Serum triglycerides determined colorimetrically with an enzyme that produces hydrogen peroxide.
In this direct colorimetric procedure, serum triglycerides are hydrolyzed by lipase, and the released glycerol is assayed in a reaction catalyzed by glycerol kinase and L-alpha-glycerol-phosphate oxidase in a system that generates hydrogen peroxide. The hydrogen peroxide is monitored in the presence of horseradish peroxidase with 3,5-dichloro-2-hydroxybenzenesulfonic acid/4-aminophenazone as the chromogenic system. The high absorbance of this chromogen system at 510 nm affords useful results with a sample/reagent volume ratio as low as 1:150, and a blank sample measurement is not needed. A single, stable working reagent is used; the reaction is complete in 15 min at room temperature. The standard curve is linear for triglyceride concentrations as great as 13.6 mmol/L. Average analytical recovery of triglycerides in human sera is 100.1%, and within-run and between-run precision studies showed CVs of less than or equal to 1.6 and less than or equal to 3.0%, respectively. The method is suitable for automation.
D005991:Glycerol Kinase; D005993:Glycerolphosphate Dehydrogenase; D006735:Horseradish Peroxidase; D006801:Humans; D006861:Hydrogen Peroxide / Q000032:analysis; D008049:Lipase; D013053:Spectrophotometry; D014280:Triglycerides / Q000097:blood*
D014280:Triglycerides; D006861:Hydrogen Peroxide; D005993:Glycerolphosphate Dehydrogenase; C036140:L-alpha-glycerol-phosphate oxidase; D006735:Horseradish Peroxidase; D005991:Glycerol Kinase; D008049:Lipase