2.1 MATERIALS

2.1.1 PLANT MATERIALS FOR ANALYSIS Fresh mature plants of Acalypha wilkesiana were collected from the natural habitat in October within Ambrose Alli University, Ekpoma, Edo State, Nigeria. The plant was authenticated in the Department of Botany, Ambrose Alli University, Ekpoma, Edo State, Nigeria. The leaves were picked, washed in distilled water air dried and weighed. The weight of the leaf was 0.1kg. The dried leaves were pounded to powdered form using a motar and pestle. The leaf powder was suspended in methanol for 24hours before filtering with a white filter cloth and boiled for 30minutes. The resulting methanol extract was allowed to attain room temperature and was evaporated to dryness using a water bath as a source of heat. The percentage yield of the extract was 6.41% (w/v). The pasty residue was used to prepare a standardized solution of the leaf extract in calculated amount of distilled water with standardized concentration of 0.46 g/ml. The standardized solution (the stock solution) was stored air tight in plastic bottles and kept frozen.

2.1.2 LABORATORY ANIMALS Exactly twenty four male albino rats weighing 148 ± 30g obtained from the Animal Holding Unit of the Institute of Agricultural Research and Training (IART) Apata-Ibadan, Nigeria were used for the study. The rats were fed with Guinea grower mash (product of bendel feeds and flour mills Ltd, Ewu, Edo state, Nigeria). The rats were grouped into four (4) groups, each groups consisting of six (6) rats.

2.2   INSTRUMENTS USED The instruments used for this analysis include;

1. Beam weigh balance
2. Electronic weigh balance                                                           

2.3 METHODS

2.3.1 EXPERIMENTAL ANIMALS AND PROCEDURE The Twenty Four Male albino rats were randomly grouped into four  consisting of six rats per group. The rats were kept in cages made of wooden frames and metal iron and were fed ad libitum (access to food and water for 24hours each day). The rats were fed with guinea grower mash and tap water with 12hours light and dark cycle. The cages were cleaned every morning and disinfected every day. The rats were allowed to acclimatize for seven days before extract administration was commenced. Calculated amount of methanolic leaf extract of Acalypha wilkesiana were constituted in distilled water from the stock solution to give doses of 500,500 and 1000 mg/kg body weight of paracetamol, vitamin C and leaf extract respectively and administered to the various groups as illustrated below; GROUP A: Control received 1ml of distilled water GROUP B: Received 500mg/kg body weight of paracetamol GROUP C: Received 500mg/kg paracetamol and vitamin C GROUP D: Received 500mg/kg paracetamol and 1000mg/kg leaf extract. Prior to the administration of methanolic leaf extract of Acalypha wilkesiana at every interval of seven days, the body weights of the animals were recorded. Administration of methanolic leaf extract of Acalypha wilkesiana was performed orally once daily between 8:30am ± 30 minutes, using metal cannula attached to a 2ml syringe prior to the administration of methanolic leaf extract of Acalypha Wilkesiana at every interval of seven days, the body weights of the animals were recorded.