共振瑞利散射法在天然药物分析中的应用研究
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摘要
共振瑞利散射(RRS)是一门新兴的分析技术，因其简便、快速和高灵敏度引起人们的广泛关注，发展十分迅速。研究表明，具有相反电荷的两种离子，借静电引力、疏水作用和电子转移作用形成离子缔合物时，能引起共振瑞利散射和二级散射(SOS)、倍频散射(FDS)等共振非线性散射(RNLS)的增强，并改变光谱特性。目前，该法在核酸、蛋白质、多糖等生物大分子的测定、痕量金属(如银、铅、铬、汞、钯、镉、铜等)和非金属离子(如硒、氨)的测定、纳米微粒(如硒、金、银、硫化银、硫化镉纳米微粒等)的研究、药物分析等方面得到的广泛的应用。
药物分析具有重要的临床意义，也是分析化学的重要组成部分。随着天然药物在药学、医学上的作用日益明显，有关天然药物有效成分的分析也得到了越来越多的研究。目前，共振瑞利散射法已成功用于丹参酮、甘草酸、小檗碱等天然药物的分析。我们的研究表明，利用天然药物的有效成分与金属离子配位形成螯合物、与杂多酸或染料分子通过静电引力形成离子缔合物以及药物自身的氧化还原等性质，能够引起共振瑞利散射的增强，并引起其它光谱特征的变化，因此推广了共振瑞利散射法在天然药物分析中的应用。
本文以槲皮素、白藜芦醇、桑色素、脱水穿心莲内脂和长春新碱为研究对象，应用RRS法研究了天然药物在与金属离子、杂多酸、染料分子之间的相互作用，以及药物本身的氧化还原能力对其他化学反应的影响，研究了它们的光谱特性、适宜反应条件、影响因素等，据此建立了共振瑞利散射法测定上述药物成分的新方法，并对反应机理、RRS增强原因、散射光谱与吸收光谱之间的联系和离子缔合物、螯合物的结构等方面进行了探讨，并在药物制剂、人体血清和天然植物等实际样品中进行了应用分析。
1汞(Ⅱ)-槲皮素螯合物体系共振瑞利散射及其分析应用
在pH 4.8～6.1的BR缓冲溶液中，槲皮素(QT)和汞(Ⅱ)形成螯合物时，将引起溶液共振瑞利散射(RRS)显著增强，并产生新的RRS光谱，其最大RRS波长位于320 nm，另在450 nm处有一小的散射峰。槲皮素在0.98～7.0μg／mL范围内与散射强度(△Ⅰ)成正比；反应具有较高的灵敏度，对槲皮素的检出限为29.5ng／mL。研究了适宜的反应条件和影响因素，考察了共存物质的影响，表明方法有良好的选择性。基于上述研究，建立了一种灵敏、简便、快速测定槲皮素的新方法，并用于天然药物槐米中槲皮素的测定。
2 FeCl_3-K_3[Fe(CN)_6]体系共振瑞利散射法测定白藜芦醇
在pH 1.8～3.5的BR缓冲介质中，白藜芦醇将K_3[Fe(CN)_6]还原为K_4[Fe(CN)_6]，它再与FeCl_3生成普鲁士蓝，反应产物还能导致吸收光谱发生变化，在700 nm处产生一个新的吸收峰，可用于白藜芦醇的分光光度法测定。此时将引起溶液共振瑞利散射(RRS)显著增强，其最大散射峰位于314 nm，另在375 nm处有一小的散射峰。白藜芦醇在0.05～5.0μg／mL范围内与散射强度(△Ⅰ)成正比；反应具有较高的灵敏度，对白藜芦醇的检出限为15 ng／mL，因此更适用于痕量白藜芦醇的测定。本文实验研究了适宜的反应条件和影响因素，考察了共存物质的影响，表明方法有良好的选择性。基于上述研究，结果建立了一种灵敏、简便、快速测定白藜芦醇的新方法，并用于葡萄叶和虎杖中白藜芦醇的测定。
3 La(Ⅲ)-桑色素相互作用的共振瑞利散射光谱及其分析应用
在中性水介质中，桑色素与镧(Ⅲ)形成螯合物时，将引起溶液共振瑞利散射(RRS)显著增强，并产生新的RRS光谱，其最大RRS波长位于368 nm，另在290 nm、342 nm和460 nm处有三个小的散射峰。桑色素在0.06～5.0μg／mL范围内与散射强度(△Ⅰ)成正比；反应具有较高的灵敏度，对桑色素的检出限为18.0ng／mL。研究了该体系的RRS光谱特征、适宜的反应条件、分析化学性质和某些分析应用，并对反应的机理进行了初步的讨论。基于上述研究，建立了一种灵敏、简便、快速测定桑色素的新方法。
4磷钨杂多酸-硫酸长春新碱离子缔合物体系共振瑞利散射光谱及其分析应用
酸性介质中硫酸长春新碱被质子化后与磷钨杂多酸形成离子缔合物时，将引起溶液共振瑞利散射(RRS)显著增强，其最大RRS波长位于315 nm。硫酸长春新碱在0.013～5.0μg／mL范围内与散射强度(△Ⅰ)成正比；反应具有较高的灵敏度，对硫酸长春新碱的检出限为4.2 ng／mL。研究了适宜的反应条件和影响因素，考察了共存物质的影响，表明方法有良好的选择性。基于上述研究，建立了一种灵敏、简便、快速测定硫酸长春新碱的新方法。
5结晶紫共振瑞利散射法测定脱水穿心莲内脂
在pH 11.2的BR缓冲溶液中，脱水穿心莲内脂水解，使其内脂环裂解成带负电荷的羟基羧酸，并于带正电荷的结晶紫发生离子反应形成离子缔合物，将引起溶液共振瑞利散射显著增强，其最大RRS波长位于282 nm，另在323 nm处有一小的散射峰。脱水穿心莲内脂在0.07～4.0μg／mL范围内与散射强度(△Ⅰ)成正比；反应具有较高的灵敏度，对脱水穿心莲内脂的检出限为20.5 ng／mL。研究了适宜的反应条件和影响因素，考察了共存物质的影响，表明方法有良好的选择性。基于上述研究，建立了一种灵敏、简便、快速测定脱水穿心莲内脂的新方法。
