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The O horizon, or detrital layer, of forest soils is linked to long‐term forest productivity and health. Fuel reduction techniques, such as prescribed fire, can alter the thickness and composition of this essential ecosystem component. Developing an understanding of the changes in the chemical composition of forest detritus due to prescribed fire is essential for forest managers and stakeholders seeking sustainable, resilient, and productive ecosystems. In this study, we evaluated fuel quantity, fuel structure, and detrital chemical composition in longleaf pine (Pinus palustris Miller) forests that have been frequently burned for the last 40 yr at the Tom Yawkey Wildlife Center in Georgetown, SC. Our results suggest that frequent prescribed fire reduces forest fuel quantity (p < 0.01) and vertical structure (p = 0.01). Using pyrolysis–gas chromatography/mass spectrometry as a molecular technique to analyze detrital chemical …