| Purpose:Such as the rushengtang out of the Ming Dynasty,the fifth son of the Ming Dynasty,Zhou Dingwang presided over,Professor Teng Shuo,Chang Shi Liu alcohol and others compiled and compiled"Puji Fang",The book is the largest prescription book in Chinese history.The prescription consists of four flavors:Platycodon grandiflorus?Jacq.?A.DC.,Glycyrrhiza uralensis Fisch,Fructus Arctii,Ophitopogin japonicum?L.f?Ker.-Gawl.The book records that its efficacy is mainly to cure phlegm and heat,to throat,to treat sore throat and so on.By consulting the modern research literature,it is found that these four-flavor single-drugs have certain effects in the treatment of sore throat and bacteriostasis,and the main effect is antibacterial and anti-inflammatory.At present,there are more reports on the research on single-flavor drugs such as Shengtang in the research literature,However,there are few studies on Rushengtang,and there is no report on the research on modern preparations such as Shengtang.Therefore,this topic identifies the active components of the antibacterial and anti-inflammatory properties of four traditional Chinese medicines through literature search:Total saponins of platycodon grandiflorum,total saponins of licorice,total lignans of burdock,total saponins of wheat and honey,and polysaccharides,They were extracted and purified separately,and the in vitro drug effects before and after purification were compared.The optimal extraction process of each active component was determined by orthogonal optimization,and the optimal purification process of each effective component was determined by macroporous adsorption resin method.The components were purified before and after purification by Oxford Cup method and double dilution method.The effect of in vitro bacteriostasis was compared.The purified active ingredients were combined according to the compatibility ratio of the original prescriptions of Pujifang,and the in vitro antibacterial effects of the compatibility group and Rushengtang were compared.The compatibility group was made into Rusheng oral liquid,and the content of platycodin D in Ruan oral liquid was determined by HPLC method.At the same time,the preliminary pharmacodynamic study on Rusheng oral liquid was carried out.Provide a basis for the rational use and development of Rushengtang.Material and method:1.The orthogonal experimental design method was used to investigate the extraction process of total saponins of platycodon grandiflorum,total saponins of licorice,total lignans of burdock,total saponins of wheat,and polysaccharides,and the optimal extraction process of each component was determined.The macroporous adsorption resin method was used to optimize the purification process of the effective components of Platycodon grandiflorum,licorice,burdock and Maimendong by static,dynamic adsorption and desorption experiments.2.Using the Oxford Cup method and the double dilution method,the selected indicator is the zone of inhibition and the MIC value.The bacteriostatic effects of total saponins of platycodon grandiflorum,total triterpenoid saponins of licorice,total lignans of burdock,total saponins of meringue and polysaccharides against Staphylococcus aureus and Streptococcus pneumoniae were studied before and after purification.3.Using the Oxford Cup method and the double dilution method,the diameter of the inhibition zone and the MIC value were used as indicators,The antibacterial effect of the total extracts such as the total extract of rushengtang and the compatibility before and after purification.4.The content of platycodin D in the self-made Rusheng oral liquid was determined by HPLC method,and the preliminary standard of Rusheng oral liquid was established.5.For the preliminary pharmacodynamics study of St.Oral Liquid,the antitussive and expectorant effects of Rusheng Oral Liquid were demonstrated by the mice's ammonia water cough and mouse phenol red excretion experiments.Results:1.Preparation of total saponins of Platycodon grandiflorum:The best extraction process was to extract 3 times,each time with 6 times the amount of 60%ethanol for 1.5 hours.The best purification