| Objective:To take Ficus microcarpa L.f.as the research object,and to investigate the purification process of macroporous resin with the contents of total flavonoids,vitexin,isovitexin,and polysaccharides.The preparation of the extract provides a reference.Taking the physical and chemical parameters of the resin purification process and the target components and impurity polysaccharide content as the research object,the correlation between them is studied to provide a reference for actual production monitoring.The product purified by macroporous resin and alcohol precipitation process was taken as the research object,and the four flavonoid components were used as indicators to study its in vitro absorption characteristics.Method:NaNO2-Al(NO3)3-NaOH colorimetric method was used to determine the total flavonoids of Ficus microca,pa L.f.,with rutin as the control,the total flavonoid content was measured at the wavelength of 500 nm.And isovitexin;the phenol-sulfuric acid method was used to determine the Ficus microcarpa L.f.polysaccharide,with glucose as the standard,and the polysaccharide content was determined at 490 nm.Using static adsorption and desorption experiments,the adsorption and desorption effects of 6 different types of macroporous resins were investigated,and the adsorption time and temperature were studied.Using dynamic adsorption and desorption experiments,the main process parameters such as sample solution concentration,volume,sample solution pH,ethanol concentration,eluent dosage,and flow rate were investigated and compared with the alcohol precipitation process results.The correlation between total flavonoids,(vitexin+isovitexin),and impurity polysaccharide content was analyzed by stepwise regression and gray correlation analysis.Inverted intestinal sac method was used to compare the absorption characteristics of macroporous resin and alcohol precipitation process.Results:Rutin was in the concentration range of 0.01～0.07 mg/mL,the standard curve equation was A=11.5450C-0.0169,R2 was 0.9994;vitexin and isovitexin 0.0005～0.0500 mg/mL,0.0038～0.3799 Within the mg/mL concentration range,the linear regression equations are:Y=3.31× 106X-9.35 × 104,Y= 4.22 ×106X-1.43 × 105,and R2 are 0.9997 and 0.9994,respectively.Glucose is in the concentration range of 0.0125～0.2500 mg/mL,the standard curve equation is A=3.5370C+0.0024,R2=0.9996.Static adsorption and desorption experiments screened out D101 macroporous resin,It has good adsorption properties for the total flavonoids of Ficus microcarpa,vitexin and isovitexin.the optimal adsorption time is 5 h,and the optimal adsorption temperature is 25℃.The best process parameters for dynamic adsorption and desorption of D101 macroporous resin are:the mass concentration of the sample solution is 30 mg/mL,the sample volume is 8 BV,the sample solution pH is 4,the sample flow rate is 2 BV/h,and 5 BV is used.Pure water and 4 BV 10%ethanol for impurity removal,40%ethanol 4 BV elution,the elution flow rate is 4 BV/h,macroporous resin can be used 10 times after regeneration.After purification of D101 macroporous resin,the total flavonoid content was 67.12 mg/g,the polysaccharide content was 99.27 mg/g,the sum of vitexin and isovitexin content was 1.50 mg/g,the solid content was 8.80 g;the alcohol precipitation process was purified After that,the total flavonoid content was 61.69 mg/g,and the sum of vitexin and isovitexin content was 0.92 mg/g.Polysaccharide 82.20 mg/g,solid content 11.60 g.During the loading process,the physical and chemical parameters and the model formulas of total flavonoids,vitexin+isovitrin,and polysaccharides are:total flavonoids(mg/mL)=2.116+0.713·BV;(vitexin+isovitexin)(mg/mL)=-7.294+0.991·BV(BV>7),(Vitexin+ Isovitrin)(mg/mL)=64.567-14.262·pH+12.353·solid content;polysaccharide(mg/mL)=213.968+16.488·conductivity+27.718-pH,polysaccharide(mg/mL)=35.652+10.022·solid content+4.526-pH During the desorption process,the physical and chemical parameters and the model formulas of vitexin+isovitrin are:(Vitexin+Isovitexin)(mg/mL)=-225.067+72.628·pH,(Vitexin+Isovitexin)(mg/mL)=-11.352+510.818·Conductivity.Gray correlation analysis results show that BV and solid content in the desorption process are strongly correlated with the concentration of total flavonoids and polysaccharides.D101 macroporous resin and alcohol precipitation purified products with the same mass concentration,the absorption rate constants of the macroporous resin group of orientin,isoorientin,vitexin and isovitrin in duodenum,jejunum and ileum,mg/(min·cm2)]are 2.0×10-2±1.0×10-2,8.0×10-2±2.8×10-2,2.1×10-2±5.0×10-2,6.0×10-2=2.8×10-2;2.2×10-2±0.2×10-2,1.5×10-2±0.2x10-2,6.8×10-2±0.4×10-2,4.0×10-2±0.3×10-2;2.5 ×10-2±1.0×10-3,0.9×10-2±2.0×10-3,1.5×10-2±2.0×10-3.The absorption rate constants of orientin,vitexin and isovitexin in duodenum,jejunum and ileum in alcohol precipitation group[Ka,mg/(min·cm2)]were 0.5×10-2±3.0×10-3,0.4×10-2±1.0×10-3,0.8×10-2±6.0×10-3:0.9×10-2±7.0×10-3,0.9×10-2±7.0×10-3,0.6×10-2±2.0×10-3;1.6×10-2±6×10-3,1.8×10-2±1.4× 10-2,1.2×10-2±3.0×10-3.Conclusion:The NaNO2-Al(NO3)3-NaOH colorimetric method and the phenol-sulfuric acid method for the determination of total flavonoids and polysaccharides in Ficus microcarpa L.f.are simple and easy to operate,with good accuracy and good repeatability.HPLC method was used to determine vitexin and isovitexin in Ficus microcarpa L.f.The method was accurate and reliable,the operation was simple,and the result error was small.The D101 macroporous resin screened by static experiments has good adsorption effect.The sum of D101 macroporous resin purified Ficus microcarpa L.f.,Vitexin and isovitexin content reached the requirements of the 2015 Chinese Pharmacopoeia,and the purification effect is superior to the alcohol precipitation process.This study The established purification process is stable and feasible.In the loading stage,BV,solid content and pH have a significant correlation with vitexin+isovitexin,and in the desorption stage,pH,conductivity and vitexin+isovitexin have a significant correlation.According to the model,The actual production provides experimental basis and scientific basis.The index component is higher and more conducive to absorption. |
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