This study evaluated the effect of distillation time (DT; 1.25, 2.5, 5, 10, 20, 40, 80, 160, 240, and 360 min) on essential oil yield, composition, and the antioxidant activity of ponderosa pine essential oil. Pine essential oil yield increased with length of the DT and reached maximum at 160 min DT. The major oil constituents were alpha-pinene and beta-pinene, ranging from 17% to 40% and from 21% to 29%, respectively, of the total oil. Overall, the concentration of alpha-pinene and beta-pinene was high at the initial DT (5–20 min) and decreased with increasing DT. The concentration of myrcene (range, 0.9% to 1.5%) was lowest at 5 min DT, then increased at 10 min DT, and did not change with longer DT. Overall, the concentrations of most other constituents (delta-3-carene, limonene, cis-ocimene, alpha-terpinyl acetate, germacrene-D, alpha-muurolene, gamma-cadinene, delta-cadinene, and germacrene-D-4-ol) were low at the initial DT and increased with increasing DT. Total yields (a function of oil yield and the concentration of individual constituents) of all constituents were generally the lowest at 5 min DT, increased with increasing DT, and reached maximum at 160 min DT. The antioxidant capacity of the pine oil in this study varied between 7.0 and 14.5 μmole Trolox/g and was unaffected by DT. This study demonstrated that DT can significantly modify the essential oil yield and composition of ponderosa pine needles. Furthermore, DT could be used to obtain pine oil with targeted chemical profiles. This report can also be used as a reference point for comparing literature reports, in which different DTs are used to extract essential oil of ponderosa pine.