Resonance Rayleigh Scattering (RRS) is a new analytical technology, for its remarkable characteristics of high sensitivity, simple operation and good selectivity, this method brings to more attention and interesting and has been developed rapidly. Many studies shown that when two ions of different charges form ion association complexes by electrostatic force, hydrophobic force or charge transferring effect, RRS and RNLS (resonance nonlinear scattering, such as double frequency (FDS) or second order scattering ( SOS) )can be enhanced and the spectra characteristics can be changed. Now, these methods have been used to determine nucleic acid, protein, amylase and other biological macromolecules, trace amount of metal and its ions (Ag~+, Pb~(2+), Hg~(2+), Cu~(2+), Pa~(2+), et al.) , and non-metal such as selenium and ammonia, furthermore, RRS has been more and more used for the study of nano-particulates and the determination of some Pharmaceuticals.
Pharmaceutical analysis is an important part of analytical chemistry, with great significance for clinic. As natural medicine become more and more important, a lot of technologies and methods for the analysis of natural medicine have been proposed. Presently, Resonance rayleigh scattering method has been applied to the determination and study of some natural medicine (tanshinone IIA, berberine hydrochloride, glycyrrhizic acid, et al.) .Our research shown that natural medicine can react with metal ions to form chelate complexes, react with dye molecules and heteropolyacids to form ion-association complexes by electrostatic force, these reactions results in a significant enhancement of RRS, and the changes of others spectra characteristics. The researches have important application prospects and worth to further studies.
In this dissertation quercetin, deoxyandrographolide, resveratrol, morin and vincristine are as samples to study the interaction of alkaloids, flavones with metal ions, dye molecules and heteropolyacids, and effect on some chemical reactions by its oxidation and deoxidization ability. Their RRS spectral characteristics, optimum reaction conditions, influencing factors and analytical application have been studied. Some new methods for the determination of these natural medicine have been proposed, furthermore, the reaction mechanism, the reasons of RRS enhanced, the relationships between RRS spectra and absorption spectra and the structure of the ion-association complexes and chelate complexes are discussed.
