The invention discloses a performance testing system and method for a thermoelectric refrigeration device. The system comprises a testing cabin, a pressure stabilizing layer, a testing layer and an air return layer are arranged in the testing cabin and communicated with one another, the thermoelectric refrigeration device to be tested is arranged in the testing layer, one side of the pressure stabilizing layer is connected with a vacuum air extractor, an air supply opening is formed in the other side, an air return opening is formed in the air return layer, and a temperature and humidity control device is arranged between the air return opening and the air supply opening; a hot end heat measuring module used for conducting heat exchange on the hot end of the thermoelectric refrigeration device to be tested and a cold end heat measuring module used for conducting heat exchange on the cold end of the thermoelectric refrigeration device to be tested are arranged in the testing layer. The performance testing system has the advantages that any temperature and humidity environment working condition can be simulated, the practical working performance of the to-be-tested thermoelectric refrigeration device used on any occasion can be measured, and the system has a high simulation reappearing ability; the system can further provide a vacuum testing environment and measure theoretical work parameters of the thermoelectric refrigeration device to be tested in the vacuum environment.