— I20 
also proved useless for the quantitative estimation of methyl ester of anthranilic 
acid, it had yet demonstrated, that the acetyl compound is not again 
separated out as such, but, on the contrary, in the animal 
organism passes over into the methyl ester, with loss of the 
acetyl-group. 
The other portion of the urine of the rabbits was slightly acidified with 
hydrochloric acid, carefully and slowly evaporated to the consistency of syrup, 
extracted with alcohol, the alcohol again driven off, and the residue dissolved 
in a fews drops concentrated hydrochloric acid and a small quantity of water. 
After adding alcohol, small crystals, glittering in the liquid, were precipitated. 
Examined under the microscope, they were found to be small lancet-shaped 
laminse, of which the majority possessed a fine sen'ated edge. As soon as the 
solution was heated, the crystals dissolved again. It was probably a question 
of hydrochloric methyl ester of anthranilic acid, which was precipitated from 
the aqueous solution when alcohol was added. After again slowly evaporating 
the alcohol and water, a white mass remained which was interspersed with 
greasy constituents. These could be readily removed from the white salt by 
means of alcohol. The presence of free methyl ester of anthranilic acid could 
be recognised by a slight blue fluorescence of the alcohol. The residue was 
dissolved in a weak solution of soda, and extracted with ether. The ether 
showed a strong blue fluorescence. After driving off the ether, there remained 
no crystals of the acetyl compound, but a thin oily film with an offensive 
odour was found to adhere to the glass. The small contents of the vessel were 
washed out with I to 2 cc dilute hydrochloric acid, and diazotised with sodium 
nitrite, and with this solution a solution of ^-naphthol was titrated. As was 
to be expected, the azo-dye was now obtained insoluble. 
AVith the urine of the dogs the diazo-reaction also gave positive results. 
But, as in the case of the urine from rabbits, the dye immediately dissolved. 
A portion of the urine was submitted to distillation. The distillate in the 
receiver had an extremely strong ammonia-like odour. As in the distillation 
of a solution of methyl ester of anthranilic acid the ester also passes over into 
the receiver, it was clear that in the distillate of the dog's urine it should be 
possible to detect methyl ester. And as a matter of fact, in the diazo-test 
the same red dye occurred again, which immediately dissolved in the liquid. 
The other portion of the urine of the dog was acidified with sulphuric 
acid, evaporated to one-fourth its volume, next rendered again strongly alkaline 
with soda liquor, and a few times extracted with ether. The ether showed a 
strong blue fluorescence. Crystals of methyl ester of acetyl anthran- 
ilic acid could not be obtained from the ether; that is to say, 
the acetyl compound had been completely decomposed. 
It follows that doses of i,og of the acetyl compound of 
methyl ester of anthranilic acid are tolerated by rabbits 
and dogs without any apparent or demonstrable disturbance. 
The acetyl-group is completely split off in the animal organ- 
ism, and the methyl ester of anthranilic acid can be de- 
tected in the urine by means of the diazo-reaction after 
having been isolated by extraction with alcohol. 
On bacteria the acetyl compound had no influence, as 
was shown by experiments carried out in the same manner 
as with sodium anthranilate (compare page ii6). The developing 
capacity of Bacteriu7n coli. Staphylococcus albus and Bacillus 
