The separation in a batch vacuum column of the essential oil of common juniper berries (Juniperus communis L., from the southern part of Serbia) was analyzed. The main goal of the analyzed separation process was to isolate several fractions from the essential oil which mainly contained α-pinene, sabinene and myrcene. These compounds contain about 65 mass% of the essential oil produced by hydrodistillation from the juniper berries originated from the southern part of Serbia. The results of experimental work in a laboratory column with 36 theoretical stages under vacuum (8.0-3.35 kPa) was simulated using Aspen software, and a proposed mathematical model was used to analyze some other operating conditions for fractionation of juniper berry’s oil (number of plates: 25, 36 and 45 and reflux ratio: 2-10). According to the results of performed simulations, the most acceptable separation procedure which takes into account the prices of raw materials and distillate (α-pinene) as well as consumed energy was proposed.