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目的:建立大鼠血浆中酮洛芬的HPLC测定方法,进行酮洛芬微乳凝胶的药物动力学研究。方法:血浆样品采用甲醇沉淀蛋白方法处理,萘普生钠为内标;色谱柱为DiamosilC18(200mm×4.6mmi.d,5μm),流动相为甲醇-0.01mol·L-1磷酸二氢钾(70:30,磷酸调pH3.0),流速1mL·min-1,检测波长为255nm。进样量为20μL,柱温为室温。结果酮洛芬与内标萘普生钠完全分离且血浆中内源性物质对酮洛芬的含量测定无影响;酮洛芬在0.20~50μg·mL-1范围内线性关系良好,r=0.9998;最低定量限为0.05μg·mL-1;日内和日间精密度均(RSD)小于2.4%;回收率为91.3%~107.2%。结论:所用方法灵敏、准确、重复性好、回收率高,可用于酮洛芬微乳凝胶的血药浓度检测。
OBJECTIVE: To establish a HPLC method for the determination of ketoprofen in rat plasma and to study the pharmacokinetics of ketoprofen microemulsion. Methods: The plasma samples were treated with methanol precipitated protein. Naproxen sodium was used as the internal standard. The column was Diamosil C18 (200 mm × 4.6 mm id, 5 μm) and the mobile phase was methanol-0.01 mol·L -1 potassium dihydrogen phosphate 70:30, phosphoric acid pH3.0), flow rate 1mL · min-1, detection wavelength of 255nm. Injection volume of 20μL, the column temperature is room temperature. Results Ketoprofen was completely separated from the internal standard naproxen sodium and plasma endogenous substances had no effect on the determination of ketoprofen. Ketoprofen showed good linearity in the range of 0.20-50 μg · mL-1, r = 0.9998 ; The lowest limit of quantification was 0.05 μg · mL-1; the intra- and inter-day RSD was less than 2.4%; the recovery was 91.3% -107.2%. Conclusion: The method is sensitive, accurate, reproducible and has high recovery rate. It can be used for the determination of blood concentration of ketoprofen microemulsion.