色谱

色谱

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超高效液相色谱-电喷雾三重四极杆质谱法测定中药中马兜铃酸A和B的含量

刘萤, 韩深, 冯骞, 王金花*   

  1. 北京出入境检验检疫局检验检疫技术中心, 北京 100026
  • 收稿日期:2011-05-17 修回日期:2011-06-28 出版日期:2011-11-28 发布日期:2011-12-25
  • 通讯作者: 王金花,博士,高级工程师. Tel: (010)58619247,
  • 基金资助:

    北京市科委计划项目(D08050200310805)

Determination of aristolochic acids A and B in Chinese herbals and traditional Chinese patent medicines using ultra high performance liquid chromatography-triple quadrupole mass spectrometry

LIU Ying, HAN Shen, FENG Qian, WANG Jinhua*   

  1. Inspection and Quarantine Technical Center of Beijing Entry-Exit Inspection and Quarantine Bureau, Beijing 100026, China
  • Received:2011-05-17 Revised:2011-06-28 Online:2011-11-28 Published:2011-12-25

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摘要： 利用超高效液相色谱-电喷雾三重四极杆质谱仪建立了中药中马兜铃酸A和B的定性定量分析方法。选取柴胡、生甘草、桔梗、龙胆泻肝丸、消胖丸、减肥茶等14种代表性样品,用甲醇-水(70:30, v/v)溶液加热回流提取,经Oasis MAX固相萃取柱富集净化后,在Eclipse RP HD C18反相柱(150 mm×2.1 mm, 1.8 μm)上进行分离;流动相为5 mmol/L乙酸铵水溶液(pH 7.5)-乙腈(75:25, v/v)。采用电喷雾离子源正离子模式(ESI+)和多反应监测模式(MRM)进行质谱分析。马兜铃酸A和B的线性范围分别为0.5～200 μg/L和1～200 μg/L,相关系数(r2)均大于0.995;检出限(LODs)分别为5 μg/kg和7.5 μg/kg;定量限(LOQs)分别为12.5 μg/kg和25 μg/kg。在100 μg/kg和500 μg/kg添加水平下,马兜铃酸A和B的回收率(n=6)范围分别为60.3%～96.4%和61.3%～94.7%,相对标准偏差均不大于10.2%。该方法灵敏度高,重复性好,操作简便,适用于中药材、饮片及中成药中马兜铃酸A和B的痕量检测。

关键词: 超高效液相色谱-串联质谱, 固相萃取, 马兜铃酸, 中药

Abstract: A method for the simultaneous determination of aristolochic acids A and B in some Chinese herbals and traditional Chinese patent medicines by ultra high performance liquid chromatography-triple quadrupole mass spectrometry (UHPLC-MS/MS) was developed. Fourteen samples, including Radix Bupleuri, Radix Glycyrrhizae, Radix Platycodonis, Longdanxieganwan, Xiaopangwan, Slimming Tea, etc., were extracted with methanol-water (70:30, v/v) and purified with Oasis MAX solid-phase extraction cartridges, then analyzed on an Eclipse RP HD C18 column (150 mm×2.1 mm, 1.8 μm) using 5 mmol/L ammonium acetate solution (pH 7.5)-acetonitrile (75:25, v/v) as the mobile phase. The mass spectrometric acquisition was carried out by means of electrospray ionization in positive mode (ESI+) with multiple reaction monitoring (MRM) method. The good linearities (r2＞0.995) were achieved within the ranges of 0.5~200 μg/L and 1~200 μg/L for aristolochic acids A and B, respectively. The limits of detection (LODs) were 5 μg/kg for aristolochic A and 7.5 μg/kg for aristolochic B, and the limits of quantification (LOQs) were 12.5 μg/kg and 25 μg/kg, respectively. The recoveries of aristolochic acids A and B at the spiked levels of 100 μg/kg and 500 μg/kg ranged from 60.3% to 96.4% with the relative standard deviations (RSDs) not more than 10.2%. The results demonstrated that the proposed method is efficient, sensitive, reproducible, reliable and suitable for the trace determination of aristolochic acids A and B in Chinese herbals and traditional Chinese patent medicines.

Key words: aristolochic acid, solid-phase extraction (SPE), traditional Chinese medicine, ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS)