茜素黄R与蛋白质相互作用的线性扫描极谱法研究及分析应用

以电化学方法研究了在pH3.2的Britton Robinson(B R)缓冲溶液中茜素黄R与蛋白质的相互作用。茜素黄R在-0.43V(vs.SCE)有一个良好的极谱还原峰,加入人血清白蛋白(HSA)后,其峰电位不变而峰电流下降,利用峰电流的下降可以测定蛋白质。优化了结合反应条件和电化学测定条件,在最佳条件下,峰电流降低值同HSA的质量浓度在4.0～40.0mg L范围内呈线性关系,其线性方程为ΔI″p(nA)=-5.48 72.28ρ(mg L),r=0.993。将该方法应用于人血清样品的测定,结果与经典的考马斯亮蓝G 250光度法一致,回收率在97%～103%之间。此方法还可应用于牛血清白蛋白、牛血红蛋白、卵清白蛋白等蛋白质的测定。     

茜素红S-牛血清白蛋白配合物的凝胶层析分离制备及紫外可见吸收光谱研究

用葡聚糖凝胶层析分离制备牛血清蛋白(BSA)-茜素红S(ARS)配合物。在420和530 nm处用紫外可见吸收光谱法同时测定含有BSA-ARS配合物和茜素红S的联立方程为:A420=4.89×103cARS 3.06×104cBSA-ARS,A530=4.60×102cARS 2.29×104cBSA-ARS;正交试验选择了合适的分离条件:柱直径1.0 cm,柱长30.0 cm,凝胶用量1.3 g,最佳进样浓度为ARS:5×10-3mol/L、BSA:1.49×10-4mol/L,进样体积1.5 mL,洗脱流速0.33 mL/min,分离度为1.25;测定了纯BSA-ARS配合物紫外吸收光谱,最大吸收峰在530 nm。     

茜素红S与壳聚糖相互作用的电化学行为研究

在pH5.0NaAc—HAc缓冲溶液中,茜素红S能与壳聚糖生成一种紫红色的复合物。复合物使茜素红S在-0.408 V处的还原峰峰电流降低。在选定的最佳条件下,用循环伏安法和线性扫描极谱法测定其峰电流的降低值与壳聚糖的浓度在0.5～7．5mg/L的范围内呈线性关系,线性回归方程为△ip^#=30．92 108．4CCTS(mg／L),相关系数0.9990;检出限0．4mg/L。据此建立一种快速、简便测定壳聚糖的方法。该方法可应用于实际样品的测定。     

电化学法研究蛋白质和茜素红S的相互作用

在pH 4 .2的Britton Robinson缓冲液中 ,茜素红S(ARS)与牛血清白蛋白 (BSA)能形成一种红色的非电活性的超分子复合物。用线性扫描二阶导数极谱法和循环伏安法对该体系进行了研究 ,复合物的形成使ARS的还原峰电流下降 ,峰电流的下降值同所加的BSA浓度在一定范围内呈线性关系。用于BSA的测定 ,在 8.0× 10 -8～ 1.2× 10 -6mol/L范围内呈线性关系 ,检测限为 4 .3× 10 -8mol/L ,对结合反应机理进行初步的探讨     

聚吡咯薄膜中茜素红S的电化学特性

利用循环伏安法研究了聚合电位和聚合电量对茜素红S掺杂聚吡咯薄膜修饰玻碳电极性质的影响,考察了不同电位范围内修饰电极的电化学特性.发现在茜素红S的氧化还原过程中存在着茜素红S分子与聚吡咯链的相互作用,且这一相互作用敏感于扫描正电位限.基于实验结果,提出了可能的茜素红S在电极表面聚吡咯膜内的氧化还原机理.     

