A novel assay for triglycerides using glycerol dehydrogenase and a water-soluble formazan dye, WST-8 (original) (raw)

2019, Annals of Clinical Biochemistry: International Journal of Laboratory Medicine

BackgroundThe glycerol-3-phosphate (GPO)-peroxidase (POD) chromogenic method is one of the most widely used methods to assay triglycerides. However, it is well known that peroxidase is affected by reducing agents, and recently, it has been reported that some materials affect its activity. Moreover, there is a high possibility of non-specific reaction, as the method uses many enzymes. Against this background, we developed a simpler assay method for triglycerides without using peroxidase.MethodsTriglycerides were hydrolysed to glycerol and fatty acids by lipoprotein lipase followed by the oxidation of glycerol to dihydroxyacetone with simultaneous production of NADH by glycerol dehydrogenase. To overcome incomplete conversion of glycerol to dihydroxyacetone by glycerol dehydrogenase at equilibrium, we added 2-(2-methoxy-4-nitrophenyl)-3–(4-nitrophenyl)-5–(2,4-disulfophenyl)-2 H-tetrazolium monosodium salt (WST-8) to the reaction mixture to remove NADH, allowing the reaction to complet...

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