The effects of metronidazole (MN) on intestinal absorption properties are less investigated. This work aimed at assessing the effect of MN on piperaquine (PQ) permeability from dihydroartemisinin-piperaquine (DP) co-formulated antimalarial product, across intestinal epithelial membrane. Excised intestinal tissues from New Zealand male albino rabbits (n=2) were loaded with DP equivalent to PQ (100 mg/mL) and MN (100 mg/mL), according to animals’ body weight. Tissues were submerged in tyrode solution (TS) in an organ bath (100 mL). DP alone was similarly loaded in duodenum and ileum as control C1 and C2, respectively. Sampling (5 mL) of TS was taken at 0, 0.5, 1, 2, 4 and 6 h post immersion. Analysis of samples was performed using high pressure liquid chromatographic (HPLC) system with Zorbact Eclipse XDB column C8 (150 x 4.6 mm, 4.6 µm), mobile phase containing acetonitrile: 10 mM ammonium acetate (70:30, % v/v). The UV wavelength of detection and flow rate were 220 nm and 0.7 mL/min, respectively. The kinetics of PQ permeation was unaffected by MN. The area under the curve at 2 h (AUC0-2) and 6 h (AUC0-6) for duodenum revealed no difference (0.6285±0.0085 versus 0.6198±0.0083 µg.mLh-1, P=0.500) but lower (2.4863±0.0328 versus 3.3975±0.3638 µg.mLh-1, P=0.008) and for ileum, lower (0.1600±0.0170 versus 1.5408±0.4275 µg.mLh-1, P=0.001) and (0.9460±0.0506 versus 5.6603±0.1073 µg.mLh-1, P=0.011), respectively. The maximum concentration achieved were also lower than the respective controls (P<0.05). MN reduced the permeability of PQ across the intestinal regions. These findings will help to optimize therapeutic implications on concurrent administration