人尿中β_2-受体激动剂的液相色谱质谱检测

建立了人尿中11种β2受体激动剂的液相色谱质谱(LC/MS)定性分析方法,检测了人单剂量口服一种β2受体激动剂后不同时间点的尿样。尿样经β葡萄糖苷酸酶酶解、BondElutCertify小柱提取后,采用AgilentZorbaxSBC18柱分离,以(A)0.01mol/L甲酸铵缓冲液(pH3.5)(B)乙腈为流动相进行梯度洗脱,洗脱程序:0min时,A为95%,B为5%;8min时,A为45%,B为55%;13min时,A为10%,B为90%,并保持6min。通过液相色谱质谱以选择离子监测(SIM)方式检测,测得11种药物的检测限为0.1～60.0μg/L。采用所建立的方法对人单剂量口服治疗量特布他林、克仑特罗、丙卡特罗、福莫特罗、非诺特罗或马布特罗后不同时间点的尿样进行了检测,结果表明检测口服给药48h内尿中的相应药物,信噪比可达4以上。

Detection of β2-agonists in Human Urine by Liquid Chromatography-Mass Spectrometry

A liquid chromatography-mass spectrometry method was developed for the qualitative analysis of eleven β_2-agonists in human urine. This method was successfully applied to the analysis of the urine samples excreted by some healthy subjects over different periods after oral administration of single dose of β_2-agonists. Urine samples were hydrolyzed with β-glucuronidase followed by a solid-phase extraction procedure using Bond Elut-Certify cartridges. The analytes were separated on Agilent Zorbax SB-C_ 18column using a gradient elution of (A) 0.01 mol/L ammonium formate buffer (pH 3.5) and (B) acetonitrile. Elution started with 95% A, followed by a linear gradient to 45% A in 8 min, and decreased to 10% A in another 5 min, and then held 6 min. An electrospray ionization (ESI) source was operated in positive ion mode. The detection limits of 11 β_2-agonists were in the range of 0.1-60.0 μg/L.