Improvements in the Process of Extracting Nuclein from Yeast and in Preparing Compounds of such Nuclein with Iron Silver, and Mercury, Soluble in Water.
Year: 1899

page_text: Date of Application, 5th Apr., 1899-Accepted, 15th July, 1899
Improvements in the Process of Extracting Nuclein from Yeast and
in Preparing Compounds of such Nuclein with Iron Silver, and
Mercury, Soluble in Water.
A communication by Dr. KARL SCHWICKERATH, of the City of Detroit, County of
Wayne, and State of Michigan, United States of America, Chemist.
I, WILLIAM PHILLIPS THOMPSON, F.C.S., M.I.M.E., of the Agency for Foreign
Patent Solicitors, 6, Lord Street, Liverpool, and 6, Bank Street, Manchester, both
in the County of Lancaster, 118, New Street, Birmingham, in the County of War-
wick, and 322, High Holborn, in the County of Middlesex, Civil Engineer, do
hereby declare the nature of this invention and in what manner the same is to be
performed, to be particularly described and ascertained in and by the following
statement:
This invention relates to a process for the economical separation of pure nuclein
from yeast, and also to processes for the combination of such nuclein into new
soluble iron, silver and mercury compounds, not heretofore known; and it con-
sists in the methods and products hereinafter described and claimed.
Heretofore the separation and production of pure nuclein from yeast has been
quite difficult, especially in consequence of the existence of a slimy or gummy-like
brown substance in the yeast which adheres tenaciously to the nuclein after its
separation. My correspondent has succeeded in overcoming this difficulty in a
simple manner, namely, in its oxidation in a neutral or faintly alkaline solution
with potassium permanganate, whereby said brown slimy substance is so changed
that it combines with the separated manganese hydroxide, forming an insoluble
compound, which is readily separated by filtration, while from the filtrate pure
nuclein is precipitated by acidulated alcohol.
The procedure for the manufacture of nuclein from yeast therefore would be as
follows:-
Fifty kilogrammes of ordinary brewer's yeast are diluted with fifty litres of
water until a uniform mixture results; then are added two kilogrammes of sodium
hydroxide. Allow to stand for about one hour, with frequent stirring, then heat
to 40 or 50 degrees Centigrade, and add sufficient acetic acid until a faintly acid
reaction manifests itself, then the heating is continued until 75 degrees are
reached, whereupon so much of a concentrated sodium carbonate solution is added
until neutral reaction. The thus separated albuminous substances, etcetera, are
filtered off, and from the filtrate the crude nuclein is precipitated by acidulated
alcohol. This nuclein separates in the form of a fine, flocculent, whitish mass,
this is washed with alcohol, finally pressed and the residue dried.
The thus obtained crude nuclein still contains a considerable quantity of the
above mentioned slimy substance, which is removed by the following method:
500 grammes of crude nuclein are macerated with 10 litres of water until the

page_text: nuclein is uniformly diffused through the mass, then, under stirring, about 500
cubic centimetres of a 5 per cent potassium permanganate solution are gradually
added. The liquid is then heated to about 70 to 75 degrees Centigrade, whereupon
the insoluble manganese compound separates in floccules, which can be readily
removed by filtration. From the thus obtained clear filtrate the pure nuclein is
separated by the addition of acidulated alcohol. Subsequently the precipitate
is washed with quantum sufficit alcohol, pressed and finally dried. The latter pro-
cess is done to better advantage in vacuo, since in this way a preparation of
nearly white appearance results.
Since yeast is a product which varies somewhat in the proportions of its com-
position, acidity, etcetera, it is obvious that it is almost impossible to give in the
above described process the exact proportions.
The nuclein obtained by the above described process is employed for the manu-
facture of compounds of the same with silver, mercury and iron, soluble in water.
A. PROCESS FOR THE MANUFACTURE OF SILVER-NUCLEIN COMPOUND.
250 grammes of nuclein are dissolved, under the addition of 100 cubic centi-
metres of a 20 per cent solution of carbonate of soda in 5 litres of hot distilled
water, then are gradually added, with stirring, 220 cubic centimetres of a 20 per
cent solution of silver nitrate. After the addition of 10 litres of alcohol the new
compound of nuclein and silver is precipitated in the form of a white mass. The
latter is washed with alcohol, pressed, finally dried, and powdered. (The powder-
ing is done to the best advantage in vacuo, whereby a product of lighter appear-
ance is obtained.)
This nuclein-silver compound exists in the form of a light brownish-white pow-
der, contains about 10% silver, and is soluble in water, more readily in hot water.
Its watery solution shows a faintly alkaline reaction, is possessed of a strong
metallic taste and is not changed by boiling. The silver exists here in peculiar
combination, so that it is not precipitated by the ordinary reagents, neither does
sodium chloride nor albuminous solutions cause any change in the nuclein-silver
solution. These properties in particular might render the nuclein-silver very
valuable as an antiseptic for the treatment of local inflammations.
B. PROCESS FOR THE MANUFACTURE OF MERCURY-NUCLEIN COMPOUND.
If one dissolves nuclein prepared by the above described process in water and
adds subsequently freshly precipitated mercuric oxide, the latter enters gradually
into solution more quickly with the aid of heat. From the thus obtained solution
the new compound of nuclein and mercury can be precipitated by alcohol and the
addition of a concentrated solution of a neutral salt (sodium chloride, sodium
sulphate, etcetera) in the form of a whitish flocculent precipitate. It is, however,
not necessary to employ freshly precipitated mercuric oxide, but the same results
can be achieved if to a solution of nuclein and water is added an excess of alkali
(sodium hydroxide, sodium carbonate, etcetera) and subsequently is added a calcu-
lated amount of a mercuric salt solution (mercuric chloride). In this case the
mercuric oxide freed from the mercuric oxide solution combines in statu nascendi
with the nuclein, and from the resulting clear solution the nuclein-mercury com-
pound is precipitated in the same manner with alcohol, and the small quantity
of a neutral salt is obviated.
The manufacture of the mercury-nuclein compound is accomplished as follows:-
500 grammes of pure nuclein are dissolved in 10 litres of hot distilled water,
with the addition of about 600 cubic centimetres of a 20 per cent sodium carbonate
solution, then add gradually to the solution 90 grammes of mercuric chloride
in sufficit quantum water, with stirring. From the thus obtained clear solution
the new compound of nuclein and mercury is subsequently precipitated by the
addition of about 12 litres of alcohol and a sufficient quantity of a concentrated
sodium sulphate solution. Of the latter so much is added until the supernatant