process was purified by D101 macroporous adsorption resin.The calculated amount of the tablet and resin volume ratio was 0.75g·mL-1,and the loading concentration was 0.20g·mL-1.The amount was 50mL,80%ethanol was used as the eluent,the elution amount was 60mL,and the eluent was evaporated to dryness to obtain a purified total saponin of platycodon grandiflorum with a content of 7.40mg·g-11 and a paste yield of 1.99%.2.Preparation of total saponins of licorice:The best extraction process is to extract 2 times,each time with 8 times the amount of 60%ethanol for 1.5 hours.The best purification process was purified by HPD-300 macroporous adsorption resin.The calculated amount of the tablet and the volume ratio of the resin was 0.25g·mL-1,and the loading concentration was0.10g·mL-1.The amount of miscellaneous water was 72mL,70%ethanol was used as the eluent,the elution amount was 70mL,and the eluent was evaporated to dryness,which gave the content of 252.33mg·g-1,and the extracting rate was 11.85%.Purified.3.Preparation of total lignans of burdock:The best extraction process is to crush the burdock?shrink,but sieve?,extract it 3 times,and extract it with 12 times of 80%ethanol for 1.5 hours.The best purification process is to purify with AB-8 macroporous adsorption resin.The calculated amount of the tablet and the volume ratio of the resin is 0.35g·mL-1,and the loading concentration is 0.05g·mL-1.The amount of miscellaneous water is 60mL,70%ethanol is used as the eluent,the elution amount is 50mL,and the eluent is evaporated to dryness.The total lignan content of burdock is 205.09mg·g-11 and the ashing rate is 7.88%.Purified.4.Preparation of Ophiopogon japonicus polysaccharide:The best extraction process is to extract twice,each time with 8 times the amount of water for 0.5 hour,alcohol precipitation with 80%ethanol concentration,and the precipitate is evaporated to dryness,which is the content of 28.80mg·g-1.The purified extract of Ophiopogon japonicus was 28.94%.5.Preparation of total saponins of Ophiopogon japonicus:The best extraction process is to extract 3 times,each time with 8 times 70%ethanol extraction for 1 hour.The best purification process was purified by D101 macroporous adsorption resin.The calculated amount of the tablet and the volume ratio of the resin was 1.50g·mL-1,and the loading concentration was 0.10g·mL-1.The amount was 72mL,70%ethanol was used as the eluent,the elution amount was 60 mL,and the eluent was evaporated to dryness to obtain a purified total saponin of Ophiopogon japonicus with a content of 0.50mg·g-11 and a yield of 2.03%.6.Comparison of in vitro antibacterial effects of total saponins of Platycodon grandiflorum before and after purification:The diameter of the inhibition zone acting on Staphylococcus aureus was 9.33±0.58mm,14.17±1.61mm,the MIC values were 125.00mg·mL-1,62.50mg·mL-1,and the diameter of the inhibition zone acting on Streptococcus pneumoniae was 9.67±0.5 8mm,10.27±0.58mm,and MIC values of 31.25mg·mL-11 and 31.25mg·mL-1,respectively;Comparison of antibacterial activity of licorice total triterpenoid saponin before and after purification:the inhibition zone diameter of rusheng aureus was 10.27±0.5 8mm,24.06±0.99mm,and the MIC values were 250.00mg·mL-11 and 125.00mg·mL,respectively.The diameter of the inhibition zone acting on Streptococcus pneumoniae is 7.68±0.15mm,8.75±0.46mm,and the MIC values are 31.25mg·mL-11 and 15.63mg·mL-1,respectively;Comparison of antibacterial activity in vitro of burdock total lignans before and after purification:the inhibition zone diameter of rusheng aureus was 9.33±0.58mm,10.00±0.50mm,and the MIC values were 250.00mg·mL-1,125.00mg·mL-1.The diameter of the inhibition zone acting on Streptococcus pneumoniae is 8.00±1.00mm,9.50±0.50mm,and the MIC values are 250.00mg·mL-11 and 250.00mg·mL-1,respectively;Comparison of antibacterial activity in vitro and in vitro of wheat polysaccharides:the inhibition zone diameter of Staphylococcus aureus was 6.66±0.29mm,6.83±0.29mm,and the MIC values were 250.00mg·mL-1,250.00mg·mL-1,respectively.The diameter of the inhibition zone of Streptococcus pneumoniae is basically no,the