UPLC-Q-TOF-MS法分析苗药黑骨藤提取物中的化学成分

Analysis of Chemical Compositions in Miao Medicine Periploca forrestii Extract by UPLC-Q-TOF-MS

目的:建立黑骨藤提取物中化学成分的分析方法,为进一步阐明黑骨藤药效物质基础提供参考。方法:以70%乙醇为溶剂制备黑骨藤提取物。采用超高效液相色谱-四级杆-飞行时间质谱联用技术(UPLC-Q-TOF-MS)对提取物进行成分分析,色谱柱为Agilent Eclipse Plus C_(18),流动相为0.1%甲酸水-0.1%甲酸乙腈(梯度洗脱),流速为0.25 mL/min;采用电喷雾离子源,在负离子扫描模式下检测。通过碎片离子信息、相关文献以及对照品比对,确定黑骨藤提取物中的化学成分。结果:在黑骨藤提取物中共分离出了23个化合物,并确定了9个化合物。其中,5、8、9、16、17、18、22、23号峰通过与对照品比对分别确定为5-O-咖啡酰基奎宁酸、3-O-咖啡酰基奎宁酸、4-O-咖啡酰基奎宁酸、3,4-O-二咖啡酰基奎宁酸、3,5-O-二咖啡酰基奎宁酸、4,5-O-二咖啡酰基奎宁酸、杠柳毒苷、杠柳苷元;13号峰通过碎片离子和文献比对推断为(1S,3R,4R,5R)-3-{([2E)-3-(3,4-dihydroxyphenyl)-2-propenoyl]oxy}-1,5-dihydroxy-4-{([2E)-3-(4-hydroxy-3,5-dimethoxyphenyl)-2-propenoyl]oxy}cyclohexanecarboxylic acid。结论:建立的UPLC-Q-TOF-MS法可以快速、高效、准确地分析黑骨藤提取物中的化学成分。

OBJECTIVE:To establish a method for the analysis of chemical compositions in Periploca forrestii extract,and to provide reference for further elucidating the chemical basis of the P.forrestii.METHODS:70%ethanol was used to prepare P.forrestii extract.UPLC-Q-TOF-MS was adopted to analyze the chemical compositions of the extract.The determination was performed on an Agilent Eclipse Plus C18 column with mobile phase consisted of 0.1%formic acid water-0.1%formic acid acetonitrile(gradient eludtion)at the flow rate of 0.25 mL/min.ESI source was applied to analyze the chemical composition of P.forrestii extract in negative scanning ion mode.The chemical constituents of P.forrestii extract were confirmed by the fragment ion information,related literature and standard control comparison.RESULTS:Totally 23 compounds were separated from P.forrestii extract,and 9 compounds were indentified.The peaks of 5,8,9,16,17,18,22 and 23 were identified as 5-O-caffeoylquinic acid,3-O-caffeoylquinic acid,4-O-caffeoylquinic acid,3,4-O-dicaffeoylquinic acid,3,5-O-dicaffeoylquinic acid,4,5-Odicaffeoylquinic acid,glucoperiplocymarin and periplogenin by comparison with standard control.The peak of 13 was deduced as(1S,3R,4R,5R)-3-{[(2E)-3-(3,4-dihydroxyphenyl)-2-propenoyl]oxy}-1,5-dihydroxy-4-{[(2E)-3-(4-hydroxy-3,5-dimethoxyphenyl)-2-propenoyl]oxy}cyclohexanecarboxylic acid by comparison with fragment ion information and related literature.CONCLUSIONS:The established UPLC-Q-TOF-MS method can be used for the rapid,efficient and accurate analysis of the chemical constituents in P.forrestii extract.