An efficient and sensitive chiral analytical method was established for the determination of propiconazole stereoisomers by supercritical fluid chromatography-tandem mass spectrometry (SFC-MS/MS). Stereoisomeric separation was performed on a Chiralpak AD-3 column with CO2/ethanol (93:7) as the mobile phase. The four propiconazole stereoisomers were well separated in 4.7 min with resolutions above 2.0. The specificity, linearity, matrix effects, accuracy, precision, and stability of the developed method were evaluated. The stereoselective dissipation of propiconazole in wheat straw, grape, and soil samples was investigated according to the proposed method. The results indicated that significant stereoselective degradation occurred in wheat straw and grapes, with preferential degradation of (-)-propiconazole A and (+)-propiconazole B in wheat straw and the opposite case in grapes. No enantioselectivity was observed in soil, although diastereoisomer A degraded more rapidly than diastereoisomer B. These results could contribute to a more accurate assessment of the environmental risk and food safety of propiconazole.