基于色谱—质谱技术研究青钱柳叶化学成分

发布时间：2018-04-21 00:34

本文选题：青钱柳叶 + 固相微萃取　；参考：《南昌大学》2017年硕士论文

【摘要】：青钱柳为胡桃科青钱柳属植物,民间主要用于清热消肿、止痛,属于我国特有的珍稀树种。近年来,青钱柳的研究由大部分集中于小分子物质分离鉴定及生物活性方面,开始逐渐转向分子量较大或成分较复杂的组分如多糖、皂苷、生物碱等方面。本文分别采用SPME-GC-MS和UPLC-QTOF-MS/MS对青钱柳叶中挥发性成分及酚酸提取物中组分进行定性分析,并采用LC-QqQ-MS测定其中主要的酚酸含量。此外,采用不同极性溶剂对青钱柳叶进行提取,并测定其总酚、总黄酮、总糖含量及抗氧化活性,最后采用UPLC-QTOF-MS/MS对抗氧化活性成分进行分析鉴定。主要研究结果如下:第1章:介绍了近年来国内外有关文献报道青钱柳化学成分、分离分析方法及生物活性方面的研究进展。简要概述了青钱柳的化学成分提取技术及分析方法现状,并阐述了液相色谱-质谱联用技术在植物化学成分分析中的优势。第2章:采用固相微萃取(SPME)和气相色谱-三重串联四极杆质谱联用仪(GC-QQQ-MS)相结合的方法对两个产地(江西修水野生和浙江文成人工栽培)的青钱柳叶挥发性成分进行研究,并比较了它们在挥发性化学成分种类及含量上的差异。结果表明:在两个产地的青钱柳叶中共鉴定出91种挥发性组分,共有44种挥发性组分,且主要是碳氢化合物类、醇类、醛类、酯类、酮类、萘类和醚类化合物,但两个样品在挥发性成分的具体组成和含量上存在一定的差异。野生青钱柳叶的挥发性成分主要是β-瑟林烯(18.52%)、石竹烯(8.11%)、β-甜没药烯(6.01%)、β-榄香烯(5.36%)、顺-香叶基丙酮(4.02%)等,人工栽培青钱柳叶的挥发性成分主要是β-波旁烯(11.05%)、β-瑟林烯(10.91%)、石竹烯(7.03%)、苯甲醇(7.01%)、2,6,11-三甲基十二烷(5.16%)、2,6-二甲基-6-(4-甲基-3-戊烯基)-双环[3.1.1]庚-2-烯(5.36%)等。第3章:采用UPLC-QTOF-MS/MS定性分析青钱柳叶酚酸提取物中化学成分,并采用LC-QqQ-MS测定其中六种酚酸的含量,即没食子酸、原儿茶酸、对羟基苯甲酸、咖啡酸、对香豆酸和阿魏酸。结果显示:可溶性酚酸提取物中共初步鉴定出了22种化学成分,包括3种有机酸、9种酚酸、4种黄酮、3种酯类和3种其他类化合物,束缚性酚酸提取物中总共鉴定出26种化合物,包括8种有机酸、8种酚酸、4种黄酮和6种其他化合物;青钱柳叶中主要以羟基肉桂酸型酚酸含量较高,羟基苯甲酸型酚酸含量较少,且可溶性酚酸含量要大于束缚酚酸的含量,其中咖啡酸的含量最高,分别为583.61μg/mg和195.52μg/mg。第4章:利用比色法测定青钱柳叶不同溶剂(水、70%乙醇、乙酸乙酯和正己烷)提取物中总酚、总黄酮、总糖含量及抗氧化活性(DPPH自由基清除能力、还原能力、总抗氧化能力、Fe2+鳌合能力),考察酚类物质含量与抗氧化活性的相关性,并采用UPLC-QTOF-MS/MS分析提取物中主要活性成分。结果表明:不同提取物中总酚、总黄酮、总糖含量及抗氧化活性之间存在显著性差异且70%乙醇提取物表现出最高的总酚(219.01 mg GAE/g)、总黄酮含量(7.23 mg CE/g)及最强的DPPH自由基清除能力(35.46 mg TE/g)和还原能力(1.89 mmol FeSO4/g);总酚、总黄酮含量与DPPH自由基清除能力、还原能力之间呈显著正相关,与总抗氧化能力呈显著负相关,表明多酚类物质是青钱柳中主要的抗氧化剂。UPLC-QTOF-MS/MS分析70%乙醇提取物并初步鉴定出22种化合物,包括8种三萜皂苷类、5种黄酮、4种酚酸、3种酯类和2种有机酸,其中酚酸和黄酮类化合物是主要的抗氧化活性成分,有机酸、三萜皂苷及酯类化合物可能是潜在的抗氧化活性成分。
[Abstract]:Penicillium is a plant of Penicillium, which is mainly used in clearing heat and relieving pain and is a rare species in China. In recent years, the research of Penicillium has been mainly concentrated on the separation and identification of small molecules and biological activity, and began to gradually turn to large molecular weight or complex components such as polysaccharides, saponins, alkaloids, etc. In this paper, SPME-GC-MS and UPLC-QTOF-MS/MS were used to qualitatively analyze the volatile components and phenolic acids in the leaves of willow, and the main phenolic acids were determined by LC-QqQ-MS. In addition, different polar solvents were used to extract the leaves of Penicillium, and the total phenols, total flavones, total sugar content and antioxidant activity were determined. The main research results are as follows: the main research results are as follows: the first chapter: the first chapter: introduce the research progress on the chemical composition, the separation analysis method and the biological activity of the domestic and foreign literatures in recent years. The chemical composition extraction technology and analysis methods of Penicillium are briefly summarized. The advantages of liquid chromatography-mass spectrometry in the analysis of plant chemical composition are described. Second: the second chapter: the combination of solid phase microextraction (SPME) and gas chromatography - three heavy series quadrupole mass spectrometry (GC-QQQ-MS), the volatile components of the leaves of Penicillium in two habitats (Jiangxi wild and Zhejiang Wencheng