Process for the Synthetic Production of Compounds resembling Peptones and Albumoses.
Year: 1898

page_text: Process for the Synthetic Production of Compounds resembling
Peptones and Albumoses.
We, LEO LILIENFELD, Doctor of Philosophy, and FRIEDRICH SILBERSTEIN, Doctor
of Law, of the Firm of Dr. Lilienfeld & Co., Chemical Manufacturers, of Ottakringer-
strasse, No. 20, Vienna, Austria, do hereby declare the nature of this invention to be
as follows:-
This invention relates to the synthetic production of compounds which fulfil the
chemical tests common to all kinds of peptones and albumoses. We employ as raw
materials a compound belonging to the class of phenols, or a mixture of such
compounds, also phosphorus-oxychloride, POC, or phosphoric chloride PC, or
phosphoric oxyde P₂O, or phosphorus-sulphochloride PSC, and an amido acid, or
a mixture of amido-acids, and form a product of condensation in the manner show.
by the following example.
About 20 grammes of phenol, C,H,OH, are melted, after which approximately the
same weight of amido-acetic acid C,HO,NH, is gradually added, and then from 60
to 120 grammes of phosphorus-oxychloride POCI, is slowly added to the mixture, and
the product is heated slowly and cautiously, until a sample, when diluted with water
and rendered alkaline by the addition of potash or soda solution, gives with a sulphate
of copper solution the well-known biuret reaction in a decided manner.
The reason for this reaction lies in the fact, that a body has been formed in the
mixture, which fulfils the chemical tests peculiar to the peptones.
The said body is not, however, in a free state, but as a hydrochlorate, owing to the
use of phosphorus-oxychloride.
In order to obtain the said body uncombined, the compound obtained is first dis-
solved in the mixture by the addition of alcohol and then precipitated from the same
by the addition of ether, which process, in the absence of sufficient cooling, may be
accompanied with the formation of esters.
Finally the product thus obtained (and which is strongly hygroscopic like the
natural peptones) is dried in vacuo.
The synthetic product may be obtained in a free state by various methods, for
instance by precipitation with tungstic-phosphoric acid and treatment of the pre-
cipitate in the known manner, and by forming metallic salts, such as double salts
of mercuric chloride, and decomposing the same in the known manner.
The smoothest way of splitting the compound consists in using sulphate of silver,
as described by Pal for the salts of natural peptones (Berichte Vol. XXV., page 1202
and Vol. XXXI., page 956).
The isolated hydrochlorate, or the entire mixture produced by the reaction, is
mixed with water and heated with an excess of sulphate of silver in a water bath,
whereby chloride of silver and the sulphate of the synthetic product is formed.
The solution of the sulphate is separated from the silver chloride by filtration, and
the minute quantity of the silver sulphate which had entered into the solution is
precipitated by means of sulphuretted hydrogen, the sulphide of silver thus formed is
CD
077

