HPLC-MS/MS法测定人血浆及尿液中盐酸戊乙奎醚浓度

Determination of hydrochloric penehyclidine in human plasma and urine by HPLC-MS/MS

目的：建立高效液相色谱-串联质谱（HPLC-MS/MS）方法测定人血浆及尿样中盐酸戊乙奎醚的浓度.方法：采用AB SCIEXQTRAP 5500三重四极杆复合线性离子阱串联质谱仪及Agilent 1200高效液相色谱仪进行检测.样品经甲基叔丁基醚提取处理,以维拉帕米为内标.色谱柱为Thermo Hypersil GOLD柱（30 mm×2,1 mm,3μm）,流动相为乙腈：水（80∶20,V/V）（含10 mmol/L的乙酸铵及0.01‰的甲酸）;流速为0.3 mL/min.盐酸戊乙奎醚和维拉帕米的MRM扫描离子通道m/z分别为316.2→ 128.3,454.6→303.4.进样量：5 μL.结果：盐酸戊乙奎醚和维拉帕米分离良好,保留时间分别为1.25 min、0.84 min.盐酸戊乙奎醚血浆及尿液样品在0.05～10 ng/mL范围内线性关系良好,定量下限为0.05 ng/mL,批内、批间RSD均低于11.69％,准确度均低于-2.81％.结论：本法样品预处理简便快捷,检测结果专属性强,灵敏度好,准确度高,适用盐酸戊乙奎醚药代动力学的研究.

AIM：To establish a high performance liquid chromatography-tandem mass spectrometry （HPLC-MS/MS） method for the determination of penehyclidine hydrochloride in human plasma and urine.METHODS：Penehyclidine hydrochloride was determinated by AB SCIEX QTRAP 5500 three triple quadrupole hybrid linear ion trap tandem mass spectrometer and Agilent 1200 high performance liquid chromatography.The samples were extracted with methyl tert-butyl ether,and verapamil is the internal standard （IS）.The analytical column was a Thermo Hypersil GOLD column （30 mm＆#215;〈2.1 mm,3 μm）.The mobile phase,acetonitrile-water （80 ∶ 20,which contains 10 mmol/L of ammonium acetate and 0.01%‰ formic acid）,was used at a flow rate of 0.3 mL/min.Penehyclidine hydrochloride and verapamil were detected on multiple reaction monitoring （MRM） by the transitions from the recursor to the production （m/z 316.2→128.3 and m/z 454.6→303.4）.The injection vpolume was 5 μL and the total run time was 4.0 min.RESULTS：Penehyclidine hydrochloride and verapamil concentrations in plasma and urine were detected well by HPLCMS/MS method.The retention time of these analytes were 1.25 min and 0.84 min.Penehyclidine hydrochloride curve in plasma and urine had good linearity in the range of 0.05-10 ng/mL.Lower limit of quantification （LLOQ） was 0.05 ng/mL.The intra-batch and inter-batch precision （RSD） in plasma and urine were all less than 11.69% and the accuracy were all less than-2.81%.CONCLUSION：In this experiment,plasma and urine concentrations of penehyclidine hydrochloride were detected by the specificity,accuracy,sensitivity and reliability technical HPLC-MS/MS,reliable the study on the pharmacokinetics of penehyclidine hydrochloride.