Enzymatic determination of carbon (14C)-labeled glycerol in biological samples |
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Affiliation: | 1. Department of Hotel Management and Catering Technology, Birla Institute of Technology, Ranchi, India;2. Department of Pharmaceutics, Seemanta Institute of Pharmaceutical Sciences, Mayurbhanj, India;3. Department of Pharmacy, Palamau Institute of Pharmacy, Chianki, India;1. Institute of Municipal and Environmental Engineering, College of Civil Engineering, Huaqiao University, Xiamen, Fujian, 361021, PR China;2. State Key Laboratory of Urban Water Resource and Environment, School of Environment, Harbin Institute of Technology, Harbin, Heilongjiang, 150090, PR China |
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Abstract: | 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-β-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. |
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