HPLC-MS/MS法测定大鼠血浆中麻黄碱、伪麻黄碱浓度及药动学研究

Simultaneous determination of ephedrine and pseudoephedrine in rat' plasma and their pharmacokinetics determined by LC-MS/MS

关键词:  麻黄碱  伪麻黄碱  药代动力学  三拗汤  液相色谱-质谱联用

Key words:  ephedrine  pseudoephedrine  pharmacokinetic  Sanao decoction  LC-MS/MS

摘要:

[目的] 建立同时检测大鼠血浆中麻黄碱、伪麻黄碱的高效液相色谱-质谱联用法(HPLC-MS/MS),并对大鼠口服三拗汤后的麻黄碱、伪麻黄碱进行药代动力学研究。[方法] 血浆样品经碱化和乙酸乙酯液-液萃取,以盐酸苯丙醇胺为内标,乙腈-0.1%甲酸水(4:96)为流动相,经Agilent ZorbAx SB-C₁₈柱(4.6 ｍｍ×150 ｍｍ,5 μｍ)分离,采用电喷雾离子源(ESI),以多反应监测(MRM)模式进行正离子检测,定量分析的离子反应分别为:m/z 166.2→m/z 148.2(麻黄碱),m/z 166.2→m/z 148.2(伪麻黄碱)和m/z 152.1→m/z 134.1(盐酸苯丙醇胺)。[结果] 麻黄碱和伪麻黄碱血药浓度在20~10 000 μg/L范围内线性关系良好,批内、批间精密度RSD均小于5.3%,高、中、低3种浓度平均方法回收率大于64.7%。[结论] 该方法专属、快速、灵敏,可用于麻黄碱、伪麻黄碱的药代动力学研究。

Abstract:

[Objective] To develop a LC-MS/MS method for the determination of ephedrine and pseudoephedrine in rat's plasma simultaneously and to study the pharmacokinetics of ephedrine and pseudoephedrine of rat's plasma after oral administration of SD(Sanao Decoction).[Methods] The blood plasma samples were extracted through alkali and ethyl acetate liquid-liquid. Phenylalanine hydrochloride was used as the internal standard. Separation was performed using an Agilent Zorbax SB-C₁₈(4.6 mm×150 mm, 5 μm). The analytical column was acetonitrile-0.1% formic acid in water solution (4:96). Electrospray ionization (ESI) source was applied and operated in the positive ion mode. Multiple reaction monitoring (MRM) mode with the transitions of m/z 166.2→m/z 148.2, m/z 166.2→m/z 148.2 and m/z 152.1→m/z 134.1 were used to quantify ephedrine, pseudoephedrine and phenylalanine hydrochloride, respectively. [Results] The linearity ranged from 20 to 10 000 μg/L. RSD for inter-match and intra-match were not more than 5.3% and the average extracted recovery for all the high, middle and low concentration was more than 64.7%. [Conclusion] The analytical method is proved to be special, sensitive, rapid and suitable for the pharmacokinetic study of ephedrine and pseudoephedrine.