The Critical Parammeters Of Shuanghuanglian Preparation On The Effect Of Ethanol Precipitation Based On Floc Micromorphology

BackgroundEthanol precipitation was a refining method in the production process of traditional Chinese medicine preparations.The operation method was to extract the medicinal materials with water,then add a certain volume fraction of ethanol to the water extract to precipitate.Finally,the precipitated impurities were removed by filtration.Ethanol precipitation had the advantages of simple operation,solvent safety,low equipment cost,and high clarity of chemicals.Therefore,it was not been completely replaced by high-speed centrifugation,macroporous adsorption resin,flocculation,and other membrane separation and purification processes.The ethanol precipitation supernatant was mainly investigated,and the content of active ingredients in the ethanol precipitation supernatant was studied in previous researchs.Therefore,based on previous research,this paper not only investigated the index components of the ethanol precipitation supernatant of Shuanghuanglian preparation,but also fractal theory and quantitative image analysis technology were used to carry out micromorphological research of ethanol precipitation flocs.By examining the effects of process parameters on the micromorphological changes of ethanol precipitation flocculant and the effect of ethanol precipitation,and the internal relationship between the two,it provided a theoretical basis for fine control of ethanol precipitation process.Objective1.In order to establish the foundation for the research of ethanol precipitation process parameters on ethanol precipitation effect and floc micromorphology,the Shuanghuanglian preparation were selected for screening the critical process parameters affecting its ethanol precipitation effect.2.The method for determining the floc micromorphology of ethanol precipitation flocs was established to provide a support for accurately expressing the Chinese medicine compound preparation of the ethanol precipitation flocs.3.The influence of critical process parameters of Shuanghuanglian preparation on the floc micromorphology and effect of ethanol precipitation was researched,and the intrinsic relationship between the floc micromorphology and effect of ethanol precipitation was discussed.Then,the reason for the change of active ingredient by the critical process parameters of ethanol precipitation was analyzed.Methods1.Referring to the contents of the related preparations in the Chinese Pharmacopoeia,Shuanghuanglian preparation was selected as the research subjects.Then,the effect of process parameters such as ethanol precipitation condition,ethanol to material ratio,the standing temperature,stirring speed,the density of extract,,added ethanol concentration,flow rate of ethanol,ratio of ethanol to material ethanol,precipitation time and temperature of the extract on the index component comprehensive retention rate and solid removal rate by using single factor test design was investigated.Finally,the critical process parameters of ethanol precipitation of Shuanghuanglian preparation were screened out.2.By using optical microscopy imaging technology to obtain the original image of Shuanghuanglian preparation ethanol precipitation flocs.The effects of background correction,enhanced contrast,noise,threshold segmentation,and floc sampling methods was investigated on microscopic parameters of the image from the aspects of floc compactness,size,elongation,and regularity.Finally,the method for determining the fractal dimension of ethanol precipitation flocs was established.3.The effects of critical process parameters of Shuanghuanglian preparation on Micro parameters,index ingredient retention rate and filter cake moisture content were investigated.Based on this foundation,the intrinsic relationship between the morphology of ethanol precipitation flocs and the loss of component was explored,the reason for the change of active ingredient by the critical process parameters of ethanol precipitation was analyzed.Results1.Based on the index components comprehensive retention rate and solids removal rate,the key technological parameters of ethanol precipitation of Shuanghuanglian preparation were screened.The results showed that the density of extract and the ethanol to material ratio compared the comprehensive retention rate and solids removal of the index components of Shuanghuanglian preparation.The rate had a significant effect.And at different levels of the same key process parameters,the similarity of the chemical fingerprints of the ethanol precipitation supernatant was greater than 0.95,and the relative retention time RSD of the common peaks were less than 3%.The chemical composition difference in the ethanol precipitation supernatant was affected by the same critical process of different levels of parameters was small.2.Through experimental research,some processing on the original image,the original image of ethanol precipitation flocs could be converted into microscopic image for binarization,such as background correction,linear gray level transformation,improved contrast,setting mean filter and median filter to eliminate gaussian noise and salt and pepper noise.After the establishment of micro parameters method of the ethanol precipitation flocs,some experimental operating parameters were determined,including the sampling volume of flocs was set at 200～400 μL,the flocs was diluted to 5～10 times,the microscopic image was magnified 100 times or 200 times and selecting 7～9 image acquisition sites.Finally,the stable and reliable method for determining the fractal dimension of the ethanol floe was established.3.From the influence of the critical process parameters on micro parameters and ethanol precipitation effect,the results showed that with the increase of the ethanol to material ratio,the flocs changed from small floc to large floc and accelerated the floc collision to increase the floc compactness,reduced the encapsulation of the ethanol precipitation supernatant,thereby the effective component retention was increased.The area of the floc was changed with the density of extract,increasing the encapsulation of the active ingredient was increased.Finally,the effective ingredient retention rate was reduced.Conclusions1.The critical process parameters affecting the ethanol precipitation effect were determined for Shuanghuanglian preparation.It provided a reference for the reasonable application and equipment design of ethanol precipitation process of Shuanghuanglian preparation.2.By establishing stable and reliable method to measure the micro parameters of Shuanghuanglian preparation ethanol precipitation flocs,it provided a foundation for the introduction of fractal theory and quantitative image analysis into the basic theory of ethanol precipitation process.3.By analyzing the intrinsic relationship between the micro parameters of flocs and the effect of ethanol precipitation,the loss of active ingredient that affected by the critical process parameters of ethanol precipitation was clarified.The above results had important practical significance for solving the problems existing in the current Shuanghuanglian preparation ethanol precipitation process.