The flight ability, nocturnal activity pattern and similar morphological characteristics of Chiroptera (bats) increase the difficulty for researchers to accurately identify them in the field. Therefore, extensive investigation should be performed to identify the diversity and accurately distribution information of Chiroptera. In August 2019, we sampled six Horseshoe Bat (Rhinolophus spp.) in Chaoyang of Liaoning province in China (119°08′21″ E, 40°53′15″ N). Based on morphological data (Table 1), barcode sequence (COI) (the accession number in NCBI database is MT240893) and phylogenetic relationship (Fig. 1), a Horseshoe Bat was identified as a Big-eared Horseshoe Bat (R. macrotis). This is the first record of Big-eared Horseshoe Bat in Liaoning, and the northernmost record of this species in China.