LC-MS/MS测定人尿液中伪麻黄碱含量

Determination of Pseudoephedrine in Urine by LC-MS/MS

关键词:  液相色谱-质谱法  伪麻黄碱  尿液

Key words:  LC-MS/MS  pseudoephedrine  urine

摘要:

目的 建立测定人尿液中伪麻黄碱药物浓度的方法。方法 采用LC-MS/MS进行测定。色谱柱：美国 Agilent Eclipse plus C₁₈(4.6 mm×100 mm，3.5 μm)；流动相：甲醇-水(3︰1，含0.005 mol?L^-1甲酸铵和0.1%甲酸)；流速：0.5 mL?min^-1；柱温：40 ℃。质谱条件：电喷雾电离源(ESI)，正离子化，扫描方式为选择性离子监测，伪麻黄碱m/z 166.2→148.1，内标对乙酰氨基酚m/z 152.1→110.1。结果 尿液中伪麻黄碱浓度在500~300 000 μg?L^-1(r²=0.998)内呈良好的线性关系，其批内、批间精密度RSD分别≤4.41%与≤10.04%；伪麻黄碱和内标的提取回收率均>90%；尿液中伪麻黄碱的平均累积排泄率为44.57%。结论 本研究建立的LC-MS/MS测定人尿液中伪麻黄碱的方法准确、简便、灵敏、重复性好，可用于临床研究中伪麻黄碱的尿药含量测定。

Abstract:

OBJECTIVE To establish a method for determination of pseudoephedrine in urine samples of healthy volunteers. METHODS The pseudoephedrine was determined by LC-MS/MS. The separation was carried on an Agilent Eclipse plus C₁₈(4.6 mm×100 mm, 3.5 μm) colume with a mobile phase of methanol-0.005 mol?L^-1 ammonium formate apueous solution with 0.1% formic acid (3∶1) at the flow rate of 0.5 mL?min^-1, The column formate temperature was set at 40 ℃. ESI source was applied and operated in positive ion mode. Quantitative determination was performed by using selective reaction monitoring (SRM) of m/z 166.2→148.1 for pseudoephedrine and m/z 152.1→110.1 for paracetamol used as internal standard. RESULTS Pseudoephedrine in urine samples showed good linearity in the range of 500?300 000 μg?L^-1(r²=0.998). The intra-batch and inter-batch precisions(RSD) were ≤4.41% and ≤10.04%, respectively. The extraction recoveries of pseudoe- phedrine and internal standard were more than 90%. The average cumulative excretion of pseudoephedrine in urine was 44.57%. CONCLUSION The method that using LC-MS/MS to determine pseudoephedrine in urine is accurate, simple and sentitive, and it has a good reproducibility. This method can be applied to the clinical pharmacokinetic study and therapeutic drug monitoring of pseudoephedrine.