HPLC-MS/MS法测定人尿液中戊乙奎醚的浓度

摘要

Objective To develop an HPLC-MS/MS method for determination of penehyclidine in human u-rine. Methods The supernatant liquid obtained from the diluted urine centrifugate was subjected to sample introduction and was analyzed by the HPLC-MS/MS method under flowing chromatographic conditions:Agilent ZORBAX Bonus-RP column (4. 6 mm × 150 mm,5 μm),the mobile phase compositing with 5 mM ammonium acetate (pH was adjus-ted to 5. 5 by formic acid):methanol (35:65,v/v),with a rate of 1. 0 mL/min,column temperature at 30 ℃,and in-jected sample volume of 10 μL. The results of positive ion MRM detection of penehyclidine and internal standards were m/z 316. 2→128. 1 and m/z 256. 2→167. 1. Results The calibration curve was linear over the concentration range of 1~100 ng/mL(r=0. 997 7)with the lower limit of quantitation (LLOQ) 1 ng/mL. The intra-day RSD and inter-day RSD were all below 7. 9%. Conclusion The method is simple, rapid, specific and sensitive, it can be used for the pharmacokinetic study of penehyclidinein human urine.%目的 建立测定人尿液中戊乙奎醚浓度的HPLC-MS/MS法. 方法 尿样经稀释,离心后取上清液进行HPLC-MS/MS测定. 色谱柱:Agilent ZORBAX Bonus-RP (4. 6 mm × 150 mm,5 μm),流动相:5 mM醋酸铵(甲酸调pH 5. 5) -甲醇(35:65,v/v),流速:1. 0 mL/min,柱温:30 ℃,进样量10 μL. 采用电喷雾离子源( ESI) ,正离子模式多重反应选择离子检测( MRM). 戊乙奎醚和内标的定量离子对分别为m/z 316. 2→128. 1和m/z 256. 2→167. 1. 结果 线性范围为1~100 ng/mL(r=0. 997 7),最低定量限为1 ng/mL,日内和日间精密度( RSD)均<7. 9%. 结论 本方法简单、快速、灵敏、专属性高,适用于人尿液中戊乙奎醚药动学的研究.