Study on spectroscopic characterization of Pd porphyrin and its interaction with ctDNA

The fluorescence and solid substrate room temperature phosphorescence (SS-RTP) properties of Pd(II) meso-tetrakis (4-N-methyl-pyridiniumyl) porphyrin (Pd(II)TMPyP) were studied. The factors influencing the SS-RTP emission, such as filter type, inorganic salt sort, drying temperature, pre-drying time and drying time were investigated in detail. Strong SS-RTP signal can be induced on the slow speed filter paper in the presence of the external inorganic salt, Ca(NO₃)₂, with the maximum excitation and emission wavelengths at 421 nm and 675 nm, respectively. The interaction between calf thymus DNA (ctDNA) and Pd(II)TMPyP was investigated at pH 7.2 using SS-RTP, fluorescence and UV-vis spectroscopy. The SS-RTP intensity of Pd(II)TMPyP was enhanced efficiently with the increasing amount of ctDNA. This phenomenon demonstrates that the intercalated porphyrin is shielded by ctDNA to avoid collision quenching. This result was supported by SS-RTP lifetime measurement, SS-RTP anion quenching experiment and fluorescence polarization measurement. Furthermore, with the addition of ctDNA, the UV-vis spectra of Pd(II)TMPyP shows apparent hypochromicity (40%) at the Soret maximum of 417 nm and a red shift of Δλ = 15 nm, also indicating that Pd(II)TMPyP intercalates into ctDNA bases. The binding constant of Pd(II)TMPyP to ctDNA was calculated to be 4.41 × 10⁵ L/mol based on the derivative McGhee-von Hippel plots.     

Study on the Interaction of Pd-TMPyP with ctDNA by Solid-substrate Room Temperature Phosphorescence

Spectral analysis methods have been employed to investigate the interactions ofporphyrins with DNA including absorption, fluorescence and circular dichroism.Phosphorescence is another excellent method used in this field for its longer emissionwavelength a…     

Synchronous fluorescence determination of DNA based on the interaction between methylene blue and DNA

The fluorescence intensity of methylene blue (MB) quenched by DNA in the pH range of 6.5-8.0 was studied with synchronous fluorescence technology. A novel method for detecting single-stranded and double-stranded DNA was developed. The decreased fluorescence intensity at 664 nm is in proportion to the concentration of DNA in the range of 0.28-11.0 μmol L⁻¹ for ctDNA, 0.14-8.25 μmol L⁻¹ for thermally denatured ctDNA and 0.28-8.25 μmol L⁻¹ for hsDNA. The detection limits (S/N = 3) are 0.11, 0.04 and 0.04 μmol L⁻¹, respectively. The method is rapid, selective, and the reagents are lower toxic. It has been used for the determination of DNA in synthetic samples with good satisfaction. In addition, the interaction modes between MB and ctDNA and the mechanism of the fluorescence quenching were also discussed in detail. The experimental results from absorption spectra and fluorescence polarization indicate that the possible interaction modes between MB and DNA are the electrostatic binding and the intercalation binding.     

Luminescence Studies on Interaction of an Isoquinoline Alkaloid with Calf Thymus DNA in Aqueous Medium and on Solid Substrate

The interaction of calf thymus DNA(ctDNA) with berberine(Ber) was systematically investigated via UV-Vis, fluorescence and phosphorescence spectroscopies in aqueous solution and on solid substrate. The very weak fluorescence intensity of Ber at 525 nm in aqueous solution can be greatly enhanced by ctDNA. The UV-Vis spectrum shows that Ber could bind with DNA by intercalation. In addition, strong fluorescence of Ber alone could be observed on filter paper substrate and can be quenched by ctDNA effectively, and the increase of fluorescence polarization of Ber on the filter paper implies the intercalation binding. Ber emits phosphorescence at 619 nm in the presence of thallium(I) acetate. The lifetime of Ber increased from 38.4 ms to 43.4 ms with the increase of ctDNA. The increase of lifetime is possibly attributed to the increase of rigidity of Ber after its intercalating into the ctDNA base pairs. This explanation is also augmented by the anionic quenching results. Ber behaves as a typical intercalating agent into ctDNA.The intrinsic binding constant between Ber and ctDNA is (1.84?0.12)?10⁴ L/mol and n=0.16.     

