提取方法及毛细管电泳法在中药研究中的应用
摘要
本文对中药有效成分的提取和分离检测方法进行了研究。将超声技术和微波技术应用于中药有效成分提取的研究中,建立了三种新的提取方法。利用毛细管电泳法对中药中有效成分进行了定量测定,并研究了中药有效成分与蛋白的相互作用。
研究了三种中药有效成分提取新方法,包括超声雾化提取法、高压微波辅助提取法和微波辅助无溶剂提取法。在超声提取法的基础上,设计了一套超声雾化提取装置。该方法是在提取剂变成气溶胶的情况下完成提取,与传统的超声提取方法相比,提取时间与提取剂用量均有所改进;建立了一种利用高压微波辅助提取法提取中药蜜丸中有效成分的方法,这一方法是将蜜丸药涂在载玻片上,而后进行提取,缩短了提取时间,提高了提取效率;通过在样品中加入对微波能有强烈吸收作用的固体介质,利用介质吸收微波,加热样品,加热速度快,使得在提取干燥中草药中挥发油成分时无需任何前处理过程成为可能。
用毛细管电泳法测定了高良姜、贯叶连翘、菟丝子、地锦草、刺五加、银杏叶和满山红7 种不同中药中槲皮素,以及槐米和槐花中槲皮素和芦丁的含量。详细考察了电泳分离参数,包括缓冲剂种类、缓冲溶液添加剂、分离电压、缓冲溶液pH 值、进样量、运行温度等。总结了每种药的最佳电泳分离条件,并对样品分析的回收率和精密度进行了测定,建立了这些中药中槲皮素和芦丁的毛细管测定方法。
采用亲和毛细管电泳(ACE)技术,对木犀草素、槲皮素和山柰酚这三种黄酮类化合物与人血清白蛋白(HSA)的结合常数进行了研究,并同荧光猝灭法进行了对比。针对已有荧光猝灭法计算结合常数的公式一般采用一定程度的近似,利用Matelab6.5 软件编制了一套迭代计算结合常数的程序Fluo1.0,使荧光猝灭法计算结合常数时不需任何假定和近似,更真实地反映了实际情况。
The analytical chemistry plays an important part in traditional Chinese medicine research. The applications of analytical chemistry in traditional Chinese medicine research involve the extraction, separation and characterization of the effective constituents in the medicines.
In the thesis, three new extraction methods to extract the effective constituents from traditional Chinese medicines were developed. The first method is ultrasonic nebulization extraction (UNE) method. A experimental device was designed and assembled to extract quercetin from Rhizoma Alpiniae Officinarum. This device was refitted with the ultrasonic humidifier. The effective constitutes in the Chinese drug are extracted at aerosol state. The qucercetin was determined by capillary electrophoresis. Compared with ultrasonic extraction (UE), the amount of extraction solvent and extraction time can be reduced when the UNE was applied. The second method is pressurized microwave-assisted extraction. Pill is the very common shape in Chinese traditional patent medicine. The pill was pilled by mixing the honey with the powder of the traditional Chinese medicine. The pills have strong viscosity and
the solvent couldn’t easily infiltrate into them. So, to extract the effective constituents from the pills is a hard work. A new extraction method was developed. The pills were coated on the surface of the slide glass and then the slide glass was put in the WR-3TC special pressurized microwave-assisted extraction system to finish the extraction. The flavonoids from Huaiyu pills were extracted by this way and total flavonoids were determined by spectrophotometey. Compared with the conventional pressurized microwave-assisted extraction, the extraction time is shorter, extraction yield is higher and amount of extraction solvent is less when the new extraction method was applied. The third method is improved solvent-free microwave extraction (SFME) method. This method can be applied to extract the volatile oil from the dry traditional Chinese medicine sample without any solvent and sample pretreatment. The volatile oil components from the Zanthoxylum Bungeanum Maxim were extracted by this way and determined by GC-MS. Compared with the conventional SFME, The extraction time is shorter when the improved SFME method was applied.
The determination of the querctin in Rhizoma Alpiniae Officinarum, Hypericum Periforatum, Semen Cuscuta, Herba Euphorbiae Humifusae, Acanthopanax, Ginkgo Biloba L. Leaves and Rhododendron Dauricuml, and simultaneous determination of the querctin and rutin in Flos Sophorae Buds and Flos Sophorae Flowers were performed by capillary zone electrophoresis (CZE). In this study, the conditions of CZE, such as running voltage, running temperature, injection pressure and time and the components of the buffer solutions were examined in detail. The experimental results for recovery and precision determination showed that CZE is a accurate, fast and highly efficient method to separate and determine the effective constituents from traditional Chinese medicine.
