Abstract: (1) Background: Scorpions (Arthropoda: Arachnida) represent a diverse group of invertebrates, accounting for a significant proportion of earth’s predators and ecosystems’ modulators. Surviving mostly in hardly reachable nests, and representing key hazards to human health, they attracted major interest for characterizing their eco-, morpho-, and genotypes. (2) Methods: Four scorpion species were collected from the New Valley governorate in Upper Egypt, where a high level of scorpionism and related neurological symptoms are found, that were Leiurus quinquestriatus, Androctonus amoreuxi, Orthochirus innesi, Buthacus leptochelys. They were DNA barcoded, genetically and phylogenetically analyzed through PCR amplification and sequencing of the mitochondrial cytochrome oxidase subunit 1 (COI) gene hypervariable 50 region. (3) Results: New, morphologically authenticated scorpion barcodes could be added to the barcoding databases. However, several discrepancies and barcode database inadequacies could be revealed. Moreover, taxon-specific patterns for nitrogenous bases’ distribution could be identified, resulting in a significantly high percentage of COI barcode guanine in scorpionids, in comparison to araneids and opilions. (4) Conclusions: For a group of animals where both cryptic speciation and a high risk of human envenomation are evident, the findings of the current study strongly recommend continuous and comprehensive research efforts dealing with morphogenetic authentication for different species of scorpions.