CS结构修饰及其与MB空间定向相互作用机理

为从分子水平探讨硫酸化多糖与小分子相互作用机理,采用氯磺酸 吡啶法(Wolfrom法)对硫酸软骨 素(CS)进行结构修饰,应用光谱法研究了亚甲蓝(MB)与CS空间定向相互作用的机理,考察了硫取代度对CS抗 凝血活性及MB与CS相互作用的影响.测得MB与不同硫取代度的CS最大结合数分别为73、89、103和119, CS的抗凝血活性随硫取代度的增加而增加.认为MB与CS相互作用具有空间位阻效应,CS的抗凝血功能与磺 酸基有关.     

马钱子碱与牛血清白蛋白相互作用的研究

应用荧光光谱和紫外光谱法研究了马钱子碱与牛血清白蛋白(BSA)的结合反应,求得它们之间的结合常数K_A和结合位点数n分别为K_A=6.3×103,n=0.94(27 ℃);K_A=7.7×103,n=0.97(37 ℃)。根据Frster非辐射能量转移理论求出了马钱子碱与BSA之间的结合距离为3.99 nm(27 ℃)和4.21 nm(37 ℃)。探讨了马钱子碱的荧光猝灭机理,结果表明马钱子碱能够插入BSA内部形成基态复合物导致内源荧光猝灭,猝灭机理主要是静态猝灭和非辐射能量转移。根据热力学参数确定马钱子碱与BSA之间的作用力类型主要为疏水性相互作用。     

荧光法研究牛血清白蛋白与三羟基苯基荧光酮-钼(Ⅵ) 配合物探针的作用机理

用荧光光谱法研究了三羟基苯基荧光酮(TH-PF)-钼(Ⅵ)配合物与牛血清白蛋白的结合反应。探讨了TH-PF-Mo(Ⅵ)配合物对蛋白质内源荧光的猝灭机理,并测定了不同温度下的结合常数,温度为25 ℃时,荧光猝灭法测得该反应的结合常数为K=4.78×104 L·mol-1,温度为40 ℃时,荧光猝灭法测得该反应的结合常数为K=3.72×104 L·mol-1。根据Frster非辐射能量转移理论,确定了给体-受体之间的作用距离和能量转移效率(E=0.314),并根据热力学参数确定了TH-PF-Mo(Ⅵ)配合物与牛血清白蛋白之间的作用力类型,以静电引力为主。     

锌试剂与壳聚糖的结合反应机理的研究

为从分子水平认识多糖分子与小分子之间相互作用的机理 ,应用光谱法研究了壳聚糖 (CTS)与锌试剂(ZCN)的相互作用机理 ;测得ZCN CTS复合物吸收光谱出现新的吸收峰所需的临界ZCN/CTS摩尔比为 2 .6 7×10 3 ,CTS对ZCN的最大结合数为 6 .93× 10 3 ,实验值与理论值相吻合 ,证明了多糖与生物探针相互作用理论模型的可靠性 ;探讨了ZCN与CTS相互作用产生变色反应的机理 ,认为其是在ZCN与CTS大分子间发生静电相互作用的基础上 ,主要由ZCN与CTS大分子间的疏水相互作用所引起     

脱氧核糖核酸与刚果红化学反应的机理研究

采用UV-Vis光谱法,研究了在pH 4.56的Tris缓冲溶液中脱氧核糖核酸(DNA)与刚果红(GGH)相互作用。生成的紫色配合物最大吸光度差ΔA在600 nm,反应前后吸收光谱变化明显,反应体系对比度好。在此波长下测得配合物的表观摩尔吸光系数ε=1.41×105 L·cm-1·mol-1,最大结合数n=32,最低检出限为c＝8.04×10-8 mol·L-1 等。研究了体系的酸度、温度、时间等基本反应条件,以及不同类型物质对反应体系的干扰状况。离子强度的改变对体系的吸光度有一定影响。探讨了小分子物质与DNA作用的方式及二者的分子结构、分子构象及电子云分布之间的关系。     

小檗碱与人血清白蛋白的相互作用

利用紫外光谱和荧光光谱技术研究了中药有效成分小檗碱与人血清白蛋白(HSA)的相互作用机制。利用荧光猝灭反应测得它们之间结合常数K=1.168×105 L·mol-1,结合位点数n=5.26。依据Frster非辐射能量转移机制,测得供体-受体间结合距离R=3.44 nm和能量转移效率E=0.303。认为小檗碱在HSA的位置阻断了酪氨酸残基与色氨酸残基之间的能量转移,并使它们的荧光猝灭,并与色氨酸结合生成了较弱的荧光发光体。     

