核桃青皮成分分析及相关产品的研发

发布时间：2018-01-06 18:45

本文关键词：核桃青皮成分分析及相关产品的研发　出处：《昆明理工大学》2016年硕士论文　论文类型：学位论文

【摘要】：核桃青皮是核桃的主要副产品之一。核桃收获期间,大量的核桃青皮作为废弃物堆放,造成环境污染和资源浪费,只有少数量药用。核桃青皮有抗癌、抗氧化、抗衰老、降血糖等功能。加强其综合利用的研究与开发,具有重要的社会经济价值及生态价值。本文以核桃青皮为原料分析其营养成分,同时采用75%乙醇提取核桃青皮多酚,对其抗氧化活性、a-淀粉酶活性抑制作用及其稳定性进行初步研究,采用LC-MS技术鉴定其多酚成分,分析其不同多酚成分在α-淀粉酶活性抑制中的相互作用,探究其没食子酸与α-淀粉酶的相互作用机制。并采用常温固液发酵,制备核桃青皮果蔬酵素。主要研究内容和结果如下：1.核桃青皮的常规营养成分分析结果表明,核桃青皮具有丰富的营养。核桃青皮富含钾、钙、磷、镁、铁等多种矿质元素,且呈现高钾、低钠的特点。核桃青皮中除色氨酸外的氨基酸总含量为33.31%,其必需氨基酸与总氨基酸的比例为34.56%,必需氨基酸与非必须氨基酸的比例为52.81%,与世界卫生组织和联合国粮农组织(WHO/FAO)提出的食物蛋白氨基酸理想比例(EAA/TAA=40%, EAA/NEAA=60%)相接近。2.采用75%乙醇提取核桃青皮汁与青皮渣的总多酚,使用福林酚法测定提取物总多酚含量,通过测定粗提物清除DPPH自由基、羟基自由基和超氧阴离子自由基的能力来评价青皮多酚提取物的抗氧化能力,测定其α-淀粉酶活性抑制作用来评价青皮多酚提取物的降糖活性。结果表明,核桃青皮青皮汁与青皮皮渣总多酚含量分别为125.58mg/g和112.42mg/g,黄酮含量分别为92.60mg/g和61.72mg/g,青皮汁中总多酚与总黄酮含量明显高于青皮渣中含量。核桃青皮汁多酚对DPPH自由基,羟基自由基和超氧阴离子自由基的清除率IC50值分别为为3.4 1μg/mL,33.94μg/mL和0.47mg/mL,具有较好的自由基清除能力,并呈浓度依赖关系。核桃青皮多酚对α-淀粉酶活性有较好的抑制作用,并且呈现浓度依赖关系,其IC50值为20.70mg/mL。3.探讨不同环境条件包括温度、湿度、光照对核桃青皮粗提物多酚稳定性的影响,采用温度30±2℃、相对湿度60±5%、光照条件对核桃青皮汁粗提物进行加速氧化处理,测定其多酚物质稳定性以及抗氧化、α-淀粉酶活性抑制作用的功能稳定性。结果表明,高温、高湿和光照条件下,随着处理时间的延长,总多酚总黄酮含量降低,其DPP H自由基和羟自由基清除率以及α-淀粉酶活性抑制率的IC50随着处理时间的延长而增大,抗氧化活性及淀粉酶活性抑制活性降低。LC-MS分析结果表明,氧化处理前粗提物一共有16中多酚类物质,其中单没食子酰基葡萄糖含量最高(20.17%),没食子酸、新绿原酸等含量也较大。氧化处理后,多酚类物质组成改变,处理后粗提物少了香草酸、焦性没食子酸等五种多酚,增加了HHDP-葡萄糖异构体和2倍酰基葡萄糖两种多酚类物质,同时,一些物质含量发生变化。4.采用等辐射分析(Isobologram)法分析核桃青皮多酚物质两两混合后对α-淀粉酶活性的抑制作用,研究表明新绿原酸与焦性没食子酸、没食子酸与鞣花酸对α-淀粉酶活性抑制作用在一定范围内具有拮抗效果,前者组合的拮抗作用明显,新绿原酸与3-没食子酰葡萄糖苷、鞣花酸与焦性没食子酸对α-淀粉酶的活性抑制作用在一定范围内具有协同效果,当鞣花酸与焦性没食子酸比例为3：1时复合物对α-淀粉酶协同作用最强。5.通过紫外光谱、普通荧光光谱、同步荧光光谱、三维荧光光谱等方法探究没食子酸与α-淀粉酶的相互作用机制。研究发现没食子酸对α-淀粉酶有明显的荧光猝灭作用,其猝灭方式为静态猝灭,没食子酸与α-淀粉酶之间形成了没食子酸-α-淀粉酶复合物,同时,没食子酸的加入使α-淀粉酶蛋白分子的α-螺旋数量减少,氨基酸残基π-π*共轭系统的转变能级(ΔE)降低,电子转移增加,蛋白质内芳香族氨基酸的微环境从分子内疏水环境变为极性亲水环境。6.本文充分利用核桃资源变废为宝,采用常温液固发酵,试制了核桃青皮果蔬酵素,所得酵素口感好,香气回味长久,颜色清亮,呈橙红／棕红色。
[Abstract]:Walnut peel is one of the main products of walnut. The walnut harvest period, a large number of walnut green husk as waste dumps, environmental pollution and waste of resources, only a few amount of medicinal. Walnut peel has anti-cancer, anti-oxidation, anti-aging, reducing blood sugar and other functions. The research and development to strengthen its comprehensive utilization, has the value and ecological value the social and economic importance. Based on the analysis of the nutritional components of walnut peel as raw material, using 75% ethanol extraction of polyphenols from walnut peel, the antioxidant activity of a-, amylase activity inhibition and stability was investigated by using LC-MS technique to identify the components of polyphenols, analysis of interactions of the different components of polyphenols in inhibiting the activity of amylase in the the interaction, and explore its mechanism of gallic acid and alpha amylase. And the normal temperature solid-liquid fermentation, preparation of walnut peel fruit and vegetable enzyme. The main research contents and The results are as follows: conventional nutritional component analysis results of 1. walnut peel showed that walnut peel has rich nutrition. Walnut peel is rich in potassium, calcium, phosphorus, magnesium, iron and other mineral elements, and has high potassium, low sodium. The total amino acid content except tryptophan in the walnut peel was 33.31%. The essential amino acid and total amino acid ratio of 34.56%, the proportion of essential amino acids and non essential amino acids was 52.81%, and the WHO and the United Nations Food and Agriculture Organization (WHO/FAO) proposed food protein amino acids (EAA/TAA=40%, EAA/NEAA =60% ratio) is close to the total polyphenol extract.2. of walnut peel juice and peel residue with 75% ethanol, the determination of total polyphenol the content of extracts using Folin phenol method, through the determination of the crude extract of DPPH free radical scavenging capacity, hydroxyl radicals and superoxide anion radicals to evaluate the antioxidant capacity of polyphenols peel extract, To evaluate