The survival of Amazon stingrays is threatened due to excessive fishing and habitat degradation. To address this issue, this study developed a groundbreaking method to authenticate and differentiate Amazon stingray meats using a portable spectrophotometer and chemometrics. Samples were collected from various species, including an endangered one with a commercialization ban and no population reduction records. Principal Component Analysis (PCA), identified natural groupings based on the meat's commercial origin, while Partial Least Squares-Discriminant Analysis (PLS-DA), accurately discriminated the commercial and geographic origins with 100 % accuracy. Moreover, Data-Driven Soft Independent Modeling of Class Analogy (DD-SIMCA), effectively distinguished Amazon stingray meat from other marketable species. This approach offers a rapid, precise, and non-destructive means for monitoring and controlling the illegal trade of these species, thereby supporting decision-making in the field and promoting the conservation and sustainability of freshwater stingrays in the Amazon region.