page_text: filtered off, and the excess of sulphureted hydrogen contained in the filtrate is driven
out.
The resulting solution contains the sulphate of the desired product, from which
barium sulphate and the pure synthetic product may be obtained by cautiously
adding barium hydroxide in drops.
With great caution it is always possible, to obtain a filtrate free from sulphuric
acid and baryta, which filtrate is then concentrated, and from which the free substance
(being insoluble in alcohol like free peptones) may be precipitated by alcohol.
By repeated solution and precipitation with alcohol the product is purified,
dehydrated by means of alcohol, subsequently treated with ether and finally dried in
vacuo.
The compound thus obtained gives the colouring and precipitating effects or
reactions which are peculiar to all peptones and albumoses, and the proportion
of C,N, HO found by analysis lie within the limits of those found for peptones and
albumoses.
Dated this 5th day of August 1898.
FAIRFAX & WETTER,
Agents for the Applicants.
Process for the Synthetic Production of Compounds resembling
Peptones and Albumoses.
We, LEO LILIENFELD, Doctor of Philosophy, and FRIEDRICH SILBERSTEIN, Doctor of
Laws, of the firm of Dr. Lilienfeld & Co., Chemical Manufacturers, of Ottakringer-
strasse, No. 20, Vienna, Austria, 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 the synthetic production of compounds which fulfil the
chemical tests common to all kinds of peptones and albumoses.
We take a compound belonging to the class of phenols, or a mixture of such
compounds, and condense it with an amido-acid or a mixture of amido acids, by means
of phosphorus oxychloride POC), or phosphoric chloride PC, or phosphoric oxide
P20, or phosphorus sulphochloride PSC.
As a rule the amido-acids are mixed in equal quantities. The quantity (by weight)
of the condensing medium to be added is from three to six times the quantity of an
amido-acid, and that of the phenol may vary from being equal to to being as much
as five times the quantity of an amido-acid. An excess of phenol is harmless, because
it would be washed off in the subsequent precipitation with alcohol.
The following examples will show more clearly the details of procedure :-
About 20 grammes of phenol (CHOH) are melted, after which approximately 4
the same weight of amido-acetic acid C₂H, O₂NH, is gradually added, and then from
60 to 120 grammes of phosphorus-oxychloride POCI, is slowly added to the mixture,
which is heated slowly and cautiously, until a sample, when diluted with water and
rendered alkaline by the addition of potash or soda solution, gives with a solution of
sulphate of copper the well-known biuret reaction in a decided manner.
4

page_text: The reason for this reaction lies in the fact, that a body has been formed in the
mixture, which fulfils the chemical tests peculiar to the peptones.
The said body is not, however, in a free state, but as a hydrochlorate, owing to the
use of phosphorus oxychloride.
In order to obtain the said body uncombined, the compound obtained is first
dissolved in the mixture by the addition of alcohol and then precipitated from the
same by the addition of ether, which process in the absence of sufficient cooling may
be accompanied with the formation of esters.
The solubility of the hydrochlorate in ethyl- or methyl- alcohol agrees completely
with the properties of the salts of natural peptones, as indicated by Paal
(Berichte, vol. XXV., p. 1202, and vol. XXXI., p. 956).
The substance precipitated is separated from the liquid by filtration or decantation,
then purified by repeated solution in alcohol, and precipitated from this solution by
means of ether, as has been recommended by Paal for the purification of the salts of
natural peptones.
Finally the product thus obtained (which is strongly hygroscopic like the natural
peptones) is dried in vacuo.
The synthetic product may be obtained in a free state by various methods, for
instance by precipitation with tungstic-phosphoric acid and treatment of the
precipitate in the known manner, or by forcing metallic salts, such as double salts of
mercuric chloride and decomposing the same in the known manner.
The smoothest way of splitting the compound consists in using sulphate of silver,
as described by Paal for the salts of natural peptones (Berichte vol. XXV., page 1202
and vol. XXXI. page 956).
The isolated hydrochlorate, or the entire mixture produced by the reaction, is mixed
with water and heated with an excess of sulphate of silver in a water bath, whereby
chloride of silver and the sulphate of the synthetic product are formed in theoretical
quantity.
The solution of the sulphate is separated from the silver chloride by filtration, and
the minute quantity of the silver sulphate which had entered into the solution is
precipitated by means of sulphuretted hydrogen, the sulphide of silver thus formed
Is filtered off, and the excess of sulphuretted hydrogen contained in the filtrate is
driven out.
The resulting solution contains the sulphate of the desired product, from which
barium sulphate and the pure synthetic product may be obtained by cautiously adding
barium hydroxide in drops.
With great caution it is always possible, to obtain a filtrate free from sulphuric
acid and baryta, which filtrate is then concentrated, and from which the free substance
(being insoluble in alcohol like free peptones) may be precipitated by alcohol.
By repeated solution and precipitation with alcohol the product is purified,
dehydrated by means of alcohol, subsequently treated with ether, and finally dried
in vacuo.
The compound thus obtained gives the colouring and precipitating effects or
reactions which are peculiar to all peptones and albumoses, and the proportions
of C,N, H,O found by analysis lie within the limits of those found for peptones and
albumoses,
2 parts of phenol and
3 parts of glassy phosphoric acid, previously pulverized, are intimately mixed and
heated over a paraffin or oil bath.
The molten mass thus obtained will after a short time commence to form bubbles
and become frothy, which indicates the beginning of the reaction, small samples of

