酸性铬蓝K在蛋白质大分子上Langmuir吸附聚集及应用

用微相吸附 -光谱修正 (MPASC)技术研究蛋白质 (BSA)与酸性铬蓝K(ACBK)探针分子间的相互作用。通过pH3.5的介质中蛋白质与酸性铬蓝K反应的光谱研究 ,测定了结合产物的物理化学参数 :结合比为n(ACBK)∶n(BSA)=62∶1,平衡常数KBSA -ACBK=2.2×105。BSA -ACBK摩尔吸收系数ε584=3.9×105L·mol -1·cm -1。该吸附反应用于蛋白质样品测定 ,结果良好。     

茜素红与蛋白质分子Langmuir吸附聚集反应研究

在pH4.3的B-R缓冲体系中,用微相吸附-光谱修正技术[1]研究了茜素红(ARS)与牛血清白蛋白(BSA)、人血清白蛋白(HSA)的结合反应。其吸附结合常数分别为:KBSA-ARS=3.950×104,KHSA-ARS=4.377×104。染料与蛋白的最大结合数分别为NARS∶NBSA=9∶1,NARS∶NHSA=7∶1。经光谱修正技术计算结合产物的实际摩尔吸光系数分别为εBSA-ARS(537nm)=2.517×104L.mol-1.cm-1,εHSA-ARS(519nm)=2.051×104L.mol-1.cm-1,检出限BSA为19mg/L,HSA为23mg/L。经探讨该结合反应机理符合Langmuir吸附聚集反应方程。     

探针体耐尔蓝(NB)与DNA结合反应研究

应用微相吸附-光谱修正(MPASC)新技术研究DNA与耐尔蓝(NB)探针分子间的相互作用,分析生物大分子内静电场的形成与Langmuir吸附的关联性,测定了结合产物结合比、平衡常数等.通过在pH=10.38介质中对DNA-NB反应的光谱分析,结果表明产物结合比NB∶ DNA-P=3∶ 1、平衡常数K=3.33×105,摩尔吸收系数ε660 nm=4.81×103 L/mol cm.样品分析表明DNA回收率95.6%～108%,相对标准偏差RSD=2.8%.     

荧光桃红B在CTMAB上Langmuir吸附及阳离子表面活性剂测定新技术

在离子型表面活性剂溶液中 ,电荷均匀分布在分子长链并形成点静电场 ,容易吸附带异性电荷的探针色体。研究了利用微相吸附 光谱修正 (MPASC)技术分析溴化十六烷基三甲胺(CTMAB)与荧光桃红B(PB)的结合反应 ,并分析了表面活性剂在微量分析中的增效机理。结果表明 ,PB CTMAB作用符合Langmuir单分子层吸附 ,产物结合比为 2∶1,结合常数为 3.4 6× 10 5(15°) ,摩尔吸光系数 (5 6 5nm)ε =1.14× 10 5L·mol- 1·cm- 1,胶束聚集态 (PB2 CTMAB) 78,定量测定了样品中阳离子表面活性剂含量 ,结果良好     

Gu(Ⅱ)、Ni(Ⅱ)与酸性茜素蓝B在溴化十六烷基三甲铵胶束上的配位反应

The coordination reactions of Cu(Ⅱ) and Ni(Ⅱ) with acid alizarine blue B (AABB) in the presence of cetyltrimethylammonium bromide (CTAB) micelle were investigated using the microsurface adsorptionspectral correction technique (MSASC). The aggregation of AABB on CTAB followed the Langmuir isothermal adsorption law. The enrichment of AABB on CTAB sensitized the complexation between Cu(Ⅱ) or Ni(Ⅱ)and AABB. The binding ratio of AABB to CTAB was 1:2.5, and monomeric aggregate, AABB2CTAB5, was formed with an adsorption constant of 5.95×105 at 20 ℃ or 2.48×105 at 40 ℃. In the ternary complexation, the ratio of AABB:Cu and AABB:Ni were 1:1 and 1:2.5, respectively. Two types of aggregates, Cu2.AABB2·CTAB80 and Ni5.AABB2.CTAB80, were formed.     

