摘要
目的:优化锁阳多糖(Cynomorium songaricum polysaccharide,CSP)的提取纯化工艺。方法:回流提取法提取CSP,以CSP得率为评价指标,在单因素试验的基础上,结合正交试验对提取工艺进行优化;以蛋白脱除率和多糖保留率为考察指标,比较Sevage法、TCA法、醋酸铅法及木瓜蛋白酶法对CSP脱蛋白的效果,选择最佳脱蛋白方法;以脱色率和多糖保留率为考察指标,优选树脂,再在单因素试验的基础上利用响应面法优化大孔吸附树脂脱除CSP中色素的工艺。结果:最佳提取工艺为提取时间为120 min,料液比为1∶40,提取温度为100℃,提取次数3次,提取率为8.16%;最佳脱蛋白方法为醋酸铅法,蛋白脱除率为85.34%,多糖保留率为90.51%,综合评分为87.93;最优树脂为HPD100型大孔吸附树脂,最佳脱色条件为多糖含量为6.0 mg·mL^(-1),树脂用量为12.5%,糖液pH为5.4,脱色时间为150 min,脱色率为88.16%,保留率为81.51%。结论:经过单因素试验及正交试验、响应面试验选出CSP的最佳提取纯化工艺,条件科学合理、稳定可行,为提取纯化CSP提供实验依据。
Objective:To optimize the extraction and purification process of cynomorium songaricum polysaccharide(cynomorium songaricum polysaccharide,CSP). Methods:CSP was extracted by reflux extraction method,and the extraction process was optimized based on a single-factor test combined with an orthogonal test,using the CSP obtaining rate as the evaluation index. Protein removal rate and polysaccharide retention rate were used as indicators to compare the effect of the Sevage method,TCA method,lead acetate method and papain method on CSP deproteinization,and to select the best deproteinization method. Using the decolorization rate and the retention rate of polysaccharides as the investigated indexes,the resin was preferably selected,and then the process of removing pigments from CSP by macroporous adsorption resin was optimized based on single-factor experiments using response surface methodology. Results:Extraction time was 120 min,material-to-liquid ratio was 1:40,extraction temperature was 100℃,extraction times were 3 times,and extraction rate was 8.16%. The best method was the lead acetate method,with 85.34% protein removal and 90.51% polysaccharide retention,giving an overall score of 87.93. The optimum resin was HPD100 macroporous adsorption resin. The optimum decolorization conditions were 6.0 mg·mL^(-1) of polysaccharide,12.5% of resin,5.4 pH of sugar solution,150 min decolorization time,88.16% decolorization rate and 81.51% retention rate. Conclusion:The optimal extraction and purification process of CSP was screened by single-factor,orthogonal and response surface experiments,and the conditions were scientifically reasonable,stable and feasible. This provides an experimental basis for the extraction and purification of CSP.
作者
吕鑫
顾志荣
祁梅
张锐
郭燕
毛小文
葛斌
Lv Xin;Gu Zhirong;Qi Mei;Zhang Rui;Guo Yan;Mao Xiaowen;Ge Bin(College of Pharmacy,Gansu University of Chinese Medicine,Lanzhou 730000,China;Department of Pharmacy,Gansu Provincial Hospital,Lanzhou 730000,China)
出处
《中国野生植物资源》
CSCD
2022年第4期34-43,共10页
Chinese Wild Plant Resources
基金
甘肃省自然科学基金项目(20JR5RA154)
甘肃省青年科技基金计划项目(21JR7RA634)
甘肃省人民医院青年项目(20GSSY4-29)
“白求恩·求索-药学科研能力建设”项目(B-19-H-20200622)
甘肃省中药质量与标准研究重点实验室基金项目(ZYZL18-004)
甘肃省中药药理与毒理学重点实验室基金项目(ZDSYS-KJ-2018-010)。
关键词
锁阳多糖
提取纯化
单因素试验
正交试验
响应面试验
CSP
Extraction and purification
Single factor experiment
Orthogonal experiment
Response surface experiment
