摘要
目的:建立HPLC-MS法同时测定氯氮平及其代谢产物去甲氯氮平、N-氧化氯氮平血药浓度。方法:0.5 mL血浆用C18固相萃取小柱进行萃取。色谱柱为C18柱(5μm,3.9 mm×150 mm),柱温40℃;流动相为乙腈-水(36:64,含10 mmol·L-1醋酸铵与0.01%甲酸,pH=5.96),流速0.95 mL·min-1,进入MS检测器的分流比3:1。质谱条件采用电喷雾电离源(ESI+),电离毛细管电压3.90 kV,第一级锥孔电压38 V,用选择离子检测,定量分析检测的离子分别为m/z 326.9(氯氮平)、312.6(去甲氯氮平)、342.6(N-氧化氯氮平)。结果:氯氮平,去甲氯氮平、N-氧化氯氮平最低检测限分别为0.2,0.1,3 ng·mL-1;线性范围分别为20～2000 ng·mL-1(r=0.9994),10～1600 ng·mL-1(r=0.9990),10～800 ng·mL-1(r=0.9969)。三者的萃取回收率大于75%,方法回收率均大于95.0%。日内、日间变异(RSD)均小于10%。结论:该方法灵敏度高、快速,简便,无杂质干扰,适合于氯氮平、去甲氯氮平、N-氧化氯氮平血药浓度监测及药代动力学研究。
Objective: To establish a high performance liquid chromatography electrospray mass spectrometry (HPLC - MS/ESI) method to simultaneously determine clozapine and its metabolites desmethyl - clozapine and clozapine - N - oxide in human plasma. Methods: The extracted sample was performed on liquid chromatography u-sing a Waters XteraTMMS C18(5 μm,3. 9 mm × 150 mm)column, with water(formic acid:0. 01% ,ammonium acetate: 10 mmol· L- 1 pH =5. 96) - acetonitrile(64: 36) as the mobile phase. The flow rate was 0. 95 mL · min-1, with a split ratio of 3: 1. The compounds were ionized in the electrospray ionization(ESI)ion source, fragmented by in - source collisions and the ionized molecular and fragment ions detected in the selected ion recording (SIR) mode. The analytes were extracted with solid - phase extraction columns. Results:The calibration curves were linear in the ranges of 20 - 2000 ng · mL-1 ( r = 0. 9994 ) for clozapine, 10 - 1600 ng · mL-1 (r = 0. 9990 ) for desmethyl -clozapine, 10 - 800 ng · mL-1 ( r = 0. 9969 ) for clozapine -N- oxide, respectively. The average extraction recoveries for three analytes were above 75% . The methodology recoveries were higher than 95. 0% for the analytes. The RSD value of within -day and between -day were less than 10%. Conclusion:The method is accurate,sensitive,specific and reliable for simultaneous determination of clozapine and its metabolites. It may be applied to study on metabolism mechanism of clozapine in clinic therapeutic dose.
出处
《药物分析杂志》
CAS
CSCD
北大核心
2004年第1期74-78,共5页
Chinese Journal of Pharmaceutical Analysis
