紫外灯照射牛血清白蛋白的共振光散射光谱法测定

利用紫外灯照射后牛血清白蛋白(BSA)的共振光散射(RLS)强度的增强,建立了一种快速、简便测定BSA的RLS光谱法。考察了pH值、反应最佳时间等因素对RLS强度的影响。实验结果表明,在pH值为4.95,紫外灯照射时间为90min条件下,测定BSA的线性范围为0.5～10mg/L,检出限为0.33mg/L。同时发现DNA与BSA共同作用时,RLS强度增强幅度增加,测定BSA的线性范围为0.05～10mg/L,检出限为0.01mg/L。将该法应用于合成样品中BSA测定,结果较好。     

用荧光光谱和共振光散射光谱研究甲硝唑与牛血清白蛋白的相互作用

用荧光光谱和共振光散射光谱对甲硝唑与牛血清白蛋白的作用进行了对比研究, 测定了该反应的结合常数、结合位点数. 探讨了甲硝唑对牛血清白蛋白荧光和共振光散射猝灭的机理． 利用热力学参数确定了分子间的作用力性质；根据非辐射能量转换机制, 确定了甲硝唑-牛血清白蛋白间的结合距离. 采用同步荧光技术考察了甲硝唑对牛血清白蛋白构象的影响．     

虎红-牛血清白蛋白共振光散射光谱研究及其分析应用

基于在pH3.0的BR缓冲溶液中蛋白质的加入使虎红的共振光散射信号增强,建立了一种蛋白质测定的新方法.详细地探讨了pH、染料用量、表面活性剂、离子强度、共存物质等对信号的影响.牛血清白蛋白浓度在0～4.0μg/mL范围内与体系散射强度呈线性关系,检出限为0.01718μg/mL.该方法用于合成样品中牛血清白蛋白的测定,...     

钙试剂-蛋白质体系的共振光散射光谱研究

基于蛋白质对钙试剂共振光散射的增强作用,拟定了一种测定蛋白质的共振光散射法。在pH 4.00的水溶液中,钙试剂在595 nm处的共振光散射增强与蛋白质浓度呈线性关系。此方法对人血清白蛋白、牛血清白蛋白的检出限分别可达0.068和0.085μg/mL。同时得到钙试剂与人血清白蛋白和牛血清白蛋白的结合常数(K)以及结合位点数(n)分别为8.15×104mol/L,0.18和7.67×104mol/L,1.5。     

共振散射光谱法研究人血清白蛋白与二甲酚橙的反应

对二甲酚橙作为光散射探针测定蛋白质进行了研究,基于人血清白蛋白对二甲酚橙的共振光散射强度的增强效应,建立了测定蛋白质的共振散射光谱法。在pH 4.00的NaOAc-HOAc缓冲溶液中,二甲酚橙只有微弱的光散射强度,但它与蛋白质的缔合物却有较强的共振光散射信号,在λ=517 nm处,光散射强度达到最大,并且与蛋白质浓度在为0.0~10.0 mg.L-1范围呈线性,检出限为0.044 mg.L-1。运用摩尔比法测定二甲酚橙和蛋白质的结合数为132,用于实际样品的分析,测定值与考马斯亮蓝法的结果一致,分析结果的RSD值(n=5)均小于5%。     

牛血清白蛋白的共振光散射光谱法测定

通过测量β-巯基乙醇（β-ME）-十二烷基苯磺酸钠（SDBS）-牛血清白蛋白（BSA）体系的共振光散射强度,建立了一种测定BSA的快速、简便的共振光散射光谱法。考察了PH值、β-ME和SDBS的浓度、试剂的加入顺序等因素对共振光散射强度的影响。实验结果表明,在PH值为2.4,β-ME的浓度为0.71mol·L^-1,SDBS的浓度为6.5×10^-4mol·L^-1条件下,测定BSA的线性范围为1.0×10^-8—7.0×10^-7mol·L^-1,检出限为6.0×10^-9mol·L^-1。将该法应用到合成样品中BSA及人血清样品中总蛋白的测定,所得结果与传统的考马斯亮蓝法的测定结果基本一致。     

酸性铬兰K-OP-牛血清白蛋白体系的共振瑞利散射及共振非线性散射光谱研究及应用

在pH 3.2的邻苯二甲酸氢钾-HCl缓冲液中,酸性铬兰K(ACBK)-OP与牛血清白蛋白(BSA)形成三元离子缔合物,导致共振瑞利散射(RRS)、二级散射(SOS)和倍频散射(FDS)的显著增强,光谱最大散射波长分别位于420,678和340nm。体系的光散射强度与BSA浓度在一定范围内呈线性增强,RRS在0~3.5 mg/L,SOS在0~3 mg/L,FDS在0~3 mg/L范围内对BSA的检出限分别为0.3,0.7和0.8μg/L,据此建立了测定BSA的共振线性(RRS)和共振非线性光散射(RNLS)分析法。以RRS法考察了酸性铬兰K-OP与白蛋白形成三元缔合物的适宜条件、影响因素等。方法可用于合成样品及血清样品中蛋白含量的测定。     

