Today, large-volume mobile phone call datasets are widely applied to investigate the spatio-temporal characteristics of human urban activity. This paper discusses several fundamental issues in estimating population distributions based on mobile call data. By adopting an individual-based call activity dataset that consists of nearly two million mobile subscribers who made over one hundred million communications over seven consecutive days, we explore the relationships among the Erlang values, the number of calls, and the number of active mobile subscribers. Then, the LandScan population density dataset is introduced to evaluate the process of estimating the population. The empirical findings indicate that: (1) Temporal variation exists in the relation between the Erlang values and the number of calls; (2) The number of calls is linearly proportional to the number of active mobile subscribers; (3) The proportion between the mobile subscribers and the actual total population varies in different areas, thus failing to represent the underlying population. Hence, the call activity reflects “activity intensity” rather than population distribution. The Erlang is a defective indicator of population distribution, whereas the number of calls serves as a better measure. This research provides an explicit clarification with respect to using call activity data for estimating population distribution.