ABSTRACT.- In order to assess the effect of equine chorionic gonadotropin (eCG) administered on Day 5 or 7 of a fixed-time artificial insemination protocol (FTAI) in anestrous suckled beef cows, two experiments were performed to determine the following endpoints:Experiment 1 (n = 22), preovulatory follicle (POF) diameter, ovulation time, corpus luteum (CL) area, estradiol (E2) and progesterone (P4) concentrations; and Experiment 2 (n = 676), a field trial to evaluate conception rate using the same experimental design. In both experiments, a synchronization protocol using estradiol benzoate (EB) (Day 0), intravaginal progestin device (IVD) (Days 0 through 7), prostaglandin (PGF) (Day 7), eCG (Day 5 or 7), and GnRH (Day 9). Treatment consisted of administering 400 IU of eCG on Day 5 (T5) or Day 7 (T7 or control) concomitant with treatment with PGF2α. In experiment 1, all cows of T5 ovulated by 16 h after GnRH administration. The POF tended (P = 0.06; P = 0.07) to be larger at 1 and 2 d before ovulation in T5. The day before ovulation, E2 tended to be lower (P = 0.06) in T5 compared with T7. The CL area and the P4 concentrations were greater (P = 0.04) on day 9 in T5 compared with T7. In experiment 2, the conception rate was greater (P = 0.04) in T5 (72.2%) compared with T7 (61.0%) group. Therefore, administration of eCG on Day 5 of the designed protocol hastened ovulation of a greater follicle, which produced a larger CL and greater concentrations of progesterone by Day 9 after ovulation, resulting in 11.2% increase in cows pregnant. © 2022 Elsevier Inc.