Sub-acute toxicity test

Sub-acute toxicity assessment according to the OECD Guideline 407 programme (OECD 407, 2008) was performed using fifteen BALB/c mice. Each sex of mice was randomly separated into three groups of five mice. In the first two groups, both sexes were administered with a dose of 50, 500 mg/kg body weight of dihydro-p-coumaric acid while normal drinking water was administered to the control group. Dihydro-p-coumaric acid was orally given to the BALB/c mice at a proportion of 10 ml/kg body weight with the use of gauze bent oral gavaging needles. Maintaining under similar feed, water and environmental conditions, all the mice were administered daily with either dihydro-p-coumaric acid or normal drinking water for 14 days. Mice were checked and observed carefully for the first 4 h post administration and two times daily (8.00 a.m. and 4.00 p.m.) for a 14 days study period. Mortality, signs of toxicity and toxicity related behavioural changes were among the study parameters. Body weight of the tested BALB/c mice were measured on day-1, day-7, and day-14 whereas weight of the feed and volume of the water consumed were observed on day-1, day-4, day-7, day-11, and day-14 of the study period. The water consumption, feed consumption and change in the percentage of the body weight were also observed and recorded.

At the end of the treatment, all the treated mice were anaesthetized using isoflurane and the blood samples were collected instantly for studying serum biochemical assay and haematological parameters using cardiocentesis. Organs like liver, heart, kidney and uterus from female mice while heart, liver, kidney and testis from male BALB/c mice were collected and weighed for calculating the organ weight and sectioning for histopathological analysis.