| In this paper,the total flavonoids of Bauhinia championii(Benth.)Benth.were extracted by the method of cellulase-ethanol synergistic extraction,and the cell wall of plant was removed by the high specificity from the enzyme.The flavonoids could be dissolved into the solution,and the necessary five polyoxymethyl flavonoids can be extracted more simultaneously,then purified by silica gel column chromatography,make the content of components namely 5,6,7,5'-tetramethoxy-(PMF1),5,6,7,3 ',4',5'-hexamethoxyflavone(PMF2),5,6,7,3',4'(PMF3),4 ',5'-methylenedioxy-5,7,3'-trimethoxyflavone(PMF4)and 5,7,3',4',5'-(PMF5)more than 50%,so it meet the standard of new drug registration,and finally the above five polyoxoflavone flavonoids would be prepared for the total flavonoids of Polygonum scoparia.Through in vitro dissolution test,the use of solid dispersion technology can significantly increase the in vitro dissolution of the total flavonoids of Radix Polygoni Multiflori.Because of the low extraction efficiency of the five flavonoids,the solid dispersions can be used to save the raw materials,reduce the dosage,decrease pollution and save human resource.The single factor method is preferred for the preparation of the solid dispersions of the total flavonoids for Radix Rehmannia glutinosa,and provides some experimental support for the further application of solid dispersion technology in Traditional Chinese Medicine.Methods:(1)The total flavonoids of Bauhinia championii(Benth.)Benth.were extracted by Cellulase-Ethanol,and the Box-Behnken effect surface was optimized on the basis of single factor test.The extraction rate of flavonoids as the reference,using index Design expert7.0.0 software to analyze and deal the data with ultraviolet spectrophotometer for testing.(2)Extraction of the total flavonoids of Bauhinia championii(Benth.)Benth.from the solution of(1)was carried out with ethyl acetate to obtain ethyl acetate layer.The ethyl acetate layer was separated and purified by silica gel column chromatography to obtain 5 polyoxoflavones which I need.(3)To establish HPLC method and method validation for determination of the content for five polyoxymethyl flavonoids and their total cantent.(4)The dispersions of polyvinylpyrrolidone,polyethylene glycol 4000 and polyethylene glycol 6000 were prepared by solvent method and solvent-melting method respectively.The optimum disperse was prepared by single factor method.The content of dissolution was tested by HPLC.(5)Finally,the solid dispersions were identified and identified by infrared spectroscopy and differential scanning calorimetry(DSC).Results:(1)The optimum conditions of the total flavonoids extracted by cellulase-ethanol were as follows: 9.13 mg / g of enzyme dosage,enzymolysis time for 2.06 h,enzymolysis temperature set at 51.03 ?.The extraction rate was 19.62% under the condition of software prediction.The triplicate extraction rate was 19.61%,18.98% and 19.14% respectively,the average value was 19.24% and the RSD was 1.68%,which indicated that the method was simple and reliable.(2)the The use of silica gel column chromatography and dextran gel chromatography macroporous resin and other common separation operation,from which the extraction of the five polyoxymethyl flavonoids was achieved.(3)linear equation for PMF1: y=5259x –70054,R2=0.99986,the range from 13.888 ng to 833.28 ng.The linearity of PMF2 is: y = 5647x-105057,R2 = 0.99975,linear range: 23.865 ng~1431.9 ng;The linear equation for PMF3 is: y=3364 x-29205,R2 =0.99997,range:10.2~612 ng;The linear range of PMF4 is: y = 3895x-18524,R2 = 0.99948,the linear range is: 5.982 ng~ 359.82ng;the linear equation of PMF5 is: y= 4600x-82390,R2 = 0.99995,linear range: 19.68 ng ~ 1180.8 ng.The recoveries were 100.56%(RSD = 1.46%),99.94%(RSD = 1.02%),100.23%(RSD = 0.92%),101.87%(RSD = 2.95%)and 101.10%(RSD = 1.91%),respectively.The five flavonoids contents were 17.76%,2.98%,32.31%,0.59% and 18.01% respectively,and the total content was 71.65%.(4)The accumulative dissolution of the total flavonoids of Polygonum cuspidatum prepared with polyvinylpyrrolidone was significantly higher than that of polyethylene glycol 6000 and polyethylene glycol 4000,and the accumulative dissolution rate reached up to 81.85%.(5)The results of the characterization of the PVB K-30 solid dispersion were tested as: the peak of the characteristic hydrogen bonds for excipient and Polygoni Multiflori at the peak of 3524.45 cm-1 and the apparent peak of the carbonyl at 1642.01 cm-1 was decreased significantly.The results of the differential scanning calorimetry(DSC)characterization were as follows: the PVPK30 solid dispersion was formed,and the melting point of Polygonum cuspidatum was completely disappeared at 130 ? ~ 140 ?.Conclusion:(1)The extraction rate of total flavonoids extracted from cellulase-ethanol was higher than that of alcohol,the ultrasonic method and microwave method.As a result,this method can be used for the extraction of total flavonoids.(2)The method for detecting the content of five polyoxymethyl flavonoids from Bauhinia championii(Benth.)Benth.was established.The method has the advantages of simple operation,high precision,good reproducibility and good stability,can be used for the determination of total flavonoids from Bauhinia Championii(Benth.)Benth..(3)The solid dispersions of the flavonoids for Bauhinia Championii(Benth.)Benth.prepared by PVP K-30 were better in vitro,and the dissolution rate of solid dispersions was higher when the ratio of raw materials and excipients was 1: 6.(4)The results of infrared spectroscopy and differential scanning calorimetry showed that preparation of solid dispersions of flavonoids was successful. |
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