基于液质联用技术的白芍净制研究

Study on the clean-processing of Paeoniae Radix Alba by UPLC-MS

目的采用液质联用技术,对比研究白芍及其根皮物质基础,初步探讨白芍净制的科学内涵。方法将收集到的白芍除去头尾及细根,置于沸水中煮10分钟左右取出,刮去外皮,晾干,得到白芍饮片和白芍根皮。采用Waters UPLC HSST3色谱柱,以乙腈-0.1%甲酸水为流动相进行梯度洗脱,全扫描/数据依赖二级质谱扫描(Full MS/dd MS2)模式分析,根据碎片信息及参考文献对白芍和白芍根皮的成分进行分析鉴定。采用赛默飞的质谱分析软件(Thermo Xcalibur Qual Browser)提取白芍及白芍根皮中化学成分的峰强度,通过对比各成分的峰强度,对白芍及白芍根皮中的化学成分的峰强度(含量)进行分析。结果白芍及其根皮中共解析出38种化合物,其中白芍中未检测到脱落酸和邻苯二甲酸二丁酯,白芍根皮中未检测到没食子酸甲酯。白芍与其根皮中所含化学成分的含量差异较大,白芍中芍药苷、没食子酰基芍药苷、4-羟基白芍苷、牡丹皮苷(F、I、E、H)、氧化芍药苷、没食子酸、没食子酸甲酯等16种成分含量高于白芍根皮;芍药内酯苷、芍药内酯苷B、鞣花酸、原儿茶酸、山奈酚、脱落酸、邻苯二甲酸二丁酯等18种成分含量在白芍根皮中较高。结论白芍去皮除去了杂质和非药效成分;经水煮、去皮后,白芍与同来源的赤芍化学成分的种类及含量均发生变化,是二者药效具有差异的主要原因,初步阐释了白芍净制的科学内涵。

Objective To compare the material basis of Paeoniae Radix Alba(PRA)and its root bark(PRARB)by UPLC-MS and reveal the scientific connotation of clean-processing of PRA preliminarily.Methods The collected PRA was removed the“head”,“tail”and fine roots.Then the roots were dealt with by scraping off the barks after boiling in water for ten minutes.After drying,the PRA and PRARB were collected.Full scanning/data-dependent mass spectrometry was used for analysis by Ultra High Performance Liquid Chromatography-LTQ-Orbitrap mass spectrometer(UPLC-MS),Waters UPLC HSST3 column and a mobile phase of acetonitrile-0.1%formic acid for gradient elute analysis.The chemical components of PRA and PRARB were identified through the fragmentation behavior obtained by MS/MS,accurate molecular weight,and molecular formula with literature.By comparing the peak intensities of each component,which were extracted by Thermo Xcalibur Qual Browser(mass spectrometry software),the peak intensity(content)of the chemical components in PRA and PRARB was analyzed.Results A total of 38 compounds were identified in PRA and PRARB,among which abscisic acid and dibutyl phthalate were not detected in PRA,and methyl gallate was not detected in PRARB.The contents of chemical components contained in PRA and PRARB were quite different.The contents of 16 components in PRA,such as paeoniflorin,galloylpaeoniflorin,4-hydroxy-paeoniflorin,mudanpioside(F,I,E,H),oxypaeoniflorin,gallic acid,methyl gallate,etc.,were higher than that of PRARB.The contents of 18 components,including albiflorin,albiflorin B,ellagic acid,protocatechuic acid,kaempferol,abscisic acid,and dibutyl phthalate,etc.,were higher in PRARB.Conclusion Processing of peeling removed impurities and non-medicinal ingredients of PRA;after boiling and peeling,the types and contents of the chemical components of PRA and paeonies radix rubra of the same origin had changed,which was the main reason for the difference in medicinal efficacy.The study preliminarily explained about the scientific connotation of clean-processing of PRA.