【作者】 尚芳红；

【导师】 黄玉明；

【作者基本信息】 西南大学 ， 分析化学， 2007， 硕士

【摘要】 天然黄酮类化合物广泛存在于药用植物、水果和蔬菜中，是一类重要的天然有机化合物，其存在形式既有与糖结合成苷的，也有游离体。约有20％的中草药中含有黄酮类化合物，可见其资源之丰富。许多研究表明黄酮类化合物具有多种生物活性，除利用其抗菌、消炎、抗突变、降压、清热解毒、镇静、利尿等作用外，在抗氧化性、抗癌、防癌、抑制脂肪酶等方面也有显著效果。对天然产物中的黄酮类化合物进行药用开发和应用引起了研究者的广泛兴趣。有必要建立简单、快速和灵敏度高的测定黄酮类天然活性成分含量的方法。目前测定黄酮类天然活性成分含量的方法较多，尤其是紫外分光光度法和高效液相色谱法使用较普遍。前者适合测定总黄酮的含量，但由于一些酚类物质的干扰使得测定结果偏高；后者仪器比较昂贵，成本高。化学发光方法由于灵敏度高、仪器设备简单、速度快等特点受到人们的重视，在分析领域中已得到广泛应用。本论文包括两部分。第一部分是文献综述。介绍了黄酮类天然活性成分的主要结构分类、生物活性、提取与分离以及近三年来对黄酮类天然活性成分分析方法。第二部分是研究报告。利用流动注射化学发光技术，检测黄酮类天然活性成分的含量，并对该类活性成分的化学发光部分体系机理进行了探讨。主要内容如下：1高锰酸钾—鲁米诺化学发光体系对染料木素的测定基于染料木素对高锰酸钾氧化鲁米诺产生的化学发光的增强作用建立了一种测定染料木素的新方法。在优化的实验条件下，测定染料木素的线性范围为：0.04～4μg／mL，检出限（3a）为0.008μg／mL，对浓度为0.5μg／mL的染料木素进行11次平行测定，其相对标准偏差（RSD）为1.18％。将本法用于槐角药丸中染料木素含量的测定，结果令人满意。2硫酸铈—罗丹明6G化学发光体系对染料木素的测定在硝酸介质中，Ce（SO₄）₂氧化染料木素，罗丹明6G可以增强其化学发光强度。据此采用流动注射技术，建立了Ce（Ⅳ）-罗丹明6G-染料木素化学发光体系测定染料木素的化学发光新方法。在优化的实验条件下，该法测定染料木素的检出限为0.4μg／L，线性范围为0.002～5.0μg／ml，对0.5μg／mL染料木素进行了11次平行测定，其RSD为1.67％。将本法用于尿样及槐角中染料木素含量的测定获得满意结果。并对反应可能的机理进行了探讨。3铁氰化钾-鲁米诺化学发光体系对山萘酚的测定在碱性介质中，山奈酚对铁氰化钾氧化鲁米诺产生化学发光有很强的增敏作用，据此建立了一种简单、快速测定山奈酚的流动注射化学发光新方法。在优化的实验条件下，该法测定出的山奈酚的线性范围为0.05～10μg／mL检出限为0.01μg／mL，对0.5μg／mL山奈酚进行11次平行测定，其RSD为2.80％。将本法用于合成样品中山奈酚的测定，结果令人满意。4高碘酸钾-鲁米诺化学发光体系对木犀草素的测定碱性条件下，鲁米诺与高碘酸钾产生化学发光，木犀草素能抑制该发光强度。据此建立了一种简便、快速测定木犀草素的化学发光新方法。在优化条件下，用该法测定的木犀草素得到的检出限为2×10^-5μg／mL，线性范围为1×10^-4～3×10^-2μg／mL。将本法用于独一味胶囊及药片中木樨草素的测定，结果令人满意。更多还原

【Abstract】 The natural flavonoid widely exists in medicinal plant, fruit and vegetable, it is a kind of important natural organic compound in the form of either sugar glucoside or free molecular. It is reported that about twenty percent of Chinese medicine contains flavonoid. Many reports indicated that the flavonoids have many kinds of biological activities, such as the sterilization, reducing inflammation, anti-sudden change, step-down pressure, alleviating fever, disintoxicating, calm. Also the flavonoid shows good performance in the antioxidation, anti-cancer, guarding against cancer, lipase suppression. Researchers have paied much attention to the pharmaceutical development and practical application of the flavonoid. It is necessary to develop simple, fast, and sensitive method for flavonoid analysis. There are several methods for flavonoids determination, such as UV spectrophotometric method and HPLC. The former is suitable for determination of total flavonoid. The overestimated results are usually got due to phenolic substances interference. The latter is more expensive and high cost equipment. Due to the high sensitivity, simple instrumentation, and fastness, the chemiluminescence method obtained the widespread application in the analysis domain.The present paper includes two parts. The first part reviews the classification, extraction, separation and analytical methods of the flavonoid natural active compound. The second part is the report with foucus on chemiluminescence analysis of natural flavonoids by flow injection technology. The principal elements are as follows:1 Luminol-Potassium Permanganate System for the Determination of GenisteinA new chemiluminescence method for the determination of genistein was presented, which is based on its enhancement effect on the light emission from the luminol-KMnO4 system. Under the optimum conditions, the linear range for the determination of genistein is in the range of 0.04～4μg/mL with a detection limit of 0.008μg/mL. The relative standard deviation for 0.5μg/mL genstein is 1.18% （n=11）. The proposed method has been applied to the determination of genistein in pharmaceutical preparations and urine samples with satisfactory results.2 Cerium（Ⅳ）-Rhodamine- 6G system for the Determination of GenisteinA chemiluminescence method for the determination of genistein was presented, which is based on its oxidation with cerium（Ⅳ） in nitric acid media accompanied with chemiluminescence using Rh6G as sensitizer, Under the optimum conditions, the linear range for the determination of genistein is in the range of 0.002～5μg/mL with a detection limit of 0.4μg/L. The relative standard deviation for 0.5μg/mL genstein is 1.67% （n=11）. The proposed method has been applied to the determination of genistein in pharmaceutical preparations and urine samples with satisfactory resultsA possible mechanism has been prospoed based on the investigation CL and UV spectra of the reaction system.3 Potassium Ferricyanide-Luminol System for the Determination of KaempferolIn alkaline medium, kaempferol could greatly enchaned the chemiluminescence emitted from potassium ferricyanide oxidation on luminal. Based on above phenomena, a simple, and rapid chemiluminescence method combined with flow injection technology for determination of kaempferol is proposed. Under the optimum conditions, the liner range for kaempferol is 0.05～10μg/mL with detection limit of 0.01μg/mL. The relative standard deviation was 2.8% （n=11） for 0.5μg/mL kaempferol. The synthetic samples were analyzed by the proposed method for kaempferol determination with satisfactory results.4 Potassium Periodate-Luminol System for the Determination of LuteolinIt was found that the light emission produced by the oxidation of luminol by potassium ferricyanide in the basic medium was inhibited by luteolin. A simple flow injection chemiluminescence method for the determination of luteolin has been developed. Under the optimum conditions, the liner range for Luteolin was is 1×10^-4～3×10^-2μg/ml with detection limit of 2×10^-5μg/mL. This method has been applied to the determination of luteolin in capsules and tablets Duyiwei with satisfactory results.更多还原