Chiral pesticides are often degraded enantioselectively in soils, leading to disparity among enantiomers that may display different toxicity levels. Monitoring pesticide degradation extents and processes remains out of reach in the field using conventional bulk and enantiomer concentration analyses. Enantioselective stable carbon isotope analysis (ESIA) combines compound specific isotope analysis (CSIA) and enantioselective analysis, and bears potential to distinguish enantiomer degradation from non-destructive dissipation. We developed ESIA of the fungicide Metalaxyl, providing the C-13/C-72 ratios for S-Metalaxyl and R-Metalaxyl separately, and applied it to follow degradation in soil incubation experiments. Significant enantioselective degradation (k(S-MTY) = 0.007-0.011 day(-1) < k(R-MTY) = 0.03-0.07 day(-1)) was associated with isotope fractionation (Delta delta C-13(S-MTY) ranging from 2 to 6 parts per thousand). While R-Metalaxyl degradation was rapid (T-1/2 approximate to 10 days), concomitant enrichment in heavy isotopes of the persistent S-Metalaxyl occurred after 200 days of incubation (epsilon(S-Metalaxyl), ranging from -1.3 to -2.7 parts per thousand). In contrast, initial racemic ratios and isotopic compositions were conserved in abiotic experiments, which indicates the predominance of microbial degradation in soils. Degradation products analysis and apparent kinetic isotope effect (AKIE) suggested hydroxylation as a major enantioselective degradation pathway in our soils. Altogether, our study underscores the potential of ESIA to evaluate the degradation extent and mechanisms of chiral micropollutants in soils.