Conjugation of organic-metallic hybrid polymers and calf-thymus DNA

Strong electrostatic interaction between metallo-supramolecular polymers and DNA was confirmed by UV-vis and CD spectral measurements during titration, and cyclic voltammetry. The stable conjugation structure based on groove binding was revealed by using QM/MM computational methodology and supported by AFM.     

Enoxacin-Tb complex as an environmentally friendly fluorescence probe for DNA and its application

The fluorescence intensity of the enoxacin (ENX)-Tb³⁺ complex enhanced by DNA was studied. On the basis of this study, an environmentally friendly fluorescence probe of enoxacin-Tb³⁺ for the determination of single-stranded and double-stranded DNA was developed. Under the optimal conditions, the enhanced fluorescence intensity was in proportion to the concentration of DNA in the range of 2.0 × 10⁻⁸ to 2.0 × 10⁻⁶ g mL⁻¹ for hsDNA, 1.0 × 10⁻⁸ to 1.0 × 10⁻⁶ g mL⁻¹ for ctDNA and 5.0 × 10⁻⁹ to 1.0 × 10⁻⁶ g mL⁻¹ for thermally denatured ctDNA. The detection limits (S/N = 3) were 5.0, 9.0 and 3.0 ng mL⁻¹, respectively. The interaction modes between ENX-Tb³⁺ and DNA and the mechanism of the fluorescence enhancement were also discussed in details. The experimental results from UV absorption spectra, fluorescence spectra and the competing combination tests between the ENX-Tb³⁺ complex and EB probe indicated that the possible interaction modes between enoxacin-Tb³⁺ complex and DNA had at least two different binding modes: the electrostatic binding and the intercalation binding. Additionally, this fluorescence probe was used to study the interaction between heavy metals and DNA.     

S-异丙甲草胺与小牛胸腺DNA的相互作用

应用紫外光谱、荧光光谱、DNA热变性法以及黏度法研究了S-异丙甲草胺与小牛胸腺DNA(ctDNA)的相互作用. 结果表明, S-异丙甲草胺使ctDNA在200 nm处的吸收峰发生明显改变, 表现出红移和减色效应, 而对260 nm处的吸收峰产生影响较小, 排除了嵌插作用的可能; ctDNA对S-异丙甲草胺内源性荧光表现出很强的猝灭作用, 且随温度的升高, 其猝灭程度有所下降, 表明S-异丙甲草胺是以形成加合物的方式与ctDNA结合的, 并求得了它们在不同温度下的结合常数; 将不同离子强度条件下S-异丙甲草胺与ctDNA作用以及不同S-异丙甲草胺浓度下ctDNA的热变性温度和黏度变化的研究结果与紫外光谱和荧光光谱相结合, 可以判断S-异丙甲草胺是以沟槽作用的方式与ctDNA结合的.     

Study on spectroscopic characterization of Cu porphyrin/Co porphyrin and their interactions with ctDNA

The luminescence behaviors of Cu(II) meso-tetrakis (4-N-methyl-pyridiniumyl) porphyrin (Cu(II)TMPyP) and Co(II) meso-tetrakis (4-N-methyl-pyridiniumyl) porphyrin (Co(II)TMPyP) are investigated, and their interactions with calf thymus DNA (ctDNA) are studied by UV-vis, fluorescence and resonance light scattering (RLS) and based on the changes of UV-vis spectra, fluorescence and RLS spectra, the intrinsic binding constants of Cu(II)TMPyP/Co(II)TMPyP with ctDNA are obtained in the case of phosphate buffer solution (pH 7.2), respectively. According to the experimental results, it can be inferred that the interaction model of Cu(II)TMPyP with ctDNA is intercalative binding, while Co(II)TMPyP is the long-range assembly on the molecular surface of ctDNA.     

Determination of nucleic acid based on increased resonance light-scattering of fluorinated surfactants

Two novel surfactants perfluoroalkanesulfonyl quaternary ammonium iodides (FC134) and potassium perfluorooctanesulfonate (FC95) were successfully used as new probes for detection of DNA by resonance light-scattering (RLS) technique. Resonance light-scattering characteristics of the binding of fluorinated surfactants FC134 and FC95 to calf thymus nucleic acid (ctDNA) were studied. After DNA was added, aggregation of FC134 on the molecular surface of DNA in the pH 3.0-6.0 and aggregation of FC95 on the surface of DNA in the pH 3.5-6.0 occurred, both of which resulted in an enhanced resonance light-scattering peak at 370 nm. The intensity of resonance light-scattering was found to be proportional to the concentration of DNA. The determination limits were 3.5 and 20.0 μg L⁻¹, respectively. UV-vis spectra and IR-spectra both proved the binding of fluorinated surfactants to DNA.     