靶向分子羧甲基多糖与DNA作用机理研究

以溴化乙锭(EB)为荧光探针,结合紫外光谱法研究了壳聚糖、羧甲基壳聚糖、羧甲基纤维素与DNA的相互作用,药物分子与DNA作用程度可用作用常数D来表示,实验表明: 几种生物多糖与DNA分子的作用强弱顺序为: 壳聚糖＞羧甲基壳聚糖＞羧甲基纤维素;多糖主要以嵌插方式与DNA作用, 使EB-DNA荧光猝灭。     

复合纳米颗粒中白藜芦醇-大麦醇溶蛋白相互作用研究

为阐明大麦醇溶蛋白包覆白藜芦醇形成纳米颗粒的分子机制,采用荧光光谱、紫外吸收光谱以及傅里叶红外光谱和差示扫描量热技术等研究了白藜芦醇与大麦醇溶蛋白分子间的相互作用。结果表明：白藜芦醇与大麦醇溶蛋白分子的相互作用,对其内源性荧光具有猝灭作用,结合差示扫描量热的结果,推测二者通过静态猝灭过程形成了新的复合物;计算出两者之间的结合常数与结合位点数分别为K_A(298 K)=2.21×105 L·mol-1,K_A(310 K)=1.53×104 L·mol-1和n_{298 K}=1.23,n_{310 K}=0.94;依据热力学参数和傅里叶红外光谱判断白藜芦醇与大麦醇溶蛋白之间的作用力为范德华力和氢键相互作用;根据Frster非辐射能量转移原理,计算出白藜芦醇与大麦醇溶蛋白的结合距离为3.25 nm,能量转移效率为0.227;同步荧光检测结果表明白藜芦醇结合大麦醇溶蛋白分子后对其构象产生了明显影响,且该影响是由色氨酸残基所处环境的疏水性变化所致。     

硫酸软骨素与天青A空间定向相互作用机理的研究

应用光谱法研究了天青A (AA)与硫酸软骨素 (CS)的相互作用吸收光谱 ,考察了不同AA CS摩尔比、乙醇、羟丙基 β 环糊精、TritonX 10 0以及NaCl等实验条件对AA与CS反应体系的影响。测得CS对AA的最大结合数N =15 1,结合常数K =5 2 4× 10 4 ,并且建立了AA与CS大分子空间定向相互作用的实验研究模型。实验结果表明 ,AA与CS相互作用产生的变色反应 ,不仅与AA与CS分子间的静电相互作用有关 ,而且还与CS大分子上结合态AA的密度有关。CS大分子上结合态AA分子间的疏水相互作用是形成5 5 0nm吸收峰以及产生变色反应的主要原因。     

牛血清白蛋白与锌试剂作用机理的荧光法研究

采用荧光光谱法、紫外-可见光度法研究了牛血清白蛋白(BSA)与锌试剂(ZCN)作用的机理。牛血清白蛋白(BSA)在280 nm的光激发下能发射345 nm的荧光(即λ_ex=280 nm,λ_em=345 nm)。当BSA中加入适量的锌试剂(ZCN),BSA的荧光被部分猝灭。由Stern-Volmer方程和双倒数曲线Lineweaver-Burk方程获得了反应的动态猝灭常数、静态猝灭常数,并对猝灭的类型做出了推断。通过计算反应的热力学参数讨论了结合反应的主要作用力类型。根据Frster非辐射能量转移机理,计算了给体(BSA)与受体(ZCN)间距离r=5.07 nm和能量转移效率E=0.67。证实了该体系的荧光猝灭是通过能量转移机制产生的单一静态猝灭过程。     

荧光光谱法研究亚甲基蓝与蛋白质的结合反应

应用荧光光谱法研究了水溶液中亚甲基蓝与牛血清白蛋白分子间的结合反应 ,讨论了亚甲基蓝对蛋白质内源荧光的猝灭机理 ,测定了结合常数 (KA=3 44× 10 5L·mol-1)和结合位点数 (n =1 0 3)。依据F rster非辐射能量转移 ,理论确定了授体 受体间的结合距离 (r=1 6 7nm)和能量转移效率 ,并用同步荧光技术考察了亚甲基蓝对牛血清白蛋白构象的影响     

Analysis of interaction between the organic dye methylene blue and the surface of AgCl(I) microcrystals

