In this paper the concentration dependency of mass diffusion coefficients in binary system was investigated. We have developed a novel and accurate visualization system using a small area of transient diffusion fields by adopting a phase shifting technique. Through accurate visualization of the transient diffusion field, it is possible to determine the mass diffusion coefficient. Unlike a conventional interferometer, the proposed system provides high spatial resolution profiles of concent-ration even though the target area is less than 1.0 mm. This allows the measurement of local transient diffusion field with a high accuracy. The determination of mass diffusion coefficient of each component in multi-component system was also conducted. For the accurate and reliable measurement of mass diffusion coefficient, the experimental error should be taken into account. The experimental data usually contains unexpected accidental error and inherent errors of the measurement system. In this study, an optimization technique using conjugate gradient method is developed for the precise determination of the mass diffusion coefficients. The difference between the experimental and numerical concentration distribution is set as the objective function for the optimization method. The conjugate gradient method searches the optimal value by minimizing the objective function. For the concentration dependency evaluation, sodium chloride (NaCl) in pure water was selected as solute. For determination of each mass diffusion coefficient in multi-component system, NaCl and lysozyme in buffer solution was selected. The experiments were performed under isothermal conditions. The proposed measurement method was validated by comparing the measured data with those available in the literature. The results indicated that the concentration dependency was successfully investigated from the experimental data. The mass diffusion coefficient of each component also could be determined from the experimental data as evidenced by good agreement with the published data. The difference between the reference and determined value of mass diffusion coefficient was less than 10%. It can be said that the diffusion of each solute inside the cell progresses independently within the dilute concentration ranges and the superposition principle of concentration of NaCl and lysozyme was satisfied. The influence of concentration of solution on the diffusion process and allowable concentration range of the superposition principle are determined and discussed.