中药桑寄生、山豆根活性成分的光谱性质及荧光分析法研究
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摘要
研究了中药桑寄生有效成分槲皮素、槲皮苷和中药山豆根的吸收光谱和荧光光谱,考察了槲皮素衍生荧光以及槲皮苷转化为槲皮素的最佳实验条件,建立了不经分离测定中药桑寄生有效成分槲皮素和槲皮苷含量的新方法;通过山豆根的荧光光谱研究,利用柱色谱分离出一种强荧光物质,在365 nm紫外灯下显蓝色荧光。
论文工作主要包括两部分:
1实验发现槲皮素自身无荧光,加入AlCl_3控制pH在3.17左右时形成Al(Ⅲ)-槲皮素荧光络合物,在λ_(ex)/λ_(em)=425/485 nm处出现荧光峰,荧光量子产率为0.014。槲皮苷本身基本无荧光,在pH=1.7,100℃恒温100分钟后,可转化为槲皮素,转化率为96.21%。据此制定了槲皮素和槲皮苷的荧光分析方法,用于中药桑寄生的质量评价。在0.01～0.85μg ml~(-1)范围内,荧光强度与槲皮素浓度之间的回归方程为F=0.23+58.92c(μg ml~(-1)),相关系数r=0.999(n=8)。方法的回收率为96.6%,精密度、稳定性、重现性良好。用本法测得桑寄生中槲皮素含量为0.0755%,槲皮苷含量为0.0545%。
2研究了山豆根水浸液的荧光光谱和紫外吸收光谱。在三维荧光等高线光谱图中出现四个明显的荧光峰,分别位于λ_(ex)/λ_(em)=225/303 nm、275/303 nm、280/372 nm和330/372 nm,推测这四个荧光峰分别属于两种(类)荧光物质。在实验条件一定时,荧光强度与浓度呈良好的线性关系,可以作为定量分析的基础。采用柱色谱方法对山豆根组分进行分离,得到一种蓝色强荧光物质,研究了其三维荧光图谱,测量了~1H-NMR图谱和质谱,推测了其结构特征。
The absorption and fluorescence spectra of Chinese herbal medicine Taxillus chinensis (DC.) Danser and its active ingredients—quercetin,quercitrin,and Sophora tonkinensis Gagnep were studied in this thesis.A new method for determination of quercetin and quercitrin in Taxillus chinensis(DC.) Danser was proposed.Based on the study of fluorescence property of Sophora tonkinensis Gagnep,a fluorescent compound with blue color when irradiated with 365 nm UV-irradiation was separated by column chromatography.
This thesis mainly consists of two parts:
1) Quercetin has no fluorescence itself.When AlCl_3 was added,a fluorescent complex of quercetin-Al(Ⅲ) was formed under the condition of pH≈3.17.The fluorescence peak of the complex was located atλ_(ex)/λ_(em)=425/485 nm with fluorescence quantum yield 0.014. Quercitrin had hardly no fluorescence,when its solution with pH=1.7 was heated at 100℃for 100 min,it was transformed to quercetin,with transformation efficiency 96.21%.A fluorimitric method for determination of quercetin and quercitrin was developed for the quality evaluation of Taxillus chinensis(DC.) Danser.In the concentration range of 0.01～0.85μg ml~(-1),the regression equation was F=0.23+58.92 c(μg ml~(-1)) with correlation coefficient r=0.999(n=8)。The recovery of the method was 96.6%,and the precision, stability and reproducibility of the method was fine。Using the new method,amounts of quercetin and quercitrin in Taxillus chinensis(DC.) Danser sample were determined to be 0.0755%and 0.0545%,respectively.
2) The fluorescence and UV absorption spectra of Sophora tonkinensis Gagnep water extraction were investigated.Four fluorescence peaks,located atλ_(ex)/λ_(em)=225/303 nm,275 /303 nm,280/372 nm and 330/372 nm respectively,were found in the three-dimensional fluorescence spectra.It was deduced that the four fluorescence peaks were produced by two kinds(or types) fluorescent substances.There were good linear relationships between the fluorescence intensity and concentrations at certain experimental conditions,which can be used as the basis for quantitative analysis.A strong fluorescence constituent of Sophora tonkinensis Gagnep was separated by column chromatography.Its three-dimensional fluorescence spectra were investigated,and its molecular character was estimated based on the measurement of ~1H-NMR and mass spectrum.
