Study On The Breviscapine Chitosan Nanoparticle

Object To prepare breviscapine loaded biodegradable chitosan nanoparticles (Bre-CS-NP) using modern targeting drug delivery system preparation technique.The characterizations of nonaparticles,morphology,particles size,particle size distribution were evaluated.Breviscapine content was determined by ultraviolet spectrophotometry,which was verified with specificity and technology.The entrapment efficiency,drug load content,in vitro release and stability of Bre-CS-NP were studied.Method The Bre-CS-NP was prepared using one step ion forming gelation method. Based on the result of single factor evaluation,the main factors effecting preparation of Bre-CS-NP were PlournicF-68 content,Dextran-70 content,stirring rate,breviscapine content, CS concentration and CS/TPP proportion,and the scope of investigation is determined.The optimized formulation and preparation parameters were obtained using uniformity design with the comprehensive score of morphology,entrapment efficiency and drug load content as evaluation index.Effect of the former factors on the comprehensive score is evaluated. Ultraviolet spectrophotometry for drug content determination was established.Morphology was observed by transmission electron microscope(TEM),particles size distribution was analyzed by normal distribution,zeta potential was determined by surface potential analysator, entrapment efficiency and drug load were determined after hypothermy ultracentrifugation,in vitro release content and mechanism were studied by dynamic membrane dialysis,initial stability was evaluated by influence factor test,accelerated test and ambient temperature sample retention test.Results The main factors effecting preparation of Bre-CS-NP were PlournicF-68 content, Dextran-70 content,stirring rate,breviscapine content,chitosan concentration and chitosan/TPP proportion,and the scope of investigation is determined.The optimized formulation and preparation parameters obtained from uniformity design experiment were as follows:PluronicF-68 36mg,Dextran-70 30mg,placed in 25ml beaker,and 3.0ml 4.0mg/ml chitosan solution with pH=6.0 was added.The uniform solution was obtained by hypersound. 2ml 1.9mg/ml Bre-pH6.8PBS was added into the former solution with the electromagnetism stirring for 30min.1.0ml 2.0mg/ml TPP solution was added slowly with stirring for 15min, and then the Bre-CS-NP was obtained.Bre-CS-NP prepared of optimized formulation and artwork were mainly circular and uniform and had no adhesion.The mean diameter was 254.1±13.6nm,and zeta potential was +31.53±1.19mV.Breviscapine content was determined by ultraviolet spectrophotometry, pH6.SPBS as dissolvent,which was not interfered by excipients and convenient and sensitive. Linear correlation was good in the concentration range of 2.0～16.0μg/ml.Regression equation was A=0.06059×-0.01568,r=0.9996.Within-day and the intra-day RSD were less than 3% and the Mean recovery was more than 99%.The entrapment efficiency and drug load content were 75.8%and 58.5%,respectively.Release of Bre-CS-NP in vitro could be described by Higuchi model and could be described by the following equation:Q=10.36 t^1/2 +10.38(r=0.9990);and the crude drug could be described by zero order kinetics:lgQ=0.2326t+1.3296(r=0.9937).Stability study results showed that,Bre-CS-NP was sensitive to megatemperature and highlight.Bre-CS-NP had no obvious changes and was stable except slight decreasing of entrapment efficiency.Conclusion Bre-CS-NP was targeting drug delivery system and drug is enriched in the target position,thus the bioavailability was enhanced,dosage and administration frequency decreased,toxicity and side effect reduced and therapeutic index elevated.Bre-CS-NR obtained by one step ion forming gelation method,with high entrapment efficiency and sustained release met the requirement of experiment design.