Research And Evaluation Of The Classic Huagaisan Standard Granules

The classic Huagaisan was scattered from the Taiping Huimin Heji Ju Fang of the Song dynasty,which contains Ephedrae herba,Armeniacae semen amarum tostum.Glycyrrhizae radix et rhizome praeparata cum melle,Citri reticulatae pericarpium,Mori cortex praeparata cum melle,Poria and Perillae fructus tostum.Huagaisan has the effect of Xuanfei solution and cough expectorant.It’s mainly used in the treatment of pulmonary sense of pathogenic cold and cough on gas,chest and diaphragm dysphoria,hypertonicity of the nape and neck,stuffy nose sound of heavy,light headedness,sputum negative,sipping sound.It’s widely used in modern clinical colds,cough,asthma,asthma and other lung diseases.In this paper,we carried out the modern research of classic Huagaisan on the basis of the original prescription,the original dose,the original decocting method and the original modern disease research.Through the study of processing methods,preparation techniques,quality standards,pharmacodynamics,safety evaluation and pharmacokinetics of Huagaisan standard granules,we developed the drug which conform to laws of the prescriptions of traditional Chinese medicine,maintain the characteristics of traditional Chinese medicine prescriptions,and satisfy the standard of industrial production,and the quality is stable and controllable.And this study can provide reference for the evaluation of the rationality of the traditional Chinese medicine preparation process.Based on the original recorded in the medical books and literature textual research,we adopt the method of multi-index combined with chromatographic fingerprints to quantify the Huagaisan standard medicinal preparation process parameters and the preparation method of the standard decoction as follows:the medicinal materials were crushed into coarse powder,take a total of 18.0 g,water 600 ml,ceramic casserole decoction to 420 ml,strike four layers of cotton gauze filter filtrate.This study established HPLC fingerprints of Huagaisan standard decoction and identificated the characteristic peaks ion by the LC-MS technologyand preparationsmedicinal materials peak pattern matching.This study established the fingerprint common pattern of Huagaisan standard decoction.After that,chemometrics methods including principal component analysis and clustering analysis were utilitied to evaluate the quality of medicinal materials which from different regions.In this study,the standard decoction of volatile oil were analyzed and identified and compared by GC-MS hyphenated techniques,to determine the 70 components,accounted for 98.63%of the total oil volatile.Attribution analysis results show that volatile oil GC-MS tic chemical signals mainly comes from the contribution of prescription ephedra,bitter almond,dried tangerine peel,lay the foundation for revealing the effect substance basis and mechanism of compatibility of medicines.To standard broth based,on the name of the classic modern technology of extraction,concentration,drying and preparation of process was studied,the maximum maintain consistency with the material foundation of the traditional decoction.For multiple index component content and chromatographic fingerprints indicators,this study of the extraction process by orthogonal design method of degree of optimization,modern technology was confirmed as the following:the prescription with 10 times,8 times and 8 times water reflux extraction respectively 2 hours.1 hour,0.5 hour.This study determined the volatile oil inclusion optimum:volatile oil adding 8 times of beta －cyclodextrin and 60 times water grinding,30 minutes.grinding fluid placed in the refrigerator for 24 hours,centrifugation,filtration,and drying in 40℃ and crushing.The inclusion of the volatile oil compounds identified by UV spectrophotometry,infrared spectroscopy,gas chromatography and so on.It is shown that beta cyclic dextrin and canopy bulk oil can form stable inclusion complexes.For the first time,we have nanofiltration technique to replace the traditional concentration process,realize equal amounts of matter transfer,reduce the loss of material and the active component of the process.With the particle moisture absorption rate as index,the optimal proportion of the prescription adjuvant was selected by the D-Optional mixture design.The optimal proportion as follows:the proportion of lactose,dextrin and soluble starch were 1,3,1.To the particles forming rate,angle of repose etc.as the index,Huagaisan standard granules of one step granulating process was optimized by Plackett Burman and box-Behnken design test.the optimum granulating process for incoming extract relative density 1.138(60 ℃),the feed speed of 47 revolutions per minute,inlet air temperature of 75 ℃,material temperature 60 ℃,35 hertz fan frequency,atomization pressure 0.4 megapascals.Multiple pilot trials demonstrated that the preparation proeess was stable,feasibleand suitable for industrial mass produetion.According to the characteristics of the Chinese medicine overall composition effect,this