酸性棕NR分光光度法测定血清白蛋白

采用分光光度法研究了酸性棕NR与血清白蛋白的结合反应。在Brit ton Robinson(B-R)(pH2.50)缓冲溶液中,酸性棕NR与血清白蛋白结合生成沉淀,探讨了该结合反应的最佳条件,并据此建立了一种高灵敏度的测定血清白蛋白的新方法。牛血清白蛋白(BSA)和人血清白蛋白(HSA)分别在28.0～112.0mg/L、24.4～122.0mg/L范围内服从比尔定律,其表观摩尔吸光系数分别为:1 44×106L·mol-1·cm-1(BSA)、1.32×106L·mol-1·cm-1(HSA)。对7个人血清样品蛋白总量平行测定6次,相对标准偏差0.83%～3.02%,回收率90.90%～109.80%,且与医院双缩脲法结果基本一致。     

茜素红S与脱氧核糖核酸相互作用的电化学研究

研究了茜素红S在玻碳电极上的电化学行为,根据茜素红S与DNA作用的伏安曲线、紫外．可见光谱及溴化乙锭对茜素红S与DNA作用的影响,认为茜素红S与DNA发生了嵌插作用。考察了温度、时间及pH值对二者作用的影响。     

茜素红S与人血清白蛋白相互作用的分光光度研究

在ｐＨ４．３左右的Ｂｒｉｔｏｎ－Ｒｏｂｉｎｓｏｎ（Ｂ－Ｒ）缓冲溶液中,茜素红Ｓ（ＡＲＳ）与人血清白蛋白（ＨＳＡ）结合,生成红色的复合物,吸光度值与ＨＳＡ含量呈线性关系。复合物的吸收峰值λｍａｘ为５３０ｎｍ,比ＡＲＳ试剂本身红移１１０ｎｍ。其表观摩尔吸光系数ε＝１．０×１０４Ｌ·ｍｏｌ－１·ｃｍ－１。ＨＳＡ标准曲线在其质量浓度１０～９００ｍｇ／Ｌ间呈线性关系,最低检出限为３ｍｇ／Ｌ。研究了该配合物用于蛋白质分光光度法测定的基本条件。应用该法测定了人血清样品,回收率在９６．９％～１０２．１％范围内。实验表明该反应选择性、重现性和对照性均好,操作简便,适用测定浓度范围宽     

阻抑茜素红S褪色动力学光度法测定痕量汞

在HAc—NaAc介质中,以邻菲啰啉作活化剂,痕量Cr（Ⅵ）对H2O2氧化茜素红S褪色反应存在强烈的催化作用,而Hg（Ⅱ）对此褪色反应具有明显的阻抑作用,由此建立了测定痕量Hg（Ⅱ）的新阻抑动力学光度法。该方法线性范围为0～1．6μg／mL,检出限为2．6×10^-8g／mL,应用于污水中Hg（Ⅱ）的测定,结果满意。     

UV-Visible Spectrometric Study and Micellar Enhanced Ultrafiltration of Alizarin Red S Dye

Ultraviolet spectrometric study of alizarin red S (ARS) showed the substantial change in dye spectra by cationic CTAB as compared to anionic SDS and nonionic TX-100 surfactant. High spectral change by CTAB confirms the anionic nature of ARS dye and thus ARS-CTAB complex formation takes place due to electrostatic force of attraction. A little spectral change by SDS is the result of similarly charged repulsive forces that overcome weak hydrophobic-hydrophobic interaction between dye and surfactant micelles. TX-100 exhibited moderate spectral effect responsive to weak hydrophobic-hydrophobic interaction alone. MEUF study of ARS dye justified the spectral changes and dye rejection percentage (R) decreases in the following order: cationic > nonionic > anionic surfactant. Permeate flux (J) slightly decreases in presence of CTAB and it remains virtually constant for both SDS and TX-100. Addition of copper salt (i.e., CuCl₂) in dye-CTAB complex solution, favors rejection (%) removing dye and copper simultaneously via micellar enhanced ultrafiltration.     

