Prathipati Pavan Kumar, Mandal Subhra, Destache Christopher J
Pharmacy Practice Department, Creighton University School of Pharmacy and Health Professions, Omaha, Nebraska, USA.
Biomed Chromatogr. 2018 Apr 26:e4270. doi: 10.1002/bmc.4270.
A simple, short, and rugged LC-MS/MS method for the simultaneous determination of tenofovir, emtricitabine, elvitegravir and rilpivirine was developed and validated. Dried blood spots were prepared with 25 μL of spiked whole blood. A 3 mm punch was extracted with methanol containing labeled internal standards. Ten microliters was injected into the LC-MS/MS using isocratic mobile phase composed of 0.1% formic acid in water and 0.1% formic acid in acetonitrile (45: 55 v/v) at a flow rate of 0.25 mL/min. The method was validated in the range of 10-2000 ng/mL for all four analytes. The intra-assay accuracy (RE) of the method was -4.73-4.78, 1.35-2.89, -8.89 to -0.49 and - 1.40-1.81 for tenofovir, emtricitabine, elvitegravir and rilpivirine, respectively. The inter-assay accuracy was within ±15% of nominal and precision (CV) was <15%. The hematocrit effect on quantification was nonsignificant at the tested hematocrit levels (35-70%). The dried blood spot method showed good agreement with the plasma method, and hence can be used as an alternative to plasma method.
开发并验证了一种简单、快速且耐用的LC-MS/MS方法，用于同时测定替诺福韦、恩曲他滨、埃替拉韦和利匹韦林。用25μL加标的全血制备干血斑。用含有标记内标的甲醇提取3mm的打孔样品。取10μL注入LC-MS/MS，采用由0.1%甲酸水溶液和0.1%甲酸乙腈溶液（45:55 v/v）组成的等度流动相，流速为0.25mL/min。该方法对所有四种分析物的验证范围为10-2000 ng/mL。该方法对替诺福韦、恩曲他滨、埃替拉韦和利匹韦林的批内准确度（RE）分别为-4.73-4.78、1.35-2.89、-8.89至-0.49和-1.40-1.81。批间准确度在标称值的±15%以内，精密度（CV）<15%。在所测试的血细胞比容水平（35-70%）下，血细胞比容对定量的影响不显著。干血斑法与血浆法显示出良好的一致性，因此可作为血浆法的替代方法。
