A peroxidase-coupled method for the colorimetric determination of serum triglycerides

• Published in: Clinical chemistry (Baltimore, Md.) Vol. 29; no. 3; pp. 538 - 542
• Main Authors: McGowan, MW, Artiss, JD, Strandbergh, DR, Zak, B
• Format: Journal Article
• Language: English
• Published: Washington, DC Am Assoc Clin Chem 01.03.1983; American Association for Clinical Chemistry
• Subjects: Adenosine Triphosphate; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Colorimetry - methods; Fundamental and applied biological sciences. Psychology; Humans; Hydrogen-Ion Concentration; Lipase; Lipids; Magnesium; Magnesium Chloride; Other biological molecules; Peroxidases; Polyethylene Glycols; Spectrophotometry; Triglycerides - blood
• ISSN: 0009-9147; 1530-8561
• DOI: 10.1093/clinchem/29.3.538

We describe an enzymatic method for rapid, precise measurement of serum triglycerides with use of sample:reagent ratios as large as 1:200. Hydrolysis of triglycerides is catalyzed by lipase to produce glycerol and free fatty acids. The glycerol generated is then phosphorylated by adenosine 5'-triphosphate in the presence of glycerol kinase. Oxidation of the resulting glycerol 3-phosphate to produce hydrogen peroxide is catalyzed by L-alpha-glycerophosphate oxidase. An intense red chromogen is produced by the peroxidase-catalyzed coupling of 4-aminoantipyrene and sodium 2-hydroxy-3,5-dichlorobenzenesulfonate with hydrogen peroxide. This sensitive chromogen system not only permits use of unusually small sample volumes, it also facilitates a linear response to serum triglyceride concentrations up to at least 10 g/L while displaying good Ringbom (measure of accuracy) characteristics.