基于液相色谱质谱联用的复方鱼腥草合剂化学成分分析及特征图谱研究

Chemical constituents and fingerprint analysis of compound Yuxingcao mistura by liquid chromatography coupled with mass spectrometry

目的：建立复方鱼腥草合剂LC-MS特征图谱的质量评价方法,并对特征图谱中的主要成分进行鉴别。方法：采用Kromasil C18色谱柱（4.6 mm×250 mm,5μm）,流动相A为0.1%甲酸溶液,流动相B为甲醇,梯度洗脱（0~15 min,20%B→30%B;15~30 min,30%B→50%B;30~40 min,50%B→65%B;40~45 min,65%B→70%B）,流速1.0 m L·min-1;质谱负离子模式检测。结果：从复方鱼腥草合剂中检测到21个特征峰,通过对照品比对和文献参考,推断了其中的20个成分,包括（R,S）-告依春、3-咖啡酰奎尼酸/4-咖啡酰奎尼酸、绿原酸、马钱苷、7-O-ethyl sweroside、獐芽菜苷、异连翘酯苷、secoxyloganin、连翘酯苷A、4,5-二咖啡酰奎宁酸/3,5-二咖啡酰奎宁酸、金丝桃苷、异槲皮苷、槲皮苷、连翘苷、黄芩苷、baicalein-6-glucuronide、汉黄芩苷/baicalein 6-methylether-7-O-β-galactopyranuronoside;特征图谱方法验证结果表明精密度、重复性、稳定性良好。结论：本文建立的方法可用于复方鱼腥草合剂的整体质量评价。

Objective： To develop a fingerprint method for quality assessment of compound Yuxingcao mistura by liquid chromatography coupled with mass spectrometry（ LC- MS）. Methods： Separation was performed using Kromasil C18column（ 4. 6 mm × 250 mm,5 μm）. The mobile phase consisted of 0. 1% formic acid aqueous solution（ A） and methanol（ B） using a gradient elution of 20% B→30% B at 0- 15 min,30% B→50% B at 15- 30 min,50% B→65%B at 30- 40 min,65% B→70% B at 40- 45 min. The detection was performed by mass spectrometry using negative mode by selected ion monitoring（ SIM）. Results： Twenty one characteristic peaks were detected from compound Yuxingcao mistura,and 20 peaks,including（ R,S）- goitrin,3- caffeoylquinic acid /4- caffeoylquinic acid,chlorogenic acid,loganin,7- O- ethyl sweroside,sweroside,isoforsythiaside,secoxyloganin,forsythoside A,4,5- di- caffeoylquinic acid /3,5- di- caffeoylquinic acid,hyperoside,isoquercitrin,quercitrin,phillyrin,baicalin,baicalein- 6-glucuronide,wogonoside / baicalein 6- methylether- 7- O- β- galactopyranuronoside,were identified by comparison of the retention time and MS data with those of reference substances and reported in reference. The fingerprint method validation of compound Yuxingcao mistura showed good precision,reproducibility and stability. Conclusion： The developed LC- MS fingerprint method can be used for global assessment of the quality of compound Yuxingcao mistura.