Synthesis and investigation on the interaction with calf thymus deoxyribonucleic acid of a novel fluorescent probe 7-oxobenzo[b][1,10]phenanthroline-12(7H)-sulfonic acid

In this paper, the synthetic route of a potential antitumor reagent, benzo[b][1,10] phenanthrolin-7(12H)-one (BPO), was improved. A sulfonic group was introduced to BPO to form a new compound, 7-oxobenzo[b][1,10]phenan-throline-12(7H)-sulfonic acid (OPSA), in order to enhance its water-solubility. The molecular structure of OPSA has been confirmed by IR, UV, MS, ¹H NMR and elements analysis. It was proved in our experiments that DNA could quench the fluorescence of OPSA and the maximum quenched intensity appeared at 408 nm (λ_ex = 284 nm). The quenched fluorescence intensity was proportional to the concentration of DNA. Based on this phenomenon, OPSA had been used as the fluorescent probe for detection of calf thymus DNA (ct-DNA) and the corresponding linear response range was from 1.0 to 150.0 μg mL⁻¹ and the limit of detection (LOD) was 3.8 ng mL⁻¹. Its interaction with ct-DNA was investigated by fluorescence, absorption and viscosity measurements. When binding to ct-DNA, OPSA showed obvious fluorescence quenching and the quenched intensity was stable with the presence and absence of NaCl. The absorption spectra of OPSA had no evidence of increasing or decreasing when ct-DNA was added. The viscosity of OPSA and ct-DNA mixture showed no obvious change comparing with the viscosity of ct-DNA along. The results suggested that the interaction between OPSA and ct-DNA was groove binding in nature. Scatchard plots constructed from fluorescence titration data gave a binding constant of 8.9 × 10⁵ L mol⁻¹ and a binding site size of 0.35 base pairs per bound drug molecule.     

Solid-substrate room-temperature phosphorescence study on zinc(II) and tin(IV) protoporphyrins and their interaction with DNA

The solid-substrate room-temperature phosphorescence (SS-RTP) of two commercially available metalloporphyrin compounds, zinc(II) protoporphyrin (ZnPP) and tin(IV) protoporphyrin (SnPP) has been studied. Strong and stable RTP signals of the two metalloporphyrins in neutral to weakly basic solutions can be simply induced on filter paper without addition of external heavy atom perturbers. Their emission bands appeared at 723 nm for ZnPP and 718 nm for SnPP at an excitation wavelength of 417 nm. Compared with SnPP, ZnPP is a better RTP probe for DNA because its RTP enhancement effect is much higher under the same experimental conditions. The interaction of ZnPP with DNA at pH 8.5 gives an apparent binding constant of 9.1 x 10(3) which is similar to that of the cationic porphyrin absorption probe CuTMPyP (copper (II)- tetrakis(4-N-methylpyridyl)porphine complex). Hydrogen bonding between the monocarboxylic acid substituent of ZnPP and the base pairs of DNA plays a crucial role in the binding.     

抗癌药物4'-O-(α-L-夹竹桃糖基)柔红霉素与小牛胸腺DNA相互作用的荧光光谱法

在pH=7.4的Tris-HCl介质中,利用荧光光谱和紫外吸收光谱法,研究了一种新型蒽环类抗癌药物柔红霉素衍生物(4′-O-(α-L-夹竹桃糖基)柔红霉素,ODNR)与小牛胸腺DNA(ctDNA)的相互作用。 通过离子强度的影响、KI荧光猝灭实验和单双链ctDNA作用的比较实验,分析了ODNR与ctDNA的相互作用模式。 结果表明,ODNR通过嵌插方式与ctDNA发生作用。 ctDNA对ODNR的荧光有明显的猝灭作用,其机理属于静态猝灭。 通过Scatchard方程求得不同温度下的结合常数和结合位点数,由热力学参数确定分子间作用力为疏水作用,也可能存在静电作用。     

