Whereas many researchers still approach Terra Preta (TP) as a soil category, new evidence suggests that TP refers to a directional grading of soil property changes (i.e., color, pH, nutrients, etc.) within human-made soils, originating from human activities in pre-Columbian times. Currently, most TP research focuses on the Brazilian part of the Amazon basin, but only little information is available on TP soils in the Colombian Amazon. Here, we sampled four TP and surrounding soils in the Colombian Amazon region at different soil depths and analyzed them for (i) general soil properties such as color, pH and texture, (ii) soil organic carbon and black carbon (BC) contents, the latter using benzene polycarboxylic acids as molecular marker, (iii) phosphorus availability based on sequential fractionation, and (iv) microbial residue contents using amino sugars. Our data from Colombia’s middle Caquetá River and Leticia confirmed that SOC, BC, and total P were present in significantly higher concentrations in the TP areas than the surrounding soils, while pH values and microbial residue contents were unchanged. The enrichment of P forms comprised both easily extractable and stable P pools, which both dominated to a different degree, both in TP and adjacent soils. The different degree of SOC, BC and P enrichment suggests different amounts of waste disposal by the ancient populations at different TP sites, now warranting further research for reconstructing ancient population sizes from TP chemical analyses.