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The antidiarrheal action of EExHs was not found to be very promising for further use however, the pure compounds from EExHs could be analyzed to justify the effects. Diclofenac sodium (25 mg/kg) was used as a reference antinociceptive drug. In acetic acid induced writhing test, oral administration of EExHs at three different doses (125, 250, and 500 mg/kg) decreased writhing in dose-dependent manner while the highest dose (500 mg/kg) achieved the maximum percentages of pain inhibition (58.8%). Lethal concentration (LC50) of EExHs for brine shrimps was 50.59 µg/mL which was relatively lower than that of the standard drug vincristine sulfate. Reducing sugar, steroid, glycoside, tannin, alkaloid, saponins, and flavonoids were found to be present in EExHs. Data were analyzed by GraphPad Prism 6.0 software using Dunnett's test for multiple comparisons. EExHs was undertaken for cytotoxic assay by Brine shrimp lethality bioassay, antinociceptive action by acetic acid induced writhing test, and antidiarrheal activity by castor oil induced antidiarrheal test. Preliminary phytochemical screening was accomplished by established methods modified in experimental protocol. Therefore, we investigated the cytotoxic, antinociceptive, and antidiarrheal effects of H. Hygrophila spinosa (Acanthaceae) has been used in a wide range of ailments including flatulence, diarrhea, dysentery, gonorrhea, and menorrhagia. Synthetic drugs are going to be replaced by plant-derived traditional drugs due to their cost effectiveness, relatively less harmfulness, and efficacy against multidrug resistance organisms.

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