伏立康唑血药浓度的测定与方法学考察

目的：建立人血浆中伏立康唑HPLC测定法,为人体药动学和药效学研究提供方法学基础。方法：取血浆0.5 mL,以劳拉西泮为内标,用乙腈-水（1∶9）2 mL以及乙腈洗脱,收集1 mL洗脱液,进样为20μL;色谱柱为Semmetry C18柱（4.6 mm×250 mm,5μm）,流动相为乙腈-水（50∶50）,流速为0.8 mL/min,在激发波长为256 nm和发射波长为372 nm处进行检测。结果：伏立康唑浓度在0.208~20.8μg/mL范围内与峰面积呈良好的线性关系（Y=0.2517ρ-0.0144,r2=0.999 8）;高、中、低3种浓度质控样品的日内和日间RSD均小于5%,相对回收率均在98%~103%范围内,将其室温放置24 h以及在-20℃反复冻融3次后,检测其回收率在83%~102%。结论：该方法操作简便、快速,准确灵敏,适用于临床药动学研究。

Determination of Voriconazole in Human Plasma by HPLC and Methodology Validation

Objective： To develop a high-perfbrmance liquid chromatographic（HPLC） assay for the determination of voriconazole in human plasma. Methods： 0.5 mL of human plasma was extracted by 2 mL elution solvent （acetonitrile： water, v：v, 1：9） with lorazepam as the internal standard. 1 mL liquid was collected, of which 20 pL was analyzed through a Semmetry-C18 column （4.6 mm ）〈250 mm,5 μm）. The mobile phase consisted of acetonitrile-water（50：50）, with a flow rate of 0.8 mL/min. And sample was determined by a fluorescence detector set at an excitation wavelength of 256 nm and emission wavelength of 372. Results： A linearity of voriconazole concentration curve was obtained between 0.208 and 20.8 ng/mL（Y=0.251 7ρ-0.014 4, r2=0.999 8）. For the low, middle and high concentrations of quality control samples, the relative recovery was between 98% and 103%, inter-and intra-day RSD were both less than 5%, the recovery of stability aider 24 hours at ambient temperature and 3 times freeze thawing at - 20 ℃ was between 83%and 102%. Conclusion： This method was sensitive, specific and simple, which is practical and suitable for clinical pharmacokinetic study.