贝母中生物碱及核苷类成分分析研究
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摘要
中药贝母来源于百合科贝母属植物的鳞茎,在我国被用于止咳祛痰已有数百年的历史。药理研究表明,生物碱部分是其主要活性部位。贝母水提物具有抗菌、抗高血压及抑制血小板凝聚等作用,因而,目前也有人认为非生物碱也有可能为有效活性成分。本文主要建立高效液相色谱-电喷雾质谱、紫外联用方法,研究了浙贝母和川贝母中生物碱及核苷类成分。
本论文主要研究内容如下:
1.建立了高效液相色谱-质谱(HPLC-ESI/MS)联用技术分析贝母生物碱的方法,并对川贝母(F.cirrhosa D.Don)与浙贝母(F.thunbergii Miq.)中贝母甲素、贝母乙素的含量进行了测定与比较。此方法可用于对市场上川贝母和浙贝母的质量监控。
2.利用LC-ESI-MS~n技术能在线提供化合物结构信息、灵敏度高等特点,首次从川贝母及浙贝母中发现了腺嘌呤、尿苷和鸟苷,建立了基于核苷类成分对川贝母和浙贝母进行品种鉴别的方法。
3.LC/MS有着高灵敏度、分析时间短的优点,可用以鉴定及分析中草药中的活性成分。本实验采用HPLC-UV-ESI/MS对浙贝母和川贝母中的核苷类成分进行定量分析,本方法灵敏度高,简便快捷,线性关系良好,适用于植物中核苷类化学成分的检测与分析。
4.本文采用制备高效液相色谱法对浙贝母中的核苷类成分(尿苷、鸟苷、腺苷和胸苷)进行分离纯化。该方法简便易行,产物纯度高,且有效降低了有机溶剂的使用量。
Bulbus Fritillariae,a well-known traditional Chinese medicine (TCM).It is derived from the bulbus of various Fritillariae species.And it has been used as antitussive and expectorant drugs for centuries. Various chemical and pharmacological studies on Fritillariae have demonstrated that isosteroidal alkaloids are the main bioactive ingredients in Fritillariae.Currently,some naturally occurring non-alkaloid constituents in aqueous extracts of Fritillaria species have attracted much attention for their other various pharmacological activities such as remarkable antibacterial actions,anti-hypertension effect and platelet aggregation inhibition.This article mainly established methods for study on the alkaloids and nucleosides in the bulbus Fritillariae by high performance liquid chromatography coupled with electrospray ionization mass spectrometry or ultraviolet spectrometry.
The studies in this thesis are summarized as follows:
1.An HPLC-ESI/MS method was developed for analysis of alkaloids in Bulbus Fritillariae.And this method has been utilized to determine and compare of the content of verticine and verticinone in F.cirrhosa D.Don and F.thunbergii Miq.This method can be applied to quality supervise for F.cirrhosa D.Don and F.thunbergii Miq.in the market.
2.Using the LC-ESI-MS~n technology for discovering nucleosides in Bulbus Fritillariariae cirrhosae and Fritillariariae thunbergii Miq.. Because this technology can provide information structure of compounds on line and has high sensitivity.Adenine,uridine and guanosine in Fritillariae species are firstly ascertained.A LC-ESI-MS~n method for the species discrimination of Bulbus Fritillariariae cirrhosae and Fritillariariae thunbergii Miq.was reported based on the nucleosides components.
3.LC/MS method has advantages of high sensitivity and short analysis time,etc.So it can be utilized to identify and analyze active components in TCM.A rapid,high sensitivity and reproducible HPLC-UV-ESI/MS technique was developed for the quantitative analysis of nucleosides components in Bulbus Fritillariariae cirrhosae and Fritillariariae thunbergii Miq..It is also suitable for the detection and analysis of nucleosides components in other plants.
4.Purification and separation of uridine,guanosine,adenosine and thymidine in F.thunbergii Miq.by preparative HPLC were discussed. The method of preparation is simple,feasible and effectively economizes organic solvent.
本论文主要研究内容如下:
1.建立了高效液相色谱-质谱(HPLC-ESI/MS)联用技术分析贝母生物碱的方法,并对川贝母(F.cirrhosa D.Don)与浙贝母(F.thunbergii Miq.)中贝母甲素、贝母乙素的含量进行了测定与比较。此方法可用于对市场上川贝母和浙贝母的质量监控。
2.利用LC-ESI-MS~n技术能在线提供化合物结构信息、灵敏度高等特点,首次从川贝母及浙贝母中发现了腺嘌呤、尿苷和鸟苷,建立了基于核苷类成分对川贝母和浙贝母进行品种鉴别的方法。
3.LC/MS有着高灵敏度、分析时间短的优点,可用以鉴定及分析中草药中的活性成分。本实验采用HPLC-UV-ESI/MS对浙贝母和川贝母中的核苷类成分进行定量分析,本方法灵敏度高,简便快捷,线性关系良好,适用于植物中核苷类化学成分的检测与分析。
4.本文采用制备高效液相色谱法对浙贝母中的核苷类成分(尿苷、鸟苷、腺苷和胸苷)进行分离纯化。该方法简便易行,产物纯度高,且有效降低了有机溶剂的使用量。
Bulbus Fritillariae,a well-known traditional Chinese medicine (TCM).It is derived from the bulbus of various Fritillariae species.And it has been used as antitussive and expectorant drugs for centuries. Various chemical and pharmacological studies on Fritillariae have demonstrated that isosteroidal alkaloids are the main bioactive ingredients in Fritillariae.Currently,some naturally occurring non-alkaloid constituents in aqueous extracts of Fritillaria species have attracted much attention for their other various pharmacological activities such as remarkable antibacterial actions,anti-hypertension effect and platelet aggregation inhibition.This article mainly established methods for study on the alkaloids and nucleosides in the bulbus Fritillariae by high performance liquid chromatography coupled with electrospray ionization mass spectrometry or ultraviolet spectrometry.
The studies in this thesis are summarized as follows:
1.An HPLC-ESI/MS method was developed for analysis of alkaloids in Bulbus Fritillariae.And this method has been utilized to determine and compare of the content of verticine and verticinone in F.cirrhosa D.Don and F.thunbergii Miq.This method can be applied to quality supervise for F.cirrhosa D.Don and F.thunbergii Miq.in the market.
2.Using the LC-ESI-MS~n technology for discovering nucleosides in Bulbus Fritillariariae cirrhosae and Fritillariariae thunbergii Miq.. Because this technology can provide information structure of compounds on line and has high sensitivity.Adenine,uridine and guanosine in Fritillariae species are firstly ascertained.A LC-ESI-MS~n method for the species discrimination of Bulbus Fritillariariae cirrhosae and Fritillariariae thunbergii Miq.was reported based on the nucleosides components.
3.LC/MS method has advantages of high sensitivity and short analysis time,etc.So it can be utilized to identify and analyze active components in TCM.A rapid,high sensitivity and reproducible HPLC-UV-ESI/MS technique was developed for the quantitative analysis of nucleosides components in Bulbus Fritillariariae cirrhosae and Fritillariariae thunbergii Miq..It is also suitable for the detection and analysis of nucleosides components in other plants.
4.Purification and separation of uridine,guanosine,adenosine and thymidine in F.thunbergii Miq.by preparative HPLC were discussed. The method of preparation is simple,feasible and effectively economizes organic solvent.
引文
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