| Objective: To select optimal extraction process of Bletilla striata polysaccharides. To prepare Bletilla striata hemostatic powder with Bletilla striata polysaccharides and investigate its hemostatic properties and biocompatibilities to provide research data to develop a new clinical hemostatic material.Method: ①The single factor tests and L9(34) orthogonal tests were designed, taking ratio of liquid to material, extraction time, extraction temperature and extraction times as factors. According to the extraction rate of Bletilla striata polysaccharides measured by UV Spectrophotometry, optimal extraction process was selected. ②Hydrochloric acid method and dialysis method were used to purify Bletilla striata polysaccharides to remove protein and small molecular weight impurity. ③Ethanol solution was used to pretreat raw starch and took ratio of liquid to material, concentration of ethanol, temperature and reflux reaction time as factors to adopt the single factor tests and L9(34) orthogonal tests. According to mobility and water absorption rate of modified starch, optimal process was selected. ④Bletilla striata hemostatic powder was prepared with Bletilla striata polysaccharides and modified starch. The influence of means of vacuum drying, spray drying and freeze drying was studied for drying Bletilla striata hemostatic powder. Then we selected the optimal preparation condition according clotting time in vitro through the investigation of different ratio of liquid to material. ⑤Tail bleeding wound models of rats and liver hemorrhage models of rats were bulit to evaluate hemostatic properties of Bletilla striata Hemostatic Powder. ⑥ The tests on guinea pig skin irritation and anaphylaxis and guinea pig embedding experiments were made to investigate its biocompatibilities. ⑦The quality of Bletilla striata hemostatic powder was investigated according sensory, physicochemical and microbial index and the index components.Results: ①It’s reasonable and feasible to determinate the content of Bletilla striata polysaccharides by UV Spectrophotometric and the optimal extraction condition is as follows: ratio of liquid to material is 15, extraction times is 3, extraction temperature is 90℃,the extraction time is 1.5h every time. In this condition, the average extraction rate of polysaccharides is 74.33%. ②The content of protein of polysaccharides is 0.28% by hydrochloric acid method. Polysaccharides is more than 70% after removing protein and 3 times purification. ③The optimal modified condition is as follow: ratio of liquid to material is 4, concentration of ethanol is 30%, temperature is 60℃, reflux reaction time is 1h. In this condition, the water absorption rate of modified starch is 221.85%. Compared with the raw starch, the water absorption rate of modified starch increased by nearly 60%, the bulk density decreased to nearly 22%. ④Freeze drying method is determined to prepare Bletilla striata hemostatic powder with condition that : ratio of quality of Bletilla striata polysaccharides(g), water volume(m L) and quality of modified starch(g) is 1:40:10. ⑤In tail bleeding wound models of rats, tail bleeding could be effectively stopped after the tail were clamped for 1min in group A, but the tails had signs of bleeding and still bled after the tail were went ahead being clamped for 2min in group B, in the end, tail failed to stop bleeding in 11 min by testing repeatedly in group B. In liver hemorrhage models of rats, the bleeding time of group C and D was(265.3±2.3)s and(264.8±1.7)s respectively(p>0.05), the bleeding amount was(0.078±0.016)g and(0.075±0.018)g respectively(p>0.05). ⑥There is not skin irritation and allergic reaction on guinea pigs in group of Bletilla striata hemostatic powder and blank group. Bletilla striata hemostatic powder embedded in guinea pig inner thigh started to be degradated in 1d and was degradated completely in 10 d. ⑦The test results of quality of Bletilla striata hemostatic powder are stable.Conclusion: Bletilla striata hemostatic powder has good hemostatic properties and biocompatibilities. The tests results of quality can provide research data for subsequent functional and safety evaluation, establishment of quality standard and development of a new clinical hemostatic material. |
PDF Full Text Request