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FOOD INDUCED- OBESITY
PURPOSE AND RATIONALE
Obesity can be induced in rats by offering a diet containing corn oil and condensed milk using standard formula
Procedure
Wister rats are house hold in individual wire bottom suspended cages in rooms maintained at 22-230 c with 12 h light /dark cycles. At the age of 6 months about 450 g) the animals are divided in two groups. Group 1 is fed ordinary rodent chow, group II a special diet containing rodent chow ,corn oil and condensed milk, resulting in a composition of 14.7% protein,44.2% carbohydrate ,15.8% lipid,2.5% fiber,1.2% vitamin mixture,19% water. Body weight and food intakes are measured and diet replaced every 3 to 4 days.
Three month from the start of the experiment, the rats will be sacrificed by decapitation for determination of adipose tissue cell size and number, carcass composition and plasma lipids and hormone and glucose levels.
The dorsal subcutaneous pad, retroperitoneal pad and one epididymal fat pad are sampled for the determination of lipid content. The method consists of homogenizing the tissue with a 2:1 chloroform methanol mixture and washing the extract by addition to it of 0.2 its volume of water. The resulting mixture separates into two phases. The lower phase is the total lipid extract. Cell number in each pad is determined by the osmium fixation method using coulter counter. Total epididymalpad weights are based on doubling the weight of the individual pads sampled.
Evaluation:
Intergroup comparisons are made with the use of 2 way ANOVA method.
Toxicity study:
Toxicity is defined as the substance which is able to harm or damage the exposed organism. These toxicities are possible to effect on a whole organism like an animal, bacteria or plant. It includes the effect on substructure of organism akin a cell (cytotoxicity) and organ (organotoxicity) e.g. liver (hepatotoxicity). Acute toxicity is described as the undesirable changes or signs of toxicity occur immediately with in 24 hr or a short period of denudation to substances as toxic effect occurs within 14 days as per guidelines of OECD 423. The need of acute oral toxicity screening is to get the information on the biological mechanism & systemic effect of chemical substances. Usually long term studies starts with dose finding method under acute conditions. Additionally, the acute systemic toxicity generates information by the chemical substance which is used in risk management in the production, handling, hazard identification and use of chemical substances. Currently, the precise or approximate value of LD50 calculated is based on the toxicological classification of chemicals. The animals were observed closely after administration of substances during first 24 hr and up to 2 weeks and changes in behaviours & appearance of animals are determined, most of the clinical signs were evaluated by characterizing acute systemic toxicity and describe its amelioration1 .
Principle: The method was followed by means of OECD 423 (Acute Toxic Class Method). The acute toxic class process is a step wise procedure with three rats of a single sex per step. Depending on the mortality of the rats and the standard two to three steps may be essential to allow conclusion on the acute toxicity of the test. material. This method results in the use of rats while allowing for satisfactory data based systematic conclusion. The process is used to defined doses (2000, 300, 50 and 5 mg/kg) the reports allow a material to be ranked and classified base upon the Globally Harmonized System (GHS) for the categorization of chemicals which produce acute toxicity.
Experimental Animals Male rats 450 g is plan to use for the present study. According to CPCSEA guidelines Wistar rats are kept in a proper ventilated room at 23°C and 12:12 hr light & dark cycle in polypropylene cages.
Experimental Procedure:
The four differenrent synthesised compounds are plan to chosen for antiobesity activity study. 10 mg /kg as initial dose and 2000 mg/kg as lethal dose as been plan to fix as per OECD 423 guideline. Food was withdrawn for remaining 3-4 hr after given of c-EACA and c-ECA for determining signs of toxicity. The weights of the rats were notified before and after given. The signs of changes in skin and fur, mucous membranes, eyes, circulatory, respiratory, autonomic and central nervous system, motor activity and behavior pattern were noted. The toxicity signs of fits, excessive salivation, tremors, diarrhoea, lethargy, sleep and coma are noted.