Zhou Ting, Zhao Ting, Cheng Qing, Liu Shan, Xu Ling, Tan Wen
School of Bioscience and Bioengineer, South China University of Technology, Guangzhou, Guangdong, 510006, China.
Biomed Chromatogr. 2014 Jul;28(7):994-1002. doi: 10.1002/bmc.3107. Epub 2013 Dec 20.
A sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous determination of R-bambuterol and its active metabolite R-terbutaline in human plasma and urine was established. The inhibition for the biotransformation of R-bambuterol in plasma was fully investigated. Plasma samples were prepared on ice and neostigmine metilsulfate added as a cholinesterase inhibitor immediately after sample collection. All samples were extracted with ethyl acetate and separated on a C₁₈ column under gradient elution with a mobile phase consisting of methanol and water containing 5 mm ammonium acetate at a flow rate of 0.6 mL/min. The analytes were detected by an API 4000 tandem mass spectrometer with positive electrospray ionization in multiple reaction monitoring mode. The established method was highly sensitive with the lower limit of quantification (LLOQ) of 10.00 pg/mL for each analyte in plasma. In urine samples, the LLOQs were 20.00 and 500.0 pg/mL for R-bambuterol and R-terbutaline, respectively. The intra- and inter-day precisions were <12.7 and <8.6% for plasma and urine, respectively. The analytical runtime within 6.0 min per sample made this method suitable for high-throughput determination. The validated method has been successfully applied to the human pharmacokinetic study of R-bambuterol involving 10 healthy volunteers.
建立了一种灵敏的液相色谱-串联质谱（LC-MS/MS）法，用于同时测定人血浆和尿液中的R-班布特罗及其活性代谢物R-特布他林。全面研究了血浆中R-班布特罗生物转化的抑制作用。血浆样品在冰上制备，样品采集后立即加入硫酸新斯的明作为胆碱酯酶抑制剂。所有样品用乙酸乙酯萃取，并在C₁₈柱上进行分离，流动相为含5 mM乙酸铵的甲醇和水，梯度洗脱，流速为0.6 mL/min。通过API 4000串联质谱仪在多反应监测模式下以正电喷雾电离检测分析物。所建立的方法灵敏度高，血浆中各分析物的定量下限（LLOQ）为10.00 pg/mL。在尿液样品中，R-班布特罗和R-特布他林的LLOQ分别为20.00和500.0 pg/mL。血浆和尿液的日内和日间精密度分别<12.7%和<8.6%。每个样品在6.0分钟内的分析运行时间使该方法适用于高通量测定。该验证方法已成功应用于涉及10名健康志愿者的R-班布特罗人体药代动力学研究。
