黄蜀葵产业化过程副产物的资源化利用研究

发布时间：2018-06-09 00:18

本文选题：黄蜀葵 + 资源性物质　；参考：《南京中医药大学》2017年硕士论文

【摘要】：本论文共分四章内容。第一章文献综述。对黄蜀葵Abelmoschus manihot(L.)Medic.药用植物资源本草记载品种、药性及其功用进行了考证,并对其主要化学成分组成类型及分布、资源利用现状等进行了综述。在此基础上,基于黄蜀葵植株中多糖类成分含量较多但现有研究大都停留在粗多糖阶段的研究现状,对现有植物多糖类物质提取、分离纯化及其主要生物活性研究进展进行了分析整理,以期为提升黄蜀葵多糖类资源性化学成分的利用效率及效益提供借鉴,为黄蜀葵产业化过程中副产物的资源化利用提供支撑。第二章黄蜀葵花药材采收期不同组织器官资源性化学成分分析与评价。分别采用高效液相色谱法(HPLC)、超高效液相色谱串联三重四极杆质谱法(UPLC-TQ/Ms)、紫外可见分光光度法、粗纤维测定法(Weende)分析评价了黄蜀葵采收药用部位花冠过程中产生的不同组织器官(根、茎、叶和花)中黄酮类、核苷类、氨基酸类、可溶性多糖类、总纤维等资源性化学成分的组成及含量。结果显示,黄蜀葵花中富含黄酮类资源性化学成分,主要组成为金丝桃苷、异槲皮苷、棉皮素-8-O-葡萄糖醛酸苷、杨梅素、槲皮素-3'-O-葡萄糖苷、芦丁和槲皮素,总量为25.450 mg/g,叶片亦有少量黄酮类资源性化学成分分布。黄蜀葵不同组织器官均富含可溶性多糖类组分和总纤维,其中茎中总多糖含量较高,达10.86%,根中总纤维含量较高,可达29.88%。在黄蜀葵植物中共检出21种氨基酸类和9种核苷类资源性化学成分,其中花中氨基酸类化学成分种类及含量较为丰富,达4.737 mg/g;叶中核苷类化学成分含量较为丰富,为1.474 mg/g。研究结果为黄蜀葵植物采收花后不同组织器官的精细化利用与产业化开发提供了科学依据。第三章黄蜀葵种子中资源性化学成分的分析与评价。分别采用气相色谱-质谱联用法、UPLC-TQ/MS、紫外可见分光光度法、粗纤维测定法、BCA试剂盒法对黄蜀葵成熟种子中的主要资源性化学成分进行了分析评价。结果表明,黄蜀葵种子含有丰富的脂肪酸类、可溶性多糖类、总纤维、可溶性蛋白、游离氨基酸类、核苷及碱基类成分。其中脂肪酸总量可达10.22%,不饱和脂肪酸占脂肪酸总量的78.01%～79.40%;可溶性总多糖、总纤维和可溶性蛋白含量分别为6.53%～6.68%、12.77%～14.26%、10.36%～14.51%;检出的19种游离氨基酸类和7种核苷及碱基类成分中,氨基酸类成分较为丰富(10.08%～10.15%),其中必需氨基酸约占游离氨基酸总量的38.42%～39.40%;核苷类成分含量相对较低(3.01～3.11 mg/g)。研究结果为黄蜀葵种子的资源化利用提供了理论依据。第四章黄蜀葵资源循环利用研究。基于前文研究发现黄蜀葵茎叶中多糖类组分相对较为丰富,具有潜在的资源价值,但目前研究及利用较少。此外,黄蜀葵花深加工过程中产生的药渣大都废弃,造成资源浪费。因此为了更好地利用黄蜀葵非药用部位和花药渣资源,提升黄蜀葵资源利用效率,开展以下研究工作:(一)黄蜀葵茎叶多糖分离纯化研究及其结构表征。通过正交实验研究黄蜀葵茎叶多糖的最佳提取工艺,在单因素试验中,发现提取温度、提取时间和料液比对多糖提取率影响最大,因此设计三因素三水平表,最终得到了黄蜀葵茎叶多糖的最佳提取工艺,分别是提取时间3 h,料液比1:20,提取温度100℃C,提取两次,并在后续试验中得到验证。在最佳提取工艺的基础上,以体外免疫活性为导向,通过大孔树脂、在线膜截留系统和DEAE-52阴离子交换树脂,分步分离纯化黄蜀葵茎叶多糖,最终得到一种分子量均一,纯度较高的黄蜀葵精制免疫活性多糖,命名为HSK-JT。经过高效液相凝胶渗透色谱法(HPGPC),采用GPC分子量分布软件计算其数均分子量、重均分子量及分布系数,其重均分子量为13821Da,分布系数为1.9654。(二)黄蜀葵茎叶多糖免疫活性评价。采用免疫细胞及小鼠免疫抑制模型,分别从体外及体内两条途径对黄蜀葵精制多糖(KSK-JT)免疫活性进行了评价。体外细胞试验结果显示:黄蜀葵精制多糖可显著促进小鼠脾细胞的体外增殖,并表现出一定的双向剂量关系,也可促进小鼠单核巨噬细胞RAW264.7产生NO,其促进作用与浓度呈正相关,表明黄蜀葵精制多糖可促进免疫细胞增殖,且可直接作用于RAW264.7细胞从而激活巨噬细胞,提高其吞噬能力,促进其释放NO,从体外细胞模型提示黄蜀葵精制多糖具有免疫调节活性;基于小鼠免疫抑制模型试验结果显示,黄蜀葵精制多糖可调节环磷酰胺致免疫抑制模型小鼠免疫器官指数、外周血象中白细胞数、血清细胞免疫因子等相关免疫指标,表现出一定的免疫增强活性,且黄蜀葵精制多糖干预作用比黄蜀葵粗多糖更好。研究结果基于黄蜀葵茎叶多糖研制具有免疫增强作用的药物或保健食品提供了数据支撑。(三)黄蜀葵花药渣资源性物质评价及其生物炭化研究。在对黄蜀葵花药渣中各类资源性化学成分进行分析评价的基础上,提出了黄蜀葵药渣的资源化利用策略。此外,基于其丰富的维素类成分(总量为13.40%),开展了基于黄葵花渣制备生物炭工艺研究,并对其相关理化指标进行了测定,研究结果为黄蜀葵花药渣的再生利用提供了支撑。
[Abstract]:This paper is divided into four chapters. The first chapter is a literature review. The medicinal plant resources of the Abelmoschus manihot (L.) Medic. of the marshmallow are tested, and their main components and distribution and the utilization status of the resources are described. Based on this, the polysaccharides from the plant of the marshmallow are based on this. The research progress of the extraction, separation and purification and the main biological activity of the existing plant polysaccharides are analyzed in order to provide reference for the utilization efficiency and benefit of the resources of the polysaccharides of the marshmallow. The resource utilization of by-products in the process is supported. The second chapter analysis and evaluation of the chemical composition of different tissues and organs in the harvest period of Shu sunflower, respectively, using high performance liquid chromatography (HPLC), super high performance liquid chromatography with three heavy