犬血浆中藤黄酸HPLC测定及其药代动力学

Pharmacokinetics of Gambogic Acid in Dogs

目的: 建立HPLC法测定犬血浆中藤黄酸浓度,并用该法研究Beagle犬静注藤黄酸后的药代动力学.方法:取犬血浆0.3 mL,1 mol/L HCl酸化,加乙酸乙酯提取,取有机相空气吹干后,残留物溶解后进样.色谱柱Lichrospher C18柱(200 mm×4.6 mm ID,5 μm)流动相为甲醇-0.05%磷酸水 (94∶6,V∶V);流速1.0 ml/min;柱温35 ℃;检测波长UV 360 nm;犬静注藤黄酸0.5,1,2 mg/kg后,于不同时间点取血测定血浆中的药物浓度并估算其药动学参数.结果: 该测定方法的最低检测浓度为0.067 μg/ml,线性范围为0.067～33.3 μg/mL,回收率大于90%,日内及日间变异均小于10%.犬静注三个剂量藤黄酸后其消除半衰期为57.95～60.95 min,分布容积为0.66～0.71 L/kg.在0.5,1,2 mg/kg的剂量下AUC分别为58.95,130.46和266.37 μg·h/mL,剂量和AUC呈线性相关.结论: 建立的犬血浆中藤黄酸HPLC测定方法适合于药代动力学研究.藤黄酸给药剂量和AUC基本呈线性关系,提示藤黄酸在犬体内处置属于线性动力学.

AIM： To develop an HPLC method for the determination of gambegie acid concentration in dog plasma and study the pharmacokineties of gambegie acid in Beagle dog. METHOD： 0.3 mL plasma samples was acidified with 1 moL/L HCl, then extracted with ethyl acetate and evaporated to dryness. The residuals were dissolved with mobile phase and injected onto Liehrospher C18（200 mm × 4.6 mm ID, 5 μm）column. The mobile phase consisted of methanol - 0.05 % phosphoric acid water（ pH 3.5）（94：6 v：v）, and was delivered at the flow rate of 1 mL/min. The elute was detected at UV 360 nm. Dog blood samples were collected after iv a single dose of gambegie acid and its pharmaeokinetie parameters were estimated. RESULT： The standard curve was linear in the range of 0.067 ～ 33.3 μg/mL. The LOD was 0.067 /μg/mL. The recovery of gambegie acid was over 90%. The relative standard intra-day and inter-day deviation was less than 10%. The within-day and between-day precision was less than 10%. The pharmaeokinetie results showed that the drug was rapidly eliminated from plasma and t 1/2 was 57.95 ～ 60.95 min after iv administration of three doses of gambegie acid. The AUC was 58.95, 130.46, 266.37 μg· min/mL following iv 0.5, 1, 2 mg/kg of gambegie acid. The relationship between dose and AUC showed excellent linearity. CONCLUSION： The established HPLC method was suitable for the pharmacokinetie study of gambegie acid. Following iv administration gambegie acid was rapidly eliminated from plasma. The disposition of gambegie acid in dogs complied with liner kinetics in the dose range.