Improved Spectrophotometric Determination of Glycerol and Its Comparison with an Enzymatic Method

A chemical method for the determination of glycerol was developed and compared to an enzymatic assay for sensitivity and reproducibility. The chemical assay is based on glycerol oxidation to formaldehyde and subsequent reaction with chromotropic acid to yield a colored product. With this method, as little as 5 μg of glycerol/ml can be detected. The enzymatic assay is based on enzymatic glycerol phosphorylation followed by glycerol phosphate dehydrogenation by nadide (nicotinamide adenine dinucleotide). The reduced nadide is used to reduce iodonitrotetrazolium violet to its colored formazan product. The enzymatic method can be used to determine 50 μg of glycerol/ml in aqueous samples.