高效液相色谱-离子阱质谱法测定尿液中β_2-受体激动剂及β-受体阻断剂  

Determination of β_2-agonists and β-blockers in urine using high performance liquid chromatography-ion trap mass spectrometry

作　　者：苗虹[1] 邹建宏[2] 范赛[1,3] 甘乐文[2] 赵云峰[1] 吴永宁[1,3] 

机构地区：[1]中国疾病预防控制中心营养与食品安全所,北京100021 [2]中国人民解放军第二炮兵总医院,北京100088 [3]中国农业大学动物医学院,北京100193

出　　处：《色谱》2010年第6期572-578,共7页Chinese Journal of Chromatography

基　　金：国家自然科学基金重点基金项目(20837003);“十一五”国家科技支撑计划项目(2006BAK02A27);卫生行业公益项目(200902009)

摘　　要：建立了尿液中23种β2-受体激动剂及5种β-受体阻断剂的高效液相色谱-离子阱质谱(HPLC-IT-MS)测定方法。尿液样品采用冷冻高速离心沉淀蛋白,上清液过ExtrelutTM硅藻土柱,用乙酸乙酯洗脱后,洗脱液经旋转蒸发仪浓缩并复溶待测。HPLC分离采用AtlantisT3-150mm色谱柱,以甲醇和含0.1%甲酸的水溶液为流动相梯度洗脱,IT-MS采用电喷雾离子源在多反应离子监测模式下测定。定量分析选择9种经过氘代同位素标记的β2-受体激动剂为内标。各化合物的线性范围为0.005～0.16mg/L,尿液中的检出限均能达到0.2μg/L。空白尿液样品中不同加标水平的回收率为57.1%～127.1%,相对标准偏差为1.1%～31.1%。该方法简便快速,灵敏度高,适用于人或动物尿液中23种β2-受体激动剂及5种β-受体阻断剂的定性和定量分析。A method has been developed for the determination of 23 β2-agonists and 5 β-blockers in urine samples using high performance liquid chromatography-ion trap mass spectrometry （HPLC-IT-MS）. Urine samples were first deproteinized by high-speed frozen centrifugation, and the supernatants were loaded on an ExtrelutTM diatomite column for clean-up. The analytes were eluted by ethyl acetate and concentrated for further analysis. The analytical separation was performed on an Atlantis T3-150 mm chromatographic column with the gradient elution using methanol and water （ containing 0. 1% formic acid）. The detection was carried on a linear ion trap mass spectrometer under multiple reaction monitoring （MRM） mode with the source operated in positive mode of electrospray ionization （ ESI^ ＋ ）. Nine deuterium labeled β2-agonists were used as internal standards for quantitative analysis. The results showed that the linear ranges for 23 β2-agonists and 5 β-blockers were 0.005 - 0. 16 mg/L, and the limits of detection were all around 0.2μg/L. The mixed standard solution was added into the blank urine samples, and the recoveries of 23 β2-agonists and 5 β-blockers were ranged from 57. 1% to 127.7% with the relative standard deviations of 1. 1% -31. 1%. The results demonstrate that the method is easy, fast, sensitive, and suitable for the confirmation and quantification of 23 β2-agonists and 5 β-blockers in urine samples.

关 键 词：高效液相色谱-质谱法 Β2-受体激动剂 Β-受体阻断剂 尿液 

分 类 号：O658[理学—分析化学]