茶叶活性成分的荧光性质与分析方法研究
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摘要
研究了儿茶素和表儿茶素的荧光光谱性质,考察了实验条件对其荧光光谱的影响。对西湖龙井茶和苦丁茶进行了荧光光谱的分析研究,建立了测定茶叶中儿茶素和表儿茶素的新方法。利用柱色谱分离出两种色素物质,它们在365 nm紫外灯下均显红色荧光。
论文工作主要包括三部分:
1、研究了儿茶素和表儿茶素的内源荧光和敏化荧光性质,探讨了溶液酸度、有机溶剂、金属离子、表面活性剂SDS和β-CD对荧光光谱的影响。儿茶素水溶液在酸性条件下有稳定的荧光,最大激发和发射波长分别为280 nm和311 nm。59时,由于酚羟基质子的电离而使荧光消失。以L-色氨酸为参比,测得儿茶素和表儿茶素的荧光量子产率分别为0.062和0.047。同时对茶叶中的咖啡碱和茶碱的荧光光谱性质进行了初步的考察,发现二者自身均不发荧光。
2、研究了21种茶叶水浸液的三维荧光光谱,发现它们的图谱在短波处都有荧光峰出现,并且光谱基本相似。详细研究了两种茶叶水浸液的荧光光谱性质,佐证了茶叶中的主要成分为儿茶素类。此外,在选好的实验条件下,两种茶叶稀溶液中所含荧光组分的荧光强度都与其浓度呈良好的线性关系,据此可以对茶叶中的荧光组分进行定量分析。
3、用高效液相色谱法/荧光检测法对茶叶中儿茶素和表儿茶素同时进行定量分析,建立了选择性测定药材中二者含量的新方法。
4、采用柱色谱方法对茶叶中色素类进行分离,得到两种在硅胶板上发红色荧光的物质,研究了其三维荧光图谱,测量了1H-NMR图谱、13C-NMR图谱、红外吸收光谱和质谱,推测了其结构特征。
In this thesis, fluorescence properties of (+)-Catechin (C) and (-)-Epicatechin (EC) were studied. Influence of experimental conditions on fluorescence spectroscopy was inspected. The fluorescence spectra of West Lake Longjing tea and Kuding tea were studied. Two kinds of pigments with red color when irradiated with 365nm UV-irradiation were separated by column chromatography.
The main contents of the thesis were as follows:
1. The native fluorescence and sensitized fluorescence properties of C and EC were studied.The impacts of acidity, organic solvents, metal ions, surface active agent (SDS andβ-CD) on fluorescence spectroscopy were inspected. Under acidic conditions, catechins solution had a stable fluorescence. The maximum excitation and emission wavelengths were 280 nm and 311 nm. As the acidity of solution changes from pH 5 to pH 9, fluorescence decreases. When pH greater than 9, flurescence disearpered due to phenolic hydroxyl proton ionization. Using L-tryptophan as a reference, fluorescence quantum yields of C and EC were determined to be 0.062 and 0.047, respectively. The fluorescence character of Caffeine and Theophylline in tea was also studied, but no fluorescence was found.
2. Three-dimensional fluorescence spectra of 21 kinds of tea were studied. It was found that they had the similar fluorescence peaks at short wavelengths. The fluorescence properties of two types of tea water extract were studied in detail. It was discovered that the main components of tea were catechins. In selected conditions, a good linear relationship between fluorescence intensity and concentration was found, which can be used for the quantitative determination of the fluorescence components of tea.
3. Using a method of high-performance liquid chromatography with fluorescence detection, C and EC in the tea were determined at the same time. This new method can also be used for the determination of C and EC in Chinese herbal medicines.
4. Using column chromatography method, two kinds of red fluorescence substances in tea were separated. Its three-dimensional fluorescence spectra were investigated, and their molecular character was estimated based on the measurement of 1H-NMR, 13C-NMR, infrared absorption spectroscopy and mass spectrum.
论文工作主要包括三部分:
1、研究了儿茶素和表儿茶素的内源荧光和敏化荧光性质,探讨了溶液酸度、有机溶剂、金属离子、表面活性剂SDS和β-CD对荧光光谱的影响。儿茶素水溶液在酸性条件下有稳定的荧光,最大激发和发射波长分别为280 nm和311 nm。5
2、研究了21种茶叶水浸液的三维荧光光谱,发现它们的图谱在短波处都有荧光峰出现,并且光谱基本相似。详细研究了两种茶叶水浸液的荧光光谱性质,佐证了茶叶中的主要成分为儿茶素类。此外,在选好的实验条件下,两种茶叶稀溶液中所含荧光组分的荧光强度都与其浓度呈良好的线性关系,据此可以对茶叶中的荧光组分进行定量分析。
3、用高效液相色谱法/荧光检测法对茶叶中儿茶素和表儿茶素同时进行定量分析,建立了选择性测定药材中二者含量的新方法。
4、采用柱色谱方法对茶叶中色素类进行分离,得到两种在硅胶板上发红色荧光的物质,研究了其三维荧光图谱,测量了1H-NMR图谱、13C-NMR图谱、红外吸收光谱和质谱,推测了其结构特征。
In this thesis, fluorescence properties of (+)-Catechin (C) and (-)-Epicatechin (EC) were studied. Influence of experimental conditions on fluorescence spectroscopy was inspected. The fluorescence spectra of West Lake Longjing tea and Kuding tea were studied. Two kinds of pigments with red color when irradiated with 365nm UV-irradiation were separated by column chromatography.
The main contents of the thesis were as follows:
1. The native fluorescence and sensitized fluorescence properties of C and EC were studied.The impacts of acidity, organic solvents, metal ions, surface active agent (SDS andβ-CD) on fluorescence spectroscopy were inspected. Under acidic conditions, catechins solution had a stable fluorescence. The maximum excitation and emission wavelengths were 280 nm and 311 nm. As the acidity of solution changes from pH 5 to pH 9, fluorescence decreases. When pH greater than 9, flurescence disearpered due to phenolic hydroxyl proton ionization. Using L-tryptophan as a reference, fluorescence quantum yields of C and EC were determined to be 0.062 and 0.047, respectively. The fluorescence character of Caffeine and Theophylline in tea was also studied, but no fluorescence was found.
2. Three-dimensional fluorescence spectra of 21 kinds of tea were studied. It was found that they had the similar fluorescence peaks at short wavelengths. The fluorescence properties of two types of tea water extract were studied in detail. It was discovered that the main components of tea were catechins. In selected conditions, a good linear relationship between fluorescence intensity and concentration was found, which can be used for the quantitative determination of the fluorescence components of tea.
3. Using a method of high-performance liquid chromatography with fluorescence detection, C and EC in the tea were determined at the same time. This new method can also be used for the determination of C and EC in Chinese herbal medicines.
4. Using column chromatography method, two kinds of red fluorescence substances in tea were separated. Its three-dimensional fluorescence spectra were investigated, and their molecular character was estimated based on the measurement of 1H-NMR, 13C-NMR, infrared absorption spectroscopy and mass spectrum.
引文
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