As for the measurement of diffusion coefficient in transparent liquids by digital holographic interferometry, there are many kinds depending on the mathematical model and experimental setup. The method of using the distance of the peaks in concentration difference profile to determine diffusion coefficient by Mach-Zehnder interferometric experimental setup, is easy to handle. In order to improve the accuracy and convenience of measurement in this method, we combine procedures of hologram analysis used by Mialdun et al (2011) and those by He et al (2009). By using polynomial to fit the continuous phase difference (interference phase) of object beam, it can be more convenient and accurate to determine the distance between the two peaks. Besides, the shift of initial time has been discussed as a separated topic at the first time and two functions for minimization have been given for determination of the initial time in this paper. With the development, diffusion coefficient of KCl in water at 0.33mol/L and 25 °C has been measured. The diffusion coefficient is 1.844 × 10(-9) m2/s with the uncertainty of ± 0.012 × 10(-9) m2/s. Our measurement has more similar result with other methods than holographic interferometry. The relative uncertainty of diffusion coefficient in our experiment is less than 1% and total uncertainty of temperature is within ± 0.04 K.