液相色谱-电喷雾质谱法测定血浆中吴茱萸碱和吴茱萸次碱浓度

Simultaneous Determination of Evodiamine and Rutaecarpine in Rabbit Plasma by HPLC-MS

目的:建立家兔血浆中吴茱萸碱和吴茱萸次碱同时测定的液相色谱-电喷雾质谱联用新方法。方法:血浆样品经甲醇沉淀蛋白后,以乙腈-2%四氢呋喃水溶液(62:38)为流动相,用 ZORBAX SB-C_(18)色谱柱分离;采用电喷雾离子源,负离子模式选择性检测(SIM)模式。干燥气流速:12 L·min^(-1);温度:350℃;雾化电压:276 kPa;毛细管电压:3500 V。吴茱萸碱用于定量分析的离子:m/z302([M-H]^-),定量分析吴茱萸次碱的离子为:m/z286([M-H]^-)及 m/z595([2N+Na-2H]^-)。结果:吴茱萸碱和吴茱萸次碱分别在0.32～16.0 ng·mL^(-1)(r=0.9999)和1.0～40.0 ng·mL^(-1)(r=0.9992)浓度范围内呈良好线性关系;血浆中吴茱萸碱和吴茱萸次碱的方法回收率分别为94.3%～98.3%和97.0%～107.0%;日内、日间精密度(RSD)皆小于5%。结论:该方法简便、准确、灵敏度高,可用于血浆中吴茱萸碱和吴茱萸次碱浓度测定及药物动力学研究。

Objective ：To establish a sensitive and specific HPLC - MS method for the simultaneous determination of evodiamine and rutaecarpine in rabbit plasma. Methods： The plasma samples were deposited by methanol before analysis, The supernatant was performed on an Aglient SB C18 analytical column using acetonitrile - 2% tetrahydrofuran（62： 38）as the mobile phase, the flow rate was 0. 5 mL min^-1. The mass spectrometer was operated under the negative ion mode and selected ion mass spectral m/z 302 （ evodiamine, [ M - H ]^ - ）, m/z 286 （ rutaecarpine, [ M - H]^ - ）and 595 （ rutaecarpine, [2M ＋ Na - 2H]^ - ） to quantify, respectively. The drying gas flow was 12, 0 L · min^-1 ,the drying gas temperature was 350 ℃ ,the nebulizer pressure was 276 kPa,the capillary voltage was 3500 V. Results： The linear ranges were 0. 32 - 16. 0 ng · mL^-1 and 1.0 -40. 0 ng · mL^-1 for evodiamine and rutaecarpine. The average extraction recoveries were all over 80%. The method extraction recoveries of evodiamine and rutaecarpine were 94. 3% -98.3% and 97.0% - 107.0%. The intra - and inter - day RSD of them were also less than 5%. Conclusions ：The method is simple, rapid and sensitive. It can be used for the pharmacokinetics studies of evodiamine and rutaecarpine.