1. A study on the Resonance Rayleigh scattering spectra of the system of Hg(II)-Quercetin Chelate and its analytical application
A novel method for the determination of trace amounts of quercetin (QT) based on resonance Rayleigh scattering ( RRS ) has been developed. In pH4.8 ~ 6.1 Britton-Robinson buffer solution, the quercetin (QT) can react rapidly with Hg( II) to form a complex, which results in a significant enhancement of RRS and an appearance of a new spectrum. The maximum scattering wavelength appears at 320 run while another little peak shows at 450 ran. The optimum conditions of the reaction and the influencing factors have been investigated. The RRS intensity is directly proportional to the concentration of QT in the range of 0.98～7.0μg/mL and its detection limit is 29.5 ng/mL. This result shows that this method has high sensitivity and good selectivity. Finally, this method was applied for determining QT in sophora japonica sample with satisfactory results.
2. Resonance Rayleigh Scattering Method for the Determination of Resveratrol with FeCl_3-K_3[Fe(CN)_6]
A novel method for the determination of trace amounts of resveratrol based on resonance Rayleigh scattering (RRS) has been studied. In pH 1.8 acetate sodium and hydrochloric acid buffer solution, the reduced production K_4[Fe(CN)_6] from the reaction between resveratrol and K_3[Fe(CN)_6] can react with FeCl_3 to form Prussian blue, which results in an appearance of a new spectrum and a significant enhancement of RRS. The maximum scattering wavelength appears at 314 nm while another little peak shows at 375 nm. The RRS intensity is directly proportional to the concentration of resveratrol in the range of 0.05～5.0μg/mL and its detection limit is 15 ng/mL. In this experiment, the optimum conditions of the reaction and the influencing factors have been investigated. This result shows that this method has high sensitivity and good selectivity. Finally, this method was applied for determining of resveratrol in grape leaves and Huzhang samples with satisfactory results.
3. A study on the Resonance Rayleigh scattering spectra of the interactions of morin with La~(3+) and its analytical application
A novel method for the determination of trace amounts of morin based on resonance Rayleigh scattering (RRS) has been studied. In neutral aqueous medium, the morin can react rapidly with La~(3+)to form a complex, which results in a significant enhancement of RRS and an appearance of a new spectrum. The maximum scattering wavelength appears at 368 nm while other three little peaks shows at 290 nm,342 nm and 460 nm. The RRS intensity is directly proportional to the concentration of morin in the range of 0.06～5.0μg/mL and its detection limit is 18.0 ng/mL. The spectral characteristics, the optimum conditions of the reactions, affecting factors and the properties of analytical chemistry and their analytical applications have been investigated. A high sensitive, simple and fast method is proposed to the determination of morin.
4. A study on the Resonance Rayleigh scattering spectra of the system of H_3PW_(12)O_(40)- vincristine and its analytical application
In 1.0mol/L HCl medium, vincristine react with H_3PW_(12)O_(40) to form an ion-association complex, the RRS intensity can be enhanced greatly and the maximum scattering wavelength appears at 315 nm. The RRS intensity is directly proportional to the concentration of vincristine in the range of 0.013～5.0μg/mL and its detection limit is 4.2 ng/mL. The optimum conditions of the reaction, the effect factors, the influence of foreign substances and the reaction mechanism have been investigated, it is shown that this method has high sensitivity. A high sensitive, simple and fast method is proposed to the determination of vincristine.
