| Inonotus obliquus is one of fungal drugs,it is found that inonorus obliquus contains lots of polysaccharides,which has the effect of inhibiting the growth of tumor,lowering blood pressure and blood sugar,antioxidation and will be widely used.The content of this research was to optimize the extraction and purification process of polysaccharides of inonporus obliquus,it was prove that the drug had anti-tumor effect through cell culture in vitro,and the purified polysaccharide was to prepare sustained-release tablets,and its quality evaluation was done.Extraction process:the hot water extraction method was used,the distilled water was used as a solvent and the extraction rate of polysaccharide was used as a reference index.The factors of extracting temperature,time,ratio of material and liquid and the number of extraction times were investigated with the extraction rate of polysaccharide as reference index.The optimum extraction conditions were as follows:the ratio of material to liquid was 1:20,the extraction temperature was 95?,the extraction time was 2.25 h and the extraction times was 1 time.In this condition,the extraction rate of the polysaccharide was improved.The preparation process was simple,easy to operate and met the requirements of preparation.Purification process:(1)The protein impurities in polysaccharides were removed by the Sevage method,and the best process was:the proportion of n-butanol and chloroform was 1:5,the ratio of drug to solvent was 4:1.The speed of this method was 3000 r/min,the rotation was 20 min and the static position was 20 min.(2)In addition,the macroporous resin adsorption method was used to remove impurities further,the selection process was:the sample amount of polysaccharide solution was 70 mL,the sample concentration was 5 mg/mL,the pH of sample solution was the original solution itself and the flow rate was 2 mL/min.After loading,the polysaccharides from macroporous resin were eluted.The eluent was 50%ethanol,the volum of which was 90,and the flow rate was 2 mL/min.(3)The dialysis bag was used to remove the small molecules with a molecular weight of 3000 D.This process was easy to operate and could effectively remove impurities in polysaccharide to obtaine polysaccharides with high purity.The antitumor activity in vitro:the inhibitory effect of polysaccharides on gastric cancer cells SGC-7901 was determined by MTT,and the inhibition rates of high,medium and low dose group were 25.52%?24.14%?8.56%at 24 h,29.09%?28.24%?12.07%at 48 h,respectively.They were significant differences compared with the positive control group,and the middle and high dose groups were significant differences compared with the negative control group,it was proved that polysaccharides of inonotus obliquus could inhibit tumor.The preparation of sustained release tablets:the purified polysaccharide obtained was used as the main drug to prepare sustained-release tablets.The prescription of which was optimized by Box-Behnken test software,and the dosage of ethyl cellulose was 158.3 mg,and filler was 5.54%and PEG was 10.21%.The process method could be implemented and met the design requirements.Quality evaluation:quality evaluation in vitro was evaluated by factors such as appearance,hardness,stability,homogeneity and dissolution.The appearance was dark brown,the thickness was uniform and the surface was smooth.The hardness range was around 5-7 N;an average of polysaccharide was 57.4 mg per slice;the results of stability investigation showed that it was no effect on release degree by 60? high temperature,75%high humidity and 4000 xL glare,however the release degree was affected at 92.5%high humidity,which indicated that should be confined storage.Through the study of the appearance,hardness,content determination,cumulative release and stability of the sustained release tablets,the method of quality evaluation was established,which could effectively control the quality of polysaccharide sustained-release tablets. |
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