Here we report the first example of employing p-nitro-octadecyloxy-calix[8]arene (C8A-NO2) as stationary phase for capillary gas chromatographic (GC) separations. The statically coated C8A-NO2 column provided column efficiencies of 2554 plates/m and showed moderate polarity. Its selectivity and retention behaviour were investigated by a number of mixtures of diverse analytes and their isomers. It was found that the C8A-NO2 column exhibited high resolving capability for aliphatic and aromatic analytes from apolar to polar nature, especially for positional and structural isomers. Among them, advantageous resolving capability over the commercial polysiloxane DB-1701 and DB-17 columns was obtained for chloroaniline and bromoaniline isomers. In addition, the C8A-NO2 column was applied for the determination of isomer impurities in commercial reagents of geraniol, cis-decahydronaphthalene, and trans-decahydronaphthalene.