Most cases involving soil in criminal investigations in Brazil have focused on the chemical and mineralogical analyses of soil fractions without including the organic matter. The organic fraction contains plant-wax markers which may be useful to “fingerprint” forensic soils due to their chemical diversity, relative longevity and resistant nature. The aim of this study was to test the long- (kilometre), medium- (metre) and short- (centimetre) scale variability of plant-wax biomarkers in a forensic context in anthropised urban soils and soils developed under subtropical conditions. Two areas from the Curitiba municipality and two areas from the Colombo municipality, Paraná State, South Brazil, were selected. Soil colour analysis was carried out to obtain reflectance data over the 360–740 nm wavelength range. Furthermore, plant-wax marker compounds (n-alkanes and fatty-alcohols) were assessed by extraction and separation into different classes and an analysis of the compounds by gas chromatography (GC/MS). The compositions of the wax-marker profiles were different in samples collected side-by-side, showing sensitivity to local variations under subtropical conditions and in areas under intense human urban disturbance. Under these conditions, biomarkers may be used in real crime scenes, even on a micrometric scale of variation.