Abstract
This chapter focuses on the enzymatic determination of glycerol. The determination of glycerol in biological material, food, and industrial products usually requires an extensive purification of the sample under investigation to remove interfering contaminants. Even after this time consuming purification, the chemical methods of analysis are liable to error, especially when the amounts of glycerol present, as in a serum, body fluids and tissues, are very small. The enzymatic method for the determination of glycerol is specific. Therefore, purification of the sample is unnecessary. The enzymatic method is based on the principle that glycerol is phosphorylated by glycerokinase (GK) and adenosine triphosphate (ATP) to give L-(−)-glycerol-1-phosphate. The two reactions are presented in this chapter. L-(−)-glycerol-l -phosphate formed is oxidized with α-glycerophosphate dehydrogenase (glycerol-1-phosphate dehydrogenase, GDH) and diphosphopyridine nucleotide (DPN). The amount of DPNH formed is equivalent to the amount of glycerol present. The equilibrium of the indicator second reaction, which lies far to the left, is displaced in the required direction by working at pH 9.8 and trapping the dihydroxyacetone phosphate (DAP) with hydrazine.
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