artificial cultivation) The results showed that 91 volatile components were identified in the leaves of two habitats, including 44 kinds of volatile components, including hydrocarbons, alcohols, aldehydes, esters, ketones, naphthalene and ether, but two samples were volatile. The volatile components of the leaves were mainly beta cyenene (18.52%), carenene (8.11%), beta - myrrh (6.01%), beta elemene (5.36%), CIS leaf base acetone (4.02%) and so on. The volatile constituents of the leaves were mainly beta parenene (11.05%), beta cyenene (10.). 91%), carenenes (7.03%), benzyl alcohol (7.01%), 2,6,11- three methyl twelve alkane (5.16%), 2,6- two methyl -6- (4- methyl -3- glutenyl) - double ring [3.1.1] heptine -2- (5.36%), etc. third: qualitative analysis of the chemical composition of chloranenol acid extract with UPLC-QTOF-MS/MS and determination of the content of six phenolic acids, that is gallic acid, by LC-QqQ-MS Catechinic acid, para hydroxy benzoic acid, caffeic acid, coumaric acid and ferulic acid. The results show that the soluble phenolic acid extracts have preliminarily identified 22 chemical constituents, including 3 organic acids, 9 phenolic acids, 4 flavones, 3 esters and 3 other compounds, and 26 compounds in the bound phenolic acid extracts, including 8 organic acids, 8. Phenolic acid, 4 kinds of flavonoids and 6 other compounds; the content of hydroxyl cinnamic acid type phenolic acid is higher, the content of hydroxybenzoic acid is less, and the content of soluble phenolic acid is greater than that of acetamicylic acid, and the content of caffeic acid is the highest, 583.61 mu g/mg and 195.52 mu g/mg. fourth chapters, respectively: Determination of green money by colorimetric method. Total phenol, total flavone, total sugar content and antioxidant activity (DPPH free radical scavenging ability, reductive ability, total antioxidant capacity, Fe2+ chelating ability) in the extracts of different solvents (water, 70% ethanol, ethyl acetate and hexane) were investigated, and the correlation between phenolic content and antioxidant activity was investigated, and the main extracts were analyzed by UPLC-QTOF-MS/MS. The results showed that there were significant differences in total phenol, total flavonoids, total sugar content and antioxidant activity in different extracts, and the 70% ethanol extracts showed the highest total phenols (219.01 mg GAE/g), total flavonoids (7.23 mg CE/g) and the strongest DPPH free radical removal ability (35.46 mg TE/g) and reduction ability (1.89 mmol FeSO4/g). Total phenol, total flavonoids content was positively correlated with DPPH radical scavenging ability and reduction ability, and was negatively correlated with total antioxidant capacity. It showed that polyphenols were the main antioxidant.UPLC-QTOF-MS/MS analysis of 70% ethanol extracts and 22 kinds of compounds, including 8 kinds of three terpenoid saponins, 5 flavones and 4 species. Phenolic acids, 3 kinds of esters and 2 organic acids, of which phenolic acids and flavonoids are the main antioxidant components. Organic acids, three terpene saponins and esters may be potential antioxidant components.

【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O657.63;R284.1

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