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目的:通过4种不同提取方法用于中药生产中废弃物的多糖资源化的比较研究,优选出脉络宁注射液生产废弃物石斛药渣中提取多糖的方法,实现对中药资源的再循环利用。方法:采用酶解法、湿法超微粉碎法、超声法和加热回流法分别提取石斛药渣中石斛多糖,以苯酚-硫酸法测定其多糖提取率;通过单因素考察和正交试验对提取率最高的提取方法进行工艺优选;考察石斛药渣多糖(DDP)对小鼠脾淋巴细胞的代谢转化和小鼠单核巨噬细胞RAW264.7释放一氧化氮(NO)能力的影响。结果:4种提取方法的多糖提取率分别为0.51%,8.79%,5.34%,3.44%,确定最佳提取方法为酶解;其最佳酶解条件:p H5.3,料液比1∶55,温度50℃,酶活性4 000 U·g-1,提取时间300 min;与对照组比较,质量浓度在62.5~125 mg·L-1,DDP显著促进小鼠脾淋巴细胞增殖,DDP与脂多糖(LPS)协同促进作用显著(P<0.05,P<0.01),质量浓度为62.5 mg·L-1时,DDP与Con A协同促进作用显著(P<0.05),质量浓度在62.5~500 mg·L-1,DDP对小鼠单核巨噬细胞RAW264.7产生NO的促进作用极显著(P<0.01)。结论:酶解法提取石斛药渣中多糖的工艺简便、得率稳定,具免疫增强作用,具有推广应用价值。
OBJECTIVE: To compare and optimize the utilization of polysaccharides in the waste products of traditional Chinese medicine (TCM) by four different extraction methods, and to extract the polysaccharides from Dendrobium officinale from the production waste of Mailuoning Injection, the method of recycling polysaccharides of traditional Chinese medicine resources was realized. Methods: Polysaccharides from Dendrobium officinale were extracted by enzymatic hydrolysis, wet ultrafine grinding method, ultrasonic method and refluxing method respectively. The extraction rate of polysaccharide was determined by phenol-sulfuric acid method. The extraction rate The highest extraction method for process optimization; study Dendrobium dregs polysaccharide (DDP) on mouse splenic lymphocyte metabolic transformation and mouse monocyte-macrophage RAW264.7 release of nitric oxide (NO) ability. Results: The extraction rates of four polysaccharides were 0.51%, 8.79%, 5.34% and 3.44%, respectively. The optimal extraction method was enzymolysis. The optimum conditions for enzymolysis were p H5.3, 55, temperature 50 ℃, enzyme activity 4 000 U · g-1, extraction time 300 min; compared with the control group, the concentration of 62.5 ~ 125 mg · L-1, DDP significantly promote mouse splenic lymphocyte proliferation, DDP and (P <0.05, P <0.01). When the concentration of LPS was 62.5 mg · L-1, the synergistic effect of DDP and Con A was promoted (P <0.05) and the concentration of LPS was between 62.5 and 500 mg · L-1, DDP significantly promoted NO production in RAW264.7 cells (P <0.01). Conclusion: Enzymatic extraction of polysaccharides from Dendrobium officinale has the advantages of simple process, stable yield and immune enhancement effect, which has the value of popularization and application.