Materials and method 1-Experimental animals: A total number of 40 Sprague Dawley (S.D) male rats, 12-14 weeks old age and weighting 180-250 gm were used in the experimental study. Rats were obtained from Animal House of Nile Center for Experimental Researches, Mansoura , Egypt . Animals were housed in separate metal cages, fresh and clean drinking water was supplied adlibtium through specific nipple. Rats were kept at a constant environmental and nutritional condition throughout the period of the experiment. All animals' procedures were performed in accordance to the institutional Ethics Committee and in accordance with the recommendation for proper care and use of laboratory animals. Animals were divided into 4 groups as follows: Group …show more content…
The ratio of the absorbances of the glycosylated hemoglobin & the total hemoglobin fraction of the control and the test is used to calculate the percent glycosylated hemoglobin of the sample . Reagents: 10 Tests 25 Tests Ion Exchange Resin (Predispensed Tubes) 10 × 3ml 25 × 3ml Lysing reagent 5 ml 12.5 ml Resin separators 10 pieces 25 pieces Procedure: Wavelength: 415 nm (Hg 405 nm) Temperature: R.T Light Path: 1cm A.Hemolysate preparation : 1-Dispense 0.5 ml lysing reagent in to tube as labeled as test (T) 2-Add 0.1 ml of the reconstituted well mixed blood sample into the appropriatelylabeledtubes. Mix unit completes lysis is evident. 3-Allow to stand for 5 minutes . B.Glycosylated haemoglobin (GHb) separation : 1-Remove cap from the ion – exchange resin tubes and label as test . 2-Add 0.1ml of the haemolysate from step A in to the appropriately labeled ion exchange resin tubes . 3-Insert a resin separator in to each tube so that the ubber sleeve is approximately 1cm above the liquid level c the resin suspension . 4-Mix the tubes on a rocker , rotator or a vortex mixer continuously for 5