98 HISTOEY AND CAUSE OF THE COCONUT BUD-EOT. 



the separately funnel. The ethereal extract was evaporated to a 

 syrup and a small residue was left, thus suggesting the presence of 

 either lactic, oxalic, or succinic acid. To the extract was added and 

 thoroughly mixed 100 c. c. of water. Then an excess of zinc oxid 

 was added, and the mixture was heated nearly to boilmg and filtered. 

 To 6 c. c. of the filtrate were added 4 c. c. of concentrated sulphuric 

 acid, and the whole was warmed to 75° C. The absence of any crim- 

 son color mdicated the absence of glycocholic, taurochohc, or cholic 



acid. 



To another portion of the filtrate was added Lugol's iodin, and the 

 absence of any blue color here also indicated the absence of any 

 chloric acid. Another portion of the filtrate was acidified with hydro- 

 chloric acid. Ammonia was added in slight excess, and the excess 

 then boiled oflF. A solution of cobalt nitrate was added, and absence 

 of any lactic acid was indicated by the lack of a violet color. 



Another portion of the filtrate was evaporated to dryness and then 

 dissolved in 10 c. c. of hot water and allowed to crystallize, but there 

 resulted only a yellowish amorphous mass which indicated the absence 

 of any crystals of zinc lactate. 



The residue left from the filtermg after the addition of zinc oxid 

 was dissolved in hydrochloric acid on the filter, and then a portion 

 tested for oxalic acid as follows: 



It was neutralized with ammonia until faintly alkaline, and then a 

 solution of calcium chlorid was added. There was no resultant white 

 precipitate of calcium oxalate, which indicated the absence of oxalic 



acid. 



Another portion of this filtered residue was neutrahzed with am- 

 monia, and the excess boiled off. To a portion of this was added 

 ferric-chlorid solution on a glass rod. A distinct red-brown colora- 

 tion showed the presence of succinic acid. The absence of buff 

 coloration indicated the lack of any benzoic or hippuric acid in the 

 solution ; the absence of a violet coloration indicated the lack of any 

 salicyhc acid; and the absence of an inky coloration indicated the 

 lack of tannic or gallic acid in the solution. 



This last series of tests for oxalic, succinic, benzoic, hippuric, sali- 

 cyhc, tannic, and lactic acids was repeated, and the same results 

 obtained. 



So far as this analysis shows, only succinic acid was certainly 

 demonstrated to be present, and possibly formic. It has been shown 

 by other investigators ' that in the case of Bacillus coli, acetic, formic, 



I M. Scmel reported (Archives M6dlcales Beiges, ser. 4, vol. 1, 1893, pp. &-33) finding lactic, acetic, and 

 formic adds. 



Leo. F. Reltger (Studies from the Rockefeller Institute for Medical Kesearch , vol. 1, 1904, pp. 284-293) 

 reports finding in egg-meat cultures of Bacillus cnli, indol, skatol, phenols, aromatic oxyacids, skatol- 

 ciirbonic acid, leucln, tyrosin, trylophan, hydrogen disulphid, mercaptan, albumoscs, and peptones. 



Arthur Harden (.Tournal of Hygiene, vol. 5, lOO.I, pp. 488-493) states that he found lactic acid, acetic acid, 

 and a small amount of succinic acid present in glucose cultures of Bacillus coli. 

 229 



