Preparation And Quality Control Of Deproteinised Calf Blood Extract Gel For Buccal Delivery

ObjectiveIn this paper,the bovine blood protein extract(DCBE)was prepared by using calf blood as the raw material,and the quality standards of raw materials and preparations of DCBE oral gel were established.The therapeutic effect of DCBE oral gel on oral cavity ulcer model were analyzed.There results may provide a new product for oral cavity ulcer patients.Methods(1)Study on the DCBE raw materials: Using bovine blood as raw material,DCBE raw materials were obtained by removing fibrin,destroying erythrocytes,denaturing protein by high temperature,removing viruses,removing impurities by centrifugation and ultrafiltration,concentrating and decolorizing.The amino acids and free amino acids of DCBE were detected by automatic amino acid analyzer.The characteristic structure of DCBE was determined by infrared spectroscopy and ultraviolet spectrophotometry.The molecular weight of raw materials was determined by mass spectrometry.The isoelectric point of raw materials was determined by capillary electrophoresis.Thymosin was used as control to analyze whether DCBE contains thymosinby HPLC.The virus in the product were determined by Enzyme-linked immunosorbent assay.The quality control and quality standards were established through the influence of factors,accelerate the experiment,and long-term experimental evaluation stability.(2)Prescription and quality study of DCBE oral gel: The composition and preparation process of DCBE oral gel were established based on the properties of raw and supplementary materials,pre-prescription study,compatibility study of raw materials and excipients,pH stability of formulation and antibacterial efficacy test..At the same time,the preparation and sterilization process of the product were studied.The quality control method and quality standard of the preparation were also established,and the stability of three batches of preparations were investigated.(3)The effect of oral gel of DCBE on oral cavity ulcer model: The blank gels,DCBE oral gel and watermelon frost were used to treat the oral ulcers in rats.The pathological changes of the ulcer tissue in the three groups were observed and the pathological sections of ulcer in different groups were evaluated.Results(1)DCBE obtained by the process of removing fibrin,disrupted erythrocytes,virus inactivation,removal of macromolecular hybrid protein,concentrated and decolorized.The DCBE has high respiratory activity.The free amino acid content was 79.2(mg / 100 mL).The content of hydrolyzed amino acid was 247.9(mg / 100 mL).The results of IR and UV showed that the product had the characteristic structure of amino acids and peptides with strong absorption peak in the ultraviolet region.The isoelectric point of DCBE is 4.9.The molecular weight range of DCBE is 300 ~ 2000 Da.DCBE does not contain thymosin ingredients.The conventional cow susceptible virus could not be detected in the raw material.Then the quality standard of DCBE were established.The results of stability showed that the raw materials of DCBE could kept in refrigerat for 24 months.(2)The formulation of DCBE as follows:1% CMC-Na as a gel forming agent,0.01% sodium saccharin as a flavoring agent and 0.03% ethylparaben as a preservative.The quality standards of the preparation were established according to the Chinese Pharmacopoeia.The results of influence factors,accelerated and long term stability experiments showed that the viscosity of gel will decline under high temperature and high light.There is no any change with long-term storage at room temperature for 24 months.Pharmacodynamics results showed that DCBE gel group has better therapeutic effect on oral cavity ulcer model than the control group and watermelon cream group,indicating that DCBE gel could accelerate ulcer healing,HE staining results suggest that the mucosal epithelium and epithelial cells in DCBE group are structurally intact,indicating that DCBE has obvious repair and regeneration functions on mucosal epithelial cells and vascular endothelial cells.ConclusionThe preparation process of DCBE is feasible and the quality is controllable.The preparation of oral gel is simple,the product is stable and the therapeutic effect is definite.