反相高效液相色谱法测定人血浆中氯氮平浓度

Determination of the Levels of Clozapine in Human Plasma by RP-HPLC

目的 :以反相高效液相色谱法测定人血浆中氯氮平的浓度。方法 :血浆样品经碱化后用乙醚萃取 ,醚层用100μl0 1mol/mlKH2PO4 反萃取 ,40μl进样。色谱柱为DikmaDiamonsilC18 柱 ,保护柱为WatersNova -PakC18 柱 ,流动相为0 5%三乙胺溶液 -乙腈 (74∶26,V/V) ,流速为1ml/min ,检测波长为215nm。结果 :测定方法在0 1～20μg/ml范围内线性关系良好 ,萃取回收率在80 3%～84 8%之间 ;日内、日间精密度在3 5%～8 3 %之间 ,最低检测浓度为50ng/ml。结论 :本测定方法快速、灵敏、准确 ,适用于临床治疗药物浓度监测。

OBJECTIVE:To develop a simple,sensitive and rapid RP-HPLC with ultraviolet detector method for the deˉtermination of clozapine in human plasma.METHODS:The plasma samples(1ml)were extracted with5ml ether after alkaˉlization with0.5mol/ml NaOH(50μl).The organic phase was back-extracted with100μl of0.1mol/ml KH 2 PO 4 (adjusted pH to2.2with H 3 PO 4 ).The separation column was Dikma Diamonsil C 18 column and the protection column was Waters Nova-Pak C 18 column.The mobile phase consisted of0.5%triethylamine-acetonitrile(74∶26,v/v)with flow rate of1ml/min.The deˉtecting wavelength was215nm.RESULTS:A linearity was obtained in the range of100～2000ng/ml(r=0.999),the lowest detectable concentration in plasma was50ng/ml.The extraction recovery was between80.3%and84.8%.The relative stanˉdard deviations of intra-day and inter-day assay were within3.5%～8.3%.CONCLUSION:The established method was shown to be sensitive,accurate and simple for the determination of clozapine levels in human plasma.It was suitable for the pharmacokinetic and TDM study of clozapine.