研究了三种中药有效成分提取新方法,包括超声雾化提取法、高压微波辅助提取法和微波辅助无溶剂提取法。在超声提取法的基础上,设计了一套超声雾化提取装置。该方法是在提取剂变成气溶胶的情况下完成提取,与传统的超声提取方法相比,提取时间与提取剂用量均有所改进;建立了一种利用高压微波辅助提取法提取中药蜜丸中有效成分的方法,这一方法是将蜜丸药涂在载玻片上,而后进行提取,缩短了提取时间,提高了提取效率;通过在样品中加入对微波能有强烈吸收作用的固体介质,利用介质吸收微波,加热样品,加热速度快,使得在提取干燥中草药中挥发油成分时无需任何前处理过程成为可能。
用毛细管电泳法测定了高良姜、贯叶连翘、菟丝子、地锦草、刺五加、银杏叶和满山红7 种不同中药中槲皮素,以及槐米和槐花中槲皮素和芦丁的含量。详细考察了电泳分离参数,包括缓冲剂种类、缓冲溶液添加剂、分离电压、缓冲溶液pH 值、进样量、运行温度等。总结了每种药的最佳电泳分离条件,并对样品分析的回收率和精密度进行了测定,建立了这些中药中槲皮素和芦丁的毛细管测定方法。
采用亲和毛细管电泳(ACE)技术,对木犀草素、槲皮素和山柰酚这三种黄酮类化合物与人血清白蛋白(HSA)的结合常数进行了研究,并同荧光猝灭法进行了对比。针对已有荧光猝灭法计算结合常数的公式一般采用一定程度的近似,利用Matelab6.5 软件编制了一套迭代计算结合常数的程序Fluo1.0,使荧光猝灭法计算结合常数时不需任何假定和近似,更真实地反映了实际情况。
The analytical chemistry plays an important part in traditional Chinese medicine research. The applications of analytical chemistry in traditional Chinese medicine research involve the extraction, separation and characterization of the effective constituents in the medicines.
In the thesis, three new extraction methods to extract the effective constituents from traditional Chinese medicines were developed. The first method is ultrasonic nebulization extraction (UNE) method. A experimental device was designed and assembled to extract quercetin from Rhizoma Alpiniae Officinarum. This device was refitted with the ultrasonic humidifier. The effective constitutes in the Chinese drug are extracted at aerosol state. The qucercetin was determined by capillary electrophoresis. Compared with ultrasonic extraction (UE), the amount of extraction solvent and extraction time can be reduced when the UNE was applied. The second method is pressurized microwave-assisted extraction. Pill is the very common shape in Chinese traditional patent medicine. The pill was pilled by mixing the honey with the powder of the traditional Chinese medicine. The pills have strong viscosity and
the solvent couldn’t easily infiltrate into them. So, to extract the effective constituents from the pills is a hard work. A new extraction method was developed. The pills were coated on the surface of the slide glass and then the slide glass was put in the WR-3TC special pressurized microwave-assisted extraction system to finish the extraction. The flavonoids from Huaiyu pills were extracted by this way and total flavonoids were determined by spectrophotometey. Compared with the conventional pressurized microwave-assisted extraction, the extraction time is shorter, extraction yield is higher and amount of extraction solvent is less when the new extraction method was applied. The third method is improved solvent-free microwave extraction (SFME) method. This method can be applied to extract the volatile oil from the dry traditional Chinese medicine sample without any solvent and sample pretreatment. The volatile oil components from the Zanthoxylum Bungeanum Maxim were extracted by this way and determined by GC-MS. Compared with the conventional SFME, The extraction time is shorter when the improved SFME method was applied.
The determination of the querctin in Rhizoma Alpiniae Officinarum, Hypericum Periforatum, Semen Cuscuta, Herba Euphorbiae Humifusae, Acanthopanax, Ginkgo Biloba L. Leaves and Rhododendron Dauricuml, and simultaneous determination of the querctin and rutin in Flos Sophorae Buds and Flos Sophorae Flowers were performed by capillary zone electrophoresis (CZE). In this study, the conditions of CZE, such as running voltage, running temperature, injection pressure and time and the components of the buffer solutions were examined in detail. The experimental results for recovery and precision determination showed that CZE is a accurate, fast and highly efficient method to separate and determine the effective constituents from traditional Chinese medicine.
引文
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