A fluorometric method for the determination of triglycerides in nanomolar quantities.
A fluorometric assay for triglycerides in nanomole quantities is described. Glycerol is liberated from triglycerides with lipase from Chromobacter viscosum, then converted by glycerol kinase to glycerol-3-phosphate, which is oxidized by glycerol-3-phosphate oxidase, producing H2O2. The H2O2 ultimately forms a peroxidase-catalyzed fluorogen with p-hydroxyphenylacetic acid. The excitation and emission wavelengths of the fluorogen are 325 and 415 nm, respectively. The assay is linear in the range 0.05-35 nmol of triglycerides using triolein as standard.
D002853:Chromatography, Liquid; D005990:Glycerol / Q000032:analysis; D006863:Hydrogen-Ion Concentration; D008049:Lipase; D010648:Phenylacetates; D013050:Spectrometry, Fluorescence; D014280:Triglycerides / Q000032:analysis*
D010648:Phenylacetates; D014280:Triglycerides; C008070:4-hydroxyphenylacetic acid; D008049:Lipase; D005990:Glycerol