Our study objectives were to quantify genetic gains (GG) in yield plus macronutrient (S, P, K, Ca, Mg) and micronutrient (B, Mg, Zn, Mn, Fe, Cu, Al) contents, and investigate the relationship between harvest index (HI), nitrogen harvest index (NHI), and nutrient HI (NutHI) at physiological maturity in Bayer Crop Science legacy hybrids. Grain nutrients were analyzed in 38 maize hybrids, released between 1983 and 2020 in the US Corn Belt with nearly identical maturity groups. Yield was measured in all hybrids at two locations (Jerseyville, Illinois, and Lebanon, Indiana), while aboveground biomass was measured in a subset of 23 hybrids in 2021, representing both old and modern hybrids. The study spanned two years (2021 and 2022) and two N rates (45 kg N ha^-1 as Low-N in both years, and 205 and 246 kg N ha^-1 in 2021 and 2022, respectively, as High-N). A split-plot design was used, with main plots assigned to N rates and subplots to hybrids. While occasional increases in grain nutrient concentrations under Low-N compared to High-N (e.g., P and K) were observed, these did not alter the overall trends of increasing nutrient content due to yield improvements, plus consistently higher nutrient contents removed at High-N vs Low-N. Average NutHI at High-N was improved for P and Fe, but was lower for Zn, Mn and Cu. Relationships of NutHI to NHI were consistently negative at High-N, but neutral to positive for all nutrients except Fe at Low-N levels. Although increasing maize NHI is a desirable outcome in terms of N fertilizer balance, genetic gains in NHI were generally accompanied by lower NutHI at High-N and little to zero NutHI gain for nutrients other than N at Low-N.