The comparative biology of the hipposiderid genus Aselliscus has been little studied. Here we report studies of echolocation, diet, and phylogeny of Aselliscus stoliczkanus. The phylogenetic relationships of Aselliscus were investigated based on sequence comparisons of mitochondrial cytochrome-b and nicotinamide adenine dinucleotide dehydrogenase subunit 1 genes. Dates of divergence within the hipposiderid radiation also were estimated. The echolocation call frequency of A. stoliczkanus is quite high, with the dominant constant frequency component at 119–120 kHz, and a terminal sweep down to 104.5 kHz. The call duration is about 5.4 ms. The diet of A. stoliczkanus is mainly composed of lepidopterans (78.5%), beetles (14.9%), and hemipteran insects (6.5%) in November. Our results indicate that Aselliscus is monophyletic and is correctly classified in the Hipposideridae, and the divergence time for Aselliscus was estimated at 22 million years ago.