基于UPLC-Q-TOF-MS技术分析不同变种来源葛花的化学成分差异性

Difference Analysis of Chemical Constituents in Different Varieties of Puerariae Flos by UPLC-Q-TOF-MS

目的:比较野葛花、粉葛花及葛麻姆花3个不同变种来源葛花的化学成分差异。方法:采用超高效液相色谱-四极杆-飞行时间质谱法(UPLC-Q-TOF-MS),以0.1%甲酸水溶液(A)-乙腈(B)为流动相进行梯度洗脱(0~20 min,10%~30%B;20~30 min,30%~55%B;30~35 min,55%~95%B;35~37 min,95%B;37~40 min,95%~10%B),流速0.25 mL·min^(-1);运用电喷雾离子源(ESI),在正、负离子模式下分别进行扫描并采集MS数据,扫描范围m/z 50~1 500。结合文献信息及化学成分数据库对葛花化学成分进行鉴定,并通过MarkerViewTM1.2.1软件对所得数据归一化处理,应用SICMA-P 14.1软件对3个变种来源葛花的MS数据进行主成分分析(PCA)和正交偏最小二乘法-判别分析(OPLS-DA),筛选出不同变种来源葛花的显著差异性成分。结果:从3个不同变种来源的葛花中共鉴定35个化合物,其中异黄酮类成分22种,黄酮类成分6种,皂苷类成分7种;野葛花、粉葛花及葛麻姆花分别包含了32,35,33个。正、负离子模式下筛选得到葛花苷,鸢尾苷,6"-O-木糖鸢尾苷,黄豆黄苷,4’-甲氧基鸢尾黄素-7-葡萄糖苷,6"-O-木糖黄豆黄苷,葛花苷元,槐花皂苷Ⅲ,6"-O-丙二酰基黄豆苷,次葛花苷,鸢尾苷元,芦丁,大豆皂苷BB,牡荆素,鹰嘴豆芽素A,染料木苷,葛花亭,赤豆皂苷Ⅱ共18个差异化合物。结论:该研究首次对葛麻姆花中化学成分进行了较为系统的研究,发现葛麻姆花虽作为混伪品,但鸢尾苷和6"-O-木糖鸢尾苷的含量较高,具有较大的开发潜力。葛花苷、鸢尾苷等功效成分在3个变种来源的葛花之间具有显著差异,将这3个变种均作为葛花应用需谨慎考量。

Objective: To compare the chemical constituents of Puerariae Flos from three different varieties of Pueraria montana var. lobata,P. montana var. thomsonii and P. montana var. montana. Method:Ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS)was used with the mobile phase of 0.1% formic acid aqueous solution(A)-acetonitrile(B)for gradient elution(0-20 min,10%-30%B;20-30 min,30%-55%B;30-35 min,55%-95%B;35-37 min,95%B;37-40 min,95%-10%B),the flow rate was 0.25 mL·min^(-1). Electrospray ionization(ESI)was used to scan and collect MS data in positive and negative ion modes with scanning range of m/z 50-1 500. The chemical components from different sources of Puerariae Flos were identified in combination with the chemical composition database and literature information. After the obtained data were normalized by MarkerViewTM1.2.1,they were imported into SICMA-P 14.1 software for principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA) to select the main differentiated components among the three different varieties. Result: A total of 35 compounds were identified from three different varieties of Puerariae Flos,including 22 isoflavones,6 flavonoids and 7 saponins. The flowers of P. lobata,P. montana var. thomsonii and P. montana var. montana contained 32,35,33 compounds,respectively. And 18 differential compounds were screened under the positive and negative ion modes,including kakkalide,tectoridin,6 ″-O-xylosyl-tectoridin,4’-methyltectorigenin-7-glucoside, glycitin, 6 ″-O-xylosyl-glycitin, irisolidone, kaikasaponin Ⅲ, 6 ″-Omalonylglycitin,kakkalidone,tectorigenin,rutin,soyasaponin BB,vitexin,biochanin A,genistin,kakkatin,azukisaponin Ⅱ. Conclusion: This research is the first to systematically study the chemical constituents of the flower of P. montana var. montana,although the flower of P. montana var. montana is used as adulterants,it has high contents of tectoridin and 6 ″-O-xylosyl-tectoridin,which has great potential for development. The efficacy components such as kakkalide and tectoridin in Puerariae Flos from the three sources of varieties are obviously different,and it is necessary to carefully consider the application of these three varieties as Puerariae Flos.