The performance of the rectifying column affects the coefficient of performance (COP) of an ammonia absorption refrigerator. Ammonia mass fraction of refrigerant is one of the key parameters indicating the performance of the rectifying column. We propose a method to estimate the refrigerant mass fraction with mass and energy balance equations around the separator at the inlet of the evaporator, and describe the results of experiments which measured the refrigerant mass fraction by sampling from the refrigerant receiver. Throughout these investigations, the refrigerant mass fraction turned out to be lower than the expected value calculated from the condition of dry-saturated vapor at the top of the rectifying column. The refrigerant mass fraction can be estimated within an accuracy of 0.3% by the estimation method based on mass and energy balance equations.