Identifying and protecting shark nurseries is a common management strategy used to help rebuild overfished stocks, yet we know little about connectivity between juvenile and adult populations. By analysing trace metals incorporated into vertebral cartilage, it may be possible to infer natal origin based on nursery-specific chemical signatures. To assess the efficacy of this approach, we collected juvenile blacktip sharks (Carcharhinus limbatus; n = 93) from four regions in the Gulf of Mexico in 2012 and 2013 and analysed their vertebral centra with laser ablation–inductively coupled plasma–mass spectrometry. We observed significant regional differences in six element:Ca ratios in both 2012 and 2013. Multi-element chemical signatures were significantly different among regions and between year-classes. Year-class-specific linear discriminant function analysis yielded regional classification accuracies of 81% for 2012 and 85% for 2013, although samples were not obtained from all four regions in 2012. Combining year-classes resulted in an overall classification accuracy of 84%, thus demonstrating the usefulness of this approach. These results are encouraging yet highlight a need for more research to better evaluate the efficacy of vertebral chemistry to study elasmobranch population connectivity.