【作者】 陈留勇；

【导师】 孟宪军；

【作者基本信息】 沈阳农业大学 ， 食品科学， 2004， 硕士

【摘要】 采用水提醇沉法从黄桃果肉中提取出水溶性粗多糖，利用正交试验确定了最佳提取工艺条件：浸提温度90℃、液固比（v／w）2：1、浸提时间3h。粗多糖为黄褐色粉末，难溶于冷水，可溶于热水、稀酸、稀碱，不溶于乙醇、丙酮、乙醚、正丁醇等有机溶剂，粘度较大。黄桃水溶性粗多糖经测定，其各组分含量分别为中性多糖60％，酸性多糖36.97％，蛋白质0.91％，水分1.37％。过氧化氢氧化法除去色素后，为纯白色，7次Sevag法除蛋白，蛋白质含量为0.13％。经DEAE-Sepharose离子柱层析得到两个组分，分别出现在水洗脱部分和盐洗脱部分，收集相同峰位组分，减压浓缩后，蒸馏水透析48 h，冷冻干燥得纯白色多糖HTP1和HTP2。样品HTP1和HTP2经SephacrylS-300凝胶柱层析，均为单一对称峰。经HPSEC鉴定HTP1为单一组分，分子量为52107，HTP2也为单一组分，分子量为59855。乌式粘度计测定特性粘度HTP1为13.83ml／g，HTP2为37.16ml／g。HTP1和HTP2分别经完全酸水解、高碘酸氧化、Smith降解、紫外光谱、红外光谱和核磁共振图谱分析。HTP1主要单糖组成为D-Glc、D- Gal、D- Arb和L- Rha，摩尔比为D-Glc：D-Gal：D-Arb：L-Rha=1.0：4.2：14.5：1.5，主链为α（1→3）键连接的阿拉伯半乳糖，有部分1→6键分支。HTP2主要单糖组成为D-GalA、D-Gal、D-Man，摩尔比为D-GalA：D-Gal：D-Man=34.3：5：1，主链为α（1→3）键连接的半乳糖醛酸。 本实验同时考察了粗多糖和HTP对有害菌的抑制作用，发现粗多糖对大肠杆菌、黑曲霉和扩展青霉都有一定的抑制作用，尤其是对黑曲霉的抑制尤为明显；而HTP只对大肠杆菌有抑制作用，对黑曲霉和扩展青霉没有抑制作用。本实验研究结果表明，HTP1对羟基自由基（·OH）和超氧阴离子自由基（O₂^-）均有一定清除作用，其IC₅₀分别为2.87 mg／ml和401.3μg／ml。HTP2对·OH的清除能力远远大于HTP1，其IC₅₀为270.92μg／ml。但对O₂^-却无任何清除作用。HTP1和HTP2各分别以100mg／kg、200mg／kg剂量经腹腔注射，连续10d对足趾皮下接种Lewis肺癌的小鼠给药，同时以环磷酸胺作为抗肿瘤阳性对照、云芝多糖作为提高免疫力阳性对照、等体积的生理盐水作为阴性对照。末次给药后处死小鼠，截取小鼠足趾，计算抑瘤率分别为54.69%、43.44%及47.08%、44.48%。其中HTPI高剂量组的抗肿瘤疗效较HTPZ高剂量组更为明显。HTPI和HTPZ均能明显提高NK细胞活性，且活性提高均在47%以上，与云芝多糖52%的提高率相差不大，但与阴性对照相比差异均达极显著水平（P<0.01）。同时对荷瘤小鼠的淋巴细胞转化的活性也具有一定的促进和提高，HTPI和HTPZ的高剂量组与低剂量组的刺激指数相差不大，均在1 .70一1.74之间，无显著差异，低于阳性对照云芝多糖（1.83），但与阴性对照组相比，差异达极显著水平（P<0.01）。说明黄桃水溶性多糖能明显促进淋巴细胞的转化。更多还原

【Abstract】 The water-soluble polysaccharides of A.Persica.L.var.seleropersica were extracted in hot water, and then isolated by ethanol. The effect of three factors such as the temperature of extraction, the proportion of hot water volume, the time of extraction on the extraction of the water-soluble polysaccharides were investigated. With the orthogonal design of L9(34), the optimum conditions of extraction were obtained as follow: adding 2 times water as much as fruit weight in 90癈 for 3 hours. The crude polysaccharide was filemot, can soluble in hot water, alkali and acid, cannot soluble in ethanol, acetone, aether etc. The protein in crude polysaccharides was removed by Sevag way and the pigment in crude polysaccharides was removed by H2O2. The water-soluble polysaccharides of peach which color was pure white were got. HTP were further purified to HTP1 and HTP2 with DEAE-Sepharose column chromatography. HTP1 and HTP2 were purified with Sephacryl S-300 column chromatography respectively, however were singular peak. The high performance size-exclude chromatography proved that HTP1 and HTP2 were pure material. The average molecular weight of HTP1 was 52107 and the average molecular weight of HTP2 was 59855. Their structures were investigated by chemical methods (composition analysis, periodate oxidation, and Smith degradation) and spectroscopic methods (UV, IR, NMR). Their monosaccharide was identified by HPLC after TFA hydrolyze. HTP1 is composed of D-glucose, D-galactose, D-Arabinose and L-Rhamnose in the ratio of 1.0: 4.2: 14.5: 1.5, HTP2 iscomposed of D-galactose, D-Mannose and D-Galacturonic acid in the ratio 34.3: 5: 1. The result of periodate oxidation and Smith degradation showed that the major linkage of HTP1 is (1→3) D-galactose and the major linkage of HTP1 is (l→ 3) D-Galacturonic acid. Infrared analysis show that HTP1 and HTP2 were all alpha -glycoside linkage.The crude polysaccharides and the HTP had different effect on antibacterial. The crude polysaccharides can inhibit Escherichia coli, Penicillium citrinum and Aspergillus niger, and HTP can only inhibit Escherichia coli, has no effect on Penicillium citrinum and Aspergillus niger. HTP1 and HTP2 have evident effect in cleaning .OH, the IC50 are 2.87mg/ml and 270.92 n g /ml. HTP1 has evident effect in cleaning O2- ,the IC50 is 401.3 μg /ml. HTP2 has no effect on cleaning O2:. HTP1 and HTP2 were injected into the mice which were inoculated Lewis lung cancer with high dose 200mg/kg and low dose 100mg/kg respective. The result showed that when high dose was injected, the percentage of anti-tumors of HTP1 and HTP2 were 54.69% and 47.08%, when low dose was injected, the percentage of anti-tumors of HTP1 and HTP2 were 43.44% and 44.48%. HTP1 and HTP2 exhibited stimulating activity to the translation of lymphocyte and improving to the activity of NK. These indicated that HTP1 and HTP1 could improve the immunity of the mouse.更多还原