毛茛化学成分HPLC、UPLC/Q-TOF-MS分析
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摘要
毛茛Ranunculus japonicus Thunb.为毛茛科毛茛属植物显花植物,英文名Japan Buttercup,以带根全草入药。夏秋采集,切段,鲜用或晒干用,多年生草本生于田野、路边、沟边、山坡杂草丛中;东北至华南都有分布。别名鱼疔草、鸭脚板、野芹菜、山辣椒、老虎脚爪草、毛芹菜、起泡菜。辛、微苦,温,有毒。民间用于治疗多种疾病,如胃痛,黄疸,疟疾,淋巴结结核,等,主治黄疸,哮喘,疟疾,偏头痛,牙痛,鹤膝风,风湿关节痛,目生翳膜,瘰疬,痈疮肿毒。现代研究表明毛茛具有抗炎,镇痛,抗菌的药理作用,对心脑血管的药理作用显著,为探明毛茛药效物质基础,进一步研究毛茛的药理作用机制,本课题首次采用UPLC/Q-TOF-MS联用技术等现代分析技术开展了毛茛化学成分的分析鉴定研究,主要内容和结果如下:
(一)利用HPLC-PDA对毛茛总苷的化学成分进行分析,建立了毛茛总苷的HPLC-PDA色谱图的色谱方法;
(一)利用UPLC/Q-TOF-MS对毛茛总苷的化学成分进行分析和快速鉴定,鉴定了毛茛总苷中的20种成分,其中主要为黄酮及内酯类成分;
(二)利用HPLC-PDA对毛茛醇提物中的生物碱类化学成分进行分析,建立了毛茛生物碱的HPLC-PDA色谱图的色谱方法
(四)利用UPLC/Q-TOF-MS技术对毛茛醇提物中的生物碱类化学成分成分进行了分析和鉴定;鉴定了毛茛醇提物中的15种成分,其中12种为生物碱类成分;
本课题首次采用HPLC,UPLC/Q-TOF-MS色谱联用技术鉴定了毛茛总苷中的主要化学成分,为探明毛茛总苷药理作用的物质基础提供了依据,建立了一种快速分析鉴定中药中复杂成分的方法,以供其他药学工作者参考;另外,本课题首次采用HPLC,UPLC/Q-TOF-MS色谱联用技术分析并鉴定了毛茛醇提物中的生物碱类成分,因此类成分在毛茛中的含量较少,为其他中药中非常量成分的鉴定提出了一种快速可行的方法学参考,同时也有助于进一步探明毛茛的药效物质基础。
Ranunculus japonicus Thunb is a widely spreaded from northeast to southChina perennial herb in China,which belongs to family of Ranunculaceae used as herb in medicine.collected in summer or autumn used by cut off、fresh or open-air drying. perennial herb in field ,roadside,border, clivas. There are some synonyms for it such as Yuliaocao,Yajiaoban,Shanlajiao,Tigerfoot herb Qipaocai and Maoqincai. Acrid,subtle bitter.warm, poisonous.This plant has Pharmacological activity, such as antichloristic, analgesia, antibacterial and antitumor. And it is used as chinese fork medicine in the treatment of Jaundice, Tuberculosis, and Hepatitis. to try to find out the material foundation of pharmacodynamic action of Ranunculus japonicus Thunb and to progress studying pharmacologic action of Ranunculus japonicus Thunb, this topic carried out the study of analysis and identification by the combination technique of Ultra-performance liquid chromatography with quadruple-time-of-flight mass spectrometry (UPLC/Q-TOF-MS). Principal work and main result showed as follow:
(1) analysis the the main components in the total glycosides of Ranunculus japonicus by HPLC-PDA,to set up a method of chromatographic process of HPLC-PDA of total glycosides of Ranunculus japonicus;
(2) Rapid analysis of the main components in the total glycosides of Ranunculus japonicus by UPLC/Q-TOF-MS first time,identified 20 compounds from the total glycosides of Ranunculus japonicus,most of them are flavanoid and lactone compond;
(3) Analysis the the main components in the alkaloids extract of Ranunculus japonicus by HPLC-PDA,to set up a method of chromatographic process of HPLC-PDA of alkaloids of Ranunculus japonicus;
(4) Analysis of the main components in alkaloid extract of Ranunculus japonicus by UPLC/Q-TOF-MS first time. identifed 15 compounds from Ranunculus japonicus,12 compounds of them are alkaloids .
