The Pharmacy Education Eligibility Test (PEET) is required by most pharmacy schools for ascertaining students aptitude and skills. The aim of this study was to assess the opportunity costs for the PEET preparation based on self-report survey. The questionnaire was developed and finally verified by experts in pharmacy and education. Opportunity costs were analyzed by summing the measurement of time, as indirect costs, and direct costs required to prepare the exam. A total of 392 students from 9 universities responded to the survey, of which 245 students were included the analysis of opportunity costs for the PEET preparation. A total of 392 students from 9 universities responded to the survey, of which 245 students responded to it, including opportunity cost analysis according to PEET preparation. Most of the respondents (92.60%) were very high dependent on private education. Tuition fee for subject classes and housing expenses accounted for the largest portion of direct cost. The average period was 17.76 months, and the preparation period was significantly longer with older age and rural living. Indirect costs were calculated as the minimum amount of activity during the preparation period. Finally, the average opportunity costs were 55,057 South Korean thousand won and statistically significant as the age were higher. However, it was not related to living region, major before pharmacy school entrance, time of preparation start, household income, and economic support.