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目的建立人血浆中奥氮平、氯氮平的UPLC-MS/MS检测方法。方法用甲基叔丁基醚提取,离心后取上清液用氮气吹干,用乙腈-水(8∶2)溶解后进行质谱分析;采用BEH C18色谱柱(50 mm×2.1 mm,1.7μm),流动相A为乙腈,B相为0.01 mol·L-1甲酸铵(甲酸调p H3.5),梯度洗脱(0 min、70%B,0~1.5 min、70%→10%B,1.5~2.5 min、10%B,2.5~2.6min、10%→70%B,2.6~4 min、70%B),流速0.2 m L·min~(-1),柱温45℃,进样量5μL;电喷雾离子化(ESI),正离子模式多重反应选择离子检测(MRM),奥氮平、氯氮平及内标的检测离子对分别为:m/z313.29→256.25、198.24;327.20→192.17,270.14;386.43→122.37,109.43。结果奥氮平和氯氮平的线性范围分别为0.43~54.48、0.21~27.24 ng·m L-1(r=0.994、0.998,n=6)。在人血浆基质中待测成分高、中、低浓度的日内、日间RSD均小于15%,奥氮平和氯氮平的方法回收率为84%~113%、86%~114%;血浆样品室温下放置8 h或处理后放置8 h、-20℃放置20 d及-20℃冻融3次对测定结果无影响;血浆基质效应研究证实,该样品预处理方法对血浆中的奥氮平、氯氮平测定无干扰。结论所用UPLC-MS/MS法处理简单、灵敏、特异性高,定量准确,为奥氮平、氯氮平制剂的药动学研究及临床血药浓度的检测提供了分析测定方法。
Objective To establish a method for the determination of olanzapine and clozapine in human plasma by UPLC-MS / MS. The method was extracted with methyl tert-butyl ether, and after centrifugation, the supernatant was blown dry with nitrogen and dissolved by acetonitrile-water (8: 2). MS analysis was performed on a BEH C18 column (50 mm × 2.1 mm, ), Mobile phase A was acetonitrile, phase B was 0.01 mol·L-1 ammonium formate (pH = 3.5 for formic acid), gradient elution (0 min, 70% B, 0-1.5 min, 70% → 10% B , 1.5-2.5 min, 10% B, 2.5-2.6 min, 10% → 70% B, 2.6-4 min, 70% B) at a flow rate of 0.2 mL · min -1 (5μL); Electrospray ionization (ESI), positive ion mode multiple reaction selective ion detection (MRM), olanzapine, clozapine and internal standard ion pairs were: m / z 313.29 → 256.25,198.24; 327.20 → 192.17, 270.14; 386.43 → 122.37, 109.43. Results The linear ranges of olanzapine and clozapine were 0.43 ~ 54.48 and 0.21 ~ 27.24 ng · m L-1, respectively (r = 0.994, 0.998, n = 6). The RSDs of day, day and daytime were all less than 15% in high, medium and low concentrations of human plasma matrix. The recoveries of olanzapine and clozapine were 84% -113% and 86% -114% respectively. The plasma samples Placed at room temperature for 8 h or 8 h after treatment, placed at -20 ℃ for 20 d and frozen at -20 ℃ for 3 times had no effect on the determination results. The study of plasma matrix effect confirmed that the sample pretreatment method for the determination of olanzapine , Clozapine determination without interference. Conclusion The method of UPLC-MS / MS is simple, sensitive, specific and accurate. It provides an analytical method for the determination of olanzapine and clozapine pharmacokinetics and the determination of clinical plasma concentration.