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Bio-oils from the fast pyrolysis of switchgrass forage and two sets of alfalfa stems (from two stages of developmentsbud and full flower stages) have been analyzed by wet-chemical methods, GC-MS, and HPLC. The chemical composition of bio-oil is important because it may give insights into its quality, stability, or suitability for downstream upgrading. Pyrolysis experiments were conducted at 500 °C under a nitrogen atmosphere in 2.5 kg/h fluidized bed reactor. A total of 62 chemical species were positively identified in the liquids. Of the identified compounds, 27 have been quantified. The chemical compositions of the bio-oils are compared with what is known about the compositions of these feedstocks. While the compositions of the bio-oil from the two alfalfa stems were similar, there were numerous differences in the compositions of the alfalfa and switchgrass bio-oils. Noted are the higher levels of nitrogen, water, and aromatic hydrocarbons in bio-oils produced from alfalfa stems than from switchgrass and woody feedstocks that have been previously characterized. Also noted is a much lower concentration of levoglucosan and hydroxyacetaldehyde concentrations among bio-oils from alfalfa stems compared with bio-oil from switchgrass or woody biomass.