[1] Since the molecule has a hydroxyl (-OH) group, it is frequently bound to other lipids including fatty acids; most analytical methods, therefore, utilise a strong alkali (KOH or NaOH) to saponify the ester linkages.

Instrumental analysis is frequently conducted on gas chromatograph (GC) with either a flame ionisation detector (FID) or mass spectrometer (MS).

As well as the faecally derived stanol, two other isomers can be identified in the environment; 5α-cholestanol 5β-coprostanol is formed by the conversion of cholesterol to coprostanol in the gut of most higher animals by intestinal bacteria.

It is generally accepted that the metabolism of cholesterol to coprostanol by gut bacteria proceeds in an indirect manner via ketone intermediates, rather than direct reduction of the Δ5,6 double bond.

Since 5β-coprostanol is formed from cholesterol in the vertebrate gut, the ratio of the product over reactant can be used to indicate the degree of faecal matter in samples.

Herbivores such as cows and sheep consume terrestrial plant matter (grass) which contains β-sitosterol as the principal sterol.

In the gut of these animals, bacteria biohydrogenate the double bond in the 5 position to create 24-ethyl coprostanol and so this compound can be used as a biomarker for faecal matter from herbivores.

This reaction occurs principally in anaerobic reducing sediments and the 5α-cholestanol / cholesterol ratio may be used as a secondary (process) biomarker for such conditions.

Reducing environments are frequently associated with areas experiencing high organic matter input; this may include sewage derived discharges.

[5][6] Variations in the concentration of coprostanol over time can be used to create human population reconstructions within a specific depositional environment.