摘要
目的:建立超高效液相色谱-串联质谱(UPLC-MS/MS)法检测正天丸中马兜铃酸Ⅰ、马兜铃酸Ⅱ、马兜铃酸Ⅲa、马兜铃酸Ⅳa、马兜铃内酰胺Ⅰ、马兜铃内酰胺Ⅱ的含量。方法:采用Waters ACQUITY UPLC BEH C_(18)(100 mm×2.1 mm,1.7μm)色谱柱,以乙腈-0.1%甲酸溶液(含5 mmol·L^(-1)甲酸铵)为流动相进行梯度洗脱,流速为0.3 mL·min^(-1),柱温40℃;采用电喷雾离子源(ESI),多反应监测模式(MRM),正离子模式检测,定量离子对为m/z 359.2→298.2(马兜铃酸Ⅰ)、m/z 329.2→268.2(马兜铃酸Ⅱ)、m/z345.2→282.2(马兜铃酸Ⅲa)、m/z 375.2→312.2(马兜铃酸Ⅳa)、m/z 294.2→279.2(马兜铃内酰胺Ⅰ)、264.1→206.2(马兜铃内酰胺Ⅱ)。结果:马兜铃酸Ⅰ、马兜铃酸Ⅱ、马兜铃酸Ⅲa、马兜铃酸Ⅳa、马兜铃内酰胺Ⅰ及马兜铃内酰胺Ⅱ的线性范围分别为1.614~103.3、6.486~1038、3.293~1054、6.270~1003、6.180~988.8、3.098~991.2 ng·m L^(-1),线性关系良好(相关系数r均大于0.9996);平均回收率分别为82.8%~104.6%、90.7%~98.4%、94.4%~105.1%、91.5%~113.7%、92.3%~99.9%、85.9%~102.0%,RSD分别为2.8%~10.8%、3.0%~5.8%、1.5%~2.5%、1.7%~6.7%、3.4%~6.2%、3.3%~7.2%;12批正天丸中检出马兜铃酸Ⅰ质量分数为52.35~70.15 ng·g^(-1),马兜铃酸Ⅳa质量分数为2394.29~3281.55 ng·g^(-1),马兜铃内酰胺Ⅰ质量分数为139.85~172.20 ng·g^(-1),未检出马兜铃酸Ⅱ、马兜铃酸Ⅲa和马兜铃内酰胺Ⅱ。结论:所建立的方法专属性强,灵敏度和准确度均较高,可用于正天丸中6种马兜铃酸类成分的含量测定,为正天丸及含马兜铃酸中药制剂的安全使用和质量控制提供参考。
Objective:To establish a UPLC-MS/MS method for simultaneous detection of 6 aristolochic acids(aristolochic acidⅠ,aristolochic acid,aristolochic acidⅡa,aristolochic acidⅣa,aristolochic lactamⅠand aristolochic lactamⅠ)in Zhengtian Pills.Methods:The separation was performed on a Waters ACQUITY UPLC BEH C_(18)column(100 mm×2.1 mm,1.7μm)by gradient program with acetonitrile-0.1%formic acid solution(including 5 mmol·L^(-1)ammonium formate)as mobile phase.The flow rate was set at 0.3 mL·min^(-1)and the column temperature was 40 C.Detection and quantification were performed by mass spectrometry in multiple reaction monitoring(MRM)mode with m/z 359.2→298.2 for aristolochic acid I,m/z 329.2→268.2 for aristolochic acidⅡ,m/z 345.2→282.2 for aristolochic acidⅢa,m/z 375.2→312.2 for aristolochic acidⅣa,m/z 294.2→279.2 for aristolochic lactamⅠand m/z 264.1→206.2 for aristolochic lactamⅡ.Results:The linear ranges of aristolochic acidⅠ,aristolochic acidⅡ,aristolochic acidⅡa,aristolochic acidⅣa,aristolochic lactamⅠand aristolochic lactamⅡwere 1.614-103.3 ng·m^(L-1),6.486-1038 ng·mL^(-1),3.293-1054 ng·mL^(-1),6.270-1003 ng·mL^(-1),6.180-988.8 ng:mL^(-1)and 3.098-991.2 ng`mL^(-1)respectively.All ingredients showed good linear relationship(r≥0.9996).The average recoveries were 82.8%-104.6%,90.7%-98.4%,94.4%-105.1%,91.5%-113.7%,92.3%-99.9%,85.9%-102.0%,respectively,with RSDs of 2.8%-10.8%,3.0%-5.8%,1.5%-2.5%,1.7%-6.7%,3.4%-6.2%,3.3%-7.2%,respectively.In 12 batches of Zhengtian Pills,the contents of aristolochic acidⅠwere 52.35-70.15 ng·g^(-1),the contents of aristolochic acid IV a were 2394.29-3281.55 ng·g^(-1),and the contents of aristolochic lactamⅠwere 139.85-172.20 ng·g^(-1),while aristolochic acidⅡ,aristolochic acidⅢa and aristolochic lactamⅡwere not detected.Conclusion:The established method is specific,sensitive and accurate,and can be used to determine the contents of six aristolochic acids in Zhengtian Pills,providing reference for the safe use and quality control of Zhengtian Pills and traditional Chinese medicine preparations containing aristolochic acid.
作者
余碧莲
谢凯莉
罗晴
赵锦江
张挺
严萍
YU Bi-lian;XIE Kai-li;LUO-Qing;ZHAO Jin-jiang;ZHANG Ting;YAN Ping(Guangzhou University of Chinese Medicine,Guangzhou 511492,China;China Resources Sanjiu Medical&Pharmaceutical Co.,Ltd.,Shenzhen 518110,China)
出处
《药物分析杂志》
北大核心
2025年第6期968-977,共10页
Chinese Journal of Pharmaceutical Analysis
