As essential components of a minimal database, the hematological and biochemical blood profile are potent and significant diagnostic tools. Thus, the current study aimed to ascertain the serum biochemical and hematological characteristics, as well as any potential sex-related variations, of apparently healthy Arabian horses raised in Libya. The blood samples were collected from 65 clinically healthy horses of both sexes; males (n=26) and females (n=39) at winter season 2022 and included to analyze blood constituents and biochemical profile. The results of this study revealed that differences among Arabian horses according to gender factor not significant (p> 0.1) of all the parameters including the total white blood cells (WBC), red blood cells (RBC), hemoglobin (Hb), mean cellular volume (MCV), mean cellular hemoglobin (MCH), mean cellular hemoglobin concentration (MCHC), red cell width distribution-coefficient of variation (RDW-CV), red cell distribution width-standard deviation (RDW-SD), platelet count (PLT), mean platelet volume (MPV), platelet distribution width (PDW) and plateletcrit (PCT). Additionally, biochemical analysis showed no significant differences (p> 0.05) between stallions and mares of tested breed with exception of serum bilirubin, creatinine (Cr), urea, potassium (K) and chloride (Cl). This study shows no significant sex differences between male and female Arabian horses raised in Libya, whereas gender had only slight effect on the hematological and biochemical parameters.