.3
通讯作者：
Kashuba, A. D. M.
中科院分区：
文献类型：
--
作者：
Choi, S. O.;Rezk, N.;Kashuba, A. D. M.
建立了一种灵敏、准确、精密的液相色谱-质谱联用测定人阴道组织中替诺福韦（TNF）的方法。组织均质化后，采用Varian Bond Elut-C-18柱固相萃取进行样品净化。采用Varian Polaris 3C（18）-A反相分析柱（1150 mm x 2 mm）实现TNF和内标物（甲苯磺丁脲）的色谱分离。使用0.1%甲酸水溶液和0.1%甲酸乙腈溶液的梯度方法。TNF和甲苯磺丁脲的检测通过电喷雾电离质谱法在正离子模式下分别使用288.05和271.00 m/z实现。在1- 1，000 ng/mL之间获得线性TNF校准曲线，相关系数（r（2））大于0.999。TNF的日内和日间准确度范围分别为89.7%和109.4%以及97.3%和104.9%，精密度范围分别为1.3%和10.9%以及2.6%和9.0%。这是第一个经验证的方法，开发定量人体组织中的TNF。
A sensitive, accurate, and precise liquid chromatography-mass spectrometry assay for the determination of tenofovir (TNF) in human vaginal tissue was developed and validated. After homogenization of the tissue, solid-phase extraction on Varian Bond Elut-C-18 column was used for sample clean up. Chromatographic separation of TNF and the internal standard (tolbutamide) was achieved with a Varian Polaris 3C(18)-A reversed-phase analytical column (1150 mm x 2 mm). A gradient method using 0.1% formic acid in water and 0.1% formic acid in acetonitrile was employed. Detection of TNF and tolbutamide was achieved by electrospray ionization mass spectrometry in the positive ion mode using 288.05 and 271.00 m/z, respectively. Linear TNF calibration curves were obtained between 1-1,000 ng/mL with a correlation coefficient (r(2)) greater than 0.999. Intra- and inter-day accuracy for TNF ranged from 89.7% and 109.4% and from 97.3% and 104.9%, and precision ranged from 1.3% and 10.9% and 2.6% and 9.0%, respectively. This is the first validated method developed to quantitate TNF in human tissues.