Approximately 55% of the Rubus glaucus Benth (Andean blackberry) produced in Colombia is sold fresh and 10% is used in agroindustry. There are two types of cultivars for this species: Andean blackberries with prickles and ones without prickles. The investigations carried out in the Department of Risaralda are focused on finding the best genetic materials; however, studies on the bioprospecting that can generate an added value to these fruits are lacking. Therefore, soil fertility analysis, physicochemical characterization, and evaluation of the total phenolic content and antioxidant activity of Andean blackberries with and without prickles were carried out. The parameters evaluated for soil analysis were pH, organic matter, minor elements, phosphorus, nitrogen, aluminum, and texture, and the parameters evaluated for physicochemical characterization were titratable acidity, ash, dietary fiber, fat, moisture, maturity index, protein, and soluble solids. The total phenolic content and antioxidant activity were evaluated with the Folin–Ciocalteu and DPPH methods, respectively. Statistical analysis showed significant differences in the content of fat, proteins, total soluble solids, titratable acidity, acidity index, and ash between the materials with and without prickles. The ash content and soluble solids are highlighted: they presented differences by study municipality, The ash content and soluble solids are highlighted; in addition, a high content of phenolic compounds was observed in fruits from plants without prickles. The diagnosis of blackberry crops by the materials planted and place of cultivation in the Department of Risaralda showed that the materials without prickles are more widely distributed in the department due to their ease of handling. According to the results of the soil analysis, most Castilla blackberry crops in the Santa Rosa, Santuario, and Pereira municipalities are within the recommended parameters for blackberry production at the national level.