论文工作主要包括两部分:
1实验发现槲皮素自身无荧光,加入AlCl_3控制pH在3.17左右时形成Al(Ⅲ)-槲皮素荧光络合物,在λ_(ex)/λ_(em)=425/485 nm处出现荧光峰,荧光量子产率为0.014。槲皮苷本身基本无荧光,在pH=1.7,100℃恒温100分钟后,可转化为槲皮素,转化率为96.21%。据此制定了槲皮素和槲皮苷的荧光分析方法,用于中药桑寄生的质量评价。在0.01～0.85μg ml~(-1)范围内,荧光强度与槲皮素浓度之间的回归方程为F=0.23+58.92c(μg ml~(-1)),相关系数r=0.999(n=8)。方法的回收率为96.6%,精密度、稳定性、重现性良好。用本法测得桑寄生中槲皮素含量为0.0755%,槲皮苷含量为0.0545%。
2研究了山豆根水浸液的荧光光谱和紫外吸收光谱。在三维荧光等高线光谱图中出现四个明显的荧光峰,分别位于λ_(ex)/λ_(em)=225/303 nm、275/303 nm、280/372 nm和330/372 nm,推测这四个荧光峰分别属于两种(类)荧光物质。在实验条件一定时,荧光强度与浓度呈良好的线性关系,可以作为定量分析的基础。采用柱色谱方法对山豆根组分进行分离,得到一种蓝色强荧光物质,研究了其三维荧光图谱,测量了~1H-NMR图谱和质谱,推测了其结构特征。
The absorption and fluorescence spectra of Chinese herbal medicine Taxillus chinensis (DC.) Danser and its active ingredients—quercetin,quercitrin,and Sophora tonkinensis Gagnep were studied in this thesis.A new method for determination of quercetin and quercitrin in Taxillus chinensis(DC.) Danser was proposed.Based on the study of fluorescence property of Sophora tonkinensis Gagnep,a fluorescent compound with blue color when irradiated with 365 nm UV-irradiation was separated by column chromatography.
This thesis mainly consists of two parts:
1) Quercetin has no fluorescence itself.When AlCl_3 was added,a fluorescent complex of quercetin-Al(Ⅲ) was formed under the condition of pH≈3.17.The fluorescence peak of the complex was located atλ_(ex)/λ_(em)=425/485 nm with fluorescence quantum yield 0.014. Quercitrin had hardly no fluorescence,when its solution with pH=1.7 was heated at 100℃for 100 min,it was transformed to quercetin,with transformation efficiency 96.21%.A fluorimitric method for determination of quercetin and quercitrin was developed for the quality evaluation of Taxillus chinensis(DC.) Danser.In the concentration range of 0.01～0.85μg ml~(-1),the regression equation was F=0.23+58.92 c(μg ml~(-1)) with correlation coefficient r=0.999(n=8)。The recovery of the method was 96.6%,and the precision, stability and reproducibility of the method was fine。Using the new method,amounts of quercetin and quercitrin in Taxillus chinensis(DC.) Danser sample were determined to be 0.0755%and 0.0545%,respectively.
2) The fluorescence and UV absorption spectra of Sophora tonkinensis Gagnep water extraction were investigated.Four fluorescence peaks,located atλ_(ex)/λ_(em)=225/303 nm,275 /303 nm,280/372 nm and 330/372 nm respectively,were found in the three-dimensional fluorescence spectra.It was deduced that the four fluorescence peaks were produced by two kinds(or types) fluorescent substances.There were good linear relationships between the fluorescence intensity and concentrations at certain experimental conditions,which can be used as the basis for quantitative analysis.A strong fluorescence constituent of Sophora tonkinensis Gagnep was separated by column chromatography.Its three-dimensional fluorescence spectra were investigated,and its molecular character was estimated based on the measurement of ~1H-NMR and mass spectrum.
引文
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