共振光散射光谱研究噻嗪红R与牛血清白蛋白的作用及其分析应用

研究了噻嗪红R(Thiazine red R,TR)与牛血清蛋白(BSA)作用的共振光散射(RLS)光谱特征。考察了各种影响因素,并计算出了BSA与TR的结合比(质量之比)为1.25。结果表明,在优化条件下体系的RLS强度与蛋白质浓度在一定范围内具有良好的线性关系,据此建立了一种蛋白质测定的新方法。在最佳实验条件(TR,2.0×10-5mol/L;pH2.36)下,BSA的线性范围是0.01~5.0μg/mL,检出限为1.4ng/mL。该方法已用于合成样品及尿液的测定。     

甲基紫共振光散射光谱法测定日落黄

建立了甲基紫共振光散射光谱法测定饮料中日落黄的方法。在pH 3的B-R缓冲溶液中,日落黄与甲基紫之间相互作用形成离子缔合物,引起共振光散射强度的显著增加。共振光散射峰分别位于339 nm和650 nm处,在650 nm处,共振光散射强度与日落黄质量浓度在0.02～0.2 mg/L范围内呈现良好的线性关系(r2=0.9981)。考察了酸度、甲基紫用量、离子强度、温度等对体系光散射强度的影响,测定了缔合物的组成比。方法的检出限为7.5μg/L,相对标准偏差(RSD)为2.1%。方法用于饮料样品中日落黄的测定,结果与紫外分光光度法一致。     

免疫共振散射光谱法测定痕量人血清前白蛋白

在pH 7.6的磷酸氢二钠-磷酸二氢钠缓冲溶液中,羊抗人前白蛋白与前白蛋白发生特异性结合生成免疫复合物,导致体系的共振散射增强。体系在最大散射波长470 nm处,前白蛋白的质量浓度在0.067~2.82 mg.L-1范围内与共振散射增强强度(ΔI)呈线性关系,检出限(3S/N)为0.028 mg.L-1。应用于测定人血清前白蛋白,测定值相对标准偏差(n=6)在1.15%~3.00%之间。     

Determination of bovine serum albumin using resonance light scattering technique with sodium dodecylbenzene sulphonate-cetyltrimethylammonium bromide probe

In this paper, the anionic surfactant sodium dodecylbenzene sulphonate (SDBS) and cationic surfactant cetyltrimethylammonium bromide (CTMAB) were used as resonance light scattering (RLS) probe to determine bovine serum albumin (BSA). Based on the weak RLS intensity of SDBS-CTMAB probe and the enhancement of RLS intensity of BSA in the presence of the probe, a simple assay for BSA was developed. The experimental results showed that the formation of three component complex BSA-SDBS-CTMAB is the main reason for the enhancement of RLS intensity of BSA, in which SDBS as a bridge can interact with both BSA and CTMAB. The effects of pH value, incubation time, concentrations of SDBS and CTMAB on the enhanced RLS intensity of BSA were investigated. Under the optimum conditions, the enhanced RLS intensity is proportional to the concentration of BSA in the range from 2.5 x 10(-8) to 2.0 x 10(-6)mol L(-1). The detection limit is 9.7 x 10(-9) mol L(-1) for BSA. The study of foreign substance effect on the determination of BSA indicated that most of metal ions have little effect on the determination of BSA. The results of assay for BSA in synthetic samples were satisfactory.     

蛋白质与肝素结合平衡的共振散射光谱法研究

基于共振光散射光谱(RLS)研究了牛血清白蛋白(BSA)与肝素(HP)之间的结合平衡.结果表明,HP与BSA通过电子作用力以及疏水作用力结合,引起体系共振光散射增强.根据406nm的光散射增强信号建立了BSA与HP结合平衡的数学关系式,并考察了不同温度、pH及离子强度对它们之间结合平衡的影响.基于RLS数据计算了BSA...     

Ultrasensitive determination of serum albumin using resonance light scattering based on ZnS-polyacrylic acid nanoparticles

ZnS-polyacrylic acid (ZnS-PAA) was prepared by an in situ polymerization method using nano-ZnS as core in the presence of acrylic acid (AA), and ZnS-PAA nanoparticles was characterized by ultraviolet spectrometry (UV) and transmission electron microscopy (TEM). Based on the significant increase of the resonance light scattering (RLS) intensity with the interaction between nanoparticles and serum albumin, RLS method was developed for the sensitive determination of serum albumin (BSA and HSA). Under optimum conditions, the change of the intensity (ΔI) of the RLS spectra at λ = 392 nm was linearly proportional to the concentration of BSA and HSA. The linear range was 1–100 ng mL^?1 for HSA and 1–120 ng mL^?1 for BSA, and the limit of detection (LOD) was 0.4 ng mL^?1 for HSA and 0.5 ng mL^?1 for BSA. This method proved to be very sensitive, rapid, simple and tolerant of most interfering substances.     

水飞蓟素与牛血清白蛋白相互作用的荧光光谱研究

采用同步荧光光谱技术研究了pH=7.40的Tris-HCl缓冲体系中水飞蓟素与牛血清白蛋白(BSA)的相互作用以及水飞蓟素对BSA构象的影响.结果表明,水飞蓟素对BSA的荧光猝灭过程为静态猝灭,结合热力学参数△rH∞=-45.21 kJ·mol-1,△rSm=-61.61 J·K-1·mol-1;据此可以推断,水飞蓟素...     

