ANTI-NUTRITIONAL CONSTITUENT OF COLOCASIA ESCULENTA ...
ANTI-NUTRITIONAL CONSTITUENT OF COLOCASIA ESCULENTA ... ANTI-NUTRITIONAL CONSTITUENT OF COLOCASIA ESCULENTA ...
C.2.2.3 C.2.2.3.1 Preliminary toxicity test Animals C.2.2.3.2 Diet L. Only male, inbred, Sprague-Dawley rats, weighing between 200 and 250 g were used in this section ofthe study. The animals were randomly assigned to five groups, two per cage. The animals were housed in solid-bottom plastic cages, with wood shavings for bedding. They were kept in a well-ventilated room., with the temperature ranging between 25 and 28° C and the humidity between 50 and 55 per cent. All the animals were maintained on a stock., pelleted diet, with food and water available coustantly. The preliminary toxicity test-group animals were fed by means of a stomach pump with 10, 5 and 2.5 mg kgo!dayo! of beta-sitosterol (Table C2-1). The two control groups received oleic acid and distilled water respectively. Because it is a clinically-intended route for the test article, oral administration was selected for the present study. Table C2-1: Composition ofadministered materials for preliminary toxicity test groups Gnno .. ' p. Oleic acid (wlv)* Beta-sitosteroJ (wlv) Distilled water A Distilled water lml B Oleic acid 0.03 mglml C Beta-sitosterol (ID mg kgO'day0') 0.03 mglml 2.5 mgfml D Beta-sitosterol (5 mg kg"dayo') 0.03 mglml 1.25 mglml E Beta-sitosterol (2.5 mg kg0'day0') 0.03 mglml O.63mg1ml ! Iml * oleic acid was added to a three per cent sodium caseinate solution. 192
C.2.2.3.3 C.2.2.4 C.2.2.4.1 C.2.2.4.2 Experimental design All animals were acclimatized for five days under standard laboratory conditions. There were two rats in each group for the short-term preliminary toxicity test (Groups A-E). The animals were given their respective diets for six days: Group A received 10 mg ki1day-1 of beta-sitosterol; Group B received 5 mg kg- l day-l of beta-sitosterol; and Group C received 2.5 mg kg-1day·1 of beta-sitosterol. Group D received the equivalent amount of oleic acid and Group E received distilled water, both groups performing as controls. The body weight, food and water consumption ofeach rat were monitored daily. All animals were observed for clinical signs oftoxicity and mortality throughout the study period. Experimental test Experimental design The animals were selected and housed as descnbed in Section C2.2.3. They were fed a standard, balanced diet of pellets and water was made available ad libitum. AninIals were divided into three groups and received their respective supplemented diets for two weeks: Group A received 10 mg kg- l day-l ofbeta-sitosterol; Group B received distilled water as a control; and Group C received the equivalent anIount of oleic acid as a control. Body weight, food and water consumption ofeach rat were measured daily_All animals were observed for clinical signs of toxicity throughout the study period. After two weeks, the animals were sacrificed by a quick blow to the back ofthe neck, after overnight food deprivation. Haematology The animals were fasted overnight, prior to necropsy and blood collection. Blood SanIples were collected in ethylene diamine tetra acetic acid (EDTA) tubes by cardiac puncture, for haematological testing and in serum separator tubes (SST) for serum 193
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C.2.2.3<br />
C.2.2.3.1<br />
Preliminary toxicity test<br />
Animals<br />
C.2.2.3.2 Diet<br />
L.<br />
Only male, inbred, Sprague-Dawley rats, weighing between 200 and 250 g were used in<br />
this section ofthe study. The animals were randomly assigned to five groups, two per<br />
cage. The animals were housed in solid-bottom plastic cages, with wood shavings for<br />
bedding. They were kept in a well-ventilated room., with the temperature ranging<br />
between 25 and 28° C and the humidity between 50 and 55 per cent.<br />
All the animals were maintained on a stock., pelleted diet, with food and water available<br />
coustantly. The preliminary toxicity test-group animals were fed by means of a stomach<br />
pump with 10, 5 and 2.5 mg kgo!dayo! of beta-sitosterol (Table C2-1). The two control<br />
groups received oleic acid and distilled water respectively. Because it is a clinically-intended<br />
route for the test article, oral administration was selected for the present study.<br />
Table C2-1: Composition ofadministered materials for preliminary toxicity test<br />
groups<br />
Gnno<br />
.. ' p.<br />
Oleic acid (wlv)* Beta-sitosteroJ (wlv) Distilled water<br />
A Distilled water lml<br />
B Oleic acid 0.03 mglml<br />
C Beta-sitosterol (ID mg kgO'day0') 0.03 mglml 2.5 mgfml<br />
D Beta-sitosterol (5 mg kg"dayo') 0.03 mglml 1.25 mglml<br />
E Beta-sitosterol (2.5 mg kg0'day0') 0.03 mglml O.63mg1ml<br />
!<br />
Iml<br />
* oleic acid was added to a three per cent sodium caseinate solution.<br />
192