Application Of Analytical Detection Resonance Light Scattering Techniques In Protein

Resonance light scattering technique is a new detecting technology that originated in1990s,because of its simple operation, high sensitivity and celerity, spectrum has drawed the attentionof the broad analytical workers. Now the technology is applied in material science, astronomy,life science, environmental science, analytical chemistry and other fields of analysis detection.Protein is the basis of life. serum protein plays a transport, immune, buffer and nutritionaleffects in vivo, it also can maintain the osmotic pressure. Therefore, the accurate detection ofserum protein content has important implications for disease diagnosis in biomedical science.Study on the mechanism of small molecules and serum protein has a certain guiding significanceto reveal serum protein transport mechanism and study the transport and efficacy of drugs.The work of this paper mainly included two aspects: first, it set up three new ways todetect the serum albumin; Second, it studied mechanism of action of two kinds of smallmolecules and bovine serum albumin. The research content of this paper was as follows:（1） Three new methods for BSA detection of the malachite green and Zn （Ⅱ）, Congo redand Cu （II） and were established, the enhancement signals of resonance light scattering werelocated at372nm,532nm and340nm. The linear equations were the I_RLS=20.3+0.82ρ（μg/mL）,I_RLS=145.8+210.3ρ（μg/mL） and I_RLS=1187.9+32.16ρ（μg/mL） respectively. The linear ranges ofmethod were0.1²5g/mL,0².0g/mL and1.0⁶0.0g/mL respectively. The detection limitwere0.06g/mL,0.005g/mL and0.15g/mL respectively. The new method of RLS detection ofbovine serum albumin was established, three new detection methods were applied in compositesample standard addition recovery experiments, RSD value was less than3.0%. The result wassatisfactory.（2）Using fluorescence spectroscopy, synchronous fluorescence spectroscopy and ultravioletspectroscopy respectively studied the drug molecule ceftriaxone sodium （CS）, sodiumtetraiodofluorescein （FSS） and BSA interactions. Results showed that the fluorescencequenching of CS on BSA was static quenching and the variation in thermodynamic parametersshowed that the acting force between them was mainly electrostatic force, hydrophobicinteraction force may still exist. Synchronous fluorescence spectra showed that a binding site ofCS and BSA should be near the tryptophan of BSA, and the CS binding resulted in the BSAconformational changes. According to the dipole-dipole non radiation energy transfer theory, the distance between CS and BSA was calculated4.61nm, CS made BSA fluorescence quenching,which belonged to non radiation energy transfer. Fluorescence quenching of TSS on BSA wasstatic quenching, the variation in thermodynamic parameters showed that Δ H<0、 ΔS>0thereaction between them was the electrostatic force. Synchronous fluorescence spectrum showedthat the binding sites of TSS and BSA should be near the tryptophan of BSA, and thecombination of TSS made conformation changes of BSA. According to the dipole-dipole nonradiation energy transfer theory, the distance between TSS and BSA was calculated to be2.95nm, TSS made BSA fluorescence quenching which belonged to non radiation energy transfer.