study established Huagaisan standard granules fingerprints consistency evaluation method of Huagaisan standard granules,established the efficacy and safety with prescription correlation is high effective composition content determination method,mainly include the ephedrine hydrochloride,pseudoephedrine hydrochloride,laetrile.liquorice glycosides,glycyrrhizic acid,hesperidin,rosemary acid etc.We established a method for the analysis of chromatographic fingerprints.To evaluate the similarity of Huagaisan standard granules by cosine method,correlation coefficient method and Huagaisan standard granules ratio similarity method.The box plot method compared the contents of index components of similar system cluster analysis results show that fingerprint similarity is not equal to content similarity.The combination of the multi index component content andsimilarity of fingerprints can effectively control the quality of traditional Chinese medicine.In accordance with the national drug standards,and reference to the eu requirements for drug registration,we established a relatively perfect quality standard system of Huagaisan standard granules.According to the guiding principles of stability study technical of Chinese medicine,natural medicine,this research adopts the simulation of the commercial packaging condition factors affecting Huagaisan standard granules test and accelerated stability test for six months and six months long term stability test.The results show that the stability of the product is good.The study provides theoretical basis for packaging,storage,transportation,and determination of validity of Huagaisan standard granules,and guarantee the safe and effective clinical application.We adopt rats as experimental animals,by way of lavage were given equal crude drug doses of Huagaisan standard granules and standard decoction,with the main effective components of the classic party ephedrine hydrochloride and pseudoephedrine hydrochloride as testing indexes,different time points of rat blood samples for testing analysis,results show that the ephedrine hydrochloride had good linear relationship(0.376～24.041 μg·ml-1),r=0.9999,LLOQ(0.376 μg ml-1).The pseudoephedrine hydrochloride had good linear relationship(0.204～13.080 μg ml-1),r=0.9995,LLOQ(0.204 μg ml-1).This study of the main pharmacokinetic parameters by Winnonlin software fitting,and by the SPSS 19.0 software on the two independent samples t-test and nonparametric rank and test analysis.According to the results of Huagaisan standard granules group and the standard decoction measured the pharmacokinetic parameters including AUC(0-t),AUC(0-∞),Cmax,Tmax,t1/2z,MRT(0-t),MRT(0-∞),AUC_%Extrap_obs,Vz_F_obs and Cl_F_obs were no significant differences.The bioavailability of ephedrine hydrochloride and pseudoephedrine hydrochloride were respectively 99.72%,101.04%.Comparing with the standard decoction,Huagaisan standard granules has not been efficacy pharmacokinetic characteristics of composition change.They were bioequivalent bioavailability.This study was carried out the main pharmacodynamic comparison test of Huagaisan standard granules and standard decoction.The results showed that Huagaisan standard granules and standard decoction mice cough incubation period can reduce the frequency of cough,increased respiratory mice phenol red excretion,prolonged latent period of asthma guinea pigs,inhibited xylene-induced mouse ear acute swelling and inflammation,inhibition of granulation chronic swelling agar rats,increased serum hemolysis mouse antibody-producing hormone and reduced the amount of nitro-2,4-dichlorobenzene suppress immune function in mice induced delayed hypersensitivity and so on.The test proved that Huagaisan standard granules retention function and indications of standard decoction.the standard granules and standard decoction are equivalent.According to the requirements of the relevant supplementary provisions of the Chinese traditional medicine registration management and to ensure the safety of products,this study conducted Huagaisan standard granules acute toxicity test,the results showed that single drug lavage Huagaisan standard granules Lethal Dose 50 test is 54.309 grams per kilogram,that is about 211.2 times of clinical dosage,reveal the product is safe to use.In this paper,according to the major drug discovery projects "the research of the classical square standard granules"(N0.2013zx09508-105)of the overall deployment and the research train of thought,is compared with the standard decoction,we have established a biological method based on pharmacodynamics,toxicology,pharmacokinetic and multi-index chemical composition combined with fingerprints chemical characterization methods of evaluating the rationality of the classic square succession process,to draft "is derived from the ancient classical square of traditional Chinese medicine compound particles" provides the reference for the technical specification and registration standards of research,as well as the research and development of the classical square provides a new thought.