quadrupole mass spectrometry (UPLC-TQ/Ms), ultraviolet visible spectrophotometry, and crude fiber determination (Weende) The composition and content of the flavonoids, nucleosides, amino acids, soluble polysaccharides and total fibers in the different tissues and organs (root, stem, Ye Hehua) of the medicinal part of the anemone were analyzed and evaluated. The results showed that the flavonoids in the sunflower were rich in flavonoids, which were mainly composed of Hypericum. Glucoside, isoquercetin, anothelin -8-O- glucuronide, myricetin, quercetin -3'-O- glucoside, rutin and quercetin, with a total amount of 25.450 mg/g, and a small amount of flavonoids in the leaves were distributed. The different tissues and organs of the anemone were rich in soluble polysaccharide components and total fiber, and the total polysaccharide content in the stem was higher, up to 10.86%. There are 21 kinds of amino acids and 9 kinds of nucleoside chemical constituents in the root of the root of the marshmallow, of which the species and content of the amino acids in the flower are more abundant, up to 4.737 mg/g, and the contents of the nucleosides in the leaves are more abundant, and the results of the study for the 29.88%. are 1.474 mg/g.. A scientific basis for the fine utilization of different tissues and organs and industrial development was provided after collecting flowers. The analysis and evaluation of the resource chemical components in the seeds of the anflower in the third chapter were analyzed by gas chromatography-mass spectrometry, UPLC-TQ/MS, UV visible spectrophotometry, crude fiber determination, and BCA Kit Method for the main seeds of the mature marshmallow seeds. The results showed that the seeds of the marshmallow were rich in fatty acids, soluble polysaccharides, total fibers, soluble proteins, free amino acids, nucleosides and base groups. The total amount of fatty acids could reach 10.22%, unsaturated fatty acids accounted for 78.01% to 79.40% of the total fatty acids, and the total soluble polysaccharides were soluble. Total fiber and soluble protein content were 6.53% to 6.68%, 12.77% to 14.26%, 10.36% to 14.51%, 19 free amino acids and 7 nucleosides and base groups were found to be rich in amino acids (10.08% to 10.15%), of which essential amino acids accounted for 38.42% to 39.40% of the total free amino acids, and the content of nucleosides was relative. Lower (3.01 ~ 3.11 mg/g). The research results provide a theoretical basis for the resource utilization of the seeds of the sunflower. Fourth chapters on the recycling of the resources of the marshmallow. Based on the previous study, it is found that the polysaccharides in the stems and leaves of the sunflower are relatively rich and have potential resource value, but the study and utilization are less. In order to make better use of the resources of the non medicinal parts of the anemone and anther residue, and to improve the utilization efficiency of the resources of the anemone, the following research work is carried out: (1) study on the separation and purification of Polysaccharides from the stem and leaves of the anemone and the characterization of its structure. In the optimum extraction process, it was found that the extraction