This study first time identified 20 compounds from the total glycosides of Ranunculus japonicus by UPLC/Q-TOF-MS,supply the evidence to the material foundation of pharmacodynamic action of Ranunculus japonicus;set up a analysis method for complicated components of chinese traditional medicine to the pharmaceutical workers as a reference.Aslo this work analysis and identified alkaloids of Ranunculus japonicus by UPLC/Q-TOF-MS first time. This work introduced a rapid feasible method for analyzing and identifing the minor constituent of complicatied components.
(一)利用HPLC-PDA对毛茛总苷的化学成分进行分析,建立了毛茛总苷的HPLC-PDA色谱图的色谱方法;
(一)利用UPLC/Q-TOF-MS对毛茛总苷的化学成分进行分析和快速鉴定,鉴定了毛茛总苷中的20种成分,其中主要为黄酮及内酯类成分;
(二)利用HPLC-PDA对毛茛醇提物中的生物碱类化学成分进行分析,建立了毛茛生物碱的HPLC-PDA色谱图的色谱方法
(四)利用UPLC/Q-TOF-MS技术对毛茛醇提物中的生物碱类化学成分成分进行了分析和鉴定;鉴定了毛茛醇提物中的15种成分,其中12种为生物碱类成分;
本课题首次采用HPLC,UPLC/Q-TOF-MS色谱联用技术鉴定了毛茛总苷中的主要化学成分,为探明毛茛总苷药理作用的物质基础提供了依据,建立了一种快速分析鉴定中药中复杂成分的方法,以供其他药学工作者参考;另外,本课题首次采用HPLC,UPLC/Q-TOF-MS色谱联用技术分析并鉴定了毛茛醇提物中的生物碱类成分,因此类成分在毛茛中的含量较少,为其他中药中非常量成分的鉴定提出了一种快速可行的方法学参考,同时也有助于进一步探明毛茛的药效物质基础。
Ranunculus japonicus Thunb is a widely spreaded from northeast to southChina perennial herb in China,which belongs to family of Ranunculaceae used as herb in medicine.collected in summer or autumn used by cut off、fresh or open-air drying. perennial herb in field ,roadside,border, clivas. There are some synonyms for it such as Yuliaocao,Yajiaoban,Shanlajiao,Tigerfoot herb Qipaocai and Maoqincai. Acrid,subtle bitter.warm, poisonous.This plant has Pharmacological activity, such as antichloristic, analgesia, antibacterial and antitumor. And it is used as chinese fork medicine in the treatment of Jaundice, Tuberculosis, and Hepatitis. to try to find out the material foundation of pharmacodynamic action of Ranunculus japonicus Thunb and to progress studying pharmacologic action of Ranunculus japonicus Thunb, this topic carried out the study of analysis and identification by the combination technique of Ultra-performance liquid chromatography with quadruple-time-of-flight mass spectrometry (UPLC/Q-TOF-MS). Principal work and main result showed as follow:
(1) analysis the the main components in the total glycosides of Ranunculus japonicus by HPLC-PDA,to set up a method of chromatographic process of HPLC-PDA of total glycosides of Ranunculus japonicus;
(2) Rapid analysis of the main components in the total glycosides of Ranunculus japonicus by UPLC/Q-TOF-MS first time,identified 20 compounds from the total glycosides of Ranunculus japonicus,most of them are flavanoid and lactone compond;
(3) Analysis the the main components in the alkaloids extract of Ranunculus japonicus by HPLC-PDA,to set up a method of chromatographic process of HPLC-PDA of alkaloids of Ranunculus japonicus;
(4) Analysis of the main components in alkaloid extract of Ranunculus japonicus by UPLC/Q-TOF-MS first time. identifed 15 compounds from Ranunculus japonicus,12 compounds of them are alkaloids .
This study first time identified 20 compounds from the total glycosides of Ranunculus japonicus by UPLC/Q-TOF-MS,supply the evidence to the material foundation of pharmacodynamic action of Ranunculus japonicus;set up a analysis method for complicated components of chinese traditional medicine to the pharmaceutical workers as a reference.Aslo this work analysis and identified alkaloids of Ranunculus japonicus by UPLC/Q-TOF-MS first time. This work introduced a rapid feasible method for analyzing and identifing the minor constituent of complicatied components.
引文
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