Department of Pharmaceutical Sciences, College of Pharmacy 3039, University of Nebraska Medical Center, Omaha, NE 68198-6025, United States.
J Chromatogr B Analyt Technol Biomed Life Sci. 2011 Aug 1;879(23):2332-8. doi: 10.1016/j.jchromb.2011.06.032. Epub 2011 Jul 1.
Animal pharmacokinetic and tissue distribution assays of antiretroviral therapeutic drugs require accurate drug quantification in biological fluids and tissues. Here we report a simple, rapid, and sensitive ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for quantification of commonly used antiretroviral drugs ritonavir (RTV), indinavir (IDV), atazanavir (ATV), and efavirenz (EFV) in mouse serum and tissues (liver, kidney, lung, and spleen). These antiretroviral drugs are currently the cornerstones of common therapeutic regimens for human immunodeficiency virus (HIV) infection. Chromatographic separation was achieved using a gradient mobile phase (5% acetonitrile in methanol and 7.5mM ammonium acetate (pH 4.0)) on an ACQUITY UPLC(®)BEH Shield RP 18 column. All compounds eluted within a 7 min run time. Lopinavir was used as an internal standard. Detection was achieved by dual positive and negative ionization modes on a quadrupole linear ion trap hybrid mass spectrometer with an electrospray ionization (ESI) source. The dynamic range was 0.2-1000 ng/mL for RTV, IDV, and ATV, and 0.5-1000 for EFV. The method was validated and showed high and consistent intra-day and inter-day accuracy and precision for all analytes. This method is used to support the preclinical development studies of targeted- and sustained-release combination ART (nanoART). The current data demonstrate a 1.5-4 fold increase in serum and tissue AUC of nanoformulated ATV, RTV, and EFV administered to mice when compared to native drug. In addition, the tested formulation enhanced exposure of the same anti-HIV drugs in mouse tissues.
抗逆转录病毒治疗药物的动物药代动力学和组织分布研究需要准确测定生物体液和组织中的药物浓度。本研究建立了一种简单、快速、灵敏的超高效液相色谱-串联质谱（UPLC-MS/MS）法，用于定量检测鼠血清和组织（肝、肾、肺和脾）中的常用抗逆转录病毒药物利托那韦（RTV）、茚地那韦（IDV）、阿扎那韦（ATV）和依非韦伦（EFV）。这些抗逆转录病毒药物是目前人类免疫缺陷病毒（HIV）感染常用治疗方案的基石。采用梯度洗脱（5%乙腈-甲醇和 7.5mM 乙酸铵（pH 4.0）），在 ACQUITY UPLC(®)BEH Shield RP 18 柱上实现了色谱分离。所有化合物在 7 分钟内洗脱。洛匹那韦作为内标。采用电喷雾离子源（ESI）的四极杆线性离子阱混合质谱仪，以正负离子同时检测的方式进行检测。RTV、IDV 和 ATV 的线性范围为 0.2-1000ng/mL，EFV 的线性范围为 0.5-1000ng/mL。该方法经过验证，具有高的和一致的日内和日间准确性和精密度。该方法用于支持靶向和持续释放组合抗逆转录病毒疗法（nanoART）的临床前开发研究。目前的数据表明，与原型药物相比，当给予小鼠时，纳米制剂 ATV、RTV 和 EFV 的血清和组织 AUC 增加 1.5-4 倍。此外，该制剂还增加了相同抗 HIV 药物在小鼠组织中的暴露。
