This study aimed to investigate the diversity and distribution of loaches of the genus Cobitis in the Inner Mongolia Autonomous Region, China. Focusing on specimens tentatively identified as Cobitis choii collected during field surveys, we sought to confirm their taxonomic status through integrated morphological and molecular evidence, determine whether they represent a new regional record, and analyze their distribution characteristics and resource status in the region. In July 2024, 16 Cobitis specimens were collected from three rivers—Huihe River (3 specimens), Ursun River (4 specimens), and Bayan Wula Bridge River (9 specimens)—in Hulun Buir City, Inner Mongolia, China. The species were identified through a combination of morphological examination (15 morphometric characteristics) and molecular phylogenetic analysis based on the mitochondrial COI gene. Morphological measurements followed standards from The Loaches of China. Molecular methods included genomic DNA extraction, PCR amplification and sequencing of the COI gene, sequence alignment by BLAST, and construction of a maximum likelihood phylogenetic tree based on genetic distances to clarify phylogenetic relationships.Morphologically, all specimens exhibited diagnostic characteristics of C. choii, including a faint longitudinal band on the body side, an L5 band composed of 14 ~ 19 distinct spots, and a lamina circularis on the second branched pectoral-fin ray in males (Fig. 1). Key morphological ratios were consistent with previously reported data, though some ratios (e.g., head length/standard length, body depth/standard length) showed wider variation ranges, possibly due to the predominance of juvenile individuals or regional population differentiation (Table 2). Molecular phylogenetic analysis revealed that all obtained COI sequences clustered within a clade of known C. choii sequences, with very low intragroup genetic distances (0.000 0 ~ 0.002 5), strongly supporting their conspecificity (Fig. 2; see detailed genetic distance matrix in Appendix 1). On the basis of congruent morphological and molecular evidence, this study provides the first confirmed record of C. choii in the Inner Mongolia Autonomous Region, which represents a new species record for the area. Stable populations are found across multiple rivers in Hulun Buir City. We recommend enhanced habitat protection and systematic monitoring for this species to support future research and conservation of regional fish biodiversity.