National Institute of Environmental Health Sciences, National Health Research Institutes, Miaoli, 35053, Taiwan.
Department of Public Health, College of Public Health, China Medical University, Taichung, 40678, Taiwan.
Chemosphere. 2022 Aug;301:134778. doi: 10.1016/j.chemosphere.2022.134778. Epub 2022 Apr 30.
β-Adrenergic agonist compounds are medicines that open up the lung's medium and large airways. β-Adrenergic agonist compounds have been illegally or legally used to increase lean muscle mass in meat animals, bodybuilding, weight-loss programs, and athletes. Developing a rapid analytical approach for determining β-adrenergic agonist compounds in biological samples is crucial for individual exposure assessment. This study established an analytical method for simultaneously measuring eight β-adrenergic agonist compounds in human urine, including clenbuterol, terbutaline, salbutamol, ractopamine, zilpaterol, cimaterol, tulobuterol, and fenoterol. Two hundred microliters of a urine sample were added to eight deuterium-labeled internal standard mixtures and glucuronidase/arylsulfatase for enzymatic hydrolysis, and were then analyzed using an online clean-up system coupled with a liquid chromatography-tandem mass spectrometry system (LC-MS/MS). The limit of quantification ranged from 0.03 to 0.12 ng/mL urine for the eight β-adrenergic agonist compounds. The relative standard deviations (RSD) of the within-run and between-run precisions were less than 10%, and the relative accuracy errors were less than 17% in the three-level spiked artificial urine samples. Two hundred eighty human urine samples collected from the general population in Taiwan were assessed to demonstrate the capability and feasibility of this method. The detection frequencies were 33% for clenbuterol, 5% for ractopamine, and less than 5% for the others. We concluded that the isotope dilution-online clean-up system coupled with LC-MS/MS method is a valuable analytical method for investigating urinary β-adrenergic agonist compounds in humans and is valuable for human biomonitoring studies.
β-肾上腺素能激动剂化合物是一种能扩张肺部中、大气道的药物。β-肾上腺素能激动剂化合物曾被非法或合法地用于增加肉用动物、健美、减肥计划和运动员的瘦肉量。开发一种快速分析方法来测定生物样本中的β-肾上腺素能激动剂化合物对于个体暴露评估至关重要。本研究建立了一种同时测定人尿中 8 种β-肾上腺素能激动剂化合物（包括克仑特罗、特布他林、沙丁胺醇、莱克多巴胺、扎鲁特罗、西马特罗、妥布特罗和非诺特罗）的分析方法。将 200 μL 尿液样品加入 8 种氘标记的内标混合物和葡萄糖醛酸酶/芳基硫酸酯酶进行酶解，然后用在线净化系统与液相色谱-串联质谱系统（LC-MS/MS）联用进行分析。8 种β-肾上腺素能激动剂化合物的定量限范围为 0.03 至 0.12 ng/mL 尿液。在三种水平人工尿液样品中，日内和日间精密度的相对标准偏差（RSD）均小于 10%，相对准确度误差均小于 17%。从台湾普通人群中采集了 280 个人尿样本，以证明该方法的能力和可行性。检测到的频率为：克仑特罗 33%，莱克多巴胺 5%，其余均低于 5%。我们得出结论，同位素稀释-在线净化系统与 LC-MS/MS 方法相结合是一种有价值的分析方法，可用于研究人体尿中的β-肾上腺素能激动剂化合物，对于人体生物监测研究具有重要价值。
