Extraction And Anticoagulant Activity Of Crude Polysaccharide From Aronia Melanocarpa

Objective: Aronia melanocarpa has a variety of nutrients and a wide range of biological activities,such as anti-inflammatory,anticoagulant,antitumor,hypoglycemic and lipidlowering pharmacological activities.At present,the nutritional contribution and pharmacological activity of polyphenols are mainly focused,and there are few reports on polysaccharides of aronia melanocarpa.The purpose of this study was to optimize the extraction method and content determination of crude polysaccharide of aronia melanocarpa,and further investigate the anticoagulant activity of crude polysaccharide of aronia melanocarpa in vitro,so as to provide a theoretical basis for clinical research.Methods: In this paper,the crude polysaccharide was extracted by decocting method and ultrasonic assisted method.With the yield of crude polysaccharide as the index,the single factor optimization experiment was carried out on the determination conditions,and the optimal extraction conditions of the two extraction methods were selected by three-factor and three-level response surface analysis method.Then,phenol-sulfuric acid method was used to optimize the color development conditions of crude polysaccharide of aronia melanocarpa,and the best technological parameters for the determination of crude polysaccharide content of aronia melanocarpa were obtained.The anticoagulant activity of crude polysaccharide of aronia melanocarpa was further discussed.Results: In this paper,the crude polysaccharides from the fruit extract of aronia melanocarpa were studied,and the decocting time,decocting times and solid-liquid ratio of the two extraction methods were investigated by single factor experiment.The extraction process was optimized by response surface method with ultrasonic time,ultrasonic frequency and solid-liquid ratio.Experimental results showed that the optimal extraction parameters of decocting method and ultrasonic method were as follows: decocting time 81 min,decocting times 4,solid-liquid ratio 1:35 g:m L;The ultrasonic time was 60 min,the ultrasonic frequency was 3,the solid-liquid ratio was1:50 g:m L;The results showed that the yield of crude polysaccharide extracted by decocting method was 11% and 3%,respectively,higher than that by ultrasonic method.Then the content of crude polysaccharide was determined by phenol-sulfuric acid method,and the influencing factors of color development conditions were investigated one by one,and the best experimental conditions were obtained.In other words,the absorbance was determined at 490 nm when the color developing temperature was 50℃,the water bath time was 25 min,the concentration of sulfuric acid was 5 m L,and the standing time was 30 min.The total sugar content in the crude polysaccharide of L.sorbus was 47.14%.The results showed that the crude polysaccharide of aronia melanocarpa had a certain degree of anticoagulant activity in vitro,especially the effect of activating partial thromboplastin time(APTT)was significantly higher than that of thrombin time(TT).These results indicated that the crude polysaccharide of aronia melanocarpa exerts its anticoagulant activity in vitro through both endogenous and co-coagulation pathways.Conclusion: This experiment not only obtained the optimal extraction parameters and yield of crude polysaccharide under the optimal extraction conditions,but also further clarified the content of crude polysaccharide and the optimal color conditions of crude polysaccharide,which provided a theoretical basis for later research.In addition,the experimental results showed that the crude polysaccharide had certain anticoagulant ability in vitro.