5. Resonance Rayleigh Scattering Method for the Determination of deoxyandrographolide with crystal violet
In pH11.2 BR buffer medium, deoxyandrographolide react with crystal violet to form an ion-association complex after been hydrolyzed, which results in a significant enhancement of RRS and an appearance of a new spectrum. The maximum scattering wavelength appears at 282 nm while another little peak shows at 323 nm. The RRS intensity is directly proportional to the concentration of deoxyandrographolide in the range of 0.07～4.0μg/mL and its detection limit is 20.5 ng/mL. The spectral characteristics, the optimum conditions of the reactions, affecting factors and the properties of analytical chemistry and their analytical applications have been investigated. A high sensitive, simple and fast method is proposed to the determination of deoxyandrographolide.
药物分析具有重要的临床意义，也是分析化学的重要组成部分。随着天然药物在药学、医学上的作用日益明显，有关天然药物有效成分的分析也得到了越来越多的研究。目前，共振瑞利散射法已成功用于丹参酮、甘草酸、小檗碱等天然药物的分析。我们的研究表明，利用天然药物的有效成分与金属离子配位形成螯合物、与杂多酸或染料分子通过静电引力形成离子缔合物以及药物自身的氧化还原等性质，能够引起共振瑞利散射的增强，并引起其它光谱特征的变化，因此推广了共振瑞利散射法在天然药物分析中的应用。
本文以槲皮素、白藜芦醇、桑色素、脱水穿心莲内脂和长春新碱为研究对象，应用RRS法研究了天然药物在与金属离子、杂多酸、染料分子之间的相互作用，以及药物本身的氧化还原能力对其他化学反应的影响，研究了它们的光谱特性、适宜反应条件、影响因素等，据此建立了共振瑞利散射法测定上述药物成分的新方法，并对反应机理、RRS增强原因、散射光谱与吸收光谱之间的联系和离子缔合物、螯合物的结构等方面进行了探讨，并在药物制剂、人体血清和天然植物等实际样品中进行了应用分析。
1汞(Ⅱ)-槲皮素螯合物体系共振瑞利散射及其分析应用
在pH 4.8～6.1的BR缓冲溶液中，槲皮素(QT)和汞(Ⅱ)形成螯合物时，将引起溶液共振瑞利散射(RRS)显著增强，并产生新的RRS光谱，其最大RRS波长位于320 nm，另在450 nm处有一小的散射峰。槲皮素在0.98～7.0μg／mL范围内与散射强度(△Ⅰ)成正比；反应具有较高的灵敏度，对槲皮素的检出限为29.5ng／mL。研究了适宜的反应条件和影响因素，考察了共存物质的影响，表明方法有良好的选择性。基于上述研究，建立了一种灵敏、简便、快速测定槲皮素的新方法，并用于天然药物槐米中槲皮素的测定。
2 FeCl_3-K_3[Fe(CN)_6]体系共振瑞利散射法测定白藜芦醇
在pH 1.8～3.5的BR缓冲介质中，白藜芦醇将K_3[Fe(CN)_6]还原为K_4[Fe(CN)_6]，它再与FeCl_3生成普鲁士蓝，反应产物还能导致吸收光谱发生变化，在700 nm处产生一个新的吸收峰，可用于白藜芦醇的分光光度法测定。此时将引起溶液共振瑞利散射(RRS)显著增强，其最大散射峰位于314 nm，另在375 nm处有一小的散射峰。白藜芦醇在0.05～5.0μg／mL范围内与散射强度(△Ⅰ)成正比；反应具有较高的灵敏度，对白藜芦醇的检出限为15 ng／mL，因此更适用于痕量白藜芦醇的测定。本文实验研究了适宜的反应条件和影响因素，考察了共存物质的影响，表明方法有良好的选择性。基于上述研究，结果建立了一种灵敏、简便、快速测定白藜芦醇的新方法，并用于葡萄叶和虎杖中白藜芦醇的测定。
3 La(Ⅲ)-桑色素相互作用的共振瑞利散射光谱及其分析应用
在中性水介质中，桑色素与镧(Ⅲ)形成螯合物时，将引起溶液共振瑞利散射(RRS)显著增强，并产生新的RRS光谱，其最大RRS波长位于368 nm，另在290 nm、342 nm和460 nm处有三个小的散射峰。桑色素在0.06～5.0μg／mL范围内与散射强度(△Ⅰ)成正比；反应具有较高的灵敏度，对桑色素的检出限为18.0ng／mL。研究了该体系的RRS光谱特征、适宜的反应条件、分析化学性质和某些分析应用，并对反应的机理进行了初步的讨论。基于上述研究，建立了一种灵敏、简便、快速测定桑色素的新方法。
4磷钨杂多酸-硫酸长春新碱离子缔合物体系共振瑞利散射光谱及其分析应用
酸性介质中硫酸长春新碱被质子化后与磷钨杂多酸形成离子缔合物时，将引起溶液共振瑞利散射(RRS)显著增强，其最大RRS波长位于315 nm。硫酸长春新碱在0.013～5.0μg／mL范围内与散射强度(△Ⅰ)成正比；反应具有较高的灵敏度，对硫酸长春新碱的检出限为4.2 ng／mL。研究了适宜的反应条件和影响因素，考察了共存物质的影响，表明方法有良好的选择性。基于上述研究，建立了一种灵敏、简便、快速测定硫酸长春新碱的新方法。
5结晶紫共振瑞利散射法测定脱水穿心莲内脂