Study binding of Al-curcumin complex to ds-DNA, monitoring by multispectroscopic and voltammetric techniques

In this work a complex of Al3+ with curcumin ([Al(curcumin) (EtOH)2](NO3)2) was synthesized and characterized by UV-vis, FT-IR, elemental analysis and spectrophotometric titration techniques. The mole ratio plot revealed a 1:1 complex between Al3+ and curcumin in solution. For binding studies of this complex to calf thymus-DNA various methods such as: UV-vis, fluorescence, circular dichroism (CD), FT-IR spectroscopy and cyclic voltammetry were used. The intrinsic binding constant of ACC with DNA at 25°C was calculated by UV-vis and cyclic voltammetry as 2.1×10(4) and 2.6×10(4), respectively. The thermodynamic studies showed that the reaction is enthalpy and entropy favored. The CD results showed that only the Δ-ACC interacts with DNA and the Δ-ACC form has not any tendency to interact with DNA, also the pure curcumin has not any stereoselective interaction with CT-DNA. Fluorimetric studies showed that fluorescence enhancement was initiated by a static process in the ground state. The cyclic voltammetry showed that ACC interact with DNA with a binding site size of 2. From the FT-IR we concluded that the Δ-ACC interacts with DNA via partial electrostatic and minor groove binding. In comparison with previous works it was concluded that curcumin significantly reduced the affinity of Al3+ to the DNA.     

Solid-substrate room-temperature phosphorescence study on zinc(II) and tin(IV) protoporphyrins and their interaction with DNA

The solid-substrate room-temperature phosphorescence (SS-RTP) of two commercially available metalloporphyrin compounds, zinc(II) protoporphyrin (ZnPP) and tin(IV) protoporphyrin (SnPP) has been studied. Strong and stable RTP signals of the two metalloporphyrins in neutral to weakly basic solutions can be simply induced on filter paper without addition of external heavy atom perturbers. Their emission bands appeared at 723 nm for ZnPP and 718 nm for SnPP at an excitation wavelength of 417 nm. Compared with SnPP, ZnPP is a better RTP probe for DNA because its RTP enhancement effect is much higher under the same experimental conditions. The interaction of ZnPP with DNA at pH 8.5 gives an apparent binding constant of 9.1 × 10³ which is similar to that of the cationic porphyrin absorption probe CuTMPyP (copper (II)- tetrakis(4-N-methylpyridyl)porphine complex). Hydrogen bonding between the monocarboxylic acid substituent of ZnPP and the base pairs of DNA plays a crucial role in the binding. Received: 21 December 2000 / Revised: 9 April 2001 / Accepted: 20 April 2001     

时间分辨光谱技术研究亚甲基蓝与小牛胸腺DNA的相互作用

采用时间分辨光谱技术研究了亚甲蓝与小牛胸腺DNA（MB-ctDNA）重水混合体系中MB敏化单态氧（1O2）动力学过程,以此进一步探讨MB与DNA的相互作用。结果表明显示,低浓度ctDNA和高浓度ctDNA的单态氧磷光动力学曲线有着明显不同,这些差异被归结为MB与DNA间结合方式和作用机制的改变。在低浓度ctDNA条件下,MB分子和ctDNA之间形成离子型结合物,MB的吸收带出现显著的减色效应,敏化1O2产量随DNA浓度增加而急剧下降,但ctDNA与1O2没有发生明显的相互作用;而在高浓度DNA时,MB分子和ctDNA之间的相互作用方式转变为以嵌入式结合为主,激发态MB与ctDNA间的能量转移和介质的粘度效应,改变了1O2的动力学特性,大大降低了光敏剂MB敏化1O2的产量,但1O2不为ctDNA所猝灭。这些结果阐明,在MB与DNA的混合体系中,敏化单态氧损坏DNA的Ⅱ型反应不是主要的PDT作用机制。     

Electrochemical study on oxidation of ofloxacin at nano-ZnS/poly (Styrene sulfonic acid sodium salt) modified electrode and its interaction with calf thymus DNA

