深圳大学学报理工版

在pH值为 7.38的磷酸盐缓冲溶液(phosphate buffer saline,PBS)中,采用荧光光谱和紫外-可见吸收光谱,考察联苯胺(benzidine,BZ)与鲱鱼精双链脱氧核糖核酸(double-stranded deoxyribonucleic acid, dsDNA)的相互作用机制,运用离子强度验证结合体系(BZ-dsDNA)荧光光谱的影响和DNA熔点变化两种方法的结合模式.结果表明,BZ以嵌插方式与鲱鱼精dsDNA结合,结合作用力主要是氢键和范德华力,结合反应为自发过程.实验还发现,当dsDNA质量浓度为8.00×10^-6～48.00×10^-6 g/L时,BZ荧光强度F₀和结合体系BZ-dsDNA荧光强度F的比值(F₀/F )与dsDNA质量浓度呈线性关系,dsDNA的检测限为2.00×10^-6 g/L,且其测定不受常见共存金属离子和抗坏血酸等的干扰.因此,基于BZ与dsDNA作用的荧光光谱建立的检测dsDNA的新方法,可用于检测实验室废水中的dsDNA,平均回收率在98.00%～103.00%.

The interaction mechanism of benzidine(BZ)with herring fine dsDNA in PBS(pH=7.38)is investigated by the fluorescence spectroscopy and the UV-visible absorption spectroscopy. Their binding mode is verified by two methods. One examines the effect of the ionic strength on the fluorescence intensity of the binding system BZ-dsDNA. The other is an DNA melting experiment. The results show that BZ combines with dsDNA in the mode of intercalation mainly by hydrogen bonding and van der Waals forces, and the interaction reaction is spontaneous. In addition, the ratio of fluorescence intensity of BZ(F₀)to BZ-dsDNA(F)is linearly proportional to the concentration of dsDNA in the range of 8.00×10^-6–48.00×10^-6 g/L with a detection limit of 2.00×10^-6 g/L. The common co-existed metal ions and ascorbic acid do not interfere with the determination. A new fluorescence spectroscopy method for the determination of dsDNA is developed based on the interaction between dsDNA and BZ. This method is applied into determining dsDNA in waste water from the lab which achieves the average recovery between 98.00% and 103.00%.

引言
1 实验
2 结果与讨论
3 结语

图1 加入不同质量浓度dsDNA 的联苯胺荧光光谱曲线<br/>(T=298 K, c(BZ)=5.50×10-5 mol/L)<br/>Fig.1 Fluorescence spectra of BZ in the presence of dsDNA with different mass concentrations

图1 加入不同质量浓度dsDNA 的联苯胺荧光光谱曲线
(T=298 K, c(BZ)=5.50×10-5 mol/L)
Fig.1 Fluorescence spectra of BZ in the presence of dsDNA with different mass concentrations

表1 不同温度下 BZ-dsDNA的 Stern-Volmer方程<br/>Table 1 Stern-Volmer equations of BZ-dsDNA at different temperatures

表1 不同温度下 BZ-dsDNA的 Stern-Volmer方程
Table 1 Stern-Volmer equations of BZ-dsDNA at different temperatures

表2 BZ-dsDNA不同温度下的静态猝灭方程1)<br/>Table 2 Static quenching equations of BZ-dsDNA at different temperatures1)

表2 BZ-dsDNA不同温度下的静态猝灭方程1)
Table 2 Static quenching equations of BZ-dsDNA at different temperatures1)

表3 BZ-dsDNA不同温度下的热力学参数 <br/>Table 3 Thermodynamic parameters of BZ-dsDNA at different temperatures

表3 BZ-dsDNA不同温度下的热力学参数
Table 3 Thermodynamic parameters of BZ-dsDNA at different temperatures

图2 不同质量浓度dsDNA存在下的<br/>联苯胺紫外可见光谱曲线<br/>(T=298 K, c(BZ)= 5.50×10-5 mol/L)<br/>Fig.2 UV-vis spectra of BZ in the presence of dsDNA with different mass concentrations

图2 不同质量浓度dsDNA存在下的
联苯胺紫外可见光谱曲线
(T=298 K, c(BZ)= 5.50×10-5 mol/L)
Fig.2 UV-vis spectra of BZ in the presence of dsDNA with different mass concentrations

图3 联苯胺结构式<br/>Fig.3 Structural of benzidine

图3 联苯胺结构式
Fig.3 Structural of benzidine

图4 氯化钠浓度对两体系荧光强度比的影响<br/>(T=298 K, c(BZ)= 5.50×10-5 mol/L, <br/>ρ(dsDNA)=12.00×10-6 g/L)<br/>Fig.4 The effect of NaCl concentrations on fluorescence intensity ratio of two systems

图4 氯化钠浓度对两体系荧光强度比的影响
(T=298 K, c(BZ)= 5.50×10-5 mol/L,
ρ(dsDNA)=12.00×10-6 g/L)
Fig.4 The effect of NaCl concentrations on fluorescence intensity ratio of two systems

图5 dsDNA在有无联苯胺添加时的熔点曲线<br/>(c(BZ)=5.50×10-5 mol/L, ρ(dsDNA)=12.00×10-6 g/L)<br/>Fig.5 Melting curves of dsDNA in the presence and absence of BZ

图5 dsDNA在有无联苯胺添加时的熔点曲线
(c(BZ)=5.50×10-5 mol/L, ρ(dsDNA)=12.00×10-6 g/L)
Fig.5 Melting curves of dsDNA in the presence and absence of BZ

表4 干扰倍数<br/>Table 4 Interference of foreign substances

表4 干扰倍数
Table 4 Interference of foreign substances

表5 样品分析结果(n=6)<br/>Table 5 Analytical results of samples(n=6)

表5 样品分析结果(n=6)
Table 5 Analytical results of samples(n=6)

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备注

引言

1 实验

2 结果与讨论

3 结语

期刊信息

备注

引言

1 实 验

2 结果与讨论

3 结 语

期刊信息

1 实验

3 结语