SummaryWe present a novel study of the compositional relationships between soil organic components extractable in aqueous extractants and those in sub‐soil drainage and surface runoff waters from the soil. The surface soil (0–20 cm) of a stagnogley in long‐term grassland was sequentially and exhaustively extracted in aqueous media at pH values of 7, 10.6 and 12.6. Extracts from the soils and their runoff and drainage waters were processed by the procedures of the International Humic Substances Society (IHSS), and fractionated into humic, fulvic, and XAD‐4 acids. Elemental, δ13C, δ15N, sugar, amino acids, and solid state CPMAS 13C NMR analyses were used to identify similarities and differences between the fractions from the different extracts. There were few differences between the compositions of drainage water samples taken 1 year apart, and these had compositional features similar to those from the more highly oxidized fractions isolated from the soil at pH 7. There were significant differences between the humic components from the drainage waters and isolated from the soil at pH 7 and those of the humic fractions isolated at the higher pH values whose compositions are more clearly related to origins in plants. The compositions of the surface runoff waters indicate origins in transformed plant and animal manures on the soil surface, whereas those of the deep drainage waters originate in more extensively transformed materials, including products of microbial metabolism. The resin technique used in the fractionation allowed the isolation of novel humic acid fractions from the soil extracts, in particular at pH 7 and 12.6. These fractions clearly originated in microbial sources, were rich in saccharides and amino acids (peptides), and low in lignin‐derived components.