| Ulcerative Colitis is a chronic non-specific inflammatory bowel disease of unknown etiology,generally considered to be related to immune,genetic,infection,mental and environmental factors.The disease still can not cure,and is in higher incidence in the world and it is rated as one of the modern cureless diseases by the World Health Organization.In recent years,the incidence of UC in our country has increased remarkblely,and is the key link in the development of Ulcerative Colitis-associated Colorectal Cancer,seriously influence the patients’ quality of life.Thus,the treatment of UC has already become a focus in clinical research.Objective:Qingchang Wenzhong Formula is a clinical prescription that originates from Dongfang Hospital affiliated to Beijing University of Traditional Chinese Medicine.It has a good effect on the treatment of UC.The formula is composed of Coptidis Rhizoma,Notoginseng Radix et Rhizoma,Indigo Naturalis and other herbs,characterized by good cleaning intestinal damp and hot,warming and tonifying the spleen,mildly regulating cold and heat,removing blood stasis and hemostasis.The early clinical application of the formula was for the oral store.There existed a lot of inconvenience in oral administration and storage.Therefore,this study combined modern Chinese traditional medicine preparation technique to optimize the extraction,purification,concentration and drying process,prepare Qingchang Wenhzhong Formula into granules to improve the convenience and stability,and establish the quality standard.Methods:This study mainly includes the extraction,purification,concentration and drying process,forming process,pilot-scale experiments and quality standards of Qingchang Wenzhong Granule.The specific methods are as follows:(1)Study on the extraction,purification,concentration and drying process:the formula study was divided into two parts.Alcohol extraction part:with the extraction rate of Rhizoma coptidis alkaloids,Panax notoginosides and Indirubin as index,Coptidis Rhizoma,Notoginseng Radix et Rhizoma and Indigo Naturalis adopted orthogonal test to optimize the alcohol extraction process and adopted the single factor test to investigate the centrifugation,concentration and drying process.Water extraction part:with the extraction rate of Sophora alkaloid,Radix Sophorae Flavescentis,Glycyrrhizae Radix et Rhizoma and other three herbs adopted orthogonal test to optimize the water extraction process and adopted the single factor test to investigate the alcohol precipitation,concentration and drying process.(2)Study on the forming process:investigate related parameters of the wet granulation process.Based on the granulating process,determined the proper excipient,the amount of the excipient and the concentration of the adhesive using moisture absorption rate of particle,particle forming rate,soft material properties as the assessment indexes.In addition,the critical relative humidity of forming particles was studied,providing a reference for humidity control of preparation process.(3)Study on the scale-up:the optimized preperation process was used in the three batches pilot-scale study of Qingchang Wenzhong Granule.(4)Study on the quality standards:refering to methods under the Granule of General Principle 0104 in the Chinese Pharmacopoeia of 2015 edition,the moisture content and dissolubility examination was conducted;Refering to methods under "Anshenbao Keli"in the Chinese Pharmacopoeia of 2015 edition,the n-butanol extract content of Qingchang Wenzhong Granule was measured and its standard was formulated;Adopting TLC and HPLC methods,the qualitative and quantitative research of Qingchang Wenzhong Granule was conducted to formulate its quality standards.Results:(1)The extraction,purification,concentration and drying process:The optimal alcohol extraction process:Coptidis Rhizoma,Notoginseng Radix et Rhizoma and Indigo Naturalis was extracted with 10 folds of 60%alcohol for 3 times,1.5 hours for each time.The filtrate was centrifuged by the speed of 5000 r/min for 10 min or 7000 r/min for 5 min.The centrifugal fluid was concentrated to a relative density of 1.15-1.30(60 ℃)by recycling alcohol below 80℃.Followed with the decompression drying below 50℃,the dry extract was crushed into fine powder.The optimal water extraction process:Radix Sophorae Flavescentis,Glycyrrhizae Radix et Rhizoma and other three herbs was extracted with 12 folds of water for 3 times,1.5 hours for each time.The filtrate was concentrated to a relative density of 1.05-1.10(60℃)below 80℃,cooled to room temperature.Add suitable amount of alcohol to attain to the alcohol concentration of 60%and stand for 12 hours at 4℃.The filtrate was concentrated below 80℃.Followed with the decompression drying below 80 ℃,the dry extract was crushed into fine powder.(2)Forming process:dextrin as excipient,drug and excipient ratio was 2:1,wet granulation,95%ethanol as wetting agent,environmental critical relative humidity at below 82%.(3)Quality standard:examined the moisture content and dissolubility,which were both in line with the requirements of Granule;established the content limit of n-butanol extractive;established the TLC method of all eight herbs,which is of good specificity.Besides,set up the HPLC methods of Berberine hydrochloride,Indirubin and Sophora alkaloid in Qingchang Wenzhong Granule.This method is stable and feasible.The content of berberine hydrochloride(C20H18ClNO4)shall not be less than 11.92 mg per 1 g;the content of Indirubin(C16H10N2O2)shall not be less than 0.08 mg per 1 g;the content of matrine(C15H24N2O)and oxymatrine(C15H24N2O2)shall not be less than 3.47 mg per 1 g.Conclusion:Through the research on extraction,purification,concentration,drying and forming process,the stable and feasible preparation process of Qingchang Wenzhong Granule was established,the preparation of the qualified forming particles accomplished.Through the research on quality standard,the content limit of n-butyl alcohol extract,methods of qualitative identification and quantitative determination were established.These methods can be repeated and of good precision and accuracy,achieve the purpose of product quality control,attain to the expected goal. |
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