超高效液相-串联质谱法测定镇脑宁胶囊中7种马兜铃酸类成分  

Simultaneous Determination of Seven Aristolochic Acid Analogues in Zhennaoning Capsules by Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry

作　　者：周颖 郑成 戴忠[2] 王娟娟 李俊行 朱蓉 陈碧莲 马双成 ZHOU Ying;ZHENG Cheng;DAI Zhong;WANG Juanjuan;LI Junhang;ZHU Rong;CHEN Bilian;MA Shuangcheng(NMPA Key Laboratory for Quality Evaluation of Traditional Chinese Medicine,Zhejiang Institute for Food and Drug Control,Hangzhou 310052,China;National Institutes for Food and Drug Control,Beijing 100050,China)

机构地区：[1]浙江省食品药品检验研究院,国家药品监督管理局中成药质量评价重点实验室,杭州310052 [2]中国食品药品检定研究院,北京100050

出　　处：《中国药学杂志》2023年第21期1955-1961,共7页Chinese Pharmaceutical Journal

基　　金：国家药品标准制修订研究课题资助(2021Z16-2)。

摘　　要：目的 建立镇脑宁胶囊中马兜铃酸类成分的超高效液相-串联质谱联用检测分析方法。方法 采用Phenomemex Kinetex (2.1 mm×100 mm, 2.6μm)色谱柱,以甲醇-体积分数0.1%甲酸溶液(含1mmol·L-1乙酸铵)为流动相梯度洗脱,流速0.3 mL·min-1,以正离子多反应监测模式,建立同时测定镇脑宁胶囊中马兜铃酸Ⅲa、7-羟基马兜铃酸Ⅰ、马兜铃酸Ⅳa、马兜铃酸Ⅱ、马兜铃内酰胺BⅡ、马兜铃内酰胺Ⅰ、马兜铃酸Ⅰ共7种马兜铃酸类成分的液质联用检测方法。结果 马兜铃酸Ⅲa、7-羟基马兜铃酸Ⅰ、马兜铃酸Ⅳa、马兜铃酸Ⅱ、马兜铃内酰胺BⅡ、马兜铃内酰胺Ⅰ、马兜铃酸Ⅰ分别在1.241~1 240.572、1.034~1 034.056、2.109~2 108.709、1.070~1 070.378、1.064~1 064.000、2.154~2 154.022、1.086~1 085.922 ng·mL-1内线性关系良好;平均加样回收率分别为101.8%(RSD1.84%)、101.9%(RSD 2.67%)、102.2%(RSD 2.74%)、102.1%(RSD 2.13%)、101.8%(RSD 2.60%)、100.7%(RSD 0.96%)、104.1%(RSD 6.15%)。10批样品均检出马兜铃酸Ⅳa、马兜铃内酰胺Ⅰ和马兜铃酸Ⅰ等马兜铃酸类成分,含量范围分别为3.875~6.913,2.984~6.072及0.350~1.719 ng·g-1,而马兜铃酸Ⅲa、7-羟基马兜铃酸Ⅰ、马兜铃酸Ⅱ和马兜铃内酰胺BⅡ均未检出。结论 建立的同时测定7种马兜铃酸类成分的液质联用方法准确、可靠。镇脑宁胶囊中检出马兜铃酸Ⅳa、马兜铃内酰胺Ⅰ和马兜铃酸Ⅰ。其中,马兜铃酸Ⅳa、马兜铃内酰胺Ⅰ毒性较低或无明显毒性,而真正可能有毒性的是含硝基基团的马兜铃酸Ⅰ,因部分样品马兜铃酸Ⅰ含量超出限度,提示该制剂中需严格控制马兜铃酸Ⅰ的含量。OBJECTIVE To establish the determination method for aristolochic acid analogues in Zhennaoning Capsules.METHODS A Phenomemex Kinetex(2.1 mm×100 mm,2.6μm)column was used with methanol-0.1%formic acid(containing 1 mmol·L-1 ammonium acetate)as mobile phase gradiently eluted at a flow rate of 0.3 mL·min-1.By using multi-reaction monitoring mode(MRM),an ultra-high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS)method was established for simultaneous determination of seven aristolochic acid analogues including aristolochic acidⅢa,7-hydroxy aristolochic acidⅠ,aristolochic acidⅣa,aristolochic acidⅡ,aristolactam BⅡ,aristolactamⅠ,and aristolochic acidⅠ.RESULTS The linear relationships for aristolochic acidⅢa,7-hydroxy aristolochic acidⅠ,aristolochic acidⅣa,aristolochic acidⅡ,aristolactam BⅡ,aristolactamⅠ,and aristolochic acidⅠwere good in the ranges of 1.241-1240.572,1.034-1034.056,2.109-2108.709,1.070-1070.378,1.064-1064.000,2.154-2154.022,1.086-1085.922 ng·mL-1,respectively.And their respective average recovery rates were 101.8%(RSD 1.84%),101.9%(RSD 2.67%),102.2%(RSD 2.74%),102.1%(RSD 2.13%),101.8%(RSD 2.60%),100.7%(RSD 0.96%),104.1%(RSD 6.15%),respectively.Among ten batches of samples,aristolochic acidⅣa,aristolactamⅠand aristolochic acidⅠwere detected and the content range was 3.875-6.913,2.984-6.072 and 0.350-1.719 ng·g-1,respectively,meanwhile,there were no aristolochic acidⅢa,7-hydroxy aristolochic acidⅠ,aristolochic acidⅡand aristolactam BⅡdetected.CONCLUSION The established UHPLC-MS/MS method is accurate and reliable.Aristolochic acidⅣa,aristolactamⅠand aristolochic acidⅠwere detected in Zhennaoning Capsules.Among them,aristolochic acidⅣa and aristolactamⅠhave low toxicity or no obvious toxicity,while the toxin substance is aristolochic acidⅠwith nitro group.Because the content of aristolochic acidⅠin some samples exceeds the limit,it suggests that the content of aristolochic acidⅠin this preparation needs to be strictly contr

关 键 词：镇脑宁胶囊 马兜铃酸类成分 超高效液相色谱-串联质谱法 

分 类 号：R284[医药卫生—中药学]