Peppermint (Mentha ×piperita) is currently not a common crop in the southeastern United States. Recently, there has been an interest in peppermint as an essential oil crop for this region. In 2007, a field experiment was conducted in Mississippi to determine the effect of nitrogen (N) (0 and 80 kg·ha−1), location (Verona and Stoneville), and harvesting stage (bud formation and flowering) on peppermint productivity, oil content, and composition. Peppermint biomass yields were higher at Verona (8.12 t·ha−1) than at Stoneville (6.12 t·ha−1). However, the essential oil content was higher at Stoneville (1.1%) than at Verona (0.6%). Generally, N rate at 80 kg·ha−1 did not affect oil yield and composition compared with 0 kg·ha−1. The concentrations of the major essential oil constituents were: (-)-menthol at 26% to 30%, (-)-menthone at 14% to 21%, (+)-menthofuran at 5% to 11%, and eucalyptol at 3% to 4% of total essential oil (wt/wt) and were generally within the values previously reported. The concentrations of (-)-menthone and (+)-menthofuran were significantly higher at Stoneville, 21% and 11%, respectively, than at Verona (14% and 6%). The (-)-menthol, (-)-menthone, and (+)-menthofuran yields/ha were higher at Stoneville than at Verona. Overall oil content and yields were higher at bud formation than at flowering, 1.3 and 1.8 times, respectively. In addition, the concentration of (-)-menthone and eucalyptol and the yields of (-)-menthone, (+)-menthofuran, and eucalyptol were higher at bud formation than at flowering. The results from this study demonstrated peppermint can be successfully grown in Mississippi and provide essential oil yields and composition comparable to those in the northwestern United States, the traditional peppermint-growing region.