用自动化柱切换二维高效液相色谱建立人血浆中氯氮平测定方法

摘要

Objective To observe a two dimensional liquid chromatographic with automated column switching methods to examine clozapine in human plasma. Methods First-dimensional liquid chromatography(LC1)system used FRO SCX(20 × 4.6 mm,5 μm,ANAX)column with a mobile phase consisting of water(60 mmol/L ammonium acetate)and acetonitrile(50:50,V/V)at a flow rate of 2.0 ml/min.The temperature of central treatment system was 40 ℃.LC2 system used Ultimate C18(250 mm ×4.6 mm I.d,5 μm)column with a mobile phase consisting of water(50 mmol/L ammonium acetate)and acetonitrile at a flow rate of 2.0 ml/min and another flow rate of 1.2 ml/min.The "heart cutting" window was 1.65-2.3 minute.The sample was pretreated by acetonitrile and introduced by 200 ul sample loop. Results The assay was linear from 30.0 to 1200 μg/L(r=0.99992).The LLQD was 21.5 μg/L,The between-day precision was 4% or less(n=18).The accuracy was between 100.6%-104.4%. Conclusion This method works with few manual procedure by fully automation.Our results also show that this LC/LC-UV platform has good accuracy and precision.%目的 用自动化柱切换二维高效液相色谱(HPLC)建立人血浆中氯氮平测定方法,用于氯氮平治疗药物监测或药代动力学研究. 方法 第一维色谱(LCI)萃取柱为FRO SCX(20×4.6 mm,5 μm,ANAX),流动相为60 mmol/L醋酸铵:乙腈=50:50(V/V),萃取泵流速为2.0 ml/min,ASP-2420C控制系统温度为40 ℃,LC2系统中分析色谱柱为Ultimate C18流动相为50 mmol/L,醋酸铵:乙腈=45:65(V/V),流速为1.2 ml/min,转移辅助流速1.0 ml/min,溶液为10%乙腈溶液.检测波长为259 nm,"中心切割"窗口为1.7～2.3 min,定量环为200μl,样品用乙腈预处理. 结果 氯氮平在30.0～1200.0 μg/L线性关系良好,相关系数为0.999 92,最低定量浓度为21.5 μg/L.日内相对标准偏差(RSD)＜3%(n=6),日间RSD均＜4%(n=18),准确度在100.6%～104.4%. 结论 用自动化柱切换二维HPLC建立的人血浆中氯氮平测定方法人为干预步骤少、自动化程度高,准确性与精密度良好,适合于氯氮平血药浓度测定及动力学研究.