Wiriyakosol Narit, Puangpetch Apichaya, Manosuthi Weerawut, Tomongkon Sasinapha, Sukasem Chonlaphat, Pinthong Darawan
Department of Pharmacology, Faculty of Science, Mahidol University, Bangkok, Thailand; National Doping Control Centre, Mahidol University, Bangkok, Thailand.
Division of Pharmacogenomics and Personalized Medicine, Department of Pathology, Faculty of Medicine, Ramatibodi Hospital, Mahidol University, Bangkok, Thailand.
J Chromatogr B Analyt Technol Biomed Life Sci. 2018 May 15;1085:89-95. doi: 10.1016/j.jchromb.2018.03.045. Epub 2018 Mar 29.
Tenofovir disoproxil fumarate is a pro-drug of the active metabolite tenofovir widely used against the HIV1, HIV2, and Hepatitis B virus. Several studies have been conducted and found kidney injury associated with tenofovir exposure. High tenofovir plasma concentration correlated with kidney injury in tenofovir-exposed patients. The present study developed and validated a simple and cost-effective LC/MS/MS method to determine tenofovir level in human plasma. A small plasma volume of 80 μl is utilized for the sample preparation. The samples were separated by Luna C18 (100 mm × 2.0 mm, 3 μm) using gradient elution with a mobile phase consisting of water (containing 0.1% formic acid) and acetonitrile (90:10, v/v). The detection was achieved through multiple reaction monitoring using positive ionization mode on the triple quadrupole mass spectrometer with a run time of 10 min. The monitoring transitions were set at m/z 288.0 → 176.1 and 136.1 for tenofovir and m/z 226.1 → 152.0 for acyclovir (as the internal standard). This standard curve was linear from 10 to 640 ng/ml, with the lower limit of quantification of 10 ng/ml. The inter- and intra-day precision results were less than 12.3% and their accuracies were within the acceptable range of 84.9-113.1%. The validated method was successfully applied to the study of tenofovir induced kidney injury in HIV-1 infected patients taking 300 mg once daily for more than 4 weeks.
富马酸替诺福韦二吡呋酯是活性代谢物替诺福韦的前体药物，广泛用于抗HIV-1、HIV-2和乙型肝炎病毒。已经进行了多项研究，发现替诺福韦暴露与肾损伤有关。在暴露于替诺福韦的患者中，高血浆替诺福韦浓度与肾损伤相关。本研究开发并验证了一种简单且经济高效的液相色谱/串联质谱法，用于测定人血浆中的替诺福韦水平。样品制备仅需80μl的少量血浆。使用Luna C18（100mm×2.0mm，3μm）色谱柱，以水（含0.1%甲酸）和乙腈（90:10，v/v）组成的流动相进行梯度洗脱分离样品。通过在三重四极杆质谱仪上采用正离子模式的多反应监测进行检测，运行时间为10分钟。替诺福韦的监测离子对设置为m/z 288.0→176.1和136.1，阿昔洛韦（作为内标）的监测离子对设置为m/z 226.1→152.0。该标准曲线在10至640ng/ml范围内呈线性，定量下限为10ng/ml。日间和日内精密度结果均小于12.3%，准确度在84.9 - 113.1%的可接受范围内。该验证方法成功应用于对每天服用300mg超过4周的HIV-1感染患者中替诺福韦诱导的肾损伤的研究。