A PSS/nano-ZnS thin film modified electrode has been fabricated on the surface of glassy carbon electrode (GCE). In pH 5.0 phosphate buffer solution (PBS), Ofloxacin (OFX) appeared as an anodic peak with the peak potential of 1.28 V at PSS/nano-ZnS film modified electrode. In comparison with a bare GCE or a nano-ZnS modified electrode, the PSS/nano-ZnS film modified electrode exhibited an enhanced effectiveness for the oxidation of OFX. Cyclic voltammetry (CV) coupled with fluorescence and UV-vis absorbance spectra techniques were used to study the interaction of OFX with Calf thymus DNA (ctDNA). The interaction of OFX and ctDNA could result in a considerable decrease in the peak currents and positively shift in the peak potential, as well as changes of fluorescence, UV-Vis adsorption spectra. All acquired data showed that the new adduct between OFX and ctDNA was formed. Published in Russian in Elektrokhimiya, 2009, Vol. 45, No. 3, pp. 289–295. The article is published in the original.     

Absorption,Fluorescence and Resonance Rayleigh Scattering Spectra of Interaction of Papain with Calf Thymus DNA

In weak acidic medium, interaction between papain and calf thymus DNA (ctDNA) resulted in absorption spectral change, fluorescence quenching of papain and remarkable enhancement of resonance Rayleigh scattering (RRS). The interaction types and binding modes were discussed by characteristics of RRS, absorption, fluorescence and circular dichroism spectra combining thermodynamic data. Four interaction types include electrostatic attraction, hydrophobic force, hydrogen bonding and aromatic stacking interaction. Papain interacted with the major groove of ctDNA. Aromatic stacking interaction is the main reason of change of absorption spectrum and fluorescence quenching of papain. Surface enhanced scattering effect, resonance energy transfer effect, increase of molecular volume and conformational change make contribution to RRS enhancement. The enhanced RRS intensity (ΔI) is directly proportional to the concentration of ctDNA or papain. The detection limit (3σ) is 5.2 ng·mL^?1 for ctDNA and 5.6 ng·mL^?1 for papain. This creates conditions for determination of papain and ctDNA.     

Solid substrate-room temperature phosphorimetry for the determination of residual clenbuterol hydrochloride based on the catalysis of sodium periodate oxidizing eosine Y

Clenbuterol hydrochloride (CLB) could catalyze NaIO₄ oxidation of eosine Y (R), which caused the room temperature phosphorescence (RTP) signal of R to quench sharply. The ΔI_P (=I_P2 − I_P1, I_P2 was RTP intensities of reagent blank and I_P1 was RTP intensities of test solution) of the system was directly proportional to the content of CLB. According to that academic thought, a new solid substrate-room temperature phosphorimetry (SS-RTP) for the determination of trace CLB has been established. This method has high sensitivity (detection limit (LD): 0.021 zg spot⁻¹, corresponding concentration: 5.2 × 10⁻²⁰ g mL⁻¹) and good selectivity (Er = ±5%, interfering species were of no interference). It has been applied to the determination of residual CLB in the practical samples. The results were verified using HPLC and GC/MS methods. The reaction mechanism of catalytic SS-RTP for the determination of residual CLB was also discussed.     

一种新型发光探针与DNA相互作用的研究

本文以稳态荧光光谱、紫外-可见吸收光谱、荧光偏振、热变性、阴离子猝灭等手段,研究了一种具有强电荷转移能力的化合物2,7-二[(N-乙基咔唑-3-基)丙烯-1-酮基]芴与DNA的相互作用。研究结果表明,2,7-二[(N-乙基咔唑-3-基)丙烯-1-酮基)芴与DNA的作用方式是混合模式,以嵌插作用为主,同时存在沟槽相互作用,其咔唑基团可能插入到DNA的碱基对之间,结合常数K为8 123.48mol/L,结合位点n为0.71。该发光探针灵敏度高,结合稳定。     

三种不同元素掺杂的碳点与DNA相互作用及其机制研究

碳点(carbon dots,CDs)在材料学及生物医学相关领域的研究已引起科研人员的广泛关注.本文采用一步水热法,制备了三种具有不同元素掺杂的CDs、N-CDs、N,S-CDs,并结合多种光谱法、熔链温度法和黏度法,系统地研究了三种不同元素掺杂的碳点与ctDNA结合模式以及结合能力的差异.荧光实验证明三种不同性质的碳...