The influence of chirality in calixarene threading has been studied by exploiting the “superweak anion approach”. In particular, the formation of chiral pseudorotaxanes bearing a classical stereogenic center in their axle and/or wheel components has been considered. Two kind of pseudorotaxane stereoadducts, the “<i>endo</i>-chiral” and “<i>exo</i>-chiral” ones, having the stereogenic center of a cationic axle inside or outside, respectively, the calix-cavity of a chiral calixarene were preferentially formed with specifically designed chiral axles by a fine exploitation of the so-called “<i>endo</i>-alkyl rule” and a newly defined “<i>endo</i>-α-methyl-benzyl rule” (<i>threading of a hexaalkoxycalixarene with a directional (α-methyl-benzyl)benzylammonium axle occurs with an</i> endo<i>-α-methyl-benzyl preference</i>). The obtained pseudorotaxanes were studied in solution by 1D and 2D NMR, and in the gas-phase by means of the enantiomer-labeled (EL) mass spectrometry method, by combining enantiopure hosts with pseudoracemates of one deuterated and one unlabeled chiral axle enantiomer. In both instances, there was not a clear enantiodiscrimination in the threading process with the studied host/guest systems. Possible rationales are given to explain the scarce reciprocal influence between the guest and host chiral centers.