HPLC 法测定猪体内双氯芬酸的血药浓度

摘要

In order to establish a HPLC method for the determination of plasma concentrations of diclofenac in piglets,hexane-isopropanol solution (90∶ 10,V/V)was used for extracting diclofenac of plasma.The plasma concentrations of diclofenac were measured by HPLC UV-Detector.The chromatography was car-ried on a C18 column,the mobile phase was composed of methanol-15 g/L acetic acid (20∶80,V/V),the detection wavelength was set at 276 nm for diclofenac,and analysis based on the external standard method. Under the condition of this test,diclofenac in plasma was well separated,and no interference from plasma matrix was observed.The linear ranges for diclofenac were 0.025 μg/mL-20.0 μg/mL(R2 =1),the lowest limit of quantitation was 0.05 μg/mL,and the ingredients showed good relationships between the peak area and the concentration.The inter-day and intra-day RSD was less than 4% and 6%.The recovery was be-tween 85% and 97%.The method could be used for the determination the diclofenac levels of piglets,and applied to study on the pharmacokinetics and bioavailability researches of the preparation containing di-clofenac sodium.%建立猪血浆中双氯芬酸含量的 HPLC 测定法。猪血浆中双氯芬酸采用正己烷-异丙醇混合溶液（90∶10，V／V）提取，HPLC 紫外检测器检测。色谱柱为 C18柱，流动相为甲醇-15 g／L 乙酸水溶液（80∶20，V／V），测定波长为276 nm。外标法定量。结果表明，血浆中杂质不干扰双氯芬酸的测定，双氯芬酸血药浓度在0．025μg／mL～20．0μg／mL 范围内线性关系良好（R2＝1），最低定量限为0．05μg／mL，高、中、低空白血浆添加样品浓度的日内变异系数和日间变异系数分别小于4％和6％，回收率为85％～97％。结果表明，该方法适用于猪血浆中双氯芬酸含量的测定，可用于含双氯芬酸钠的制剂在猪体内的药物代谢动力学和生物利用度研究。