Study On The Preparation And Quality Control Of Chitosan And The Property Of Insulin-loaded Chitosan Nanoparticles

Chitosan is honored as the sixth life elements after proteins, fats, carbohydrates, vitamins and inorganics in academic circles, which has non-toxic, non-stimulation, non-allergenic and non-mutagenic effect, and is good biocompatibility and biodegradable polycation compound. Chitosan is extremely active in the pharmaceutical fields, and is natural cationic drug delivery. Chitosan as drug carriers is to control drug release, to prolong therapeutic effects of drugs and to reduce side effects of drugs, which can enhance membrane permeability of the hydrophobicity drugs and drug stability and change administration routes, but also can enhance the capacity of targeted drug delivery. The purpose of this study involved chitin and chitosan which are prepared from red tail prawn shell, resource utilization, the establishment of quality control and detection methods of molecular weight and deacetylation degree of chitosan, prepared different degree of deacetylation and molecular weight of chitosan, studied on performance of insulin-loaded chitosan nanoparticles and water-soluble chitosan nanoparticles, which are established the foundation for the development of a new type of oral insulin preparations.In this study, red tail prawn shells are the east of GuangDong province as the raw materials, which was adopted a relatively simple process to prepare chitosan, and recovered the useful components which were produced in the preparation process of the waste water, which included fat, protein, calcium carbonate, the alkali solution was reused in the preparation process, which was reduced the cost of production in order to achieve the use of resources. The COD value was decreased 19% after took off fat from wastewater than not took off fat, the fat was recovered from the preparation process, which can reduced COD of the wastewater and the degree of pollution, and finally, the COD meeted national wastewater discharge standards at the three levels of standards after preliminary treatment of the wastewater. We utilized the high degree of deacetylation and high molecular weight of chitosan as raw materials, to obtain the low molecular weight, water-soluble chitosan through the oxidative degradation, to study various conditions on single-factor experiment, using orthogonal experimental design to study the best experimental conditions of preparation of low molecular weight chitosan, and finally, to prepare a series of different molecular weight chitosan at optimized experimental conditions. Another we utilized the low degree of deacetylation of chitosan as raw material, to prepare the high degree of deacetylation of chitosan at the best experimental conditions through single-factor and orthogonal experiment, and finally, preparing a series of different degree of deacetylation of chitosan at optimized experimental conditions.Aalkalimetry is standard method of determining degree of deacetylation of chitosan generally applied in aquaculture and other industry. The method adopts helianthine- triphenylrosaniline as indicator, which end-point color is difficult to observe, making poor reproducibility and wide error when low degree of deacetylation of chitosan is determined, and triphenylrosaniline has toxic, causing environmental pollution and harm to human health. In this paper, we have studied different types of indicators which can be used in alkalimetry, the results suggested that helianthine-methylene blue (1:1) mixed indicator can instruct color changes of titration end significantly, the accuracy and reproducible are good. This indicator is likely to become common indicator of determining different degree of deacetylation of chitosan, which can resolve current common and poison problem of alkalimetry.In order to study the advantages of chitosan which is loaded drugs, we studied the preparation technology, the impact factors of insulin-loaded, evaluation indexes and in-vitro release performance of chitosan nanoparticles and water-soluble chitosan nanoparticles, respectively. The encapsulation efficiency and loading capacity of insulin-loaded chitosan nanoparticles was 66.31±1.79% and 10.31±1.52%, respectively, the average size of nanoparticles was 292nm. The encapsulation efficiency and loading capacity of insulin-loaded water-soluble chitosan nanoparticles was 48.28±0.90% and 9.52±1.34%, respectively, the average size of nanoparticles was 153nm. We deeply researched the determination method for the encapsulation efficiency and loading capacity of insulin-loaded chitosan nanoparticles, the improved Bradford method eliminated the interference of supernatant of blank chitosan nanoparticles in order to determine content of insulin of insulin-loaded chitosan nanoparticles, and the methodological investigation showed sensitivity, accurate and reliable. A simple, rapid and economy improved Brandford method for the determination of insulin from nanoparticulate carriers made from chitosan is validated.