(R,R)- and (S,S)-enantiomers of anthracene-containing pyridino-18-crown-6 ether having tert-butyl groups at the stereogenic centers were prepared with the aim of achieving higher enantioselectivity than for the reported (S,S)-analogue having isobutyl groups. The enantiomeric recognition abilities of the new sensor molecules toward chiral protonated primary amines and amino acid esters were studied in acetonitrile by UV–vis and fluorescence spectroscopies. The pKa values of these pyridino-crown ethers and their reported (S,S)-analogues having methyl or isobutyl groups have also been determined in acetonitrile.