Yadav Manish, Singhal Puran, Goswami Sailendra, Pande Umesh C, Sanyal Mallika, Shrivastav Pranav S
Bioanalytical Research Department, Veeda Clinical Research, Ambawadi, Ahmedabad 3800015, India.
J Chromatogr Sci. 2010 Oct;48(9):704-13. doi: 10.1093/chromsci/48.9.704.
A selective, sensitive, rugged, and high throughput liquid chromatography tandem mass spectrometry method is developed for the determination of one nucleotide tenofovir (TFV) and two nucleosides emtricitabine (FTC) and lamivudine (3TC) reverse transcriptase inhibitors in human plasma. Plasma samples were prepared by solid-phase extraction of the analytes and acyclovir (ACV) as internal standard using Waters Oasis MCX cartridges. The chromatographic separation is achieved in a run-time of 3.0 min on an ACE 5 CN column (150 mm × 4.6 mm, 5 μm) under isocratic conditions. The mobile phase consisted of 0.5% formic acid in water and acetonitrile (55:45, v/v). The protonated precursor --> product ion transitions for TFV, FTC, 3TC, and internal standard were monitored on a triple quadrupole mass spectrometer operating in the multiple reaction monitoring (MRM) and positive ion mode. A linear dynamic range of 4.0-802 ng/mL, 15.0-3006 ng/mL, and 20.1-4023 ng/mL is established for TFV, FTC, and 3TC, respectively, using 0.2 mL plasma sample. The method is fully validated for its sensitivity, selectivity, accuracy and precision, ion suppression, matrix effect, recovery, stability, and dilution integrity. It is successfully applied to a bioequivalence study of [300(TFV) + 200(FTC) + 300(3TC)] mg tablet formulation in 43 healthy human subjects under fasting conditions.
建立了一种选择性好、灵敏度高、耐用且高通量的液相色谱串联质谱法，用于测定人血浆中的一种核苷酸类药物替诺福韦（TFV）以及两种核苷类药物恩曲他滨（FTC）和拉米夫定（3TC）这三种逆转录酶抑制剂。血浆样品通过使用Waters Oasis MCX柱以阿昔洛韦（ACV）作为内标对分析物进行固相萃取来制备。在ACE 5 CN柱（150 mm×4.6 mm，5μm）上，于等度条件下3.0分钟内完成色谱分离。流动相由含0.5%甲酸的水和乙腈（55:45，v/v）组成。在以多反应监测（MRM）模式和正离子模式运行的三重四极杆质谱仪上监测TFV、FTC、3TC和内标的质子化前体→产物离子转换。使用0.2 mL血浆样品时，分别为TFV、FTC和3TC建立了4.0 - 802 ng/mL、15.0 - 3006 ng/mL和20.1 - 4023 ng/mL的线性动态范围。该方法在灵敏度、选择性、准确度和精密度、离子抑制、基质效应、回收率、稳定性以及稀释完整性方面均得到充分验证。它已成功应用于43名健康人类受试者在空腹条件下对[300（TFV）+ 200（FTC）+ 300（3TC）]mg片剂制剂的生物等效性研究。
