The Establishment And Optimization Of Pretreatment Technology For Determination Of Chinese Herbal Medicine By HPLC

With the development of information technology,High-performance liquid chromatography?HPLC?is becoming one of the foremost analytical techniques used for scientific purposes and laboratory measurements.It is widely used in chemical,pharmaceutical,and biomedical analysis,as well as in drug treatment monitoring,due to its inherent ability to analyze,separate,and purify a variety of chemical samples,including,but not limited to,acidic,basic,and neutral analytes.However,effective research and reasonable mechanism can availably ensure that the resource is not only taken full advantage but also can meet the needs of the medicine at every stage to develop.Also,high-performance liquid chromatography?HPLC?has grown into a mature and wide technique due to its superior high-pressure infusion pump,high sensitivity,together with the excellent selectivity.Simultaneously,the application of an ultraviolet detector can effectively detect and analyze the organics in the process of separation.The mechanism of the method is that the difference of interaction between the sample to be tested and the stationary phase along with the mobile phase of the chromatographic column to make relative movement in the two phases.After repeated adsorption-desorption-distribution processes,the components have different retention time,aiming to realize the detection of the sample when each component successively flows out of the chromatographic column and enters the detector.On the basis of the method,the study of Chinese herbal medicine is the most representative,mainly because the flavonoids and organic acids in Chinese herbal medicine have many important physiological and biochemical effects on humans and animals.Besides,this method can accurately and quickly determine the identity,purity and physical characteristics of the sample.Thus,the way equipped with the preconcentration of micelle extraction and ultrasonic-assisted homogeneous ionic liquid microextraction has made great progress,suchas purification,enrichment and extraction,compared with other pre-treatment technologies.Meanwhile,it is good for flavonoids and organic acids to quickly detect and apply in clinical application.To date,more and more researchers are devoting to taking note of environmental protection and seeking an effective method to study medicine.In this paper,the flavonoids and organic acids in yidianhong,hawthorn and drunken herbs were extracted and determined by HPLC.The research report mainly consists of three parts as follow:Part 1 Determination of Eight active components content in Emilia sonchifolia by HPLCTo establish a method for the simultaneous determination and separation of eight flavonoids,organic acids and their contents of protocatechuic acid,coffeic acid,chastein,rutin,quercitrin,quercetin,luteolin and apigenin in the extracts of red medicinal material.Chromatographic analysis was carried out by ultrasound-assisted extraction coupled with high-performance liquid chromatography?HPLC?.Samples and reference materials were dissolved in the mobile phase for a constant volume.After filtration,20 microliters were absorbed and injected into high-performance liquid chromatography?HPLC?.Finally,the peak area is calculated by the external standard method.The experiment shows that the method is simple,fast,accurate,and has good reproducibility and stability.It provides an experimental basis for the quality control of medicinal plant Emilia sonchifolia.Part 2 Simultaneous Determination of Nine Phenolic Acids and Flavonoid Compounds in Hawthorn by Micelle Extraction-Cloud point preconcentration-High Performance Liquid ChromatographyTo establish a simple and rapid micellar extraction method combining a non-ionic surfactant,turbidimetric preconcentration,and ultrasonic extraction technology for the simultaneous separation and determination for the content of 9 active ingredients in hawthorn,including protocatechuic acid,caffeic acid,vitexin,ferulic acid,hypericin,quercetin,luteolin,kaolin,and apigenin.Methods: non-ionic surfactant Genapol X-080 was used as the extraction agent,and a certain amount of sodium chloride was added to perform turbidity point preconcentration.Factors such as surfactant concentration,solid-liquid volume,electrolyte concentration,pH value of solution extraction,ultrasonic time,equilibrium time and centrifugal time were investigated to optimize the parameters of micellar extraction and turbidity point preconcentration.The samples and reference materials were dissolved in the mobile phase for a constant volume.After filtration,20 microliters were absorbed andinjected into the high-performance liquid chromatography.After repeated separation by chromatographic column,the results were analyzed by the detector.It shortens the time of preconcentration and treatment before sample extraction is simple,fast and environmentally friendly,and provides a new and reliable analytical method for the extraction of flavonoids and organic acids from hawthorn.Part 3 Ultrasound-assisted Homogeneous Ionic Liquids Microextraction-High Performance Liquid Chromatography for the Determination of Five Flavonoids in Buddleja lindleyana FortuneTo establish an Ultrasound-Assisted Homogeneous Ionic Liquids Microextraction-High Performance Liquid Chromatography for the simultaneous separation and determination of the content of 5 active ingredients in Buddleja lindleyana Fortune,including quercetin,hesperidin,rutin,apigenin,acacin.Methods: With water-soluble ionic liquids as extractant and water as the transfer medium,the target was extracted by ultrasound-assisted extraction.Water-insoluble ionic liquids were formed by adding ionic pair reagents to the extract.Phase separation occurred after centrifugation,and the ionic liquids were detected and analyzed.Factors such as Ionic liquid concentration,solid-liquid volume,pH value of solvent extraction,the concentration of NH₄PF₆,ultrasonic time,ultrasonic power and ultrasonic temperature were investigated to optimize the parameters.This method integrates extraction,concentration,separation and purification.It does not use volatile organic solvents.It has the advantages of short extraction time and less sample consumption.It can be widely used in the extraction and analysis of active components in medicinal plants.