A automated test tube cap removal apparatus includes, a cap remover, a test tube rack which holds a plurality of capped test tubes upright in a row, a clamping mechanism which is disposed in the cap removing position and clamps the plurality of test tubes, and a cap removal unit disposed in the cap removing position and configured to move upward to remove caps from some of a plurality of test tubes and move upward to remove caps from the remaining test tubes. The cap removal unit includes engaging members which are capable of advance and retreat with respect to the caps and clamp each cap, a lift mechanism which raises the engaging members while alternately moving the engaging members upward, thereby removing the cap from each test tube, a cap guide member which separates the cap and guides the cap for dropping.