Preparation of heme control for cellulose acetate electrophoresis

ABSTRACT

To avoid laboratory errors in the detection of hemoglobinopathies, a control sample containing hemoglobin [Hb] A,F,S or C should run with each set of samples.The aim of this project was to prepare a lyophilized heme control for the cellulose acetate electrophoresis.After lysing the red blood cells, the hemolysate was centrifuged at 25000 rpm. The clear red supernatant was diluted with drabkin reagent to 20-30 gr/L and Hb was converted to cyanomethemoglobin. After adding sucrose and preservatives, the hemolysate was passed through milipore filter [0.45 ?m] and then aliqouted and lyophilized.In this project three heme control samples were prepared. The first sample contains Hbs A,F,D or G the second one contains Hbs A,S, and the third contains Hb A,F. Although the amount of potassium cyanide used is 10 times less than other methods, the stability is more than 6 months at 4oC; moreover, and the electrophoretic patterns have good resolution and the obtained CV and SD show good reproducibility.The preparation method in this project is simple, reliable, cost effective and in comparison with other methods has less amount of potassium cyanide; therefore, it is safe for the laboratory staff