Abstract The objective of this study was to determine the effects of a flaxseed oil-based supplement (FlaxLic, New Generations, SD) on reproductive parameters, feeding behavior, and lipid profile in beef heifers. Crossbred Angus heifers [n = 60 body weight (BW) = 397kg ± 5.2 kg) were blocked by BW and antral follicle count and randomly assigned to two supplementation treatments: free-choice commercial lose mineral supplement (CON; n = 30) or free-choice flaxseed oil-based supplement (FLAX; n = 30) for 7 wk. Heifers were individually fed via the Insentec feeding system (Insentec RIC, Hokofarm, Marknesse, Netherlands), where basal diet (TMR) and supplements were provided in separate feeders to measure supplement intake and feeding behavior. Further, water intake and behavior were measured via Insentec water system. Individual intake was compiled over 24 h for each animal. Body weight, transrectal ultrasonography, and blood samples were collected weekly. On wk 5, heifers were enrolled in a fixed-time artificial insemination (FTAI) commercial protocol. Largest follicle diameter (LF) and corpus luteum (CL) presence were recorded weekly and on d of the FTAI protocol (d 0, d 7, and d 0). Pregnancy diagnosis was performed 30 d and 60 d following AI. Blood samples were analyzed for progesterone (P4) by radioimmunoassay (RIA) to determine puberty attainment before wk 5, and fatty acid profile was determined via gas chromatography-flame ionization detection. Heifers with a CL or P4 ≤ 1.0 ng/mL were considered pubertal. Data were analyzed using SAS 9.4. ADG was similar between groups (P = 0.90) but was affected by week (P < 0.01). TMR intake was similar in duration and quantity (P > 0.18) between groups. Supplement intake was greater (P < 0.01) for FLAX (0.73 ± 0.03 kg/d) compared with CON (0.18 ± 0.02 kg/d) and from wk 2 to wk 8 (P = 0.01). Time spent consuming the supplements was greater for FLAX compared with CON heifers (62.65 ± 2.11 vs 0.10 ± 0.11 min/d, respectively) over the entire trial (P < 0.01). Further, FLAX heifers consumed less water than CON heifers (32.2 ± 0.39 vs 33.93 ± 0.35 kg/d, respectively; P = 0.03) and from wk 2 to wk 7 (P = 0.03). LF was not different between groups on any of the days of the protocol (d 0, d 7, d 0; P > 0.20). Additionally, P4 (P ≤ 0.20), puberty attainment by wk 5 (P = 0.24), and pregnancy rates (P > 0.48) were similar between groups. Concentrations of arachidonic acid (3.00 ± 0.24 vs 2.32 ± 0.19 mg/mL; P = 0.04), linoleic (60.11 ± 4.34 vs 40.35 ± 4.04 mg/mL; P = 0.002), αLinolenic (16.73 ± 0.26 vs 15.86 ± 0.23 mg/mL; P = 0.02), and total polyunsaturated fatty acids (PUFA; 130.52 ± 5.97 vs 111.19 ± 5.56 mg/mL; P = 0.02) were greater on wk 8 for FLAX compared with CON heifers. Collectively, these findings did not show evidence that the flaxseed oil-based supplement affects variables that impact reproductive performance. However, it altered the plasma lipid profile, increasing omega-3, arachidonic, and linoleic fatty acids, as well as total PUFA.