二甲苯蓝FF在阳离子表面活性剂溴化十六烷基三甲胺上的Langmuir吸附及应用

利用微相吸附 光谱修正 (MPASC)技术研究了溴化十六烷基三甲胺 (CTMAB)与二甲苯蓝FF(XCFF)的结合反应 ,分析表面活性剂在微量分析中的增效机理。结果表明 :XCFF CTMAB间作用符合Langmuir单分子层吸附 ,产物结合比 1∶1 ,胶束产物聚集形式 :(XCFF·CTMAB) 78;结合常数等于 1 .0 2× 1 0 5;摩尔吸收系数 (λ=43 0nm)ε=9.0 5× 1 0 5L·mol- 1 ·cm- 1 。在阴离子表面活性剂存在下 ,聚集产物中XCFF灵敏脱附 ,籍此定量分析了实际样品中阴离子洗涤剂。回收率 93 .0 %～ 1 1 1 % ;RSD小于 5 .8%。     

耐尔蓝硫酸盐在核酸分子表面的长距组装及核酸的三波长共振光散射测定

在pH7.20～7.60及离子强度低于0.012的介质中,耐尔蓝硫酸盐(NBS)在核酸分子表面进行长距组装后,产生293.4nm、349.4nm和560.4nm的共振光散射(RLS)增强峰.根据RLS增强的Scatchard数据分析表明,NBS在小牛胸腺DNA、鱼精DNA和酵母RNA上的组装数分别是6.4、6.6和3.9,组装常数分别为7.1×10~6mol/L、4.6×10~6mol/L和1.7×10~6mol/L.应用三个波长处的RLS增强之和可以测定0～0.8mg/L的小牛胸腺DNA、鱼精DNA和0.20～0.60mg/L的酵母RNA,检测限分别是0.4、1.2和10.5μg/L.方法已成功的应用于合成样测定.     

耐尔蓝硫酸盐在核酸分子表面的长距组装及核酸的三波长共振光？…

在ｐＨ７．２０－７．６０及离子强度低于０．０１２的介质中,耐尔蓝硫酸盐在核酸分子表面进行长距组装后,产生２９３．４ｎｍ,３４９．４ｎｍ和５６０．４ｎｍ的共振光散射（ＲＬＳ）增强峰。根据ＲＬＳ增强的Ｓｃａｔｃｈａｒｄ数据分析表明,ＮＢＳ在小牛胸腺ＤＮＡ,鱼精ＤＮＡ和酵母ＲＮＡ上的组装数分别是６．４,６．６和３．９,组装常数分别为７．１×１０＾６ｍｏｌ／Ｌ,４．６×１０＾６ｍｏｌ／Ｌ和１．７×１、０＾     

亚甲基蓝在云母表面吸附状态的研究

利用XPS测定吸附前后亚甲基蓝（MB）各原子的电子结合能的变化,以判断原子化学环境的改变,从而确定MB在云母表面的吸附位点是二甲胺基上的氮原子.通过AFM测量得到吸附于云母表面的MB分子的平均高度为0.820 nm,这证实了Hähner吸附模型的正确性,即MB分子的最大横截面以65～70°倾斜在云母的（001）表面上.     

尿嘧啶在银电极表面吸附状态的研究

综合分析了尿嘧啶在银电极表面不同条件的表面增强激光Ｒａｍａｎ光谱（ＳＥＲＳ）,利用扩展的Ｈüｃｋｅｌ分子轨道法（ＥＨＭＯ）对尿嘧啶分子在银电极表面的吸附状态进行了计算．结果表明：最稳定的吸附构型能较成功地阐明尿嘧啶吸附在银表面的ＳＥＲＳ谱;尿嘧啶在银表面主要是以Ｎ（３）去质子化的形式吸附;尿嘧啶在银表面的吸附取向是由其浓度和银表面电位共同决定;在较低浓度和较高电位时,Ｎ（３）去质子化的尿嘧啶离子在银表面主要是通过Ｃ（２）及整个嘧啶环上的π电子体系与银表面的相互作用平躺地吸附;在较高浓度和较低电位时,吸附取向发生改变,即主要是通过Ｎ（３）与Ｏ（１０）以及Ｎ（３）与Ｏ（８）垂直地共同吸附在银表面上     

