The mechanism of the optical resolution of gamma-valerolactone (VAL) enantiomers by enclathration in cholic acid (CA) channels was investigated. C-13 cross-polarization magic-angle spinning spectra of CA/ VAL inclusion compounds show four methyl C-13 peaks of VAL with different intensities depending on the.)-enantiomers (S) in the S enantiomeric ratios. The four peaks were assigned to the inner and end (S)-(domain and the inner and end (R)-(+)-enantiomers (R). The relative intensities of the four methyl C-13 peaks cannot be explained by the random process model for inclusion but are successfully reproduced by assuming the first-order Markov process, in which the inclusion probabilities of S and R depend on which enantiomer has precedingly entered the CA channel. The probability p(S/S) that two S enantiomers successively enter a channel is thus found to be 83%, and p(R/R) is 50%. The large probability of ps/s indicates that once an S enantiomer enters a channel, it become easy for other S enantiomers to successively enter the channel, and thus the large enantiomeric excess of S is obtained. The inclusion probabilities of S and R were confirmed by 1D C-13-C-13 polarization-transfer experiments among the four methyl carbons of VAL in the CA channel. Further, we found that the C-13 line widths and peak positions of the CA tail group change depending on the enantiomeric ratio. We concluded that once S is included, it changes the conformation of the CA tail group so that other S enantiomers become easy to successively enter the channel.