A deoxycholic acid-based macrocycle CDTB was synthesized and showed selective recognition of Hg2+ ion utilizing 1,2,3-triazole motifs as binding sites. The resultant [CDTB·Hg2+] complex could further discriminate amino acids enantioselectively through the difference in fluorescence enhancement on account of the chiral scaffolds of deoxycholic acid and the macrocylic structure. This line of research not only enriches the library of cascade recognition, but also opens a door for chiral natural products in the field of the enantioselective discrimination.