Enzymatic determination of carbon (14C)-labeled glycerol in biological samples.
A method for determination of glycerol-specific-radioactivity in biological samples is presented. It is based on the following steps: (a) enzymatic conversion of glycerol to dihydroxyacetone-phosphate, (b) quantitative trapping of dihydroxyacetone-phosphate in SPE amino (NH2) columns, (c) eluation with HCl 0.5 N of dihydroxyacetone-phosphate followed by radioactivity counting and (d) estimation of the radioactivity thus trapped compared with that of enzymatically untreated aliquots of the same samples. No interferences from other 14C-labeled materials tested such as D-glucose, L-alanine, L-glutamine and D-beta-hydroxybutyrate were observed. This inexpensive and high-speed method can be applied in routine multiple estimations of glycerol-specific-radioactivity in biological samples in tracer metabolic studies.
D000818:Animals; D001711:Biotransformation; D002250:Carbon Radioisotopes; D004099:Dihydroxyacetone Phosphate / Q000032:analysis / Q000378:metabolism; D005990:Glycerol / Q000032:analysis* / Q000493:pharmacokinetics; D005993:Glycerolphosphate Dehydrogenase / Q000378:metabolism*; D051381:Rats; D012680:Sensitivity and Specificity
D002250:Carbon Radioisotopes; D004099:Dihydroxyacetone Phosphate; D005993:Glycerolphosphate Dehydrogenase; D005990:Glycerol