Humic substances are formed by the microbial degradation of dead plant matter, such as lignin. They are very resistant to further biodegradation. The precise properties and structure of a given sample depend on the water or soil source and the specific conditions of extraction. Nevertheless, the average properties of humic substances from different sources are remarkably similar.

Humic substances in soils and sediments can be divided into three main fractions: humic acids, fulvic acids, and humin. The humic and fulvic acids are extracted as a colloidal sol from soil and other solid phase sources into a strongly basic aqueous solution of sodium hydroxide or potassium hydroxide. Humic acids are precipitated from this solution by adjusting the pH to 1 with hydrochloric acid, leaving the fulvic acids in solution. This is the operational distinction between humic and fulvic acids. Humin is insoluble in dilute alkali. The alcohol-soluble portion of the humic fraction is, in general, named ulmic acid. Gray humic acids (GHA) are soluble in low-ionic-strength alkaline media; brown humic acids (BHA) are soluble in alkaline conditions independent of ionic strength; and fulvic acids (FA) are soluble independent of pH and I.

Chromatography and liquid-liquid extraction can be used to separate the components that make up a humic substance. Substances identified include mono-, di-, and tri-hydroxy acids, fatty acids, dicarboxylic acids, linear alcohols, phenolic acids, and terpenoids.

Chemical characteristics of humic substances

Example of a typical humic acid, having a variety of components including quinone, phenol, catechol and sugar moieties

A typical humic substance is a mixture of many molecules, some of which are based on a motif of aromatic nuclei with phenolic and carboxylic substituents, linked together; the illustration shows a typical structure. The functional groups that contribute most to surface charge and reactivity of humic substances are phenolic and carboxylic groups. Humic acids behave as mixtures of dibasic acids, with a pK1 value around 4 for protonation of carboxyl groups and around 8 for protonation of phenolate groups. There is considerable overall similarities among individual humic acids.[4] For this reason, measured pK values for a given sample are average values relating to the constituent species. The other important characteristic is charge density. The molecules may form a supramolecular structure held together by non-covalent forces, such as Van der Waals force, π-π, and CH-π bonds.

The presence of carboxylate and phenolate groups gives the humic acids the ability to form complexes with ions such as Mg2+, Ca2+, Fe2+ and Fe3+. Many humic acids have two or more of these groups arranged so as to enable the formation of chelate complexes.[6] The formation of (chelate) complexes is an important aspect of the biological role of humic acids in regulating bioavailability of metal ions.