摘要
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 simuhaneous determination of Olanzapine and Clozapine in human plasma using UPLC - MS/MS. METHODS Plasma sample was extracted by methyl tert - butyl ether. The resulting supernatant liquid after centri- fuge was dried by nitrogen and was then analyzed by the UPLC - MS/MS method after it was diluted quantitatively by acetonitrile - water（8 ： 2）. The following chromatography conditions were used as the following：BEH C 18 （50 mm × 2.1 mm, 1.7 μm）, the mobile phase consisting of acetonitrile （A） ; 0.01 mol·L ^-1 ammonium formate （ formic acid adjust pH3.5 ） （ B ） ; gradient elution （ 0 rain,70% B ; 0 - 1.5 min ,70% →10% B ; 1.5 - 2.5 min, 10% B ;2.5 - 2.6 rain, 10% →70% B ;2.6 - 4 min,70% B at flow rate of 0.2 mL·min^- 1 ）. Column temperature was 45 ℃, and inject volume 5 μL. With positive ion MRM detection of Olanzapine （ m/z 313.29→256.25, 198.24） ,Clozapine（327.20→192.17,270. 14） ,using buspirone hydrochloride（m/z 386.43→122.37,109.43 ） as internal standard, respectively. RESULTS A good linearity was shown in the range of 0.43 - 54.48 ng - mL^- 1 （ r = 0. 994, n = 6） for Olanzapin, and 0. 21 - 27.24 ng·mL^ -1 （ r = 0.998, n = 6） for Clozapine. Intra - day RSD and inter - day RSD of the two compoent was all less than 15%. The recovery was within 84% -113% ,86% -114%. Plasma matrix effect test showed that the endogenous matrix has no effect on quantification of the two components. CONCLUSION A rapid,sensitive and accurate UPLC - MS/MS method was developed and validated for the determination of Olanzapine and Clozapine in human plasma.
出处
《华西药学杂志》
CAS
CSCD
2016年第6期600-603,共4页
West China Journal of Pharmaceutical Sciences