在pH 11.2的BR缓冲溶液中，脱水穿心莲内脂水解，使其内脂环裂解成带负电荷的羟基羧酸，并于带正电荷的结晶紫发生离子反应形成离子缔合物，将引起溶液共振瑞利散射显著增强，其最大RRS波长位于282 nm，另在323 nm处有一小的散射峰。脱水穿心莲内脂在0.07～4.0μg／mL范围内与散射强度(△Ⅰ)成正比；反应具有较高的灵敏度，对脱水穿心莲内脂的检出限为20.5 ng／mL。研究了适宜的反应条件和影响因素，考察了共存物质的影响，表明方法有良好的选择性。基于上述研究，建立了一种灵敏、简便、快速测定脱水穿心莲内脂的新方法。
Resonance Rayleigh Scattering (RRS) is a new analytical technology, for its remarkable characteristics of high sensitivity, simple operation and good selectivity, this method brings to more attention and interesting and has been developed rapidly. Many studies shown that when two ions of different charges form ion association complexes by electrostatic force, hydrophobic force or charge transferring effect, RRS and RNLS (resonance nonlinear scattering, such as double frequency (FDS) or second order scattering ( SOS) )can be enhanced and the spectra characteristics can be changed. Now, these methods have been used to determine nucleic acid, protein, amylase and other biological macromolecules, trace amount of metal and its ions (Ag~+, Pb~(2+), Hg~(2+), Cu~(2+), Pa~(2+), et al.) , and non-metal such as selenium and ammonia, furthermore, RRS has been more and more used for the study of nano-particulates and the determination of some Pharmaceuticals.
Pharmaceutical analysis is an important part of analytical chemistry, with great significance for clinic. As natural medicine become more and more important, a lot of technologies and methods for the analysis of natural medicine have been proposed. Presently, Resonance rayleigh scattering method has been applied to the determination and study of some natural medicine (tanshinone IIA, berberine hydrochloride, glycyrrhizic acid, et al.) .Our research shown that natural medicine can react with metal ions to form chelate complexes, react with dye molecules and heteropolyacids to form ion-association complexes by electrostatic force, these reactions results in a significant enhancement of RRS, and the changes of others spectra characteristics. The researches have important application prospects and worth to further studies.
In this dissertation quercetin, deoxyandrographolide, resveratrol, morin and vincristine are as samples to study the interaction of alkaloids, flavones with metal ions, dye molecules and heteropolyacids, and effect on some chemical reactions by its oxidation and deoxidization ability. Their RRS spectral characteristics, optimum reaction conditions, influencing factors and analytical application have been studied. Some new methods for the determination of these natural medicine have been proposed, furthermore, the reaction mechanism, the reasons of RRS enhanced, the relationships between RRS spectra and absorption spectra and the structure of the ion-association complexes and chelate complexes are discussed.
