An Improved Process for the Production of Purified Sterilised Liquid Hæmoglobin.
Year: 1894

page_text: Date of Application, 27th June, 1894
Complete Specification Left, 13th Mar., 1895-Accepted, 24th May, 1895
PROVISIONAL SPECIFICATION.
An Improved Process for the Production of Purified Sterilised
Liquid Hæmoglobin.
I ADOLF HOMMEL of Industriequartier, Zurich in the Republic of Switzerland
Doctor of Medicine do hereby declare the nature of this invention to be as
follows:--
This invention is based upon the observation by me that blood only exhibits its
well-known and marked tendency to decompose, so long as it contains free oxygen,
ammoniacal decomposing agents (such as urea, hippuric acid and the like) as well
as the usual volatile fatty acids; but that, whenever these components are removed
therefrom, the mass of the red blood corpuscles in solution are no more liable to
decomposition than any other organic bodies. Having ascertained this fact, I have
taken it for a basis for my improved method of treating blood, which consists in
adding an antiseptic medium to it, before it commences to change its condition into
that of hæmoglobin and is freed from gas, such antiseptic agent being volatile, and
capable of combining with the foreign decomposing agents or bodies above referred
to, of retarding their decomposing action, and of eventually separating from the .
blood again in conjunction with the said decomposing agents. To accomplish this
object, it is necessary that the antiseptic agent employed should be capable of
becoming volat:lized in racun together with the said foreign bodies, in such a
thorough manner that there should remain no vestige or trace thereof, when the
bæmoglobin has attained the desired state of perfect purity and solubility.
The process is carried out in the following inanner: Fresh fibreless animal blood
is operated upon either in its original complete state, or after having been treated
in centrifugal apparatus, in which case the red blood corpuscles, settled at the
bottom of such apparatus, are finally separated from the serun by very carefully
decanting off the latter. The mass under treatment is then to as full an extent as
possible freed from gas at a high pressure by means of a vacuum but without the
assistance of any artificial heating. This process take a considerable time to
complete, but by such preliminary removal of the gas, the subsequent treatment of
the blood is greatly facilitated. On being treed from gas, the blood has about
20 per cent. of water (preferably boiled, and then allowed to cool down to about 20°
Centigrade) added to it, such water being mixed with a volatile antiseptic substance.
The object of this addition of water is more quickly to dissolve the red blood
corpuscles, with the addition of the antiseptic agent is made, as before stated, with
a view to preventing the decomposition of the blood. Among the antiseptics
suitable for this purpose creosote should be mentioned in the first instance (it being
diluted to a very considerable extent, say, in the proportion of about one to three
hundred of the diluent. Another suitable agent is diluted alcohol. In the case of
alcohol, a convenient proportion is 35 parts of alcohol to 100 parts of water, so that
the final mixture may contain in all, say, 7 per cent. of alcohol. The mixture is"
then evaporated in vacuo at a high pressure and at as low a temperature as possible,
varying between, say, 30 and 40 degrees Centigrade.
After the removal of the
20 per cent. of water, the operation is interrupted. Generally speaking at this
juncture, the red blood corpuscles have not had time to completely dissolve, and
therefore it is usually found necessary to renew the admixture of water. In fact.
this admixture has, in certain cases, to be repeated several times.
:

page_text: The blood being more liable to decompose at the beginning of the evaporating
process, than at any other period, that is to say, while the ammoniacal decomposing
agents (urea, hippuric acid and the like) and the volatile fatty acids are still
contained in the blood, the temperature may be raised after the first break in the
operation, above referred to. Where creosote is added, it is even desirable to
attain a state of coagulation, such as that of albumen; vict. from 55 to 60 degrees
Centigrade, inasmuch as it is far more difficult to remove creosote from the mixture,
than it is to separate alcohol. The removal of the urea and hippuric acid from the
blood is effected by hydration, or the conversion of the mixture under treatment
into volatile compounds. The urea is thus decomposed into neutral carbonate of
CHNO + H₂O = C₂H₂O₂ + C₂H,NO,
Benzoic acid is volatile in the vapours rising from the water, while glycol is
retained in the mixture. Its action is a preserving one, though of course, the
quantity, of glycol, present is generally so small that its action is scarcely worth
considering. The product is evaporated down to about 60 per cent. of the original
quantity of blood (the mixture above referred to being understood to consist of
80 1er cent. of blood and 20 per cent. of water). If, on attaining this percentage
traces of the antiseptic agent should still be found water should be added over and
over again until the last remnants of the antiseptic agent are completely
eliminated.
The final product thus obtained will be a sufficiently liquid, clear, deep red
substance, which, as has been ascertained by bacteriological tests, is perfectly free
from the micro-organisms circulating in the blood. The insipid taste peculiar to
blood may be counter-acted by the addition of suitable chemicals, chemically pure
glycerine being one of the best, since it is of considerable value also as a means for
preserving the preparation in an unimpaired condition.
Dated the 27th day of June 1894.
JOHNSONS & WILLCOX,
47, Lincoln's Inn Fields, London, Agents.
COMPLETE SPECIFICATION.
;
An Improved Process for the Production of Purifled Sterilised
Liquid Hæmoglobin.
I, ADOLF HOMMEL of Industriequartier Zurich in the Republic of Switzerland
Doctor of Medicine do hereby declare the nature of my 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 is based upon the observation by me that blood only exhibits its
well known and marked tendency to decompose so long as it contains free oxygen
and ammoniacal decomposing agents (such as urea, hippuric acid and the like) as
well as the usual volatile fatty acids, but that whenever these components are
reinoved therefrom, the mass of the red blood corpuscles in solution are no more
liable to decomposition than are any other organic bodies. Having ascertained this

