Using a 3-ml syringe with a 16-G, 3-in. gavage needle attached, administer indomethacin orally. Similarly administer test compound at three concentrations and vehicle control to appropriate animals. Check each animal twice daily for discomfort, diarrhea, lethargy, ruffled fur or other general signs of illness.
Indomethacin (the reference compound) is given at a dose of 4.75 mg/kg in a volume of 1.0 ml/100 g body weight. It is prepared in vehicle (see recipe).
Experimental design for testing a compound is determined based on the chemistry of the compound and known in vitro and pharmacokinetic data. Typically three to four doses of the test compound are used, enough doses to produce a linear response curve. Vehicle for the test compound is defined by the experimental protocol or is selected based on physiochemical properties of the test compound.
If any animal dies or needs to be euthanized during the experiment, it should be necropsied to verify that the cause of death is due to perforation of the small intestine.