Genetic species determination has become an indispensable tool in forensics, archaeology, and ecology. However, current technology requires a tradeoff between speed and versatility. Fast and cost-efficient methods like PCR or immunoassays can only detect single taxa, whereas global species identification by DNA or protein sequencing requires more time and resources. Although a good compromise was found with collagen-based peptide mass fingerprinting, the approach is not broadly used due a lack of accessible data interpretation software, missing confidence of fragment-based peptide identification, and the low variability of collagen sequences between close species. Here, we introduce “Species by Proteome INvestigation” (SPIN), a fast and automated proteomics workflow capable of querying over 150 mammalian species in 7.2 minutes of mass spectrometry (MS) analysis.