Peris-Vicente Juan, Villarreal-Traver Mónica, Casas-Breva Inmaculada, Carda-Broch Samuel, Esteve-Romero Josep
Dep. de Química Física i Analítica, ESTCE, Universitat Jaume I, Castelló, Spain.
J Sep Sci. 2014 Oct;37(20):2825-32. doi: 10.1002/jssc.201400574. Epub 2014 Sep 25.
Danuravir, ritonavir, emtricitabine, and tenofovir are together prescribed against AIDS as a highly active antiretroviral therapy regimen. Micellar liquid chromatography has been applied to determine these four antiretroviral drugs in plasma. The sample preparation is shortened to the dilution of the sample in a micellar solution, filtration, and injection. Clean-up steps are avoided, due to the solubilization of plasma matrix in micellar media. The drugs were analyzed in <20 min using a mobile phase of 0.06 M sodium dodecyl sulfate/2.5% 1-pentanol (pH 7) running under isocratic mode through a C18 column at 1 mL/min at room temperature. Absorbance wavelength detection was set at 214 nm. The method was successfully validated following the ICH Harmonized Tripartite Guideline in terms of selectivity, limit of detection (0.080-0.110 μg/mL), limit of quantification (0.240-0.270 μg/mL), linearity between 0.25 and 25 μg/mL (r(2) > 0.995), accuracy (89.3-103.2%), precision (<8.2%) and robustness (<7.5%). Real plasma sample from patients taking this therapy were analyzed. This is the first paper showing the simultaneous detection of this four drugs. Therefore, the methodology was proven useful for the routine analysis of these samples in a hospital laboratory for clinical purposes.
达努那韦、利托那韦、恩曲他滨和替诺福韦联合使用作为一种高效抗逆转录病毒治疗方案用于治疗艾滋病。胶束液相色谱法已被应用于测定血浆中的这四种抗逆转录病毒药物。样品制备被简化为将样品在胶束溶液中稀释、过滤和进样。由于血浆基质在胶束介质中的增溶作用，避免了净化步骤。在室温下，使用0.06 M十二烷基硫酸钠/2.5% 1-戊醇（pH 7）的流动相，以1 mL/min的流速通过C18柱在等度模式下运行，在<20分钟内对药物进行分析。吸光度波长检测设置在214 nm。该方法按照国际人用药品注册技术协调会（ICH）三方协调指南在选择性、检测限（0.080 - 0.110 μg/mL）、定量限（0.240 - 0.270 μg/mL）、0.25至25 μg/mL之间的线性关系（r(2)>0.995）、准确度（89.3 - 103.2%）、精密度（<8.2%）和稳健性（<7.5%）方面成功得到验证。对接受该治疗的患者的实际血浆样本进行了分析。这是第一篇展示同时检测这四种药物的论文。因此，该方法被证明对于医院实验室出于临床目的对这些样本进行常规分析是有用的。
