POISONOUS ALKALOIDS AND ALBUMINOIDS. 365 



separate from anthrax cultures, in addition to a poisonous 

 alkaloid, two albumoses, which apparently represent slightly 

 different stages in the transformation of albumen into pep- 

 tone) : it may be well also to give the characteristic reactions 

 that are obtained with these albumoses. 



Hankin's method is as follows : A I per 1,000 pure solution of Liebig's 

 extract of meat is carefully sterilized by being heated in a small sterilizer for 

 two or three hours on two or three successive days ; to the fluid so sterilized 

 a quantity of pure fibrin is added and the whole is again sterilized "by 

 repeated heating to boiling point for a short time only on each occasion " ; 

 if this is heated for a longer period a considerable quantity of the fibrin is 

 digested and converted into peptone, a substance that would interfere very 

 considerably with the after-examination, in addition to which the anthrax 

 bacillus would have little material left on which to exert its " peptonizing " 

 function. This is inoculated with blood from an animal that has died from 

 anthrax and is "kept at the ordinary temperature. " The cultivation is 

 allowed to go on for a week, at the end of which time the albumose is 

 extracted. If the flask be kept at the temperature of the body, 37 C, the 

 transformation of the albumose into the peptone goes on much more rapidly. 

 To separate the albumose the culture fluid is first acidulated with acetic acid, 

 and then thoroughly saturated with ammonium sulphate, when there is thrown 

 down a bulky precipitate of albumose. In order to concentrate the solution, 

 instead of using Brieger's method of evaporation in vacuo or under pressure at 

 a low temperature he resorted to the method of diffusion or dialysis. A quan- 

 tity of thymol, to prevent putrefaction, is added to a watery solution of the 

 albumose, and the whole is placed in a parchment sausage skin which is im- 

 mersed in a foot glass full of methylated spirit. The spirit can be changed 

 after some hours if it is necessary to prolong the process, but this is not 

 usually necessary. " In this way," says Hankin, " I have been able to bring 

 400 cubic centimetres of albumose solution down to 100 c.cm. in the course 

 of a single night, at the ordinary temperature without risk to the albumose 

 or trouble to myself. The concentrated solution is then poured into 

 absolute alcohol, which precipitates the albumose and removes any im- 

 purities that might be derived from the methylated spirit. This prolonged 

 treatment with alcohol will tend to remove any free ptomaines or other 

 substances soluble in alcohol." In order to remove any ferments that are 

 capable of acting along with the albumose, Hankin, following Roux and 

 Yersin, adds a quantity of lime water to his solution, so that, on the addi- 

 tion of a solution of phosphoric acid, a gelatinous precipitate of calcium 

 phosphate is produced, in the formation of which ferments are usually 

 entangled and carried down, and on filtration a purer solution of albumose 

 is obtained. 



Brieger and Fraenkel have adopted Hankin's method in the 

 preparation of their toxalbumens, but instead of dialyzing, 

 they evaporate down in vacuo at a low temperature until 

 the liquid has been reduced to less than a fourth of its 

 original quantity. They again wash in alcohol and filter. 



