中药材中乙烯利残留量检测方法的研究

Research on the Determination Method of Ethephon Residue in Traditional Chinese Medicine

应用超高效液相色谱-串联质谱技术(UPLC-MS/MS)建立了中药材中乙烯利残留量的检测方法。中药样品经粉碎后以含1%甲酸的50%甲醇溶液提取和冷冻处理,使用乙腈稀释后,经十八烷基键合硅胶(C_(18))、硅胶(Silica)、石墨化炭黑(GCB)净化,以1.2%甲酸溶液和0.5%甲酸乙腈为流动相,采用Anionic Polar Pesticide Column(2.1 mm×100 mm,5μm)在15 min内实现分离。在电喷雾负离子模式下,采用多反应监测(MRM)模式检测,以结构相似物作为内标定量。结果表明,9种中药材中乙烯利在相应质量浓度范围内线性良好,相关系数(r)均大于0.99,检出限(LOD)和定量下限(LOQ)分别为0.5~1.0μg/kg和10~20μg/kg。在3个加标水平下,乙烯利在9种中药材中的平均回收率分别为71.6%~91.5%、77.1%~97.6%、84.7%~99.5%,日内相对标准偏差(RSD)均不大于6.5%。将该方法应用于93批实际样品检测,乙烯利的检出率为68.8%。该方法省时便捷、灵敏度高,能够满足多数中药材中乙烯利残留量的检测要求。

An ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)was developed for the determination of ethephon residue in traditional Chinese medicine.The traditional Chinese medicine samples were extracted and frozen with 50% methanol solution containing 1% formic acid after crushing,diluted with acetonitrile,purified by octadecyl bonded Silica gel(C_(18)),silica gel,graphitized carbon black(GCB),with 1.2% formic acid aqueous solution and 0.5% formic acid acetonitrile as mobile phase.Anionic Polar Pesticide Column(2.1 mm×100 mm,5μm)was used to separate the pesticide in 15 min.Multiple reaction monitoring(MRM)mode was used in electrospray negative ion mode,and structural similarity was used as internal standard.The results showed that ethephon in 9 kinds of traditional Chinese medicine were linear(r>0.99)in the corresponding mass concentration ranges,and the limits of detection(LODs)and limits of quantitation(LOQs)were 0.5-1.0μg/kg and 10-20μg/kg,respectively.The average recoveries with three concentration levels were 71.6%-91.5%,77.1%-97.6% and 84.7%-99.5%,with the intra-day relative standard deviations(RSDs)less than 6.5%.The method was applied to 93 batches of actual samples,and the detection rate of ethephon was 68.8%.The method is time-saving,convenient and sensitive,and can meet the requirements of detecting ethephon residue in most traditional Chinese medicine.