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作 者:向维[1,2] 丁馨[1] 张薛磊[2] 吴媛媛[1] 陈松华[1,2]
机构地区:[1]中国人民解放军南京军区南京总医院疗养区(全军医学康复中心),江苏南京211131 [2]南京大学医学院临床学院,江苏南京210002
出 处:《浙江大学学报(医学版)》2012年第5期569-575,共7页Journal of Zhejiang University(Medical Sciences)
基 金:南京军区医学科学研究"十一五"课题(10MA133);国家自然科学基金(30872367)
摘 要:目的:优化彩虹明樱蛤酸性多糖的制备工艺。方法:采用碱提醇沉法,运用正交试验对彩虹明樱蛤多糖的分离过程进行优化,包括时间、温度、pH值和乙醇浓度4个因素;所获多糖经去蛋白、去色素、去离子后,用DEAE-纤维素离子交换柱纯化,以Sephadex G-100和醋酸纤维薄膜电泳鉴定及红外光谱初步研究。结果:优化的多糖提取制备工艺为:时间6 h、温度70℃、pH 8.0、乙醇浓度70%,获得纯白略带暗褐色的颗粒状固体;DEAE-纤维素离子交换柱分离洗脱后得到靶多糖组分彩虹明樱蛤酸性多糖(Moerella iridescens acidic polysaccharide,MIAP),经Sephadex G-100和醋酸纤维薄膜电泳鉴定,MIAP为均一多糖组分,呈乳白色绵柔絮状;红外光谱扫描,MIAP含α-D型吡喃葡萄糖。结论:正交试验法优化了MIP的提取工艺,分离纯化了MIAP。Objective: To optimize extraction and purification methods of acidic polysaccharide from Moerella iridescens(MIAP).Methods: With alkali extraction process and orthogonal experiment,the time consumption,temperature,pH value of the solution and alcohol concentration during the extraction were optimized.The crude products were deprived of protein,pigment and ion,then were purified with DEAE-cellulose ion-exchange chromatography and verified with Sephadex G-100 and cellulose acetate membrane electrophoresis,and examined with infrared spectrum.Results: The optimized extraction conditions were as follows:extraction time 6 h,extraction temperature 70℃,the solution pH 8.0 and the concentration of alcohol precipitation 70%.Intuitive features showed that the MIAP was pure white crystalline granular with slight dark brown color.The purification results demonstrated that the target MIAP was eluted and identified as a homogeneous components by DEAE-cellulose ion exchange column,Sephadex G-100 and cellulose acetate membrane electrophoresis.Infrared spectral scanning suggested that MIAP was α-D-type terminated glucopyranose.Intuitive features showed that MIAP was soft and cottony white.Conclusion: The extraction process with orthogonal test has been optimized and the acidic polysaccharide from Moerella iridescens is successfully isolated.
关 键 词:贝类 多糖类/分离和提纯 彩虹明樱蛤多糖/分离和提纯 正交试验/方法
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