高效液相色谱-质谱法快速鉴定复方毛冬青冲剂中三萜皂苷活性成分

Rapid Identification of Triterpene and Saponin in Compound Radix Ilicis Pubescentis Medicinal Granules by High Performance Liquid Chromatography-Mass Spectrometry

建立了高效液相色谱-质谱法(HPLC-MS)快速鉴定复方毛冬青冲剂中三萜皂苷活性成分的方法。以甲醇为萃取剂超声萃取复方毛冬青冲剂30min。采用高效液相色谱-离子阱质谱(HPLC-IT-MS)和高效液相色谱-飞行时间质谱(HPLC-TOF-MS)对萃取液进行分析,选用Agilent Zorbax Eclipse XDB C18色谱柱(250mm×4.6mm,5μm),以水(含0.1%甲酸)-乙腈为流动相进行梯度洗脱,柱后流出液采用电喷雾离子阱质谱(ESI-IT-MS)和电喷雾飞行时间质谱(ESI-TOF-MS)的正、负离子模式进行检测。检测结果经离子阱一级质谱(IT-MS1)、离子阱二级质谱(IT-MS2)和分析时间质谱(TOF-MS)信息分析,并与相关文献报道进行比较,鉴定出1种三萜酸和8种三萜皂苷成分,并推测了其它3种可能的三萜皂苷化学成分,通过对照品对比分析,三萜酸确证为Ilexgenin A,其中一种三萜皂苷确证为Ilexsaponin A1。本方法无需对照品即可快速有效地鉴定出复方毛冬青冲剂中的三萜皂苷活性成分,为建立冲剂的质量标准提供了依据。

A method was developed for the rapid identification of triterpene and saponin in compound Radix ilicis pubescentis medicinal granules by high performance liquid chromatography-mass spectrometry（HPLC-MS）.The sample was extracted with methanol by ultrasonic wave extraction for 30 min.The extract was analyzed by high performance liquid chromatography-ion trap-mass spectrometry（HPLC-IT-MS） and high performance liquid chromatography-time of fight-mass spectrometry（HPLC-TOF-MS）.An Agilent Zorbax Eclipse XDB C18 column（250 mm×4.6 mm,5 μm） was applied in the HPLC analysis with water（0.1% formic acid） and acetonitrile as mobile phase.The elutes were detected by IT-MS and TOF-MS to obtain the positive-and negative-ion MS1 and MS2 spectra together with extract molecular weights.A triterpenic acid and eight triterpenoid saponins were identified,and three structures of other triterpenoid saponins were deduced by comparison of that reported in literatures.The triterpenic acid was confirmed to be ilexgenin A and one of the triterpenoid saponins was confirmed to be ilexsaponin A1 by reference substances.Consequently,the method can be performed to identify the triterpene and saponin constituents in compound Radix ilicis pubescentis medicinal granules quickly and effectively even without the reference substances,and provided essential scientific data and information for its quality control.