【作者】 周洁；

【导师】 姚卫蓉；

【作者基本信息】 江南大学 ， 营养与食品卫生学， 2009， 硕士

【摘要】 本论文以酒浸杨梅为研究目标,通过体内抑菌实验探索其活性特征,并通过体外抑菌实验研究其抑菌活性成分的结构,为充分开发利用杨梅资源奠定理论基础。首先,以不同酒精度的白酒浸泡杨梅,验证了杨梅浸酒后具有体外抑菌活性；定量比较发现酒液的抑菌活性强于果肉的抑菌活性,且浸泡方式(整粒、破碎)、白酒的种类及其酒精度(25%、35%、56%)对抑菌性的影响不大。接着,本文研究了杨梅果实提取物的抗小鼠腹泻作用,包括预防和治疗作用。研究治疗作用时先给小鼠腹腔注射甲型副伤寒沙门氏菌造成急性腹泻模型,再经口灌胃杨梅果实提取物；研究预防作用时,先给小鼠灌胃杨梅果实提取物,再给小鼠注射菌液致泻。分别给小鼠灌胃杨梅醇提物和水提物,通过观察腹泻次数发现,灌胃杨梅果实提取物后,小鼠的腹泻情况明显缓解,醇提物的抗腹泻效果优于水提物。然后,利用氯仿、石油醚(30-60℃)、乙醚、乙酸乙酯对杨梅果实的醇提物逐级纯化,通过抑菌圈实验发现氯仿相和石油醚相没有抑菌活性,乙醚相、乙酸乙酯相和最后的水相都有抑菌活性,乙酸乙酯相的活性最强；薄层层析(展开剂为丙酮：水=7：5)后经生物自显影法初步证实其中至少有三个抑菌活性成分。利用聚酰胺柱层析法,以10%、40%、60%的乙醇溶液为梯度洗脱剂,对经氯仿、石油醚(30～60℃)纯化后的粗提物进一步分离,得到4个馏分T1,T2,T3和T4。T3没有抑菌活性,粗提物、T1、T2、T4的最低抑菌浓度(MIC)分别为：0.625、0.625、2.5、0.156 mg／ml；经生物活性追踪后,得率为65.62%,T1的总活性占98.91%,是杨梅果实中抑菌活性最强的部分。进一步将T1通过硅胶柱层析和Sephadex LH-20柱层析纯化得到纯物质T1-A,经13C核磁共振进行结构鉴定,初步确定T1-A是一种由5个单糖分子组成且分子量为840的低聚糖,其MIC值与T1的一致。更多还原

【Abstract】 Firstly, the antibacterial activity was qualitatively identified, and the antibacterial intensity of the solution was quantitively stronger than that of the flesh. The activity could not be influenced by alcohol drink species, alcohol concentration or the methods of immersing.The prevention and antidiarrheal activities were studied. When the antidiarrheal activity was studied, the acute mice diarrhea models were induced by intraperitioneal injection of Salmonella, then the extracts were administrated orally to the mice, and the number of diarrhea was recorded; When the prevention activity on the mice diarrhea was studied, the extracts were administrated orally to the mice, then the mice diarrhea was induced. It showed that the bayberry extracts had both prevention and antidiarrheal activities on diarrhea of mouse induced by Salmonella paratyphi A, and the effect of the ethanol extract was stronger than that of water extract.The ethanol extract of bayberry was fractional extracted by chloroform, petroleum ether (30～60℃), ethyl ether and ethyl acetate. And the ethyl ether, ethyl acetate and water phase were found to have antibacterial activity. Five components were separated by TLC with dilution solvent of acetone:water(7:5), and three components were identified to have significant antibacterial activity through bio- autography detection.The ethanol extract followed by chloroform, petroleum ether purification were further isolated by polyamide column chromatography, with 10%,40%,60% ethanol as mobile phase and 0.8ml/min velocity. Four fractions were obtained and only the third fraction T3 did not have the anti-biological activity. The MIC values of the crude extract and the other three fractions (T1,T2,T4)were 0.625,0.625,2.5,0.156mg/ml. By bio-tracking method, the primary antibacterial compound was the first fraction.The first fraction was further purified by silica and Sephadex LH-20 column, a oligosaccharide compound T1-A with 840 Dalton molecular weight was identified by UPLC-MS, IR and 13C-NMR, and its MIC was the same with the first fraction’s.更多还原