高效液相色谱法测定兔血浆中盐酸氯胺酮质量浓度

目的 建立测定兔血浆中盐酸氯胺酮质量浓度的高效液相色谱法.方法 采用安捷伦1200型液相色谱仪,色谱柱为Hypersil ODSC18柱(250 mm×4.6 mm,5μm),流动相为甲醇-乙腈-磷酸盐缓冲液(50:15:35),流速为1.0 mL/min,柱温为30℃,检测波长为220 nm.以布比卡因为内标检测血浆中盐酸氯胺酮的质量浓度.结果 盐酸氯胺酮的质量浓度在0.25～25 μg/mL范围内与峰面积线性关系良好(R2=0.999 5),盐酸氯胺酮和布比卡因的保留时间分别为5.868 min和10.776 min,最低检测浓度为0.1 μg/mL,日内日间RSD均小于5%,其中低、中、高(0.5,2.5,25μg/mL)质量浓度的提取回收率分别为84.91%,79.80%,80.63%,方法回收率分别为97.19%,100.97%,99.62%.结论 提取方法可靠、专属性理想、稳定性好,适用于盐酸氯胺酮血药浓度的测定.

Determination of the Plasma Concentration of Ketamine Hydrochloride in Rabbits by HPLC

Objective To establish a high performance liquid chromatographic method for the determination of ketamine hydrochloride in rabbit plasma.Methods Using agilent 1200 liquid chromatography,the analytical column was packed with Hypersil ODS C18（250 mm×4.6 mm,5 μm）.The mobile phase was methanol-acetonitrilephosphate buffer（50：15：35） at a flow rate of 1.0 mL/min,and the detective wave-length was 220 nm.Bupivacaine was used as internal standard.Plasma concentrations of ketamine hydrochloride was determined by HPLC.Results The calibration curve showed good linearity in a concentration range of 0.25～25 μg/mL（R^2=0.999 5）,the retention times of ketamine hydrochloride and bupivacaine were 5.868 min and 10.776 min,respectively.Lowest detectable limit was 0.1 μg/mL,the intra-day and inter-day assay variability values were all less than 5.0%,The average recovery was 84.91%for 0.5 μg/mL,79.80%for 2.5 μg/mL,80.63%for 25 μg/mL,the relative recovery was 97.19%for 0.5 μg/mL,100.97%for 2.5 μg/mL,99.62%for 25 μg/mL.Conclusion The method is reliable,accurate and sensitive for determination of ketamine hydrochloride in rabbits plasma.It is suitable for determining the concentration of ketamine hydrochloride in plasma.