1. A study on the Resonance Rayleigh scattering spectra of the system of Hg(II)-Quercetin Chelate and its analytical application
A novel method for the determination of trace amounts of quercetin (QT) based on resonance Rayleigh scattering ( RRS ) has been developed. In pH4.8 ~ 6.1 Britton-Robinson buffer solution, the quercetin (QT) can react rapidly with Hg( II) to form a complex, which results in a significant enhancement of RRS and an appearance of a new spectrum. The maximum scattering wavelength appears at 320 run while another little peak shows at 450 ran. The optimum conditions of the reaction and the influencing factors have been investigated. The RRS intensity is directly proportional to the concentration of QT in the range of 0.98～7.0μg/mL and its detection limit is 29.5 ng/mL. This result shows that this method has high sensitivity and good selectivity. Finally, this method was applied for determining QT in sophora japonica sample with satisfactory results.
2. Resonance Rayleigh Scattering Method for the Determination of Resveratrol with FeCl_3-K_3[Fe(CN)_6]
A novel method for the determination of trace amounts of resveratrol based on resonance Rayleigh scattering (RRS) has been studied. In pH 1.8 acetate sodium and hydrochloric acid buffer solution, the reduced production K_4[Fe(CN)_6] from the reaction between resveratrol and K_3[Fe(CN)_6] can react with FeCl_3 to form Prussian blue, which results in an appearance of a new spectrum and a significant enhancement of RRS. The maximum scattering wavelength appears at 314 nm while another little peak shows at 375 nm. The RRS intensity is directly proportional to the concentration of resveratrol in the range of 0.05～5.0μg/mL and its detection limit is 15 ng/mL. In this experiment, the optimum conditions of the reaction and the influencing factors have been investigated. This result shows that this method has high sensitivity and good selectivity. Finally, this method was applied for determining of resveratrol in grape leaves and Huzhang samples with satisfactory results.
3. A study on the Resonance Rayleigh scattering spectra of the interactions of morin with La~(3+) and its analytical application
A novel method for the determination of trace amounts of morin based on resonance Rayleigh scattering (RRS) has been studied. In neutral aqueous medium, the morin can react rapidly with La~(3+)to form a complex, which results in a significant enhancement of RRS and an appearance of a new spectrum. The maximum scattering wavelength appears at 368 nm while other three little peaks shows at 290 nm,342 nm and 460 nm. The RRS intensity is directly proportional to the concentration of morin in the range of 0.06～5.0μg/mL and its detection limit is 18.0 ng/mL. The spectral characteristics, the optimum conditions of the reactions, affecting factors and the properties of analytical chemistry and their analytical applications have been investigated. A high sensitive, simple and fast method is proposed to the determination of morin.
4. A study on the Resonance Rayleigh scattering spectra of the system of H_3PW_(12)O_(40)- vincristine and its analytical application
In 1.0mol/L HCl medium, vincristine react with H_3PW_(12)O_(40) to form an ion-association complex, the RRS intensity can be enhanced greatly and the maximum scattering wavelength appears at 315 nm. The RRS intensity is directly proportional to the concentration of vincristine in the range of 0.013～5.0μg/mL and its detection limit is 4.2 ng/mL. The optimum conditions of the reaction, the effect factors, the influence of foreign substances and the reaction mechanism have been investigated, it is shown that this method has high sensitivity. A high sensitive, simple and fast method is proposed to the determination of vincristine.
5. Resonance Rayleigh Scattering Method for the Determination of deoxyandrographolide with crystal violet
In pH11.2 BR buffer medium, deoxyandrographolide react with crystal violet to form an ion-association complex after been hydrolyzed, which results in a significant enhancement of RRS and an appearance of a new spectrum. The maximum scattering wavelength appears at 282 nm while another little peak shows at 323 nm. The RRS intensity is directly proportional to the concentration of deoxyandrographolide in the range of 0.07～4.0μg/mL and its detection limit is 20.5 ng/mL. The spectral characteristics, the optimum conditions of the reactions, affecting factors and the properties of analytical chemistry and their analytical applications have been investigated. A high sensitive, simple and fast method is proposed to the determination of deoxyandrographolide.
引文
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