Using NAD+ as hydrogen acceptor, LDH catalyzes the conversion of both lactate and NAD+ into pyruvic acid and NADH respectively. 1-Methoxy-5-methyl phenazine methyl sulfate (PMS) transfers hydrogen from NADH to NBT which deoxidize into purple chromogenic substrate. Lactic acid content can be calculated by measuring the OD value at 530 nm.
Lipoproteins (except LDL) such as HDL, CM, and VLDL change structure and dissociate under the action of surfactants. The released micronized cholesterol molecules react with cholesterol enzyme reagents, and the generated hydrogen peroxide is trapped in the absence of coupling agent. It is consumed without color development. At this time, the LDL particles are still intact, and then the reagent containing coupling agent is added, which can dissociate the LDL particles to release cholesterol, which is catalyzed by cholesterol esterase (CE) and cholesterol oxidase (CO) and produce hydrogen peroxide. Hydrogen peroxide is catalyzed by oxidase (POD) in the presence of 4-aminoantipyrine (4-AA) and phenol (T-OOS) to form a red quinone compound. Because the cholesterol molecules of other lipoproteins have been removed, the shade is proportional to the amount of LDL-C.
The coloured substance have a maximum absorption peak at 546 nm. Measure the OD value at 546 nm and the LDL-C content in the sample can be calculated.
Lipoproteins (except low-density lipoprotein cholesterol (LDL)) such as high density lipoprotein (HDL), chylomicron (CM), and very low density lipoprotein (VLDL) change structure and dissociate under the action of surfactants. The released micronized cholesterol molecules react with cholesterol enzyme reagents, and the generated hydrogen peroxide is trapped in the absence of coupling agent. It is consumed without color development. At this time, the LDL particles are still intact, and then the reagent containing coupling agent is added, which can dissociate the LDL particles to release cholesterol, which is catalyzed by cholesterol esterase (CE) and cholesterol oxidase (CO) and produce hydrogen peroxide. Hydrogen peroxide is catalyzed by oxidase (POD) in the presence of 4-aminoantipyrine (4-AA) and phenol (T-OOS) to form a red quinone compound, the shade of color is directly proportional to the LDL-C content.
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