We have used IR spectroscopy to study the nature of interaction between molecules of the organic dye methylene blue (MB) and the surface of AgCl(I) microcrystals. We recorded the IR absorption spectra of the organic dye methylene blue in the gas phase, adsorbed on the surface of AgCl(I) microcrystals, and also in the form of crystal hydrates. The results obtained allowed us to hypothesize that unsaturated dimethylamino groups and the sulfur atom in the heterocycle of this dye are the most active in the interaction between methylene blue molecules and the surface of AgCl(I) microcrystals. With the help of IR spectra of methylene blue crystal hydrates, we confirmed the significant role of OH groups during adsorption of molecules on the surface of AgCl(I) microcrystals. We suggest the possibility of formation of hydrogen bonds between the anions of the crystal lattice and the unsaturated dimethylamino groups as well as the sulfur atoms in the heterocycle, by formation of hydrogen bonds with the OH groups. __________ Translated from Zhurnal Prikladnoi Spektroskopii, Vol. 74, No. 6, pp. 731–737, November–December, 2007.     

Catalytic reduction of methylene blue by sulfide ions in the presence of nanoAlMCM-41 material

This article reports the reduction of methylene blue (MB) by sulfide ions (S²⁻) in aqueous solution by a nanoAlMCM-41 catalyst. The results obtained showed that, under acidic conditions (pH=2) MB was reduced to the Leuco methylene blue (LMB) form by sulfide ions and by changing the pH between 2.0 and 7.0 a reduction of the MB dye to the MBH₂⁺ form was observed. In alkaline media (pH=8), a reduction of methylene blue by sulfide ions to the MBH₂⁺ form and demethylation of the dye was observed simultaneously. The demethylation of MB leads to a thionine dye product. Using nanoAlMCM-41 but with encapsulated CoS nanoparticles only causes demethylation of MB in aqueous solution.     

美兰光化学法对红细胞损伤的NMR研究

运用³¹P NMR研究红细胞中高能磷酸化合物的代谢变化,探讨美兰光化学法对红细胞的损伤. 采用MSL-300 MHz型超导核磁共振仪分别测量健康成人红细胞对照组、经美兰（亚甲基蓝）光化学处理组及预先加入L-组氨酸组的高能磷酸化合物,通过分析比较红细胞中2,3-二磷酸甘油酸（2,3-DPG）、三磷酸腺苷（ATP）及磷酸化糖（SP）相对含量,了解美兰光化学法对红细胞代谢的影响. 经亚美兰光化学法处理的红细胞,葡萄糖进入戊糖磷酸途径的分量增加,使红细胞中2,3-DPG和ATP含量显著减少,而SP含量大幅上升,在某种程度上影响红细胞的理化性质和功能指标;L-组氨酸对此有一定的保护作用.     

荧光光谱滴定法研究葫芦脲[7]对亚甲基蓝的包结作用

采用荧光光谱滴定法研究了不同温度下及醋酸缓冲溶液中,葫芦脲[7](CB7)对亚甲基蓝(MB)的包结作用。在一定的浓度范围内,体系的荧光强度随着CB7浓度的增大而增强,且荧光峰位有一定的蓝移,结果显示二者形成了1∶1的包结配合物,同时计算了它们的包结稳定常数以及相关热力学常数ΔG,Δh和ΔS,热力学常数数值说明反应是焓变驱动的。此外,利用1H NMR谱图,结合葫芦脲[7]以及客体亚甲基蓝的结构特征,对其包结机理及主客体作用模式进行了探讨。     

金属钴树状高分子与DNA相互作用的光谱法研究

通过氯化钴、乙二醛和第五代树状高分子反应合成了金属钴树状高分子配合物。并以亚甲基蓝为荧光探针,通过紫外-可见光谱、荧光光谱和同步荧光方法研究了金属钴树状高分子配合物与鲱鱼精DNA(hsDNA)的相互作用。结果显示,此配合物与hsDNA作用时,其紫外吸收产生明显增色效应,荧光强度增强。NaCl不同程度抑制金属钴树状高分子与hsDNA的结合。配合物也以竞争方式抑制亚甲基蓝与hsDNA作用,而亚甲基蓝可以插入金属钴树状高分子配合物的内部。这些结果证明,配合物主要通过与hsDNA链上带负电荷的磷酸基静电相吸形式结合而堆积在双螺旋hsDNA分子表面,减弱了结合位点附近亚甲基蓝分子与hsDNA的静电作用,而钠离子中和了hsDNA上磷酸基团上的负电荷,削弱了该配合物与hsDNA的静电结合。     

