A new simple separation method was proposed for the laboratory-scale separation of asphaltene using columns packed with Teflon-beads. At first, a column packed with Teflon-beads having an inner diameter of 10-30 mm was washed with heptane, and then heavy oil-heptane solution was introduced into the column, followed by eluting remaining maltene using heptane. After the maltene recovery, dichloromethane was introduced from the bottom of the column to elute trapped asphaltene from the top of the column. The maximum asphaltene amount of the column under operational conditions was nearly proportional to the cross-section area of the column, for example, 0.8 g using a column having an inner diameter of 20 mm. The recoveries and H/C atomic ratio of maltenes and asphaltenes were almost the same as those obtained by conventional centrifugation methods, indicating the proposed method was equivalent to centrifugation methods. The proposed method was also useful for the recovery of small amounts of asphaltene such as 50 mg.