Study On The Preparation Process And Quality Control Research About Film-forming Gel Of Antibacterial Peptide From Rana Temporaria

A kind of small molecule active peptides in the skin of Rana—antimicrobial peptides,known as “super naturalantibiotic ”.Antimicrobial peptides of Rana have many biological activities,such as broad spectrum antibacterial activity,antiviral activity,their capacity to kill a wide range of fungi,viruses and parasites,also have antitumoral activity.In this paper,based on the excellent antibacterial activity of the antibacterial peptide of Rana Temporaria,a formulation applied to the external wound site was developed to prevent further infection of the wound.The film-forming gel has the advantages of high stability,less skin irritation,good appearance,also has sustained release and prolonged drug effect.In this paper,we prepare the Rana-antibacterial-peptides film-forming gel which has the broad spectrum antibacterial effect and long-term antibacterial effect,also with the excellent ability of anti-friction.This article includes the following:1.Antimicrobial peptides of Rana in vitro establishment of analytical methodology.The in vitro analysis method of the antibacterial peptide of Rana was established by BCA reagent method.Use this method to get the antimicrobial peptides of Rana precision,repeatability and recovery rates are in line with the requirement in viro assay.The BCA reagent method was used to determine the content of antibacterial peptide,and the drug content of Rana-antibacterial-peptides film-forming gel.2.Preparation of Rana-antibacterial-peptides film-forming gel.The single-factor method was used to screen the type of matrix and film former of the Rana-antibacterial-peptides film-forming gel,and also preliminary test on the dosage.Under the conditions that the ratio of MC,PVP and the ratio of antimicrobial peptides of Rana to absolute enthanol were used as the independent variable,while used the filming time as the response value,screen for the best prescription by using response surface analysis.The optimal prescription was: MC 2.8 %,PVP 0.8 %,antibacterial peptide of Rana 57.4 %,absolute ethanol 40 %,glycerol 1 %,at the same time,the optimal preparation process was selected.3.Quality standard of antimicrobial peptides of Rana.The Rana-antimicrobial-Peptides film-forming gel is yellow transparent,uniform and fine texture gel.That film-forming gel has an antibacterial peptide content of 1.628 mg/ml,a viscosity of 9.21 Pa·s,a pH of 3.82,and a particle size of less than 180 ?m,which is in line with the quality standards of the Chinese Pharmacopoeia gel.After the gel was applied to the skin,there was no irritating reaction and a transparent film was formed within 78 s.4.Study on the stability of Rana-antibacterial-peptides film-forming gel.It was conducted influencing factors test(temperature,humidity,light),accelerated test and long-term test on the Rana-antibacterial-peptides film-forming gel.It was found that the appearance,pH,viscosity and content of the antimicrobial peptides of Rana were basically unchanged under high temperature and high humidity conditions.However,under the condition of high light intensity,the appearance and content have changed,indicating that the Rana-antibacterial-peptides film-forming gel is suitable for preservation in the dark.Afterwards,the accelerated test and long-term test of that film-forming gel were carried out,and there was no significant change,which indicated that this film-forming gel had good stability.5.Study on the efficacy and the anti-friction of Rana-antibacterial-peptides film-forming gel.It was found that the antibacterial activity of the preparation against Staphylococcus aureus,Escherichia coli,Agrobacterium tumefaciens and Candida albicans was as high as 100 %,also the inhibition zone is 20.98 mm,20.29 mm,20.72 mm,and 20.56 mm,respectively.Meanwhile,it was found that the bacteriostatic duration of all four bacteria was above 10 h.Then,the preparation was also tested for transdermal accumulation.It was found that after 24 hours,the transdermal amount of the drug was only 15.58 %,and the rest remained on the skin,that resulting in a long-lasting antibacterial effect.Through the anti-friction test of the film,it was found that the film was still intact after being wiped by paper,palm and clothing,which shows that it has strong anti-friction.