高效液相色谱法测定氯硝西泮血药浓度

 目的 应用高效液相色谱法测定血清中氯硝西泮血药浓度及对癫痫患儿的血药浓度进行监测。方法 采用Nova-Pak C₁₈色谱柱，乙腈-0.1 mol·L^-1醋酸钠=35∶65作流动相，紫外检测波长254 nm，苯妥英钠为内标，用叔丁基甲醚提取，常温下空气吹干，重溶进样分析。结果 氯硝西泮在5.0～100.0 μg·L^-1浓度范围内线性关系良好，相关系数为0.9990，在3种不同浓度下，平均相对回收率为99.80%，99.34%，99.62%，绝对回收率为91.96%，92.16%，90.06%，日内RSD为1.41%，3.76%，3.30%，日间RSD为1.58%，1.71%，4.73%，最低检出限为2.0 μg·L^-1血清。该方法监测儿科临床口服氯硝西泮达稳态的癫痫患儿11例，血药浓度为13.04～39.17 μg·L^-1。结论 该方法为临床治疗的有效及安全性提供了保证和依据。

Studies on high performance liquid chromatographic determination of clonazepam inserum

OBJECTIVE To establish a HPLC method for determining the serum clonazepam concentrations in epileptic children.METHODS With Nova-Pak C₁₈column,the mobile phase was consisted of 35% acetonitrile and 65% 0.01 mol·L^-1 sodium acetate.Phenytoin was used as the internal standard.It was extracted by tertbutylanisole and drying at room temperature.The residue was reconstitued by the mobile phase and then analyzed at UV 254 nm.RESULTS The good linearity was obtained over the range of 5.0～100.0 μg·L^-1 with correlation coefficient r=0.9990.At three concentrations,the average recoveries were 99.80%,99.34% and 99.62%,respectively.RSD of within-day were 1.14%,3.76% and 3.30%,respectively.RSD of between-day were 1.58%,1.71% and 4.73%,respctively.The minimal detectable concentration was 2.0 μg·L^-1.The steady state serum concentrations were 13.04～39.14 μg·L^-1 in 11 epileptic children after oral administration of clonazepam.CONCLUSION This method can provide the basis for clinical effective and safe treatment of clonazepam.