RP-HPLC法测定癫痫患者氯硝西泮血药浓度

Determination of clonazepam in serum by RP-HPLC

目的:建立RP-HPLC法测定氯硝西泮(CZP)血药浓度。方法:血清样品加入磷酸盐缓冲液(pH=5.8),以二氯甲烷提 取后在37℃经氮气吹干,残渣用甲醇溶解后进样分析。色谱条件:Sphrisorb ODS(250mm×4.0mm,5μm)分析柱;流动相为甲醇 -水(53:47,V/V);流速0.9mL·min^(-1);检测波长320nm;柱温27℃;进样量20μL。结果:CZP在0.01～0.40μg·mL^(-1)浓度范围内 线性关系良好(r=0.9999,n=6),最低检测浓度0.5ng·mL^(-1)(r_(SN)≥3)。平均回收率为100.8％,日内及日间RSD<5％。结论: 该方法较为灵敏、准确,适用于临床氯硝西泮血药浓度监测。

AIM: To develop a RP-HPLC method for the determination of clonazepam in serum. METHODS: The drug was extracted with dichloromethane from serum in which added phosphate buffer (pH 5.8) and dried by nitrogen flow in 37 ℃ water bath. The residue was dissolved with methanol and then analyzed. Chromatographic conditions included Spherisorb ODS (250 mm × 4.0 mm, 5μm) column and the mobile phase consisting of methanol: water (53: 47, V/V) at a flow rate of 0.9 mL·min-1 , UV detector was set at 320 nm and column temperature was controlled about 27 ℃, injection volume was 20μL. RESULTS: The calibration curve was linear within the range of 0.01 - 0.40 μg· mL-1( r = 0.999 9, n =6). The minimal detectable concentration of this method was 0.5 ng·#mL-1( rSN≥3). The average recovery was 100.8% . Both coefficient of variation for within-day and between-day were lower than 5% . CONCLUSION: The method is sensitive, accurate and suitable for therapeutic drug monitoring.