page_text: the molten mass are now tested by the biuret reaction, and as soon as the reddish
colour (colour of onions) ceases to increase in intensity on the addition of copper
sulphate and lye, the reaction may be considered as being completed.
The molten mass is now dissolved in water and supersaturated with a strong
solution of barium hydroxide until the reaction reinains intensely alkaline, after
which the mass is allowed to stand a considerable time (from 12 to 24 hours) for
completely precipitating the barium phosphate.
The solution is now filtered off the separated bariura phosphate, and the excess of
the barium hydrate contained in the filtrate, is exactly precipitated by means of
sulphuric acid, until a filtered-off sample will not give any precipitate either with
barium hydroxide or with sulphuric acid.
The mixture is again filtered, then concentrated by evaporation over a water bath,
and alcohol is added, to precipitate the peptone which has been produced.
The purification of the latter is effected by repeatedly forming a concentrated
aqueous solution and precipitating by the addition of alcohol, finally the product is
thoroughly washed with absolute alcohol, exhausted with ether and dried in vacuo.
As regards the use of phosphoric acid, it should be observed that the ordinary
glassy phosphoric acid of commerce yields a product rich in ashes, owing to the
contents of sodium derived from the porcelain vessels, which defect may be avoided
by using metaphosphoric acid reduced in platinum crucibles and therefore free from
sodium.
As this metaphosphoric acid has a very thick consistency, it must be introduced
first in the retorts used for the reaction, after which the other ingredients, intimately
mixed together, are added.
It should be observed, moreover, that asparagin alone and various other amido-acids
in combination with each other may be similarly treated with phosphoric oxychloride,
glassy phosphoric acid, and their substitutes, in which case similar albuminous
products are obtained, but as the process remains the same, it will not be necessary
to describe further examples.
Instead of the amido-acids used in the two examples mentioned above, it is possible
to employ separately or in combination those amido-acids which are produced by the
splitting of proteids, such as tyrosin, leucin, amido-valerianic acid, in short all amido-
acids of the fatty and of the aromatic series.
Experiments on the decomposition of the new products by means of hydrochloric
acid, sulphuric acid, and barium hydroxide have given the following results:-
A compound or preparation obtained from asparagin and para-amido-benzoic acid
was split up by means of hydrochloric acid and stannous chloride according to the
well-known method of Hlasiwetz and Habermann. The product of decomposition
contained large quantities of asparagin acid and a small quantity of tyrosin, the latter
being recognisable by the shape of its crystals, the tyrosin reactions, and elementary
analysis.
When the compound was split up with barium-hydroxide according to Schützen-
berger, the presence of the above-mentioned amido-acids in addition to other products
of oxidation, such as oxalic acid, benzoic acid, carbonic acid, and ammonia, was
recognisable in the product of decomposition.
Finally the compound was decomposed by melting it with caustic potash, whereby
indol was split off, which could be recovered in a perfectly pure condition.
The amido-compounds obtained by splitting up the new products depend, of course,
on the amido-acids which have been employed for the condensation.
Having now particularly described and ascertained the nature of our said inven-
tion, and in what manner the same is to be performed, we declare that what we
claim is
:
1. A process for the synthetic production of compounds resembling natural peptones
and albumoses, which consists in condensing a compound belonging to the class of
phenols, or a mixture of such compounds, with an amido-acid or a mixture of amido-

page_text: acids, by means of phosphorus oxychloride POCI, or phosphoric chloride PC or
phosphoric oxide P₂Os, phosphorus sulphochloride PSC,, or ineta-phosphoric acid.
2. The production and separation of a compound resembling natural peptones or
albumoses in the manner substantially as described.
3. The new products obtainable substantially as described.
FAIRFAX & WETTER,
Agents, 37 and 39, Essex Street, Strand, London.