page_text: arcononic quid appears ainiost clear. Ine separated mercury uciein is then
washed with alcohol, pressed and dried, (the latter process preferably in vacuo).
The mercury-nuclein compound thus obtained contains about 10 per cent. mer-
cury, is of a metallic taste, perfectly soluble in water, more readily so in hot
water. Albuminous liquids or alkalies do not cause any change when added to a
solution of nuclein-mercury. The nuclein-mercury might prove of great value
in the treatment of syphillis and local infectious diseases.
C. PROCESS FOR THE MANUFACTURE OF IRON-NUCLEIN COMPOUND.
The method of preparing this compound is similar to that of the nuclein-mer-
cury compound. To a solution of nuclein containing an excess of alkali is added
a calculated quantity of a ferric salt. The iron hydroxide formed by the presence
of the alkali combines in statum nascendi with the nuclein, a dark red brown solu-
tion of iron-nuclein results, from which solution, by means of alcohol and a small
quantity of neutral salt (sodium chloride, sodium sulphate, etcetera) the new iron
compound is precipitated in the form of a cinnamon-brown flocculent mass. Hence
the process would be as follows;-
-
300 grammes of nuclein are dissolved in 6 litres of hot distilled water, with
the addition of about 250 cubic centimetres of a 20 per cent sodium carbonate
solution, then is added gradually, and under stirring, a calculated amount of an
iron chloride solution (105 grammes of iron chloride solution United States Phar-
macopoeia) containing 13 per cent. metallic iron) upon the addition of which amount
of iron chloride solution a precipitate is formed, which, however, subsequently
dissolves. The final result is a dark-reddish brown solution, from which the new
compound of nuclein and iron is separated by the addition of about 12 litres of
alcohol and a small quantity of sodium chloride solution. The flocculent, cinna-
mon-brown precipitate is washed with alcohol, pressed and finally dried, prefer-
ably in vacuo.
This iron-nuclein compound exists in the form of a cinnamon-brown powder
and contains from 4 to 5 per cent metallic iron. It is tasteless, readily soluble in
hot water, with neutral reaction, and is scarcely affected by artificial gastric juice.
No reaction is caused in a solution of the iron-nuclein and water by the addition of
alkalies, potassium sulphocyanide, or ferrocyanide, ammonium sulphide colors
the solution green but does not cause a direct precipitation of black iron sulphate.
The peculiar organic combination of the iron and nuclein compound might
render it a valuable therapeutic agent.
Having now particularly described and ascertained the nature of the said
invention and in what manner the same is to be performed, as communicated to
me by my foreign correspondent, I declare that what I claim is;-
1. The herein described process for the manufacture of nuclein, consisting sub-
stantially in
(a) The separation of crude nuclein from yeast, consisting in extracting
the impure nuclein from yeast by bringing into solution the albuminous sub-
stances and the crude nuclein by means of an alkali, and precipitating the
albuminous substances by the addition of acetic acid and subsequently heating
to 75 degrees Centigrade; removing the albuminous substances by filtration
and by precipitating the crude nuclein from the thus obtained filtrate by
acidulated alcohol.
(6) In treating the crude nuclein in a watery neutral or faintly alkaline
solution with dilute potassium permanganate solution, separating the thus
obtained precipitate by filtration, and the separation of the pure nuclein from
the filtrate by acidulated alcohol.
2. The herein described process for the manufacture of compounds of nuclein
with silver, mercury and iron, soluble in water, consisting substantially in
(a) The addition of a certain quantity of a silver oxide salt solution, mer-
curic oxide salt solution, or iron oxide salt solution, respectively, to a solu-
tion of nuclein in water in the presence of a certain amount of free alkali.

page_text: (b) The separation of the thus formed silver, mercury or iron nuclein con-
pound respectively, by precipitating with alcohol under the addition of a
small amount of a neutral salt.
W. P. THOMPSON & Co.,
Agents.