Total cholesterol includes free cholesterol and cholesterol esters. Cholesterol ester can be hydrolyzed by cholesterol esterase to produce cholesterol and free fatty acid. Cholesterol is oxidized by cholesterol oxidase to produce △4-cholestenone and hydrogen peroxide. In the presence of 4-aminoamylpyridine and phenol, hydrogen peroxide catalyze peroxidase to form red quinone compounds of benzoquinone imine phenizone.The color depth of the generated quinone is directly proportional to the cholesterol content. The absorbance values of the standard tube and the sample tube are measured respectively, and the cholesterol content in the sample can be calculated.
GSSG is reduced to GSH by glutathione reductase, and GSH can react with DTNB to produce GSSG and yellow TNB. The amount of total glutathione (GSSG+GSH) determines the amount of yellow TNB. Thus the total glutathione can be calculated by measuring the OD value at 412 nm. The content of GSSG can be determined by first removing GSH from the sample with appropriate reagent and then using the above reaction principle.
The excess iron is added to the serum to bind all the transferrin in the serum, and the excess iron is adsorbed by adding the iron adsorbent. The iron bind with the transferrin is separated from the protein by the action of acid solution and reductant. Fe3+ in serum is reduced to Fe2+, Fe2+ binds with bipyridine to form pink complex. In a certain range, the amount of iron ions is positively correlated with the color, and the measured iron content is TIBC. Subtracting serum iron value from TIBC is called unsaturated iron binding capacity (UIBC).
The amount of iron bound to transferrin in the blood is called serum iron. The maximum ability of transferrin to bind iron is called total iron binding capacity (TIBC), and iron saturation refers to the percentage of serum iron binding capacity to total iron binding capacity. The detection principle of this kit: add an excess of iron to the sample and detect the total iron content. The content of unbound iron was detected when transferrin was saturated with iron. The total iron binding capacity of the sample is obtained by subtracting the content of unbound iron from the total iron content. The sample is not added with iron, and the total content of iron bound to transferrin and free iron is detected. Meanwhile, the content of free iron that has not bound to transferrin is detected. The serum iron content is obtained by subtracting the latter from the former.
The excess iron is added to the serum to bind all the ferritin in the serum, and the excess iron is adsorbed by adding the iron adsorbent. The iron bind with the ferritin is separated from the protein by the action of acid solution and reductant. Fe3+ in serum is reduced to Fe2+, Fe2+ binds with bipyridine to form pink complex. In a certain range, the amount of TIBC is positively correlated with the depth of color. The iron content measured is, minus serum iron value, which is called unsaturated iron binding force. Total iron binding capacity minus serum iron value is unsaturated iron binding capacity (UIBC).
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