高效液相色谱-质谱法测定伪麻黄碱血药浓度

Determination of Pseudoephedrine in Human Plasma by HPLC-MS

目的建立高效液相色谱-质谱(HPLC-MS)法测定伪麻黄酸碱的血药浓度。方法将血浆样品采用甲醇沉淀后,以格列吡嗪为内标进行测定。色谱柱为Agilent Zorbax SB-C18柱(150mm×4.6mm,5μm),流动相为5mmol/L醋酸铵-甲醇(10∶90,V/V),流速为1.0mL/min,离子检测方式设为选择性离子检测(SIM),检测离子极性为正离子,采用气动辅助电喷雾离子化(ESI)方式,检测对象为伪麻黄碱(M+H离子,m/z148.4),内标格列吡嗪(M+Na离子,m/z468.1)。结果线性范围为5.01～1002μg/L(r=0.9997),最低检测限为5μg/L。方法回收率为89.6%～95.1%,日内、日间精密度RSD分别小于5.2%和7.1%。结论该方法选择性好,灵敏度高,简便易行,适用于伪麻黄碱药代动力学和生物等效性研究。

Objective To establish HPLC- MS method for determination of pseudoephedrine in human plasma. Methods The extraction solvent was methanol. Liquid-liquid extraction was used to treat with human plasma. Agitent Zorbax SB-C18 （150 mm×4.6 mm, 5 μm） column was used with methanol - 5 mmot/L ammonium acetate water solution （90 ： 10, V/ V） as the mobile phase and a flow rate of 1.0 mL/min. A Waters ZQ 2000 HPLC- MS system with an atmospheric pressure chemical ionization （APCI） source was used as the detector and was operated in a positive ion mode. Selected ion monitoring （SIM）was performed and the m/z of ions selected for quantification were 148.4 （pseudoephedrine） and m/z 468. 1 （internal standard, gtipizide）. Results Calibration curve for pseudoephedrine was linear over the range of 5.01- 1 002 μg/L. The lowest limit of quantitation was 5 μg/L. The method recovery rates were 89,6%-95. 1%.The inter and intra day precisions were less than 5.2% and 7. 1%. Conclusion The method was proved to be sensitive, accurate and convenient. It can be applied to study the pharmacokinetics of pseudoephedrine in human plasma.