Quantitative gas-liquid chromatographic determination of free glycerol in blood serum.

Quantitative gas–liquid chromatographic determination of free glycerol in blood serum was accomplished through the use of butane-1,4-diol as internal standard. After removal of proteins with phosphotungstic acid, the glycerol and butanediol were acetylated for 1 hr with acetic anhydride. The acetates thus formed were extracted with diethyl ether, the solvent was removed, and the butanediol diacetate and glycerol triacetate were separated on a column with butanediol succinate as stationary phase at temperatures programmed from 150 to 190°. Normal sera were found to contain 0.4–1.2 mg of glycerol per 100 ml, the estimation having a range of ±5–10%. The method can be exploited to determine as little as 0.01 mg of glycerol per 100 ml when a sensitive flame ionization detector is used.