The distribution of chemicals in forest soil layers is poorly understood. Hence, using GCMS we identified the occurrence of main chemical compounds and their relative composition in 3-different soil layers (litter, humus and subsoil layers) of rhododendron forest. We found that the total contents of main chemical compounds in rhododendron forest followed the order litter > humus > subsoil layers. In all these 3-layers, the primary chemical was palmitic acid. The higher fatty acids and alcohols significantly differed among the layers. The mannitol and pentadecanoic acid showed the largest differences and their contents were highest in humus layer. This study showed that the litter layer has more allelochemicals than humus and subsoil layers. These results can be very important for ecosystem management and protection of rhododendron forest.