To investigate impact of hyperlipidemia on clarithromycin pharmacokinetics in Wistar rats, hyperlipidemia was successfully induced by providing high fat diet (rich of cholesterol) for 6-8 weeks. Pharmacokinetic parameters of clarithromycin in induced hyperlipidemia and control group (non-hyperlipidemia) were determined in rats after oral (10, 20, and 100 mg/kg) single administration of clarithromycin. Group allocation for in-vivo study (n=6) as follows: G1: Vehicle control, G2-G4: Non-hyperlipidemic rats treated with clarithromycin, G5-G7: hyperlipidemic rats treated with clarithromycin. For In-vitro study (n=4): G8: Nonhyperlipidemic rats pooled liver homogenate, G9: Hyperlipidemic rat pooled liver homogenate for investigation of clarithromycin metabolism. For biliary excretion studies (n=4): G10: nonhyperlipidemic rats treated with clarithromycin and G11: Hyperlipidemic rats treated with clarithromycin. The area under the plasma concentration–time curve (AUC0–∞) and the peak plasma concentration (Cmax) of clarithromycin (non-hyperlipidemic group) was 1.77 ± 0.33 μg. h/mL and 0.47 ± 0.06 μg/mL for 10 mg/kg; 5.63 ± 1.51 μg. h/mL and 1.21 ± 0.11 μg/mL for 20 mg/kg; 58.97 ± 12.74 μg. h/mL and 12.48 ± 3.8 μg/mL for 100 mg/kg, respectively. In hyperlipidemic group the AUC0–∞ and Cmax of clarithromycin was 3.47 ± 0.82 μg.h/mL and 0.84 ± 0.09 ug/mL for 10 mg/kg; 12.21 ± 2.83 μg.h/mL and 2 ± 0.24 μg/mL for 20 mg/kg; 141.88 ± 54.03 μg.h/mL and 20.57 ± 7.6 μg/mL for 100 mg/kg, respectively. Over all exposure was significantly increased in hyperlipidemic groups at across the tested doses. The percentage remaining of clarithromycin in hyperlipidemic rat’s liver homogenates and biliary excretion was significantly (P<0.05) high as compared with non-hyperlipidemic rats. The % dose excretion of clarithromycin in urine was lower as compared with bile in hyperlipidemic rats. Conclude that the clarithromycin was considerably affected by hyperlipidemia. The enhanced area under the curve of clarithromycin in hyperlipidemic rats might be mainly due to inhibition of the CYP mediated metabolism in the liver by hyperlipidemia which was reproduced in bile and urine.