How does HDAOS detect serum total cholesterol

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The new Trinder’s reagent is a type of chromogen substrate commonly used in biochemical reagents and in many types. The detection method is basically similar to the TMB in the ELISA kit, and both belong to the enzyme-catalyzed spectrophotometric detection. Different Trinder’s reagents have more or less differences in the detection of different biochemical indicators. Here is how HDAOS detects serum total cholesterol levels.

Chromogen substrate HDAOS for biochemical detection

Vacuum blood collection tube to collect samples

Before using HDAOS to detect serum total cholesterol, the collection and processing of samples is also very critical. If errors occur in the collection of samples, the subsequent detection will not be accurate. Generally, venous blood is collected with a vacuum blood collection tube anticoagulant tube, and anticoagulated with EDTA dipotassium or heparin. The test specimen can be serum (or plasma can also be used). If an anticoagulant sample is used for testing, please gently invert and mix 5-8 times after collection, so that the sample is in full contact with the anticoagulant.

HDAOS detects cholesterol in samples

Cholesterol is also called cholesterol. A derivative of cyclopentane polyhydrophenanthrene. Total cholesterol (TC) includes free cholesterol (FC) and cholesterol ester (CE). Cholesterol esters in serum can be hydrolyzed by cholesterol esterase CHE into free cholesterol and free fatty acids (FFA). Cholesterol is oxidized by cholesterol oxidase CHO to generate 4-cholestenone and hydrogen peroxide. In the case of peroxidase catalysis, the generated hydrogen peroxide couples the chromogen substrate HDAOS and 4-AAP to oxidize, which can generate red quinone imines. The absorbance is measured with a spectrophotometer, and the absorbance is compared with the TC in the object. The content is directly proportional.

Advantages of using HDAOS to detect cholesterol

There are more than hundreds of methods for determining serum total cholesterol, which can be divided into five categories: chemical reagent colorimetry, enzymatic analysis, fluorescence method, gas phase and high performance liquid chromatography. Among them, the enzymatic determination of total cholesterol using chromogen substrates such as HDAOS is the most common at present. Its advantages are rapid and accurate, strong specificity, small sample volume, no extraction, and convenient for automated analysis and batch determination (gas phase and High performance liquid chromatography conditions require high).

In the chromogen substrate HDAOS test for serum total cholesterol, the quality of the substrate, HRP or POD and enzyme preparations cholesterol esterase and cholesterol oxidase will directly affect the test results. It is recommended to find a manufacturer such as Desheng to purchase the corresponding chromogen in bulk Substrates, enzyme preparations and buffers will also be more affordable.