An alfalfa (Medicago sativa L) biomass energy production system would produce two products. Leaves would be separated from stems to produce a protein feed for livestock while stems would be processed to produce ethanol. Therefore, maximum yields of both leaves and stems are essential for profitability of this biomass production system. Our objective was to evaluate the impact of growth environment (locations, years and plant density) and harvest maturity stage (early bud (4 annual cuts) and late flower (3 annual cuts)) on leaf crude protein and potential ethanol yields for four alfalfa germplasms, two with high forage quality, and two non-lodging biomass types. Potential ethanol yield was greater at late flower compared to early bud, while leaf crude protein concentration was similar at the two harvest maturity stages at both locations. Leaf crude protein yield was greater at the Minnesota (MN) site compared to Wisconsin (WI) site. The two non-lodging biomass germplasms had greater potential ethanol yield compared to the high forage quality cultivars in WI, but no differences among the alfalfa germplasms were found for ethanol yield at MN. In WI, no differences were found among the germplasms for leaf crude protein yield, but the high quality cultivars had greater leaf crude protein yield than the non-lodging germpLasms in MN. While germplasm differences were found for leaf crude protein and potential ethanol yields, the environmental influences of harvest date and locations had the greatest impact on these two alfalfa biomass energy products. Published by Elsevier Ltd.