The determination of glycerol concentration and specific radioactivity in rat blood
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Add time:09/06/2019 Source:sciencedirect.com
A method has been developed to measure the specific radioactivity of blood or plasma glycerol under conditions when glycerol label is only a very small fraction of total nonionic label. Glucose is removed from a deproteinized extract by glucose oxidase (EC 1.1.3.4.) and catalase (EC 1.11.1.6.), and the concentration of other sugars is reduced using hexokinase (EC 2.7.1.1.). The products are removed by ion-exchange chromatography. The glycerol is then estimated by conversion to glycerol-1-phosphate which is separated on a borate column for measurement of radioactivity, using [3H]glycerol as a marker and to estimate recoveries. The method was tested on blood from rats given [14C]glucose or [14C]bicarbonate. In the initial blood samples less than 0.1% of the nonionic label was in glycerol. Reproducibility was 4%.
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