page_text: fact, I have taken it for a basis for my improved method of treating blood which
consists in adding an antiseptic medium to it, before it cominences to change its
condition into that of hæmoglobin and when freed from gas; such antiseptic agent
being volatile and capable of combining with the foreign decomposing agents or
bodies above referred to of retarding their decomposing action, and of eventually
separating from the blood again in conjunction with the said decomposing agents or
bodies. To accomplish this object, it is necessary that the antiseptic agent employed
should be capable of becoming volatilized in vacuo together with the said foreign
bodies, in such a thorough manner that there should remain no vestige, or trace,
thereof when the hæmoglobin has attained the desired state of perfect purity,
tastelessness and solubility.
The process is carried out in the following manner: Fresh animal blood from
which the fibre has been extracted or removed is operated upon either in its original
complete state, or after having been treated in centrifugal apparatus, in which case
the red blood corpuscles, settled at the bottom of such apparatus are finally
separated from the serum by very carefully decanting off the latter.
The mass is to as full an extent as possible, freed from gas by means of a vacuum
but without the assistance of any artificial heating.
This process takes a considerable time to complete but by such preliminary
removal of the gas, the subsequent treatment of the blood is greatly facilitated.
On being freed from gas, the blood has about 20 per cent. of water (preferably water
which has been boiled and then allowed to cool down to about 20° Centigrade)
added to it, such water being mixed with a volatile antiseptic substance. The
object of this addition of water is more quickly to dissolve the red blood corpuscles,
while the addition of the antiseptic agent is made, as before stated, with a view to
preventing the decomposition of the blood. Among the antiseptics suitable for
this purpose creosote should be mentioned in the first instance (it being diluted to
a very considerable extent say in the proportion of about one of creosote to three
hundred of the diluent). Another suitable agent is diluted alcohol. In the case
of alcohol a convenient proportion is 35 parts of alcohol to 100 parts of water, so
that the final mixture may contain in all, say, 7 per cent. of alcohol. The mixture
is then evaporated in vacuo at a high relief of pressure and at as low a temperature
as possible varying between, say, 30 and 40° Centigrade. After the removal of
the 20 per cent. of water the operation is interrupted. Generally speaking, at this
juncture the red blood corpuscles have not had time to completely dissolve, and
therefore it is usually found necessary to renew the admixture of water. In fact,
this admixture has in certain cases, to be repeated several times.
The blood being more liable to decompose at the beginning of the evaporating
process, than at any other period; that is to say, while the ammoniacal decomposing
agents (urea, hippuric acid and the like) and the volatile fatty acids are still
contained in the blood, the temperature may be raised after the first break in the
operation, above referred to. Where creosote is added it is even desirable to
attain a state of coagulation such as that of albumen (viz.: temperature
from 55° to 60° Centigrade) inasmuch as it is far more difficult to remove creosote
from the mixture, than it is to separate alcohol. The removal of the urea and
hippuric acid from the blood is effected by hydration, or the conversion of the
mixture under treatment into volatile compounds. The urea is thus decomposed
into neutral carbonate of ammonia :

page_text: Benzoic acid is volatile in the vapours rising from the water, while glycocoll is
retained in the mixture. Its action is a preserving one though of course the
quantity of glycocoll present is generally so small that its action is scarcely worth
considering. The product is evaporated down to about 60 per cent. of the original
quantity of blood (the mixture above referred to being understood to consist
of 80 per cent. of blood and 20 per cent. of water). If on attaining this percentage
traces of the antiseptic agent should still be found water should be added over and
over again until the last remnants of the antiseptic agent are completely
eliminated.
The final product thus obtained will be a sufficiently liquid, clear, deep-red
substance which, as has been ascertained by bacteriological tests is perfectly free
from the micro-organisms circulating in the blood. The insipid taste peculiar to
blood may be counteracted by the addition of suitable chemicals, chemically pure
glycerine being one of the best, since it is of considerable value also as a means
for preserving the preparation in an unimpaired condition.
Having now particularly described and ascertained the nature of my inven-
tion and in what manner the same is to be performed 1 declare that what I
claim is.―
1. In the production of purified sterilised liquid hæmoglobin from animal blood
the addition to the blood of an antiseptic substance and subsequently eliminating
in vacuo the said antiseptic together with the septic foreign bodies so that the cause
of putrefaction and the antiseptic disappear from the hæmoglobin.
2. The production of purified sterilized liquid hæmoglobin from animal blood by
removing the gas therefrom then adding to the blood water and an antiseptic
substance and subsequently eliminating the said antiseptic together with the septic
foreign bodies so that the cause of putrefaction and the antiseptic disappear from
the hæmoglobin all substantially as hereinbefore described.
Dated this 13th day of March 1895.
JOHNSONS & WILLCOX,
47, Lincoln's Inn Fields, Agents.