The results obtained from the different fractions of metabolites were selected for in vivo acute toxicity studies. The toxicological study revealed that the test samples are safe for animal use up to 800 mg/kg. The test sample showed dose dependent anti-nociceptive potentials, the significant effect was observed with 500 and 800 mg/kg b.w dose. The analgesic effect of diclofenac sodium (standard drug) was greater than test sample. The significant (P<0.01) anti-nociceptive potentials was noticed at 800 mg/kg b.w. of the animal for normal saline (35.50 ± 0.84), chloroform (29.67 ± 1.22), ethyl acetate (30.67 ± 1.22) and (29.00 ± 0.93) chloroform fraction of L. plantarum secondary metabolites. The test sample reduced the motility of GIT in mice. The reduction in GIT motility is dose dependent and maximum effect was produced by 800 mg/kg dose. The test sample caused different degree of movement of charcoal meal in the intestine i.e. 24.17, 20.17 and 17.83 cm at the tested doses of 200, 500 and 800 mg/kg, respectively.