498 Dr. A. Harden and Mrs. D. Norris. Production of [Nov. 22, 



added. At the end of this time and occasionally during the progress of the 

 experiment sub-cultures were made to show that the organism in question 

 was still alive and uncontaminated. In two experiments the whole of the 

 acetaldehyde had not been used up, so that the liquid obtained by distillation 

 of the culture medium was strongly reducing to Fehling's solution. In two 

 other cases sufficient time was allowed to elapse between the last addition of 

 aldehyde and the examination of the products to ensure the complete 

 removal of this factor. 



In these two cases the distillates obtained had no reducing power. In 

 every experiment the distillate was tested by means of the Yoges and 

 Proskauer reaction for acetylmethylcarbinol, and in no case could any trace 

 of this be detected. 



The liquid was further examined for butylene glycol, which was in every 

 case found to be present, though in extremely small quantities. It was 

 detected as described above by the formation of diacetyl, this substance 

 being proved to be present in every case by the positive results given by the 

 Voges and Proskauer reaction after oxidation of the culture distillate. 



In one case sufficient osazone was prepared from the diacetyl for a deter- 

 mination of the melting point, which was found to be 244° C. Among the 

 products of the reaction were also found ethyl alcohol, acetic acid, and some 

 succinic acid. Lactic acid was not present. These products were estimated 

 in the manner previously described by Harden (13) with a few slight 

 modifications. In the estimation of the alcohol any unchanged acetaldehyde 

 present was removed by oxidation with moist silver oxide. The acetic acid 

 was determined by the method of Alacnair (14). 



The table below shows the results obtained in three typical experi- 

 ments : — 



Table III. 





1. 



2. 



3. 





6 -07 grm. 



8 -0 grm. 



7 -6 grm. 



Total formic acid added (as Ca formate) ... 



6-07 ,„ 



Nil 



7-6 „ 





grm. 



grm. 



grm. 





6-5 



4-02 



4-5 





2 5 



1 -38 



1-3 





Not estimated 



1 "25 



0-12 





+ 



0-677 



0-109 



Prom the point of view of the production of butylene glycol the presence 

 of calcium formate appears to be detrimental, and it is also interesting to 

 note the somewhat large quantity of succinic acid produced in Experiment 2, 

 which also gave the largest yield of glycol. 



