The operation of an anti-collision RFID system is characterized by the interrogation zone which should be estimated in any direction of 3D space for a group of electronic transponders. The interrogation zone should be as large as possible. However, the many problems in this area are due to the fact that energy can be transferred to transponders only on a limited distance. The greatest flexibility in developing RFID applications and shaping the interrogation zone can be achieved using the system with an antenna multiplexer. Therefore the problem of the interrogation zone determination in HF RFID systems with two orthogonal RWD antennas is presented in the paper. The perceived issues have been effectively dealt with and the solution has been proposed on the basis of the elaborated model. Conducted studies have been used to develop the software tool JankoRFIDmuxHF in the Mathcad environment. The research results are analysed in an example system configuration. The specialized measuring stand has been used for experimental verification of the identification efficiency. The convergence of the measurements and calculations confirms a practical usefulness of the presented concept of interrogation zone determination in anti-collision systems. It also shows the practical utility of the developed model and software tools.