In this study, we conducted the hot box tests to compare the changes in thermal insulation for the four types of multi-layer thermal screens by the used period after collecting them from the greenhouses in the field when they were replaced at the end of their usage. The main materials for these four types of multi-layer thermal screens were matt georgette, non-woven fabrics, polyethylene (PE) foam, chemical cotton, etc. These materials were differently combined for each multi-layer thermal screen. We built specimens (70×70 cm) for each of these multi-layer thermal screens and measured the temperature descending rate, heat transmission coefficient, and thermal resistance for each specimen through the hot box tests. With regard to the material combinations of multi-layer thermal screens, thermal insulation can be increased by applying a multi-layered PE foam. However, it is considered that the multilayered PE foam significantly less contributes to heat-retaining than chemical wool that forms an air-insulating layer inside multi-layer thermal screens. For the suitable heat-retaining performance of multi-layer thermal screens, basically, materials with the function of forming an air-insulating layer such as chemical cotton should be contained in multi-layer thermal screens. The temperature descending rate, heat transmission coefficient, and thermal resistance of multi-layer thermal screens were appropriately measured through the hot box tests designed in this study. However, in this study, we took into consideration only the four kinds of multi-layer thermal screens due to difficulties in collecting used multi-layer thermal screens. This is the results obtained with relatively few examples and it is the limit of this study. In the future, more cases should be investigated and supplemented through related research.