Studies On The Interactions Between Dye Probe And Protein Or Nucleic Acid

Rosaniline and Chlorophosphonazo I were proposed as probes of protein. Rosaniline and Aurintricarboxylic Acid were proposed as probes of nucleic acid. Results showed that:1. Interaction of Rosaniline with Bovine Serum Albumin and Its Analytical Applications : The formation constant of RL with BSA was obtained at different temperatures. According to the thermodynamic parameters, the main sort of binding force was found to be hydrophobic force. A new method for the quantitative analysis of Bovine serum albumin (BSA) was established by triphenylmethane cationic dyes rosaniline (RL) used as fluorescent probe. The results revealed that there is strong interaction between RL and BSA in the near neutral medium. The determination conditions were optimized. The detection limit of this method is 19.6 μg/L with the linear range of 0.02~5.0 μ g/mL. The method has many advantages such as fastness, high sensitivity and simplicity. It was applied to analysis synthetic and practical samples with satisfactory results. The interaction of RL with BSA was in accord with Pesavento model.2. Fluorimetric Method of DNA with Rosaniline as the Probe and the Study on the Reaction Mechanism : A new method for the quantitative analysis of deoxyribonucleic acid (DNA) was established by using triphenylmethane cationic dyes rosaniline as fluorescent probe. Our results revealed that there was strong interaction between RL and DNA in the near neutral medium. The increase in the fluorescence intensity of the complex was linear with the amount of DNA with the linear range of 0.048~33.0 μ g/mL(r=0.9904). The determination conditions were optimized. The detection limit of this method was 39.0 μ g/L. The method has many advantages such as fast, simple and high sensitivity. It was applied to analyze synthetic and practical samples withsatisfactory results. And we have had a preliminary discussion on relative mechanism, indicating that intercalation model exists between RL and DNA.3. Study of Chlorophosphonazo I fluorescence Quenching by Proteins and Application: At pH9.25 Britton-Robinson buffer solution, the interaction between Chlorophosphonazo I (CP I ) and proteins has been studied by using fluorescence spectroscopy method. The emission spectrum of CP I -BSA shows a bathochromic shift from 578nm to 610nm. The developed method was successfully applied to the determination of protein in sample, the formation constant at different temperatures and the thermodynamic functions(such as AG, AH and AS) of CP I with BSA were obtained, the mechanism of the fluorescence quenching was also explored, it is considered that the quenching is mainly due to generating quenchable complex and that interaction of CP I and proteins was static electricity forces. In addition, the optimum condition of fluorimetric determination of protein was studied. The linear equation was AF=20.8C-6.8 (fig/mL). The detection limit of this method was 0.031 g /mL with the linear range of 0.04~30.0g /mL (r=0.9956) .4. Fluorescence Spectrometry Study on the System of Aurintricarboxylic Acid with DNA : The enhancement of the fluorescence due to the interaction of aurintricarboxylic acid (ATA) with DNA at pH4.04 Britton-Robinson was investigated. It is considered that the interaction between ATA and DNA generates complex, at the same time that the emission spectrum of ATA-BSA show a red shift. The increase of the complex of fluorescence intensity is linear with amount of DNA. The determination conditions were optimized and the linear equations were determined respectively to deoxyribonucleic acid, yeast ribonucleic acid, fish sperm deoxyribonucleic acid and calf thymus deoxyribonucleic acid. A sensitive and simple method of fluorescence was established for the determination of DNA. This method has been applied to determination of DNA in samples with satisfactory results.