Two experiments were conducted to evaluate the effect of in ovo amino acid (AA) injections in broiler breeder eggs on AA utilization of embryos. All AA used in these experiments were pure crystalline AA in free-base form. Treatments in Experiment 1 comprised 1) control eggs (no injection), 2) 0.5 mL sterile-distilled water injected eggs, and 3) eggs injected with an AA solution suspended in 0.5 mL sterile-distilled water. Injections were administered into the yolk at Day 7 of incubation. At hatch, chicks were killed and bled, and plasma AA concentration was determined. Plasma AA concentration of hatched chicks decreased (P < 0.05) when water was injected. In addition, all AA from eggs injected with AA, except Glu and Lys, were decreased (P < 0.05) at hatch as compared to control eggs. However, AA pattern was not affected by in ovo water injection, but the AA ratio to Lys was reduced by in ovo AA injection. Experiment 2 was conducted to evaluate whole internal egg AA concentrations over incubation time in the presence or absence of in ovo AA administration. Treatments in Experiment 2 comprised 1) control eggs (no injection), and 2) eggs injected with a AA solution at Day 7 of incubation. The AA contents of embryo, yolk, albumen, and allantoic and amnion fluids were analyzed over time during incubation (Days 0, 7, 14, and 19 of incubation). On Day 14 of incubation, there were no differences in AA contents of all tissues between the control group and the group injected with AA on Day 7 of incubation. On Day 19 of incubation, AA contents of embryo, yolk, albumen, and allantoic and amnion fluids were increased (P < 0.05) as mediated by in ovo administration of AA at Day 7 of incubation. These results suggest that in ovo administration of AA may increase AA concentrations in chicken embryos and other egg contents.