| As a traditional Chinese medicinal material,licorice is often used as the main ingredient in prescriptions.It is widely distributed in our provinces such as Inner Mongolia,Xinjiang,Gansu and Ningxia.The Pharmacopoeia stipulates that the dried roots and rhizomes of Glycyrrhiza Uralensis,G.inflata Bat.and G.glabra L.can be used as medicine.Among them,Ural licorice has the widest distribution and the highest yield,which is the main species used.The main active ingredients of licorice are glycyrrhizic acid and licorice flavonoids.Among them,the fat-soluble isoprenoflavones in licorice are active in anti-tumor,anti-oxidation,liver protection,blood lipid reduction and other aspects,which are attracting widespread attention.However,due to the large amount of such ingredients present in the dregs after the licorice extract is extracted from licorice,they are discarded as waste and pollute the surrounding environment.So far,no method has been established to extract fat-soluble licorice flavonoids from these medicinal residues.At the same time,the poor water solubility of these ingredients results in low oral bioavailability and affects its efficacy.The purpose of this study is to establish a process for extracting fat-soluble total flavonoids from the dregs of Uralkan herbal medicine.On this basis,research and establish a total flavonoids nano self-microemulsion drug delivery system to improve its oral bioavailability and prolong its action time.Increase the curative effect,and explore its protective effect on liver injury through in vivo experiments in mice,in order to expand its use range,turn waste into treasure,and create greater economic and social benefits.The main research contents and results are as follows:（1）Study on the preparation process of total fat-soluble flavonoids in the dregs of Ural Glycyrrhizae:Using ethanol reflux to extract the fat-soluble flavonoids,based on single factors（ethanol concentration,reflux time,material-to-liquid ratio）,the extraction rate is used as the judgment standard,and positive According to the experiment,the optimal extraction method is optimized as follows:70%ethanol,reflux three times at a material-to-liquid ratio of 1:5,and extract the fat-soluble total flavonoids from the decocting residue in 2 hours each time.This method is used to extract the fat-soluble total flavonoids of Ural Glycyrrhiza uralensis.The total fat-soluble flavonoids in the residue of licorice,the content of fat-soluble flavonoids in the obtained sample was 28.5%,and the yield was 1.89%.（2）Enrichment of fat-soluble flavonoids in Glycyrrhiza uralensis:In order to increase the content of fat-soluble flavonoids in the extracted samples,the purification effects of DM-130 column chromatography and polyamide column chromatography on the extraction of fat-soluble flavonoids were compared.2 The adsorption,desorption and purification efficiency of two resins on the fat-soluble flavonoids of licorice,and finally determined the use of DM-130 macroporous adsorption column chromatography to enrich the fat-soluble flavonoids in the ethanol extract,which can make the content of the fat-soluble flavonoids obtained by the alcohol extraction From 28.5%to 48.3%,further extraction with dichloromethane can make the content of flavonoids reach 51.27%.（3）The preparation process of the fat-soluble total flavonoids of glycyrrhiza uralensis nano-self-microemulsion and nanoemulsion:compare the solubility of the total flavonoids of glycyrrhiza uralensis in different oil phases,different emulsifiers and co-emulsifiers,using the method of drawing pseudo-ternary phase diagrams,Select the best oil phase,emulsifier and co-emulsifier;On the basis of this experimental result,continue to optimize the formula,use the computer software Design-Expert 8 to design a two-factor,five-level star point design experiment,with the content of the oil phase,the mass ratio of emulsifier and co-emulsifier Km as independent variables,and nanometer The particle size and Zeta potential of the milk are the inspection indicators,and the total OD value is calculated as the reaction variable,and the best self-microemulsion composition and ratio are selected.TThe experimental results show that the optimal prescription of the fat-soluble total flavonoids of glycyrrhiza uralensis nano-self-microemulsion is ethyl oleate（23%）,EL-35 and PEG-400（2.6:1,77%）.and the prescription has the largest load.The drug amount is about 12.91%,the particle size of the formed emulsion is about 92.17 nm,and the zeta potential is 6.02m V,which is a stable nanometer self-microemulsion drug-carrying system.On the basis of the preparation process of glycyrrhiza uralensis fat-soluble total flavonoids nanoemulsion,through in vitro emulsification,a preliminary establishment of the preparation process of glycyrrhiza uralensis fat-soluble total flavonoids nanoemulsion oral liquid,using Zeta potential and particle size as parameters,examines the nano-self-microemulsion and The proportion of water and the stirring time are finally determined to be 60 mg per milliliter of self-microemulsion.The ratio of self-microemulsion to deionized water is 1:9.When preparing nanoemulsion,the stirring time should not be less than 1.5 h at 40°C.The fat-soluble total flavonoids nanoemulsion of Laer licorice prepared by the process is initially observed to be a stable nanoemulsion stable system.（4）Stability experiment of glycyrrhiza fat-soluble total flavonoids nanoemulsion:Place the prepared nanoemulsions at 4°C,room temperature（20°C）,and 60°C acceleration for 0,15,30,45,and 60 d.Take samples,measure the particle size and the change of Zeta potential,and examine the stability of the nanoemulsion.The results show that the particle size of the nanoemulsion gradually increases as the temperature increases,and the Zeta potential gradually decreases.At 4°C,60 Within a day,the particle size of nanoemulsion increased from 93.22±4.25 to 121.07±5.13,and the zeta potential decreased from5.28±0.68 to 4.02±0.46;the particle size of nanoemulsion increased from 92.71±5.93 to180.52±7.61 at 20℃,and the zeta potential increased from 5.31±0.44,down to 3.33±0.21;under the acceleration of 60℃,the particle size of the nanoemulsion increased from93.59±3.98 to 380.13±14.67,and the zeta potential decreased from 5.15±0.29 to1.78±0.37.The preliminary stability test proved that the nanoemulsion was 4℃Under refrigerated conditions,it is basically stable within 60 days.（5）The protective effect of liposoluble total flavonoids of ural licorice nano-self-microemulsion on liver injury in mice:establish a mouse model of acute liver injury with CCl4,use silibinin as the positive drug,and compare the glycyrrhiza uralensis fat-soluble total flavonoids nano-self-microemulsion The protective effect of milk and non-nano total flavonoids on acute liver injury,taking the reduction of alanine aminotransferase（GPT）and aspartate aminotransferase（GOP）as the inspection indicators,the results showed that under the condition of 300 mg/kg,the fat-soluble total flavonoids of ural Both microemulsion and non-nano total flavonoids can reduce GPT and GOP,but after nanometerization,the effect is increased by about 20%,which proves that the fat-soluble flavonoids of licorice uralensis are made into nanoemulsion,which improves the bioavailability of the drug,thereby increasing its bioavailability. |
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