Cholesterol and triglyceride levels were measured in plasma samples of control and HFD rats. The enzymatic method to determine cholesterol level is based on the cleavage of cholesterol esters by cholesterol esterase to yield free cholesterol and fatty acids. Cholesterol oxidase then catalyzed the oxidation of cholesterol to cholest-4-en-3-one and hydrogen peroxidase. In the presence of peroxidase, the hydrogen peroxide formed affected the oxidative coupling of phenol and 4-aminophenazone to form a red quinone-imine dye. The color intensity of the dye formed is directly proportional to the cholesterol concentration and was determined by measuring the increase in absorbance (Cobas CHO2I and CHO2A, Roche Diagnostics, Mannheim, Germany).
The enzymatic triglyceride assay is based on using a lipoprotein lipase for the rapid and complete hydrolysis of triglycerides to glycerol followed by oxidation to dihydroxyacetone phosphate and hydrogen peroxide. The hydrogen peroxide produced then reacted with 4-aminophenazone and 4-chlorophenol under the catalytic action of peroxidase and a red dye was formed. The color intensity of the red dye formed is directly proportional to the triglyceride concentration and was measured photometrically (Cobas TRIGL, Roche Diagnostics, Mannheim, Germany).
The enzymatic triglyceride assay is based on using a lipoprotein lipase for the rapid and complete hydrolysis of triglycerides to glycerol followed by oxidation to dihydroxyacetone phosphate and hydrogen peroxide. The hydrogen peroxide produced then reacted with 4-aminophenazone and 4-chlorophenol under the catalytic action of peroxidase and a red dye was formed. The color intensity of the red dye formed is directly proportional to the triglyceride concentration and was measured photometrically (Cobas TRIGL, Roche Diagnostics, Mannheim, Germany).