A Colorimetric Determination of Blood Acetoin” by W. W. Westerfeldt

Most of the methods described for the determination of acetoin and biacetyl are based on the original Lemoigne-van Niel procedure (1, 2), which depends upon the formation of insoluble nickel dimethylglyoxime when biacetyl is treated with hydroxylamine in the presence of a nickel salt; acetoin is similarly determined after oxidation to biacetyl by heating with ferric chloride. In a recent modification Stotz and Raborg (3) have employed a calorimetric determination of the nickel in the precipitate instead of the usual gravimetric analysis, and the sensitivity of the method has thereby been greatly increased. A polarographic method for the determination of acetoin and biacetyl has also been described by Greenberg (4). The method herein described was developed because none of the previous methods was sufficiently sensitive for our purpose; its chief advantages are simplicity and sensitivity. It is based on the color reaction originally discovered by Voges and Proskauer (5), and later shown by Harden and Norris (6, 7) to be due to the reaction between biacetyl and a guanidino group in the presence of alkali. Attempts to increase the sensitivity of this reaction led to the addition of creatine by O’Meara (8) and a-naphthol by Barritt (9). While the original Voges-Proskauer reaction could not be developed into a satisfactory quantitative method, the Barritt modification was found to be suitable. In this laboratory it was first tried by W. E. Knox, and a similar method has recently been described by Eggleton et al. (10). The chemical reactions involved in the development of the red color have been studied (10-12) but not fully elucidated. Details of the procedure for color development and the method for determining acetoin in blood are described.