The invention discloses a platelet-rich plasma separation gel, a vacuum blood collection tube which is prepared by utilizing the separation gel and used for separating platelet-rich plasma, and a separation method. The platelet-rich plasma separation gel particularly comprises a resin, and the resin is prepared from the following raw materials in parts by weight: 120 parts of butyl acetate, 75-80 parts of phenethylene, 20-25 parts of butyl acrylate, 5-12 parts of acrylic acid, 2-2.5 parts of azodiisobutyronitrile (AIBN), and 0.5-0.8 parts of dodecyl mercaptan, and the specific weight ranges from 1.068 to 1.080 at the temperature of 25 DEG C and can be accurately adjusted according to the raw material ratio. The separation gel prepared by use of the resin and the manufactured vacuum blood collection tube are also included. When the blood collection tube is used for preparing platelet-rich plasma, the process is simple, convenient and easy, and the required time is short; repeated tube transfer is not required, so that cross contamination is avoided; and the operation is simple and convenient, the preparation cost is greatly reduced, the practicability and the feasibility are greatly improved, and a wide market prospect is achieved.