【作者】 王群；

【导师】 马承慧；

【作者基本信息】 东北林业大学 ， 森林植物资源学， 2016， 硕士

【摘要】 松科植物广泛分布于我国各地区,松科植物松针中具有多种活性成分,其中多酚类物质是重要的天然抗氧化剂,具有很大的应用价值。本文比较了10种松科植物松针中活性物质含量及多酚类物质的抗氧化活性,优化了油松松针多酚提取物的纯化工艺,为松科植物的开发利用提供理论基础。10种松针提取物中蛋白质平均含量在1.89mg/g鲜重左右,多糖平均含量在54.93mg/g左右,叶绿素平均含量在1.11 mg/g鲜重左右,总多酚平均含量在41.45mg/g,黄酮平均含量约为815mg/g,原花青素平均含量约为4.3mg/g。10种松针中蛋白质、多糖、叶绿素、黄酮、多酚和原花青素含量最高的分别为雪松、蓝粉云杉、乔松、雪松、油松、雪松；含量最低的分别为红皮云杉、樟子松、萌芽松、红皮云杉、红皮云杉、红皮云杉。HPLC分析测定10种松针多酚提取物特征化合物主要为没食子酸、儿茶素、咖啡酸、表儿茶素、花旗松素和对香豆酸。10种松针多酚提取物都含有儿茶素,其中油松含量最多,萌芽松松针中没食子酸含量远远大于其他松针,雪松松针提取物咖啡酸含量最高,油松松针提取物表儿茶素含量最高,红松松针提取物花旗松素含量最高,日本冷杉松针提取物对香豆酸含量最高。以DPPH自由基清除率、羟自由基清除率、脂质过氧化抑制率和总还原能力为抗氧化评价指标,比较10种松科植物松针多酚类物质抗氧化活性。实验结果表明在清除DPPH自由基方面上,雪松具有最大清除率,其次是萌芽松和油松,萌芽松清除DPPH的IC50值最小,其次是油松和蓝粉云杉；清除羟自由基方面上,油松对OH·清除率最大,其次是红松和雪松,樟子松清除OH·的IC50值最小,其次是雪松和油松；在样品浓度较高的情况下,雪松的总还原能力最大,其次是蓝粉云杉和油松,雪松松针多酚总还原能力的EC50值最低,其次是油松和蓝粉云杉；在抑制脂质过氧化体系中,油松具有最大的抑制率,其次为雪松和蓝粉云杉,油松对脂质过氧化抑制能力的IC50值最小,远远小于其他松针多酚。因此综合看来,油松的松针多酚抗氧化性优于其他几种松科植物松针多酚。相关性分析表明松科植物松针中多酚类物质是其主要抗氧化物质。以油松松针多酚提取液为原料,比较D101、AB-8、X-5和D101大孔树脂对油松松针多酚的吸附率和解析率,经过树脂筛选,发现AB-8大孔树脂对油松松针多酚具有有较高的吸附率和解析率,因此选择AB-8大孔树脂为油松松针多酚纯化的最佳树脂。通过对油松松针多酚纯化的单因素试验及正交试验优化,纯化油松松针多酚的最佳条件为上样浓度25mg/mL,上样体积为2OmL,洗脱速度16mL/min,乙醇浓度60%。经AB-8大孔树脂纯化,油松松针多酚纯度由29.95%提升到69.78%。更多还原

【Abstract】 Pinaceae plants widely distributed in China.Pine needles contain of multiple active ingredients.Polyphenols from Pinaceae plants are important natural antioxidants and have tremendous application value. This paper compared the contents of active ingredients and the antioxidant activity of polyphenols from 10 species of pinaceae plants pine needles.Optimize the purification technology of Pinus tabulaeformis needles polyphenols extract to providing theoretical basis for the development and utilization of Pinaceae plants.The main research contents and conclusions are as follows:(1) Using spectrophotometric method to determination the contents of protein, polysaccharide, chlorophyll, flavonoids, polyphenols and procyanidins in the 10 kinds of pine needles. The protein average content in the 10 kinds of extracts from pine needles is 1.89 mg/g fresh weight, the average content of polysaccharides is about 54.93 mg/g, the average content of chlorophyll is about 1.11 mg/g fresh weight, the average content of polyphenol is about 41.45 mg/g, the average content of flavonoids is about 8.15 mg/g, the average content of procyanidins is about 4.3 mg/g. The highest contents of protein, polysaccharide, chlorophyll, flavonoids, polyphenols, procyanidins in 10 kinds of pine needles are Cedrus deodara, Picea pungens, Pinus griffithii McClelland, Cedrus deodara, Pinus tabulaeformis Carr., Cedrus deodara, respectively. The highest contents are Picea koraiensis, Pinus sylvestris Linn. var. mongolica Litv., Pinus echinata, Picea koraiensis, Picea koraiensis, Picea koraiensis.(2) Using HPLC to analysis and measure 10 kinds of pine needles extract characteristic compounds, which mainly are gallic acid, catechin, caffeic acid, epicatechin, taxifolin and coumaric acid.They all contain catechin, among them the most contents is Pinus tabulaeformis Carr., the gallic acid content of Pinus echinata is greater than other pine needles. Cedrus deodara has the the highest content of caffeic acid, Pinus tabulaeformis Carr. has the the highest content of epicatechin, Pinus koraiensis Sieb. et Zucc. has the the highest content of taxifolin, Abies firma S. et. Z. has the the highest content of coumaric acid.(3) Based on the index of scavenging rate of DPPH free radicals, hydroxyl free radicals, inhibition rate of lipid peroxidation and the total reducing power, comparing the 10 species of pinaceae plants pine needles polyphenols antioxidant activity.The experimental results show that in scavenging DPPH free radicals, Cedrus deodara has the maximum scavenging rate, followed by Pinus echinata and Pinus tabulaeformis Carr.. Pinus echinata has the lowest 1C50 to scavenging DPPH free radicals, followed by Pinus tabulaeformis Carr. and Picea pungens. In scavenging hydroxyl free radicals, Pinus tabulaeformis Carr. has the maximum scavenging rate, followed by Pinus koraiensis and Cedrus deodara. Pinus sylvestris Linn. var. mongolica Litv. has the lowest IC50 to scavenging hydroxyl free radicals, followed by Cedrus deodara and Pinus tabulaeformis Carr. Under the condition of high concentration, the highest total reducing power is Cedrus deodara, followed by Picea pungens and Pinus tabulaeformis Carr. Cedrus deodara has the lowest EC 50 of total reducing power, followed by Pinus tabulaeformis Carr. and Picea pungens. In inhibiting the lipid peroxidation, Pinus tabulaeformis Carr. has the maximum inhibition rate, followed by Cedrus deodara and Picea pungens. IC50 of inhibiting the lipid peroxidation, Pinus tabulaeformis Carr. is far less than other pine needles polyphenols. So overall, the oxidation resistance of Pinus tabulaeformis Carr. pine needles polyphenol is better than several other pinaceae plant pine needles polyphenols.Correlation analysis showed that pinaceae plant polyphenols in pine needles is the main antioxidant compounds.(4) Pinus tabulaeformis Carr. needles polyphenols extract as raw material, comparing D101, AB-8, X-5 and D101 macroporous resin adsorption and resolution rate. AB-8 macroporous resin have high adsorption and resolution rate for Pinus tabulaeformis Carr. needles polyphenols extract. So AB-8 macroporous resin is the best resin for Pinus tabulaeformis Carr. needles polyphenols.更多还原