Determination Of Trace Amount Of Bovine Serum Albumin By Resonance Light Scattering

Protein is an important constituent of the cell. It plays a vital role in the biologicalphenomena and the life course. The quantitative analysis of protein has a very importantpractical significance in Pharmacology, Biochemistry, Biotechnology, Clinical Medicineand Food Nutrition field, whose research results have guiding significance to explainthe relationship between protein structure and function, and the mechanism of actionbetween protein and substances or protein and drugs.The traditional protein analytical methods mainly have Kjecdahc、Coomassie BlueStaining、Spectrophotometry method、Fluorimetric method、Chemiluminescence、Nearinfrared reflectance spectroscopy、Biuret method and so on. However, most of thesemethods have some limitations including low sensitivity, poor selectivity weak stability,and complex and time-consuming operation. Resonance scattering spectrometry, aburgeoning spectrum analysis technique, has been widely used because of its highsensitivity, good selectivity and user-friendly operation and so on. In this paper,Resonance scattering spectrometry applied to detect trace protein is studied. The papermainly includes:Part one Study on Resonance Light Scattering, Second-order Scattering andFrequency Doubling Scattering Spectral of Acid chrome blue K-Bovine SerumAlbumin-OP systemIn buffer solution of buffer (pH3.2), negatively charged acid chrome blue K(ACBK) could react with positively charged proteins of bovine serum albumin (BSA)by virtue of electrostatic attraction hydrogen bonding and hydrophobic force to formion-association complexes. And the complexes would combine with the non-ionicsurface active agent OP through the intermolecular interactions form the bigger particles.As a result, the intensities of Resonance light scattering (RLS), second-order scattering(SOS) and frequency doubling scattering (FDS) were enhanced greatly and themaximum scattering peak is located at422nm for RLS,680nm for SOS and340nm forFDS respectively. The scattering intensities (ΔIRLS, ΔISOSand ΔIFDS) are directlyproportional to the concentrations of protein in certain ranges. Each method has highsensitivity and the detection limits (3σ) are0.3μg/L (RLS),0.7μg/L (SOS) and0.8μg/L(FDS) μg/L for proteins. These methods can be used for the determination of traceamounts of protein. In this work, the effects of interaction of Acid chrome blue K with proteins on RLS, SOS, FDS spectral characteristics, and the optimum conditions havebeen investigated. Meanwhile, the influences of coexisting substances were tested andthe results show that the method exhibits a good selectivity.Pare two Determination of trace amount of proteins by Resonance lightscattering based on HCl-mebendazole (MEB)-BSA systemA novel method for the determination of trace amount of proteins in blood byresonance light scattering was developed. The HCl-mebendazole (MEB)-BSA systemwas investigated by resonance light scattering(RLS), second-order scattering(SOS) andfrequency doubling scattering(FDS) method. Their maximum wavelengths were locatedat296nm(RLS),680nm(SOS) and340nm(FDS), respectively. The three scatteringintensity (ΔIRLS, ΔISOSand ΔIFDS) were proportional to the concentrations of protein in acertain range. The linear ranges were0.4-2.0mg/L (RLS method),0.4-2.4mg/L (SOSmethod) and0.4-2.0mg/L (FDS method), and their detection limits(3σ) were1.059μg/L(RLS),1.324μg/L (SOS) and4.743μg/L (FDS), respectively. The method is simple,highly sensitive and selective, and was practical for determination of protein contents inserum samples.Part three Investigation and application of Mo(Ⅵ)-Quercetin-BSA system byresonance light scattering and resonance non-linear scatteringIn buffer solution of HAc-NaAc (pH4.5), Mo(Ⅵ) and Quercetin was combinedwith bovine serum albumin (BSA) to form ion-complexes, which resulted in significantenhancement of resonance light scattering (RLS), Second-Order scattering (SOS) andFrequency-Double scattering (FDS). The maximum scattering wavelength is located at489nm for RLS，560nm for SOS and320nm for FDS, respectively. The light scatteringintensity of the system was proportional to the concentration of BSA in certainranges.The linear ranges were0.2-1.0mg/L for RLS method,0.2-0.8mg/L for SOSmethod and0.2-1.0mg/L for FDS method, and the detection limits of BSA were0.207μg/L (for RLS method),0.248μg/L (for SOS method) and0.700μg/L (for FDSmethod), respectively. Based on the enhancement of light scattering，the determinationof BSA by the proposed methods (resonance linear and nonlinear scattering analysismethods) were established. The proposed methods can be applied to the determinationof protein contents in Actual Sample, and the results were satisfactory．Part four Determination of trace amount of proteins by Resonance lightscattering based on Ce3+-SDBS-BSA systemA novel method for the determination of trace amount of proteins by resonance light scattering was developed. The Ce3+-SDBS-BSA system was investigated byresonance light scattering(RLS), second-order scattering(SOS) and frequency doublingscattering(FDS) method. Their maximum wavelengths were located at288nm(RLS),500nm(SOS) and320nm(FDS), respectively. The three scattering intensity (ΔIRLS, ΔISOSand ΔIFDS) were proportional to the concentrations of protein in a certain range. Thelinear ranges were1.0-2.2mg/L (RLS method),1.0-2.6mg/L (SOS method) and1.0-2.6mg/L (FDS method), and their detection limits(3σ) were0.0045mg/L (RLS),0.0258mg/L (SOS) and0.0372mg/L (FDS), respectively. The method is simple, highlysensitive and selective, and was practical for determination of protein contents.Part five Investigation and application of AO-SDS-BSA system by resonancelight scatteringIn buffer solution of Tris-HCl (pH7.2), AO-SDS-BSA to form ion-complexes, Itresulted in significant enhancement of resonance light scattering (RLS). The maximumscattering wavelength is located at309nm for RLS. The light scattering intensity of thesystem was proportional to the concentration of BSA in certain ranges.The linear rangeswas0.4～2.0mg/L, and the detection limits of BSA was1.728μg/L. The reactionconditions and influence factors of the AO-SDS-BSA system were investigated by RLSmethod. The proposed methods can be applied to the determination of protein contentsin Actual sample, and the results were satisfactory．...