HPLC-MS/MS测定动物源性中药中的3种β2-受体激动剂残留

Detection of 3 β2-agonists Residues in Animal-derived Chinese Medicine by HPLC-MS/MS

关键词:  动物源  阿胶  人工牛黄  三重四级杆  高效液相色谱法  β₂-受体激动剂

Key words:  animal-derived  Asini Colla Corii  Bovis Calculus Artifactus  triple quadrupole  HPLC  β₂-agonists

摘要:

目的 建立动物源性中药中的克伦特罗、沙丁胺醇和莱克多巴胺3种β₂-受体激动剂残留的高效液相色谱-串联四级杆质谱（HPLC-MS/MS）检测方法。方法 以Agilent XDB-C₁₈（50 mm×4.6 mm，5.0µm）色谱柱分离，串联四级杆质谱仪检测，MRM模式进行定性定量检测分析，样品经酶解后，经MCX固相萃取柱净化，检测阿胶和人工牛黄中的克伦特罗、沙丁胺醇和莱克多巴胺。结果 3种β₂-受体激动剂质谱检测的线性范围宽，相关性好，R² ≥ 0.997 1；方法精密度RSD为1.4%~2.3%（n=6）；方法回收率范围为89.4%~102.8%；定量限范围为0.25~5.50µg·L^-1；日内精密度为2.4%~4.6%（n=5），日间精密度为1.9%~3.7%（n=9）。结论 本方法专属性强，操作简单、快捷，可作为动物源性中药中克伦特罗、沙丁胺醇和莱克多巴胺3种β₂-受体激动剂残留的定量、定性的有效检测方法。

Abstract:

OBJECTIVE To develop an high performance liquid chromatography with triple quadrupole mass spectrometry (HPLC-MS/MS) method to determine 3 β₂-agonists including clenbuterol, salbutamol and ractopamine in animal-derived Chinese medicine. METHODS The analysis was performed by a HPLC-MS/MS system with Agilent XDB-C₁₈(50 mm×4.6 mm, 5.0 µm) column. Multiple-reaction monitoring (MRM) was performed to identify and quantify 3 β₂-agonists, which were extracted from Asini Colla Corii and Bovis Calculus Artifactus, purified with MCX SPE columns. RESULTS three linear calibration curves were obtained with R² ≥ 0.997 1. The precision of the method were showed by RSD (n=6) ranged from 1.4% to 2.3%. The recoveries were determinated at three concentrations and ranged from 89.4% to 102.8%. The ranges of LOQ were from 0.25 µg·L^-1 to 5.50 µg·L^-1, and the RSDs of intra-day precision (n=5) and inter-day precision (n=9) were from 2.4% to 4.6% and from 1.9% to 3.7%. CONCLUSION The method is specific, simple and fast to detect the 3 β₂-agonists in animal-derived Chinese medicine.