[关键词]
[摘要]
该研究建立了超高效液相色谱-串联三重四极杆串联质谱法(UPLC-QQQ-MS/MS)同时测定细辛中4种马兜铃酸的分析方法。样品用70%甲醇溶解,通过超声提取,采用Waters ACQUITY UPLC HSS T3 (2.1 mm×100 mm×1.8 μm)色谱柱,以0.1%甲酸水溶液-甲醇为流动相梯度洗脱分离,流速0.3 mL/min,柱温40 ℃,采用电喷雾离子源(ESI),正离子模式扫描,多反应监测模式(MRM),对马兜铃酸A,马兜铃酸B,马兜铃酸C,马兜铃酸D进行检测。结果表明马兜铃酸A,马兜铃酸B,马兜铃酸C,马兜铃酸D在4~100 μg/L浓度范围内线性关系均良好(r≥0.995),检出限分别为3.00 μg/kg、5.00 μg/kg、25.00 μg/kg、20.00 μg/kg,加标回收率在91.97%~106.40%之间,RSD≤4.24%(n=9)。该方法专属性强,准确度和精密度高,线性范围广,适用于细辛中马兜铃酸A,马兜铃酸B,马兜铃酸C,马兜铃酸D四种物质的同时检测,检测结果表明,细辛及含细辛的中成药中含有一定量的马兜铃酸A和马兜铃酸D,均不含有马兜铃酸B和马兜铃酸C,该数据对细辛日常食用及药用安全、以及细辛类中成药的质量控制提供科学依据。
[Key word]
[Abstract]
In this study, an analytical method for the simultaneous determination of four aristolochic acids in asarum by ultra-high performance liquid chromatography-tandem triple quadrupole mass spectrometry (UPLC-QQQ-MS/MS) was established. The samples were dissolved with 70% methanol, extracted ultrasonically, and separated on a Waters ACQUITY UPLC HSST3 chromatographic column (2.1 mm×100 mm×1.8 μm) using 0.1% formic acid in water -methanol as the mobile phase in gradient elution. The flow rate was 0.3 mL/min, and the column temperature was 40 ℃. Aristolochic acid A, aristolochic acid B, aristolochic acid C and aristolochic acid D were analyzed using electrospray ionization (ESI), positive ion mode scanning and multiple reaction monitoring mode (MRM). Results showed that aristolochic acid A, aristolochic acid B, aristolochic acid C and aristolochic acid D showed good linear relationships within the concentration range of 4~100 μg/L (r≥0.995). The detection limits were 3.00 μg/kg, 5.00 μg/kg, 25.00 μg/kg and 20.00 μg/kg, respectively, with the spiked recoveries ranging from 91.97% to 106.40% and RSD≤4.24% (n=9). The method had high specificity, accuracy, precision and wide linear range, and is suitable for simultaneous determination of aristolochic acids A, aristolochic acid B, aristolochic acids C, aristolochic acids D in asarum. The test results show that asarum and asarum-containing Chinese traditional medicine had certain amounts of aristolochic acid A and aristolochic acid D, but did not contain aristolochic acid B and aristolochic acid C. The data provide a scientific basis for safe daily consumption and medicinal use of asarum, and the quality control of asarum-containing Chinese traditional medicines.
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[基金项目]
广州市科技计划项目(202102080001);广东大学科技创新战略专项(pdjh2020b1153);广东省药品监督管理局科技创新项目(2021TDZ09;2021YDZ06)