Soil labile organic carbon (C) and nitrogen (N) pools play a central role in nutrient cycling, while fire is a key driver of biogeochemical cycle, shaping ecosystem structure and functioning. However, how soil labile organic C and N responds to the long-term repeated prescribed fire is largely unknown. In this study, a prescribed fire field experiment in a wet sclerophyll forest established in 1972 in southeast Queensland was used to evaluate the long-term impacts of different fire frequency regimes on labile organic C and N measured by different extraction methods. The fire frequency regimes included long unburnt (NB), burnt every two years (2yrB) and burnt every four years (4yrB). Results revealed that the 2yrB treatment had significantly lower C and N concentrations in hot water and K2SO4 extracts and in density fractions (LFD<2.3 and HFD>1.6) compared with the NB treatment. Concentrations of carbohydrate-C in hot water extracts and acid soluble and insoluble organic matter-C in cold-water extracts followed a similar trend. The maximum reduction was observed for carbohydrate‑C (72%) and the hot water extractable N (54%) in the 2yrB treatment compared with the NB treatment, showing these parameters are most sensitive indicators. However, there was no significant difference in most of the above parameters between the 4yrB and the NB treatments, indicating that less frequent fire (4yrB) allows the ecosystem to have sufficient time to recover from fire disturbance and may be a sustainable practice for fire management in this wet sclerophyll forest ecosystem.