Interaction Between Methylene Blue and CalfThymus Deoxyribonucleic Acid by Spectroscopic Technologies

Characterization of the interaction between methylene blue (MB) and calf thymus deoxyribonucleic acid (ctDNA) was investigated by UV absorption spectra, fluorescence spectra, fluorescence polarization and fluorescence quenching experiments by ferrocyanide. The above results indicated that the binding modes of MB to ctDNA were relative to the molar ratio γ (γ=[DNA]/[MB]). At low γ ratios (γ < 4), remarkable hypochromic effect with no shift of λ_max in the absorption spectra of MB was observed in the presence of increasing amounts of ctDNA, the fluorescence of MB was efficiently quenched by the ctDNA bases and the fluorescence polarization of MB was slightly increased, which indicated that MB cations bound to phosphate groups of ctDNA by electrostatic interaction and then stacked on the surface of ctDNA helix. While at high γ ratios (γ > 6), besides the fluorescence of MB was quenched efficiently by the ctDNA bases, a red shift (about 3 nm) in the absorption spectra of MB was observed and the fluorescence polarization of MB was obviously increased, which indicated the intercalation binding that MB molecules were intercalated into the space of two neighbouring DNA base pairs was the preferred mode. Effects of K₄Fe(CN)₆ on the fluorescence quenching of the MB-ctDNA system at low and high γ ratios were also performed. The results showed that at γ = 1.7, the quenching effect by ferrocyanide was higher than that of pure MB, while at γ = 13.6 a decreased quenching of the fluorescence intensity was observed as compared with that of pure MB, which further proved the above conclusion. In addition, the mechanisms of the hypochromic effect and the fluorescence quenching were also discussed in detail.     

Aggregation of 1,3,7-trimethylxanthine with methylene blue in aqueous solution

We have studied self-association of aromatic molecules of the thiazine dye methylene blue in aqueous solution, using a dimer model. We have determined the dimerization equilibrium constant for the dye molecules K_D = 3900 ± 800 M⁻¹ at T = 293 K. We have decomposed the experimental spectrum into dimer and monomer components. Using the ratio of the molar absorption coefficients for two absorption bands of the dimer spectrum, we obtained the “average” value of the angle between the electronic transition moments of the molecules in the dimers, α = 48°. We have studied heteroassociation of methylene blue (MB) and 1,3,7-trimethylxanthine (caffeine) molecules in aqueous solution. We have calculated the heteroassociation constant as 200 ± 34 M⁻¹. We conclude that heteroassociation of methylene blue and caffeine molecules leads to a lower effective dye concentration in solution, which hypothetically may affect its biological activity. We have determined the values of the Gibbs free energy, the enthalpy, and the entropy for dimerization of methylene blue molecules: ΔG₂₉₃ = −(20 ± 3) kJ/mol, ΔH = −(25 ± 9) kJ/mol, Δ S₂₉₃ = −(17 ± 6) J/mol·K; and for methylene blue-caffeine heteroassociation: ΔG₂₉₃ = −(13 ± 3) kJ/mol, ΔH = −(14 ± 10) kJ/mol, ΔS₂₉₃ = −(2.4 ± 0.2) J/mol·K, respectively. We have shown that the methylene blue aggregates and the heteroassociates with caffeine are predominantly stabilized by dispersion interactions between the chromophore molecules in the associates. __________ Translated from Zhurnal Prikladnoi Spektroskopii, Vol. 73, No. 2, pp. 158–163, March–April, 2006.     

One single-step synthesis of multifunctional methylene blue-coated magnetite nanoparticles

This work reports the preparation of hybrid nanoparticles with magnetic and fluorescent properties. The material is based on magnetite nanoparticles (NPs) coated with fluorophore methylene blue (MB). The synthesis of a multifunctional material with magnetic and fluorescent features is carried out in a single step by electrooxidation. The effect of the presence of methylene blue in the synthetic medium is discussed. The presence of MB polymer at the NP surface is demonstrated with visible UV, infrared and Raman spectroscopy. The NPs morphology, structure and size are determined by transmission electron microscopy (TEM) and X-ray diffraction. The magnetic properties are measured with a vibrating sample magnetometer (VMS). In overall, the results show that magnetite NPs generated electrochemically in the presence of MB present a core/shell structure, being the NP at the core surrounded by methylene blue polymer, leading to a nanocomposite or hybrid material.