以溴甲酚紫为电化学探针测定血清白蛋白

常见的蛋白质定量分析方法、电化学行为及其分析应用已有报道．本文基于酸性条件下蛋白质可与溴甲酚紫(BP)结合生成一种超分子复合物,使溴甲酚紫还原峰电流降低,建立了一种测定蛋白质的新方法,并用于测定人血清样品中的白蛋白含量．     

刚果红与血清白蛋白相互作用的极谱分析

在pH4．7HAc-NaAc缓冲溶液中,刚果红与蛋白质(BSA或HSA)作用形成络合物,使刚果红-0．35V(vs SCE)的还原峰电流下降,峰电流的降低值同所加的BSA或HSA质量浓度在一定范围内呈线性关系;BSA和HSA的线性范围分别为0．5～12mg／L和0．5～11mg／L,检出限分别为0．25和0．20mg／L;运用该法测定了人血清白蛋白样品,结果令人满意。     

Site-selective probe for investigating the asynchronous unfolding of domains in bovine serum albumin

A convenient method is proposed for precise investigation of the asynchronous structural transition of the domains in bovine serum albumin (BSA) during unfolding process. The method is based on a site-selective probe, alizarin red S (ARS), which has a high affinity to the subdomain IIA of BSA. BSA-ARS complex was formed and gradually unfolded by urea from 0 to 8.0 M. The unfolding occurred in different domains of BSA resulted in distinct alterations of the microenvironment of the bound ARS. The spectral response of BSA-ARS complex, including the color, the UV absorption at 530 and 432 nm, and the intrinsic fluorescence at 342 and 310 nm with the excitation wavelength of 280 nm, showed slight changes in the urea concentration from 0 to 4.5 M, drastic changes from 4.5 to 6.0 M, and almost no changes from 6.0 to 8.0 M. The redox behavior of bound ARS between 0.3 and 0.8 V also showed the same trend. Consequently, a two-step, three-state transition process was monitored by naked eyes, UV-vis spectroscopy and electrochemistry. It is the first report to realize the indicator of the intermediate state during the unfolding process of BSA through convenient methods instead of expensive approaches. The work provides a facile method for the investigation of the unfolding process of multidomain proteins.     

钪 钇茜素红S异多核络合物极谱吸附波的研究

报道了钪钇茜素红Ｓ异多核络合物的形成及该异多核络合物极谱吸附波测定钪的最佳实验条件。用此方法测定了纯氧化钇中的微量钪, 并探讨了该极谱波的性质和电极反应机制。在单扫描示波极谱上用单纯形法和改良的平衡移动法测得该异多核络合物组成的摩尔比为： ｘＳｃ３＋ ∶ｘＹ３＋ ∶ｘＡＲＳ＝１∶２∶５ 。     

Adsorptive Stripping Voltammetric Determination of Antimony by Using Alizarin Red S

Abstract

A sensitive and selective voltammetric method for determination of antimony(III) using Alizarin Red S (ARS) as complexing agent is described. The method is based on the monitoring the oxidation peak of antimony(III)-ARS complex at ?520 mV in ammonium-ammonia buffer (pH = 7.5). The peak current was measured by scanning the potential from ?700 mV versus Ag/AgClto more positive potentials without accumulation in the presence of 1 × 10^?6 mol L^?1 of ARS. The limit of detection (3 s) and limit of quantification (10 s) of the method were calculated from calibration curve as 1.45 µg L^? and 4.8 µg L^? respectively. The calibration plot for antimony(III) was linear in the range of 4.8–30 µg L^?. The interference of various ions was examined. Serious interference from Al(III), Fe(III), Cu(II), Pb(II), and Zn(II) was eliminated by addition of EDTA to the solution. The method was applied to drinking water samples. The recoveries were in the range 94% – 105%. The results obtained from the developed method were compared with those from the differential-pulse anodic-stripping method and no statistically significant difference was found.     

Cathodic Adsorptive Voltammetry of the Gallium‐Alizarin Red S Complex at a Carbon Paste Electrode

The adsorptive voltammetric behavior of the gallium‐alizarin red S (ARS) complex in NH₄OAc‐HCl buffer at a carbon paste electrode(CPE) was investigated. The results showed that the complex can be adsorbed on the surface of the CPE, yielding one reduction peak at ?0.52 V(vs. SCE), corresponding to the irreversible reduction of the ligand, ARS, bonded in the complex. The optimal experimental conditions include the use of 0.10 mol L^?1 ammonium acetate buffer(pH 4.5), 1.0×10^?5 mol L^?1 ARS, an accumulation potential of ?0.05 V, an accumulation time of 180 s ,a rest time of 10 s, a scan rate of 200 mV s^?1and a second‐order derivative linear scan mode. The peak current is proportional to the concentration of gallium(III) over the range 0.02–6.0 μg L^?1, with the detection limit of 0.01 μg L^?1 for an accumulation time of 180 s. The method was applied to the determination of gallium in food samples with satisfactory results.     