the hypoglycemic activity of peel polyphenol extracts inhibition of alpha - amylase activity and its determination. The results showed that the walnut peel peel juice and peel pomace polyphenol content were 125.58mg/g and 112.42mg/g, flavonoids content is respectively 92.60mg/g and 61.72mg/g, and the content of total polyphenol and total flavonoid content in peel juice was significantly higher than that of peel slag. Walnut peel juice polyphenols on DPPH free radical, hydroxyl radical and superoxide anion free radical rate of IC50 were 3.41 g/mL, 33.94 g/mL and 0.47mg/mL, with free radical scavenging ability better, and in a concentration dependent relationship. Polyphenol walnut peel on amylase activity had good inhibitory effect, and in concentration dependence of 20.70mg/mL.3. on different environmental conditions including temperature, humidity and its IC50 value, illumination effect on Walnut Seedcases extracts polyphenol stability, the temperature of 30 - 2 DEG C, phase The humidity of 60 + 5%, light conditions of crude extracts of walnut peel juice accelerated oxidation treatment, determination of polyphenols and antioxidant function of stability, stability of alpha amylase activity inhibition. The results showed that high temperature, high humidity and light conditions, with the prolonging of treatment time, reduce the total flavonoids content of total polyphenols the DPP, H and hydroxyl free radical scavenging rate and amylase activity inhibition rate of IC50 increased with the prolonging of treatment time, the amylase activity of antioxidant activity and inhibitory activity decreased the.LC-MS analysis results show that the oxidation treatment before the crude extract of a total of 16 polyphenols, including single galloyl glucose content was the highest (20.17%) neochlorogenic acid, gallic acid, the content is larger. After oxidation treatment, polyphenols composition changes after the treatment of crude extract less vanillic acid, pyrogallic acid and other five kinds of polyphenols, increased by HHD P- glucose isomers and 2 times acyl glucose two kinds of polyphenols, at the same time, some material content changes.4. by isobolographic analysis (Isobologram) method to analyze the inhibitory effect on alpha amylase activity of the mixture of walnut Peel Polyphenols 22, studies show that chlorogenic acid and pyrogallol, gallic acid and ellagic acid on alpha amylase the inhibitory activity of the antagonistic effect in a certain range, the combination of obvious antagonism, neochlorogenic acid and 3- galloyl glucoside, ellagic acid and gallic acid on the activity of alpha amylase inhibition has a synergistic effect in a certain range, when ellagic acid and pyrogallic acid ratio of 3:1 complexes the synergistic effect of the strongest.5. alpha amylase by ultraviolet spectroscopy, fluorescence spectroscopy, synchronous fluorescence, three-dimensional fluorescence spectroscopy method study of gallic acid and alpha. The interaction mechanism of powder enzyme. It was found that the fluorescence quenching of gallic acid had obvious effect on alpha amylase, the quenching mechanism is static quenching, gallic acid and alpha amylase formed between gallic acid alpha amylase complex, at the same time, adding gallic acid alpha amylase protein alpha helix to reduce the number of change the level of amino acid residues pi pi * conjugate system (E) is reduced, the electron transfer increases, micro environment within the protein of aromatic amino acids from the intramolecular hydrophobic environment for polar hydrophilic.6. environment by utilizing the resources of walnut waste to treasure, using solid fermentation liquid, preparation of walnut peel fruit enzyme the enzyme has good taste, aroma, long aftertaste, color is clear, orange / red brown.

【学位授予单位】：昆明理工大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：TS209;O657.63

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