RP-HPLC法测定犬血浆中司他夫定的血药浓度

Determination of Stavudine in dogs plasma by RP-HPLC

目的 建立一种RP-HPLC方法测定犬血浆中司他夫定的浓度。方法 血浆样品经碱化处理,再用有机相提取后吹干,残渣用流动相溶解进样分析。色谱柱：Diamonsil C18柱（150 mm×4.6 mm,5μm,ID）,流动相：甲醇-水（20∶80）,流速：1.0 mL/min,柱温：室温,检测波长：266 nm。以脱水胸苷为内标,测定了犬口服司他夫定口服溶液后的血药浓度-时间过程。结果 标准曲线线性范围为0.02-5.0μg/mL,最低检测限为0.02μg/mL,回收率均〉50%,日内、日间精密度RSD均〈15%。结论 此方法快速、可靠,可用于司他夫定血药浓度分析及药代动力学研究。

Objective To develop a RP-HPLC method for the determination of stavudine in the plasma of dogs. Methods Samples were treated with phosphate buffer （pH 6.0）, and then were extracted by ethylester. The extract was injected into the HPLC system with an analytical column of Diamonsil C18（150 mm× 4.6 mm, 5 μm, ID） with the column temperature at room temperature. The mobile phase consisted of a mixture of water-methanol （80：20, v/v） with a flow rate of 1.0 mL/min. The stavudine was detected by UV absorption at 266 nm. The plasma concentration of stavudine was determined after po dose of 4 mg/kg in dogs. Results The assay was linear in the range of 0.02 - 5.0 μg/mL（r= 0.998 9） with the limit of quantification at 0.02 μg/mL. The recoveries of stavudine in plasma were more than 50 % .Within-day and between-day RSD were all less than 15 %. Conclusion The method is rapid and reliable for the assay of stavudine concentration in dog plasma and relevant pharmacokinetics studies.