The objective of the present study was to evaluate plasma metabolites of blood of Jersey heifers supplemented with chitosan or omega-6 fatty acids (FA). Eight Jersey heifers (average body weight of 158.62 ± 1.75 kg, mean ± SD) were used in replicated 4 × 4 Latin square experimental design, balanced and contemporary, in 2 × 2 factorial arrangement. The experimental period consisted of 18 d (12 d for adaptation and 6 d for data collection) and 5 d were used for wash out. The experimental diets were (1) control (CO), without omega-6 FA and chitosan supplementation; (2) whole raw soybean (WRS, source of omega-6 FA) with 200g/kg of DM of WRS); (3) chitosan (CHI), with 2 g/kg of DM of chitosan; (4) chitosan and WRS (CHWS), with chitosan and WRS in the same level used in previously cited diets .The diets were formulated according to the NRC (2001) to achieve weight gain of 700 g/day. Blood samples were collected on d 15 of each period before feeding, by puncture of coccygeal vein and immediately centrifuged at 2000 g × 15 min, supernatant serum was transferred do tubes and submitted to analyses of cholesterol (CHO), low density cholesterol (LDL), high density cholesterol (HDL) urea and blood urea nitrogen (UN) by enzymatic colorimetric method. Data were submitted to PROC MIXED of SAS 9.3. The CHI diet decreased (P < 0.05) and WRS increased (P < 0.05) blood concentrations of TC, LDL, urea and UN. Heifers presented blood concentration (mg/dL) of TC (78.12 and 131.25), LDL (44.15 and 92.35), urea (35.50, 43.12) and UN (16.54 and 20.09) for fed CHI and WRS, respectively. Interaction effect for HDL blood concentration was observed (P < 0.05) between CHI and WRS. The animals fed CHWS presented higher concentration (46.37 mg/dL), than WS (32.87 mg/dL), CO diet (28.00 mg/dL), and CH (23.75 mg/dL).Chitosan and omega 6 fatty acid influenced plasma metabolites of Jersey dairy heifers.