Background-free,fast protein staining in sodium dodecyl sulfate polyacrylamide gel using counterion dyes,zincon and ethyl violet

A background-free, fast protein staining method in polyacrylamide gel electrophoresis using an acidic dye, zincon (ZC) and a basic dye, ethyl violet (EV) is described. It is based on the counterion dye staining technique that employs two oppositely charged dyes to form an ion-pair complex in staining solution. The selective binding of free dye molecules to proteins in acidic solution produces bluish violet-colored bands. It is a rapid and end-point staining procedure, involving only fixing and staining steps that are completed in 1-1.5 h. The detection limit of this method is 8-15 ng of protein that is comparable to the sensitivity of the colloidal Coomassie Brilliant Blue G (CBBG) stain. Due to its sensitivity and speed, this stain may be more practical than any other dye-based stains for routine laboratory purposes.     

钴-锌试剂络合吸附伏安法测定中草药中钴

研究了测定痕量钴的一种灵敏的吸附伏安法 ,利用钴和锌试剂络合物在悬汞电极上的吸附性 ,有效地提高了分析的灵敏度。在 0 .0 0 5mol·L-1NH3 和 0 .0 6mol·L-1NH4Cl及 2 .5× 1 0 -6mol·L-1锌试剂底液条件下 ,该体系 1 .5 ,2 .5阶微分吸附伏安法测定钴的线性范围为 1× 1 0 -10 ～ 1× 1 0 -7mol·L-1,用 2 .5阶微分吸附伏安法 ,富集 3min的检出限为 5× 1 0 -11mol·L-1。用此法直接测定了中草药中痕量钴     

测定痕量锰的锰(Ⅱ)-锌试剂-重铬酸钾体系催化分光光度法

在Na2B4O7-NaOH缓冲介质及加热条件下 ,锰 (Ⅱ )对重铬酸钾氧化锌试剂引起褪色 ,有明显的催化作用 ,据此建立了测定痕量锰的新方法 ;该法的线性范围为0.69～40μg/L,检出限为0.22μg/L ,反应表观活化能为87.1kJ/mol,反应表观速率常量1.10×10-3/s,用该法测定蒙药样品中锰含量 ,结果令人满意     

Colorimetric detection of spermine and spermidine by zincon in aqueous solution

The anionic organic dye, zincon, was found to be a colorimetric probe for spermine and spermidine at pH 4.0 in pure aqueous solution through its aggregation by electrostatic interactions with them. The addition of spermine and spermidine led to bathochromic shifts with color changes from yellow to purple and dark brown, respectively, while other amines and basic amino acids induced no or minimal spectral and color changes. From Job’s plot and titration experiments, zincon interacted with spermine (+4 charges) and spermidine (+3 charges) in 4:1 and 3:1 stoichiometric ratios, respectively, indicating that each positive charge was bound to one zincon molecule. The detection limits were determined to be 25.1 for spermine and 30.7 nM for spermidine, which were sufficient below the critical concentration for cancer diagnosis. The results indicated that zincon can be a good candidate for simple, convenient colorimetric detection of spermine and spermidine in aqueous solution.     

Indirect Determination of Trace Copper(II) by Adsorptive Stripping Voltammetry with Zincon at a Carbon Paste Electrode

Traces of copper(II) can be determined by adsorptive stripping voltammetry using 2‐carboxy‐2′‐hydroxy‐5′‐sulfoformazyl benzene (Zincon) as complex forming reagent. First in phosphate buffer pH 6.4, copper(II)‐Zincon complex was adsorbed on carbon paste electrode (CPE) with an accumulation potential of 0.6 V. Following this, adsorbed complex was oxidized and detected by differential pulse voltammetric (DPV) scan from 0.6 to 1.0 V. The effective parameters in sensor response were examined. The detection limit (DL) of copper(II) was 1.1 μg/L and relative standard deviations (RSDs) for 10 and 200 μg/L Cu(II) were 1.81 and 1.03%, respectively. The calibration curve was linear for 2–220 μg/L copper(II). The resulting CPE does not use mercury and therefore, has a positive environmental benefit. The method, which is reasonably sensitive and selective, has been successfully applied to the determination of trace amount of copper in water and human hair samples.     

以锌试剂显色法测定蛋白质的研究

有关蛋白质染色测定的分光光度研究已有不少报道,其中有些已用于蛋白质的分析测定［１～４］．锌试剂常用于金属离子的分光光度法测定,但作为生物大分子分析试剂的研究,目前还未见报道．实验表明,在ｐＨ４左右的缓冲溶液中,锌试剂与ＢＳＡ反应生成有色复合物,吸收光...     

双波长光度法同时测定头发中微量铜和锌

研究了用双波长光度法同时测定头发中微量铜和锌。在pH9．0条件下，以锌试剂为显色剂，与铜、锌络合分别形成稳定的有色络合物．测得铜络合物的最大吸收波长为615nm．锌络合物的最大吸收波长为630nm。采用双波长光度法，选用铜的测定波长对为615／648nm，铜锌合量在波长627nm的等吸光点进行测定。铜含量和铜、锌合量的线性范围都在O～30μg／25ml范围内符合比耳定律。试验结果表明，该方法准确，简便适用，方法回收率在92％～108％之间。用于同时测定头发中微量铜和锌，结果满意。     

锌试剂与牛血清白蛋白相互作用机理

用可见光区分光光度法研究了锌试剂与牛血清白蛋白之间相互反应的机理,在pH 4.2的条件下,随着牛血清白蛋白浓度的逐渐增加,反应体系在锌试剂吸收峰465 nm波长处的吸光度逐渐降低而在新出现的两吸收峰564 nm及600 nm波长处的吸光度逐渐增加.据此得出结论,锌试剂与牛血清白蛋白相互反应生成了复合物,而此复合物的形成乃由于两反应物之间的静电吸引作用及相互之间的疏水作用,而以后者为主导作用.在锌试剂与牛血清白蛋白的反应中与不同浓度的壳聚糖进行了比较试验,其结果也证实了上述解释.对这反应的其他一些影响因素也作了试验并给出了结果.     

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。     

镍,钴与zincon的络合吸附波研究：铜精矿中微量镍,钴的同时测定

有关利用络合物的极谱催化波连续测定镍,钴的报道较少。在研究铜-Zincon的极谱行为时,我们发现NH_4Cl-Na_2SO_3(pH=6.5～7.5)底液中当Zincon浓度>4×10~(-5)·mol/L时,单扫描示波极谱仪上有两个峰,其峰电位分别为-0.42V和-0.56v(vs.