Determination of proteins with Alizarin Red S by Rayleigh light scattering technique

A new protein determination method by enhanced Rayleigh light scattering (RLS) technique has been developed. In acid condition (pH=3.60), RLS of 1,2-dihydroxyanthraquinone-3-sulfonate (Alizarin Red S) can be greatly enhanced by addition of proteins, resulting in two characteristic peaks, 360 and 505 nm, respectively. The new protein assay is based on the RLS enhancement and spectrum change. The optimum condition for the reaction was investigated. The linear range is 0.20-24.9 μg ml⁻¹ for BSA and 0.20-15.5 μg ml⁻¹ for HSA. The detection limits (S/N=3) are 9.59 ng ml⁻¹ for BSA and 9.51 ng ml⁻¹ for HSA. The results of determination for human serum samples were comparable to those obtained by Bradford method. The binding stoichiometry was determined.     

负载茜素红S的纳米磁性碳粉修饰电极测定牛白蛋白

采用化学沉积法在纳米碳粉上合成磁性Fe3O4颗粒,纳米碳粉吸附茜素红S后通过磁力附着在石墨电极上,制成了负载茜素红S的纳米磁性碳粉修饰电极。研究了该电极在0.014 mol/L H2SO4-0.2 mol/L KC l溶液中对ARS-牛白蛋白的电还原作用,考察了多种实验条件对峰电流的影响。在-0.45 V电位下,牛白蛋白浓度在1.5×10-8~9×10-7mol/L范围内与线性扫描伏安法还原电流成反比,从而实现对牛白蛋白的测定,并初步研究了电化学反应机理。此方法制成的磁性纳米颗粒修饰电极集分离、富集、测定和更新于一体,具有更新方便的特点。     

Spectrophotometric determination of molybdenum with Alizarin Red S in the presence of poly(sulfonylpiperidinylmethylene hydroxide)

The present work describes a selective and rapid method for the determination of molybdenum with Alizarin Red S (ARS) in the presence of a water soluble polymer, poly(sulfonylpiperidinylmethylene hydroxide) (PSPMH). The ARS modified by PSPMH reacts with molybdenum(VI) in the solutions of pH 3.4-4.0 to produce a red complex. The composition of the complex is 1:4:1 mol ratio of Mo(VI): ARS:PSPMH. The complex obeys Beer's law from 0.05 to 5.50 μg ml⁻¹ with an optimum range. The molar absorptivity is 2.1×10⁴ l mol⁻¹ cm⁻¹ at 500 nm. The interference effects of the foreign cations have been examined and it has been determined that only Cu(II), Al(III) and Fe(III) have to be masked by EDTA and tungsten can be tolerated till 4-fold of molybdenum in case of masking by citrate. The method has been applied to the determination of geological samples without solvent extraction or separation steps.     

Voltammetric Behavior of the Alizarin Red S Interaction with DNA and Damage to DNA

The electrochemical behavior and the interaction of alizarin red S (ARS) with calf thymus DNA was investigated on a bare glassy carbon electrode (GCE) and DNA modified GCE (DNA/GCE), respectively. ARS showed a pair of redox peaks at ?0.445 V and ?0.414 V on a bare GCE. On addition of DNA into the ARS solution, the peak current of ARS decreased and the peak potential positively shifted, but without new redox peaks appeared. The ARS reduction peak current increased with immersion time on a DNA/GCE. The results showed that ARS could interact with DNA molecules by intercalative binding mode. The equilibrium constant, binding number and the ratio of binding constant for oxidized and reduced ARS forms were obtained. The DNA damage was directly detected by appearance of guanosine and adenosine bases oxidation signal. The influence of experimental conditions on DNA damage extent was discussed in detail.