Pharmaceutical Research On The Extracts And Preparations Of Scorzonera Austriaca Willd

Background:As one of the five internal organs,the liver occupies an irreplaceable position in the human body.The liver can regulate the flow of blood and qi,and it can be used to store blood and regulate blood supply.It coordinates with the spleen,stomach,lungs and gallbladder to form an organic whole that maintains the balance and development of the body.So far,studies have shown that the incidence of liver cancer in China ranks ninth in the world.Taking into account the large base factor,China has the largest number of cases in the world.In addition,about 25%of the population in China suffers from liver diseases,including hepatitis B,hepatitis C,alcoholic liver disease,drug-induced liver injury,and cirrhosis.With the development of medical business and science and technology,a large number of traditional Chinese medicines and western medicines for the treatment of liver disease have also appeared,but the incidence of liver disease is still high.Therefore,it is urgent to discover a new medicinal material that has a long-term use and has a therapeutic effect on liver disease.It has long been recorded in ancient books that the Scorzonera austriaca Willd has the effect of reducing swelling and detoxification,and after long-term research in our laboratory,it has been found that the extract of Scorzonera austriaca Willd has obvious liver-protecting effect.Therefore,the extract of Scorzonera austriaca Willd is made into capsules to achieve the effect of convenient medication.Purpose:According to the existing experimental data,a complete isolation and extraction method of the extract of Scorzonera austriaca Willd was screened out.The pharmacodynamics study of the obtained extract of Scorzonera austriaca Willd was conducted to verify the protective effect of extract of Scorzonera austriaca Willd on CCl₄-induced acute liver injury in mice.At the same time,establish the quality standard for the extract of Scorzonera austriaca Willd.For the convenience of medication,a complete capsule preparation process is used to make Yacong capsules.Check the quality of the obtained samples and study the stability,and find reasonable production,transportation and storage conditions.Method:1.Weigh 10g of the cleaned and dried Scorzonera austriaca Willd,immerse it in an ethanol solution for ultrasonic extraction,and conduct a comprehensive investigation using ethanol concentration,liquid-to-material ratio,extraction times,and extraction time as single factors.Analyze and record the peak area by HPLC.Find the selected conditions for response surface analysis to determine the final high-efficiency and suitable production conditions.2.According to the selected optimal preparation process,the quality standard for the extract of Scorzonera austriaca Willd was established.The extract of Scorzonera austriaca Willd are checked to clarify the eligibility standards of each index.And to identify the extract of Scorzonera austriaca Willd thin layer,observe the spot position.In addition,a method for the determination of the extract of Scorzonera austriaca Willd should be established,and the method should be validated.3.Using CCl₄to induce acute liver injury in mice as a model,to test whether the extract of Scorzonera austriaca Willd has a protective effect on acute liver injury in mice.A normal control group,a model control group,a high-dose group,a middle-dose group,a low-dose group and a positive drug control group were established respectively,and they were administered by gavage for 7 days.Except for the blank group,2 hours before the last injection,CCl₄ was injected to induce acute liver injury in mice.Blood was collected to detect the levels of ALT,AST and LDH,and the liver tissue pathological sections were analyzed.4.Carry out the corresponding preparation process research on the extracts of Scorzonera austriaca Willd with clear pharmacological effects obtained above,screen out suitable wetting agents and binders with granulation conditions as indicators to make granules by wet method,dry them at a suitable temperature,and fill the capsules Shell,packaging.Routine inspections of Yacong Capsules were carried out,including inspections of properties,water content,weight differences,disintegration time limit,and microbial limit.TLC identification was carried out according to the quality standards of medicinal material extracts,and the spot positions among capsule contents,Scorzonera austriaca Willd extract,and monomer were compared.The content determination uses the established HPLC method to determine the content of orientin and isoorientin in the capsule contents.5.Carry out stability studies to investigate the changes of various indicators of Yacong Capsules under influencing factor tests,accelerated tests,and long-term tests,and analyze the production environment of the preparations.Result:1.Determine the investigation level of the 4 factors in response surface experiments based on the results of single factor experiments.Integrating the three evaluation index results of extract content,orientin and isoorientin,the optimal conditions for the preparation process were selected as follows:Dried Scorzonera austriaca Willd was immersed in an 18 times 60%ethanol solution,ultrasonically extracted 3 times,each extraction time was 2h.After the resultant is passed through D101 resin to remove impurities,spin-dried,and grind into powder to obtain the extract of Scorzonera austriaca Willd.2.In accordance with relevant regulations,the extracts of Scorzonera austriaca Willd were identified,inspected and determined.Thin-layer identification showed that the spots contained the characteristic spots of Scorzonera austriaca Willd.According to the experimental data,the qualified standards for the properties,moisture and ash content of the extract of Scorzonera austriaca Willd were determined.Established an HPLC method for the determination of the characteristic peak content of Scorzonera austriaca Willd,and investigated the linearity,precision,reproducibility,stability,and sample recovery of the method.The results showed that the method has high sensitivity and can be used as a method for the determination of the extract of Scorzonera austriaca Willd.3.Scorzonera austriaca Willd extract can significantly protect the acute liver injury induced by CCl₄ in mice,and reduce the content of AST,ALT and LDH in the serum.Among them,the low-dose group has liver-protective activity,and the middle and high-dose groups can significantly reduce the the activity of three enzymes in the serum compared with the model group.The observation results of liver tissue pathology biopsy also showed that the administration group had a significant improvement effect on liver tissue.The results show that the extract of Scorzonera austriaca Willd does have a protective effect on acute liver injury in mice.4.Add an appropriate amount of 3%PVP 98%ethanol to the obtained Scorzonera austriaca Willd extract powder for wet granulation,drying,granulation,and filling the capsule shell.In the related inspections of Yacong Capsules,the properties,water content,disintegration time limit,difference in filling quantity,and microbial limit inspection were all qualified according to the pharmacopoeia.The position of the thin layer spots is consistent with the extract,and also contains characteristic compound spots.The content determination result shows that the isoorientinontent is within the allowable range.Yacong capsules comply with the regulations.5.Influencing factors in the test,high temperature and light have no effect on the contents of the capsule,while high humidity will cause the particles to absorb water and become black and stick,but it has no effect on the composition of the capsule.The accelerated test and the long-term test used appearance and properties,difference in filling quantity,disintegration time limit,thin-layer identification,orientin content,isoorientin content and microbial limit as evaluation indicators.The results showed that all inspection items of Yacong capsules were qualified.To sum up,the environment should be kept relatively dry during the production,transportation and storage of Yacong capsules.Conclusion:The method for preparing the extract of Scorzonera austriaca Willd is optimized,the quality standard is improved,and it is confirmed that the extract of Scorzonera austriaca Willd prepared by this method does have a protective effect on the acute liver injury induced by carbon tetrachloride in mice.On the basis of the preliminary experiments,the extract of the Scorzonera austriaca Willd was made into capsules,and the establishment of quality standards and stability studies were also completed.