The Study On Systematic Separation Strategy And Equipment For Traditional Chinese Herb Compounds

The complicated of the chemical compounds in traditional Chinese herb is reflected in the polarity difference,the compound species,the structure similarity of the homologous compounds and so on.It has a great significance to study active components in Chinese herb and create a new type of Chinese medicine through Systematic separate of chemical components in Chinese herbs.Currently,the separation of chemical components in Chinese herb is the focal point and difficulty in the chemical compounds of traditional Chinese herb.Multidimensional chromatography,which has a strong separation capability,is widely used in the separation and analysis of components of herb.Construct the appropriate two-dimensional liquid chromatography will be possible for the systematic separation of traditional Chinese herb.At present,Reverse phase chromatography is the most popular separation technology,while Hydrophilic chromatography is an effective complementary for Reverse phase chromatography,which is used in the separation of strong polar components.Since the difference of the two separation mechanism,the construction of two-dimensional liquid chromatography system is in good orthogonality.Constract a two-dimensional chromatographic separation system to provide a direction for traditional Chinese herb.The significance of this study is to achieve the systematic separation of herb components.The object of the study are nine normal methanol extract of Chinese herb.The purpose is to construct two-dimensional liquid chromatography system with high separation efficiency.Through screening and analysis of samples,Analysis the rule of separation of samples under the two modes,constructing hydrophilic/reverse phase two-dimensional liquid chromatography and choosing the water phase sample of licorice under off-line hydrophilic/reverse phase separation mode.This mode has a good orthogonality.And design and build the automatic two-dimensional liquid chromatography device.The contents and results are as follows :?1?Using 27 samples in water solution,extracted with petroleum ether and ethyl acetate of Chinese herb as objects of the study.Under hydrophilic phase and reverse phase separation mode,and the samples' DAD detection wavelength is 254 nm.The wholewavelength ranges from 190 to 400 nm.With the retention ability of petroleum ether phase samples in hydrophilic phase chromatographic is bad.The resolution of samples in Ethyl acetate phase and water phase is good.Hydrophilic chromatography can separate homologous composition while reverse phase chromatography get a higher peak capacity.The two chromatography have different separation features and they are suitable for constructing two-dimensional liquid chromatography.So we can construct hydrophilic/reverse phase two-dimensional liquid chromatography to achieve the Systematic separation of herb components.?2?The separation strategy was validated by selecting the aqueous phase of licorice.Construct XIon/C₁₈ 2D off-line separation mode.Choosing Chromatographic condition and the cutting time of elution fractions to achieve the separation of homologous composition under the hydrophilic phase,the reverse phase chromatography could separate the covered small peak from the main components and raise the peak capacity.Combining XIon with C₁₈.The two-dimensional chromatography of XIon and C₁₈ is more targeted than pure seperation.Besides,the two dimensional liquid chromatography system has a higher orthogonality,the orthogonal degree is 60.57%.It is proved that the method of the hydrophilic/reverse phase two-dimensional liquid chromatography is feasible for the systematic separation of herb components.?3?Finally,developed and producted the two-dimensional liquid chromatography device,named Preparative Chromatography Platform,which is combined with separation system,testing system and automatic control system.The separation system is combined with one separation column,which is composed of Hydrophilic filler,three two-dimensional separation columns,which is composed of reversed phase filler,twenty primary enriched columns and ten secondary enriched columns.Hydrophilic chromatography can separate homologous composition while reverse phase chromatography get a higher peak capacity.The control system is combined with Siemens S7₃00.Chromatography Monitoring System utilize UV230 ? UV detector online monitoring,Wavelength ranges from 190 nm to 600nm.Online two dimensional liquid chromatography has high peak capacity,high resolution,high separation efficiency,low sample loss and contamination and other obvious advantages.