| Chapter one: The application in determination of proteins by spectrophotometric, fluorospectrophotometric and resonance light scattering methods were reviewed with 86 references.Chapter two: The spectrum of the Acid Brown SR(ASR) and the interaction of ASR with bovine serum albumin(BSA) was investigated by spectrophotometric method. In acid aqueous solution, ASR can interact with BSA by electrostatic force. However the production is precipitation, and the structure of precipitation is unstable. Surfactants can greatly influence the interaction of BSA with ASR. Gumwater can make the precipitation change into soluble substance which structrue is stable. Based on this fact, ASR can be used as a new and cheap dye for quantitative analysis of protein.Chapter three: A new method for the determination of albumin in human serum and mouse serum has been developed by spectrophotometry coupled with acid brown SR(ASR) as probe molecule. The maximum absorption wavelength of ASR under acidic condition was at 445nm, while maximum absorption wavelength of their product was at 610nm. The bathochromic shift of 165nm was sufficient to be regarded the left ASR in the system, which had not reacted with albumin, did not interfere with the assay of albumin. However, the reaction between ASR and albumin such as BSA or HSA was so strong that parts of their product were undissoluble in water. The addition of gumwater into the system effectively eliminated the deposition. Under the optimum reaction conditions, the ranges of working lines for BSA and HSA were 0-91.00mg/L and 0-95.20mg/L, respectively. The detection limits were 5.72mg/Lfor BSA and S.15mg/L for HSA, respectively. The relatively standard derivation and the recovery of the method for the determination of total proteins in 6 human serum samples were 1.84%-4.41% and 93.47%-109.18%, respectively. The proposed method has been employed in the assay of protein of human serum and mouse serum. The results of this work were in agreement with those by Biuret method.Chapter four: A method of determination of contents of proteins in human serum was firstly developed by means of the resonance light scattering (RLS) produced by the interaction of proteins and tri-zao dye acid brown SR (ASR). In acidic aqueous medium, the weak RLS of ASR was greatly enhanced by the addition of proteins, and this enhancement was linear with the concentration of protein. Based on this, a novel quantitative method was developed for the determination of proteins in aqueous solution. Under optimal conditions (Casr=2.40X 10-6mol/L; pH=2.48; Britton-Robinson buffer volume 1.5mL), the linear ranges of calibration curves for the determination of bovine serum albumin (BSA), human serum albumin (HSA) and egg albumin (Alb) were 0-1.13mg/L, 0-1.17mg/L and 0-1.13mg/L, respectively. The detection limits were 15.74礸/L for BSA, 23.34礸/L for HSA and 20.75礸/L for Alb. The method was applied to analysis of total protein in human serum samples collected from the hospital and the results were in good agreement with those reported by a local hospital.Chapter five: In acidic Britton-Robinson solution, acid brown NR (ANR) could combine with albumin such as bovine serum albumin (BSA) or human serum albumin (HSA), and the reaction between ANR and albumin was so strong that their products were undissoluble in water. This led to the reduction of ANR absorbance. Based on this, a new method for determination of albumin has been developed by spectrophotometry. Under the optimum reaction conditions, ?BSA=1.4?106L/mol-cm, HSA=l3 X 10 L/mol-cm, the ranges of working lines for BSA and HSA were 27.95-111.80mg/L and 24.40?22.00mg/L, respectively. The relatively standard deviation and the: recovery of the method for the determination of total proteins in 7 human serum samples were 0.83%-3.02% and 90.90% -109.80%, respectively. And the results of this work were in agreement with those by Biuret method.Chapter six: The interaction of acid brown SG (ASG) with serum albumin has been studied by resonance light scattering (RLS) method.
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