Preparation,formulation And Evaluations Of Compound Calculus Bovis Sativus Film

Research ContextThe purpose of this study was to prepared compound Calculus Bovis Sativus film with great properties of adhesivity and flexibility by using solvent casting method.In this study,chitosan,polyvinyl alcohol,and hydroxypropyl methyl cellulose were used as film-forming agent,and calculus bovis sativus and ornidazole were used as main drugs.This film can produce effect in oral locally and maintain effective drug concentration within a period of time.At the same time,it can enhance the bioavailability of drugs and can protect the ulcer surface from the bacterial infections.It is easy to use and can reduce times of medication.Moreover,it can improve the compliances of patients.It is convenient for storage and transportation and provide a new method of using calculus bovis sativus.Main contents1.Single factor analysis was used in screening polymer film forming material with property of adhesivity;2.Solvent casting method was used to prepare compound Calculus Bovis Sativus film and prescriptions were optimized by uniform design;3.HPLC method was established for the content determination of bilirubin and ornidazole in compound Calculus Bovis Sativus film;4."Chemical burning metohd",which was recognized in domestic and foreign,was used in the establishment of rabbit model of oral ulcer.By measuring the diameter of ulcer surface,histologic characteristics and biochemical indicator of the ulcer,the efficacy of the film was evaluated;5.The drug concentrations of compound Calculus Bovis Sativus film in the saliva of health volunteers were also measured.Methods1.Single factor analysis was used in screening polymer film forming material with property of adhesivityCommonly used polymeric materials were selected and film-forming property,mechanical property,and adhesivity were evaluated to screen appropriate film materials and determine the appropriate concentration range of them.2.Solvent casting method was used to prepare the film and prescriptions were optimized by uniform design.Film appearance,adhesivity,mechanical property,physical property,swelling propert,in vitro release characteristic and drug permeability were used as evaluation indexes to choose the optimal prescription.3.HPLC method was established for the content determination of bilirubin and ornidazole in the filmBilirubin and ornidazole were separated on Hypersil ODS2 column?4.6mm×250 mm,5?m?;The mobile phase was acetonitrile-1%acetic acid with gradient elution;Detection wavelength of bilirubin and ornidazole was 450nm and 315 nm,respectively;the column temperature was 30?and the injection volume was 10?L.4.Effect of the film in rabbit model of oral ulcer2².5kg healthy male New Zealand rabbits were selected as experimental animals.Oral ulcer rabbit model was established by the method of chemical burning with a cotton swab dipped in a concentration of 50%?v/v?acetic acid solution.And then it was pressed on rabbit cleft lip at the continued60s,cleft lip at a white ulcers after 24h.It was determined at the date of the maximum diameter of cleft lip ulcers and ulcer area of observation;and reserve tissue for pathological examination,examine morphological changes.Simultaneously the SOD activity and MDA content of ulcer tissue was determined.5.Drug concentrations of the film in the saliva of health volunteersThree healthy male volunteers were selected randomly,before the experiment with distilled water rinse to remove residue mistakes inside the mouth,respectively,per 1cm².Compound Calculus Bovis Sativus film is pressed against the cheek 60s,at various time points and the saliva was collected within 4h.Then the drug concentration in saliva was determined by HPLC.Results1.The linear range of bilirubin was within 1.58⁵1?g·ml^-1?r=0.9992?,the average recoveries were 97.64%,98.59%and 98.34%,RSD were 1.33%and1.10%,0.18%.The linear range of ornidazole was within 3.22¹03?g·ml^-1?r=0.9990?,the average recoveries were 99.27%,98.37%and 99.03%;RSD were 1.67%,0.22%and 0.73%.With high specific,precision,accuracy and good stability,this method can be used for the assay of Ornidazole and Bilirubin in Compound Calculus Bovis Sativus film.2.The suitable film-forming concentration range for chitosan,Hydroxypropyl methyl cellulose and Poly?vinyl alcohol?were separately 1%¹.5%,0.5%²%and 1%¹.5%.When they were used simultaneously,the prepared film has good film-forming properties,adhesiveness and flexibility.3.Preferred formulation was:chitosan?1.2%?,polyvinyl alcohol?1.5%?,hydroxypropyl methyl cellulose?1.2%?,glycerol?2.5ml?,ICCB?0.2g?and ornidazole?0.24g?,then the volume of water was added to 40ml.The thickness of prescription was 426.2?m;the weight of film was 46.16mg;the surface pH was 6.2;the swelling index at the point of 2h was 44.60%;the in vitro adhesion force of 436g/cm² and the in vitro adhesion time was 4.84h.4.Compared to the control group,a single group and single calculus bovis group,rabbit of compound Calculus Bovis Sativus film group cleft lip at the mouth ulcers and a maximum diameter of more significant decrease?p?0.05?,compound Calculus Bovis Sativus film healing faster than other three groups have a significant effect.5.As a preliminary study,OD concentrations rose in a regular way,reaching the highest level?33.07?g/mL?at 75 min and starting to decrease after 120min.Notably,the OD level of film F5 was maintained higher than the minimum inhibitory concentration against Candida albicans?7.8?g/mL?for about 2 h.Moreover,the films,which did not irritate or discomfort the cheek mucosa,showed a good comfortability and tolerability in the oral cavity.Conclusions1.Chitosan,polyvinyl alcohol,and hydroxypropyl methylcellulose was used in this film with good flexibility,suitable adhesion.2.By using solvent casting method,the film was successfully prepared with good flexibility,adhesion and in vitro drug release and permeability.3.The HPLC method is specific,precision,high accuracy,good stability,can be used to test the content of bilirubin and ornidazole in this file.4.This film had a good effect for oral ulcer on rabbits.5.This film can be maintained at a certain oral effective concentration locally,with good compliance and well tolerance in healthy volunteers and without irritation.