Provided are a microfluidic apparatus and a method for separating a target cell using the same. The microfluidic apparatus for separating a first material in a biological sample from the biological sample, according to an embodiment of the disclosure, includes: a body rotatable on a rotation axis; a mixing chamber included in the body, wherein the biological sample and magnetic beads that are combined with a second material in the biological sample are mixed in the mixing chamber; a separation chamber included in the body and connected to the mixing chamber, wherein a mixed sample of the first material and the second material combined with the magnetic beads is separated in the separation chamber; and a magnetic member positioned on one side of the body outside the separation chamber, wherein, in the separation chamber, the first material is separated from the second material combined with the magnetic beads by a centrifugal force and a magnetic force.