A sensitive radioenzymatic assay for glycerol and acylglycerols.
A sensitive radioenzymatic assay for glycerol and acylglycerols is described. The assay depends on the quantitative phosphorylation of glycerol to glycerophosphate by glycerol kinase using [gamma-32P]ATP as a substrate. The 32P content of the formed glycerophosphate is determined and gives a measure of the original glycerol content. Acylglycerols can be determined by prior hydrolysis to glycerol. The assay is sensitive to about 0.1 nmol of glycerol and can be extended to 100 nmol. The assay can be applied to the determination of acylglycerols separated by thin-layer chromatography in amounts as low as 0.5 nmol. The assay is particularly useful in the determination of the specific activity of 14C- or 3H-labeled glycerol moeities.
D000818:Animals; D002855:Chromatography, Thin Layer; D004926:Escherichia coli / Q000201:enzymology; D005989:Glycerides / Q000032:analysis*; D005990:Glycerol / Q000032:analysis*; D005991:Glycerol Kinase; D008099:Liver / Q000032:analysis; D008722:Methods; D008832:Microchemistry; D010761:Phosphorus Radioisotopes; D051381:Rats; D014280:Triglycerides / Q000032:analysis; D014316:Tritium
D005989:Glycerides; D010761:Phosphorus Radioisotopes; D014280:Triglycerides; D014316:Tritium; D005991:Glycerol Kinase; D005990:Glycerol