Pyrogenic carbon (PyC), the combustion residues of fossil fuel and biomass, is a versatile soil fraction active in biogeochemical processes. In this study, the chemo-thermal oxidation method (CTO-375) was applied to investigate the content and distribution of PyC in 30 Australian agricultural, pastoral, bushland and parkland soil with various soil types. Soils were sampled incrementally to 50 cm in 6 locations and at another 7 locations at 0–10 cm. Results showed that PyC in Australian soils typically ranged from 0.27–5.62 mg/g, with three Dermosol soils ranging within 2.58–5.62 mg/g. Soil PyC contributed 2.0–11% (N = 29) to the total organic carbon (TOC), with one Ferrosol as high as 26%. PyC was concentrated either in the top (0–10 cm) or bottom (30–50 cm) soil layers, with the highest PyC:TOC ratio in the bottom (30–50 cm) soil horizon in all soils. Principal component analysis - multiple linear regression (PCA-MLR) suggested the silt-associated organic C factor accounted for 38.5% of the variation in PyC. Our findings suggest that PyC is an important fraction of the TOC (2.0–11%, N = 18) and chemically recalcitrant organic C (ROC) obtained by chemical C fractionation method accounts for a significant proportion of soil TOC (47.3–84.9%, N = 18). This is the first study comparing these two methods, and it indicates both CTO-375 and C speciation methods can determine a fraction of recalcitrant organic C. However, estimated chemically recalcitrant organic carbon pool (ROC) was approximately an order of magnitude greater than that of thermally stable organic carbon (PyC).