MIC values are 500.00mg·mL-1,500.00mg·mL-1,respectively;Comparison of antibacterial activity in vitro and in vitro before and after purification of total saponins of Ophiopogon japonicus:the inhibition zone diameter of Staphylococcus aureus was 7.30±0.47mm,8.35±0.15mm,and the MIC values were250.00mg·mL-1,125.00mg·mL-1,respectively.The diameter of the inhibition zone of Streptococcus pneumoniae was 7.26±0.35mm,7.55±0.10mm,and the MIC values were250.00mg·mL-11 and 125.00mg·mL-1,respectively.7.Such as the antibacterial effect of ruShengtang decoction in vitro:The diameter of the inhibition zone acting on Staphylococcus aureus and Streptococcus pneumoniae was14.00±1.73mm,9.33±1.53mm,and the MIC values were 250.00mg·mL-11 and 125.00mg·mL-1,respectively;In vitro antibacterial effect of the compatibility group before and after purification:the inhibition zone diameter of Staphylococcus aureus was 16.33±1.53mm,17.67±2.52mm,and the MIC values were 125.00mg·mL-1,62.50mg·mL-1,respectively.The diameter of the inhibition zone of Streptococcus pneumoniae was 8.33±0.58mm,13.67±0.58mm,and the MIC value was 15.63mg·mL-1,7.80mg·mL-11 respectively.The results showed that the antibacterial effect of the compatibility components before and after purification The effect was stronger than that of Rushengtang,and the antibacterial effect of the purified group was better than that of the pre-purification compatibility group.8.The determination of the content of platycodin D in Rusheng oral liquid showed that the linear relationship of platycodin D injection in the range of 0.1514.45?g was good,r=0.9995,and the content of platycodin D was 18.69mg/mL.The methodological study showed good precision,repeatability and stability,and the recovery rate of the sample was RSD=1.78%.9.The pharmacodynamic study in mice showed that the high-dose group of rusheng.Oral Liquid can prolong the incubation period of cough in mice with cough and reduce the number of coughs in a certain period of time.The phenol red excretion in the high-dose group of Rusheng oral liquid was higher than that in the positive control group,indicating that the amount of phenol red excreted in the respiratory mucosa of mice increased under the action of Rusheng oral solution,such as ruSheng oral liquid can affect the passage of airway.Permeability and secretion volume.Pharmacodynamic experiments in mice have shown that such as oral liquid has antitussive and expectorant effects.Conclusion:1.Through orthogonal extraction experiments and dynamic and static investigations,the macroporous adsorption method was used to extract and purify the effective components in Rushengtang.The process parameters were determined and the process method was advanced and reasonable.It was suitable for industrial production and other advantages.It was platycodon grandiflorum,licorice,burdock and wheat.The theoretical basis for the further research and development of Dongsiwei drugs.2.Through the diameter of the inhibition zone and the MIC value before and after the purification of the effective components,the extraction rate is reduced during the purification of the active ingredient of the unit drug,and the comparison test is carried out before and after the purification,indicating that during the purification operation,There was no obvious loss of antibacterial active ingredients,indicating that the purification process was reasonable and the dosage of the compound was reduced.3.Comparing the antibacterial effects of the compatibility group and the sacred soup,the antibacterial effect of the compatibility components on the diameter of the inhibition zone and the MIC was stronger than that of the sacred soup.It shows that the compatibility group not only reduces the dosage,but also has an advantage in efficacy.4.Determination of the content of platycodin D in Rusheng oral liquid,established a preliminary quality standard for Rusheng oral liquid.5.The preliminary pharmacodynamic experiments in vivo showed that the oral liquid has the effects of antitussive and expectorant,which provides a basis for the pharmacological aspects of Rusheng oral liquid. |
PDF Full Text Request