Langmuir Aggregation of Thionin in Nucleic Acids and Its Application

Introduction　　Nucleicacidchemistryhasbeenattractingtheattentionofmanybiochemists.1 7Understandingtheinteractionofachromophorewithnucleicacidscanhelpustorealizethefunctionofsuchgeneticbiopolymersandanalyzethetransmis sionofthegeneticinformation .Thequantitativedeterminationofnucleicacidsisalsosignificantinclinicaltestsandlabora torypractice .However,theinteractionhasnotbeenelucidat edsatisfactorilyandearlierobservationshavenotbeenex plainedclearlyandreasonably ,e.g .,thePesaventoequa tion8an…     

Interaction of Trypan Blue with Protein and Application

The microphase adsorption ‐ spectral correction (MPASC) technique has been described and applied to the aggregation of trypan blue (TB) in proteins. The formation of the microelectrostatic field in protein causes the Langmuir monolayer aggregation of TB. The adsorption ratio of TB to bovine serum albumin (BSA), ovalbumin (OVA), hemoglobin (Hb) and human γ‐globulin (γ‐G) was determined to be 14.8, 8.4, 2.8 and 27.6, respectively, and the adsorption constant of the aggregates to be 7.17 × 10⁵, 4.88 × 10⁶, 4.85 × 10⁶ and 2.99 × 10⁶. The adsorption ratio of TB to proteins interestingly indicates almost no relation to the array sequence of amino acid residues. The interaction of TB with proteins is sensitive at pH 3.29, and the reaction was applied to the determination of protein trace with satisfactory results.     

Interactions of Nile Blue Sulphate with Nucleic Acids as Studied by Resonance Light-Scattering Measurements and Determination of Nucleic Acids at Nanogram Levels

ABSTRACT

The interactions of nile blue sulphate (NBS) with nucleic acids, including calf thymus DNA, fish sperm DNA and yeast RNA, were characterized with resonance light-scattering (RLS) measurements by using a common spectrofluorometer. Accordingly a method for the determination of nucleic acids at nanogram levels was established. At pH's of 7.20～7.60 and ionic strengths lower than 0.012, the interactions of NBS with nucleic acids result in three characteristic RLS peaks at 293.4 nm, 349.4 nm and 560.4 nm. Mechanism study shows that these peaks are ascribed to the long range assembly of NBS on the molecular surface of nucleic acids, which depends on pH, ionic strength and the stranded structure of nucleic acids. A Scatchard plot was constructed by using the RLS data, yielding the assembly number and assembly constant being 6.4 and 7.13x10⁶ mol^?1 1 for NBS assembly on the molecular surface of calf thymus DNA. The same parameters are 6.6 and 4.58x10⁶ mol^?1 1 for the assembly on that of fish sperm DNA, 3.9 and 1.67x10⁶ mol^?1 1 on that of yeast RNA, respectively. Linear relationships were found between the enhanced RLS intensity at 293.4 nm and nucleic acid concentration. If 1.2x10^?5 mol I^?1 NBS was employed, 0～0.80 μg ml^?1 calf thymus DNA and fish sperm DNA, 0.20～0.60 μg ml^?1 yeast RNA can be determined with the determination limits being 3.2 ng ml^?1 for calf thymus DNA, 11.5 ng ml^?1 for fish sperm DNA and 38.3 ng ml^?1 for yeast RNA, respectively. Four synthetic samples were determined with satisfaction.     

