Characterizing inter- and intra-population levels of variability in age at sexual maturation for long-lived fishes provides insight into year-class strength and recruitment dynamics, allowing for more effective management practices. Here we analyzed the ontogenetic chronology of pectoral fin ray annuli trace elemental concentration profiles as well as changes in annuli growth in Lake Sturgeon (Acipenser fulvescens) to determine onset of sexual maturity (OSM). Elemental concentrations and growth-zone width were used to build a random forest classification model to discriminate year-specific signatures to before or after OSM from 98 individual fin rays from both sexes across multiple populations and watersheds. The model demonstrated an overall accuracy of 98.8%. Ba was the most important variable related to OSM discrimination success followed by zone width, Pb, Mn, Mg, Zn, Cu, and Sr. Fin ray elemental concentrations began to increase at approximately age 24 (± 4.7) in females and 15 (± 3.1) in males while zone width decreased. This study implies that new applications for fin ray microchemistry are possible and could benefit future fisheries management strategies specifically by adding nonlethal or less invasive sampling techniques for evaluating OSM in sturgeon.