Among the volatiles isolated by solvent extraction from cactus pear fruit (Opuntia ficus indica) and identified by HRGC-MS, selected chiral and achiral constituents were characterized by enantiodifferentiation and 13 C/ 12 C isotope ratio analysis, using multidimensional gas chromatography-mass spectrometry (MDGC-MS) and on-line gas chromatography-combustion isotope ratio mass spectrometry (HRGC-C-IRMS), respectively. While the enantiodistributions determined for methyl 2-methylbutanoate (98 : 2%, S : R), methyl 3-hydroxybutanoate (73 : 27%, R : S), 1-phenyl-ethanol (74 : 26%, R : S), linalool (68 : 32%, R : S), -nonalactone (65 : 35%, S : R), -deca-lactone (79 : 21%, R : S) and -dodecalactone (>99% R) can be discussed in relation to data previously provided for these compounds from various plant origins, the υ 13 C values of 22.0‰, 21.8‰, 19.1‰ and 20.9‰, measured for 1-hexanol, E-2-hexenol, E-2-nonenol and E,Z-2,6-nonadienol, respectively, are clear-cut indicators for their origin from plants with Crassulaceae acid metabolism (CAM), and thus may helpful for authenticity evaluations.