Langmuir Aggregation of Bordeaux R on a Cationic Surfactant and Its Application to Sensitive Quantitative Determination of Copper

The microphase adsorption-spectral correction (MPASC) technique has been applied to the study of the interaction between cetyl trimethylammonium bromide (CTMAB) and the dye Bordeaux R (BR) at pH 9.6. The aggregation of BR on the CTMAB surface obeys the Langmuir adsorption isotherm. The aggregation of BR on CTMAB accelerates the complexation between Cu(II) and BR. Results at 25°C show that the adsorption constant of the CTMAB–BR 1:1 aggregate is 6.80 × 10⁴. In the presence of CMTAB, the Cu–Br complex with a mole ratio of 2:1 has a cumulative stability constant of 1.08 × 10¹¹. The cooperative adsorption and complexation have been applied successfully in a sensitive determination of trace amounts of copper.     

探针体在蛋白质大分子上Langmuir吸附聚集及应用──蛋白质/荧光桃红B(PB)结合反应研究

蛋白质化学是生物化学家当今感兴趣的前沿研究领域. 研究小分子(离子)在生物大分子上的聚集形式以及机理, 帮助人们弄清其污染物及毒物在生物大分子上的结合, 在病理分析、临床检测以及基因变异有重要意义. 常用光谱分析方法包括分光光度法[1～6]、荧光法[7,8]和共振光散射光谱技术(RRS)等[9～12]. 但是, 大分子与探针分子间的结合机理仍存在一些尚未解决的问题[13,14]. 蛋白质分子由于复杂的立体构象形成弱微静电场[15,16], 在其作用下, 带电荷的小分子以单分子层形式被吸附到微相电场表面.     

Langmuir Aggregation of Stain on Ionic Surfactant and its Application: Interaction of Bromo‐Pyrogallol Red with Cetyl Trimethylammonium Bromide (CTAB) and Determination of Molybdenum Traces

The microphase adsorption‐spectral correction (MPASC) technique was de scribed and applied to study the interaction of bromo‐pyrogallol red (BPR) with cetyl trimethylammonium bromide (CTAB). The synergism mechanism of micelles was analyzed and discussed. The large electrostatic micelle adsorbs the chromophore in only the monolayer, and the adsorption obeys the Langmuir adsorption. The formation of the (BPR₂·CTAB)₇₈ aggregate accelerates the Mo‐BPR complex reaction. Experimental results showed the adsorption constant of the adsorption aggregate 6.20 × 10⁵ and its molar absorptivity ?_r^610nm = 1.40 × 10⁶ lmol^?1cm^?1. For analysis of samples, the recovery of Mo was between 97.4 and 107% and the RSD was less than 4.39%.     

Interactions of Night Blue with Nucleic Acids and Determination of Nucleic Acids Using Resonance Light Scattering Technique

The noncovalent interactions of night blue (NB) with several nucleic acids in buffer medium of Britton‐Robinson at pH 4.1 have been studied by spectroscopic methods. It is shown that the binding of NB with nucleic acids involves the J‐aggregation of NB molecules on the surface of nucleic acids. The aggregation was encouraged by polyanions nucleic acids, in which nucleic acids served for acting templates. In this connection, a new method of nucleic acids with sensitivity at nanogram level is proposed based on the measurement of enhanced resonance light scattering (RLS). The linear range of ctDNA, fsDNA and yRNA is 0.01—2.5, 0.03—2.5 and 0.04—1.0 μg/mL, respectively, and the corresponding detection limits (3s?) are 9.4, 7.3 and 5.7 ng/mL at 2.5 × 10–⁵mol/L of NB. Synthetic and real samples were analyzed with satisfactory results.     

langmuir aggregation of alkali blue 6b in proteins: Study and application

The formation of microelectrostatic fields in biopolymers causes the adsorption of stain molecules in protein. The interactions between the dye alkali blue 6B (AB6B) and four proteins, bovine serum albumin (BSA), human γ-globulin (γ-G), horse myoglobin (Mb) and ovalbumin (OVA), were studied by the microphase adsorption/spectral correction technique. It was observed that the interactions obey the Langmuir isothermal adsorption. The Langmuir equilibrium constants of all the aggregates were determined. Interestingly, the adsorption of AB6B has no relation to the array sequence of amino acid residues, and an electrostatic field consists of constant amino acid residues. The novel method has been applied satisfactorily to the determination of the total protein content in two different samples.