Size-exclusion chromatography--multiangle laser light scattering analysis of beta-lactoglobulin and bovine serum albumin in aqueous solution with added salt

The solution characteristics of beta-LGB (beta-lactoglobulin) and BSA (bovine serum albumin) are reported as determined by size-exclusion chromatography with on-line multiangle laser light scattering, differential refractive index and UV detection. The order of the three in series placed detectors as well as the interdetector volumes have been carefully pointed out. At concentrations below 2.5 mg/ml and at different values of pH the weight-average molecular mass of both proteins have been obtained. They indicate the appearance of monomers, dimers and higher order multimers. For beta-LGB the growth of self-associates could be observed at the isoelectric point over a period of days. The range of applicability of the method is discussed.     

卡维地洛的电化学测定及与牛血清白蛋白相互作用的研究

建立了测定卡维地洛(CAR)新的电化学方法.结合紫外、红外光谱分析,研究了CAR与牛血清白蛋白(BSA)的相互作用.实验发现,在pH4.0Britton-Robinson(B-R)缓冲溶液中,CAR在碳糊电极上产生3个不可逆的氧化峰.以0.92V处的氧化峰为研究对象,结果发现峰电流Ipa,1与CAR浓度在2.45×10-5～1.19×10-3mol/L范围呈良好的线性关系,CAR的检出限为5.6×10-6mol/L.当BSA加入CAR溶液后,CAR峰电流降低,氧化峰电流的降低值△Ipa,1与BSA的浓度在2.92×10-7～1.09×10-5mol/L范围内呈良好线性关系,BSA的检出限为4.1×10-8mol/L.电化学结果表明,CAR与BSA之间形成1∶1的结合物,结合常数为3.14×106L/mol.紫外光谱表明CAR的加入使BSA的吸收峰发生红移且有增色效应.红外光谱表明CAR与BSA分子中氨基酸残基的硫及氮原子形成键合作用.     

Determination of tannic acid after precipitation with bovine serum albumin by visible light scattering in a flow injection system

Tannic acid can form a precipitate with bovine serum albumin (BSA), and this was explored for its determination using a flow injection system along with detection via light scattering at 600 nm. The results on light scattering are confirmed using a particle sizing method. Scattering signals are negligible compared to the background generated by BSA in solutions with a pH of <4 or >8. Any precipitates on the wall of the flow cell pipeline were effectively removed by using a sodium dodecyl sulfate washing solution. The analytical range is from 70 to 250 µgmL^-1 of tannic acid. The results imply that increasing BSA concentrations will enhance sensitivity and analytical ranges. The system can be used in solutions of pH values between 5 and 7. The results correlated closely (r²?>?0.97) with those obtained by the ferrous tartrate method when analyzing a single tannic acid and two tea tannins.     

肌醇与牛血清白蛋白相互作用的荧光光谱研究

应用荧光光谱研究了肌醇与牛血清白蛋白(BSA)分子间的相互作用;求出了猝灭常数,讨论了肌醇对BSA构象的影响,并依据能量转移理论确定了肌醇与蛋白的最近距离.结果表明,肌醇与BSA两者间的相互作用为单一的动态猝灭过程.     

绞股蓝皂苷与牛血清白蛋白相互作用的荧光光谱研究

用荧光光谱技术研究了绞股蓝皂苷与牛血清白蛋白(BSA)在pH=7.40的Tris-HCl缓冲溶液中的相互作用;通过计算确定了绞股蓝皂苷与BSA的结合位点数和结合常数,利用热力学分析探讨了绞股蓝皂苷与BSA之间的结合方式;同时采用同步荧光技术考察了绞股蓝皂苷对BSA构象的影响.结果表明,绞股蓝皂苷对牛血清白蛋白的荧光猝灭过程为静态猝灭;二者主要靠疏水作用和静电引力结合.     

Resonance double light scattering method for the determination of proteins with morin-CTMAB

A new determination method of proteins with the limit of determination at nanogram levels is proposed by using a common spectrofluorimeter to detect intensity of resonance double line scattering (RDLS). Proteins including bovine serum albumin (BSA), human serum albumin (HSA) can combine with morin and cetyltrimethylammonium briomide (CTMAB) in the pH range 7.0-8.0 and produce enhanced RDLS signal at lambda(ex)/lambda(em) 305.0/610.0 nm. Optimization conditions for the morin-protein-CTMAB interaction were tested. In the studied system, BSA/CTMAB/morin = 1:2:3. The association constant of morin with BSA is 5.2 x 10(4). Under the optimum conditions, the linear range is 7.5 x 10(-8)-1.0 x 10(-5) g/ml for BSA, 2.5 x 10(-8)-5.0 x 10(-6) g/ml for HSA. The detection limits (S/N = 3) are 66.0 ng/ml for BSA and 23.0 ng/ml for HSA, respectively. Four synthetic samples were analyzed satisfactorily.