temperature, extraction time and liquid ratio had the greatest influence on the extraction rate of polysaccharide in single factor test. Therefore, the optimum extraction process of three factors and three levels was designed, and the extraction time was 3 h, the ratio of material to liquid was at 1:20, the extraction temperature was 100 C, and the extraction was two times in the follow-up test. On the basis of the best extraction process, the polysaccharide was separated and purified step by step by macroporous resin, on-line membrane retention system and DEAE-52 anion exchange resin, and finally got a kind of polysaccharide with homogeneous molecular weight and high purity, and named HSK-JT. after the purification. High performance liquid gel permeation chromatography (HPGPC) was used to calculate the molecular weight, weight average molecular weight and distribution coefficient of GPC molecular weight distribution software. The weight average molecular weight was 13821Da, the distribution coefficient was 1.9654. (two) of the polysaccharide immunoreactivity of the stem and leaf of orhospora. Using immuno cell and mouse immunosuppressive model, it was from two in vitro and in vivo, respectively. The immunological activity of the KSK-JT was evaluated. The results of cell test in vitro showed that the polysaccharides from the refined marshmallow could significantly promote the proliferation of mouse spleen cells in vitro, and showed a certain two-way dose relationship, and could also promote the production of NO in the monocyte macrophage RAW264.7 in mice, and the effect was positively correlated with the concentration. The polysaccharides from the mallow can promote the proliferation of immune cells, and can directly act on RAW264.7 cells to activate macrophages, improve their phagocytosis, and promote the release of NO. From the cell model in vitro, it is suggested that the polysaccharide of the mallow is immune to the immune regulation activity. The immune organ index of the immunosuppressive mice induced by cyclophosphamide, the number of white cells in the peripheral hemogram, the immune factors of the serum cell immune factors, and other related immune indexes, showed a certain immune enhancement activity, and the intervention of the refined polysaccharides from the mallow was better than the polysaccharide of the mallow. The results of the study were based on the immunization of the polysaccharide of the stem and leaf of the mallow. The powerful drug or health food provided data support. (three) the resource evaluation and biological carbonization of the medicament residue of the sunflower sunflower. Based on the analysis and evaluation of various resource chemical components in the medicament residue of the sunflower flower, the strategy of resource utilization was put forward. The total amount was 13.40%). The study on the preparation of biological carbon based on the flower residue of sunflower was carried out, and the related physicochemical indexes were measured. The results provided support for the reutilization of the medicinal residue of the sunflower.
【学位授予单位】：南京中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R284.1

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