A method for quantitative estimation of glycerol and total carbohydrate in biological samples is described. The samples, deproteinized with cold acetone or trichloroacetic acid, were treated in the cold with 10 vols. of 0.75% (w/w) anthrone in 84% (w/w) sulphuric acid, and then heated 10 min at 100 degrees C. Absorbance at 590 nm was used for evaluation of total carbohydrate content in the sample. The absorbance at 510 nm was used for the combined carbohydrate and glycerol estimation. This latter observation leads to the determination of the glycerol content of the sample because the carbohydrate interference is known from the data obtained at 590 nm. Uses of this method to determine glycerol and carbohydrate content in lipids of chicken egg-yolk samples are presented.