Choi S O, Rezk N, Kim J S, Kashuba A D M
Korea Food and Drug Administration, Department of Pharmacology.
J Chromatogr Sci. 2010 Mar;48(3):219-23. doi: 10.1093/chromsci/48.3.219.
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 (150 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.
开发并验证了一种灵敏、准确且精密的液相色谱-质谱分析法，用于测定人阴道组织中的替诺福韦（TNF）。组织匀浆后，采用Varian Bond Elut-C(18)柱进行固相萃取以净化样品。使用Varian Polaris 3C(18)-A反相分析柱（150 mm×2 mm）实现了TNF与内标（甲苯磺丁脲）的色谱分离。采用在水中含0.1%甲酸和在乙腈中含0.1%甲酸的梯度方法。分别使用288.05和271.00 m/z通过电喷雾电离质谱在正离子模式下检测TNF和甲苯磺丁脲。在1 - 1000 ng/mL之间获得了线性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分别在89.7%至109.4%以及97.3%至104.9%之间，精密度分别在1.3%至10.9%以及2.6%至9.0%之间。这是首个开发用于定量人体组织中TNF的经过验证的方法。
