Note: Descriptions are shown in the official language in which they were submitted.
~LfJR 30~
The Invention relates to contrast agents for use In
ultrasound diagnosis of the human or animal body.
The examination of organs using ultrasound (sonography)
Is a diagnostic method which has been well established and pray-
tlsed for some years. Ultrasound waves In the megahertz range
(above 2 megahertz with wavelengths of between 1 and 0.2 mm) are
reflected at the Interfaces of various types of tissue. The
resulting echoes are amplified and rendered visible. Of partlcu-
far Importance Is the examination of the heart by this method
which Is known as echocardlography (Heft, Jar. et at.: Clinical
echocardlography, Future, Mount Classic, New York 1978; Killer, E.
KlInlsche Echokardlographle, Eke, Stuttgart 1979; Stephen, G. et
at.: Echokardlographle, Theme, Stuttgart-New York 1981; G.
Blamlno, L. Lange: Echokardlographle, Hoechst AGO 1983).
Since fluids, Including blood, produce ultrasound Image
contrast only when there are differences In density with respect
to the surroundings, posslbllltles were sought of rendering the
blood and Its circulation visible for ultrasound examination and
this may be effected by Injecting extremely fine gas bubbles Into
I
the bloodstream.
Several methods of producing and stabilizing gas
bubbles have been described in the literature. They
can be produced, for example, before injection into the
bloodstream, by vigorously shaking or stirring a liquid
solution, such, for example, as sodium chloride
solution, dye solution or previously removed blood.
Although ultrasound image contrast is achieved by
these methods, they have serious disadvantages
which are manifested in poor reproducibility, greatly
fluctuating size of the gas bubbles and a certain risk
of embolism due to a proportion of large visible
bubbles. Some of these disadvantages have been
eliminated by other production processes, such as, for
example, by the process of US. Patent No. 3,640,271
in which bubbles of a reproducible size are produced by
filtration or by the use of direct current electrode
apparatus. Against the advantage of being able to
produce gas bubbles of a reproducible size is the
disadvantage of the considerable technical outlay
involved.
US. Patent No. 4,276,885 describes the
production of gas micro-bubbles of a specific size
which are surrounded by a gelatin membrane which
protects them from coalescence. The prepared bubbles
can be stored only in the "frozen" state, for example
by storing at refrigerator temperature, and they must
~;Lf'~3~0~
--3--
..
be raised to body temperature again before they can be
used.
US. Patent No. 4,265,251 describes the
production and use of gas micro-bubbles with a solid
saccharine covering, which bubbles may be filled with a
pressurized gas. If they are under normal pressure,
they can be used as ultrasound image contrast agents;
when used at an elevated internal pressure, they can be
used for measuring blood pressure. Although in this
lo case the storage of the solid gas bubbles does not
present any problem, the technical outlay involved in
their production gives rise to high-costs as a result
of the complex techniques.
- The risks involved with the hitherto known
contrast agents for ultrasound diagnosis are caused by
two factors: the size and number both of the particles
of solid material and also of the gas bubbles.
The ultrasound contrast agents prepared by the
previously described methods have, in all cases,
possessed only some of the following properties that
are required:-
1. Exclusion of the risk of embolism
(dependent on size and number of
gas bubbles and size and number of
particles of solid material).
2. Reproducibility.
3. Sufficiently long stability.
I I
4. Ability to pass through the lungs, for example In order to
obtain ultrasound Image contrast of the left-hand side of the
heart.
5. Ability to pass through the capillaries.
6. Sterility and freedom from pyrogens.
7. Easy production at reasonable cost.
8. Easy storage.
European Patent Application No. 52575 published May 26,
1982 to Ultrasound Inc. describes the production of ultrasound
contrast agents containing gas bubbles that are supposed to posy
sews these necessary properties. However, In order to produce
them, mlcropartlcles of a solid crystalline substance, such as,
for example, galactose, are suspended In a liquid carrier, and
the gas, which Is adsorbed at the particle surface and Is
enclosed In cavities between the particles or In Intercrystalllne
cavities, forms the gas bubbles. The resulting suspension of gas
bubbles and mlcropartlcles Is Injected over a period of 10 men-
vies. Although according to European Patent SpeclfIcatlon 52575
the suspension prepared by the described method Is capable, after
Injection Into a peripheral vein, of appearing both on the fight-
hand side of the heart and also, after passing through the lungs,
on the left-hand side of the heart and of rendering visible the
blood there and Its circulation during ultrasound examination, It
was found when checked that the contrast medium prepared by the
method
-- 4
'I
391~9~
described in European Application No. 52575 and
injected into a peripheral vein did not in fact produce
ultrasound echoes in the left-hand side of the heart.
An object of the present invention is to provide a
contrast agent for ultrasound diagnosis which is
capable, after being administered intravenously, of
rendering visible for ultrasound the blood and its
circulation conditions not only on the right-hand side
of the heart but also, after passing through the
capillary bed of the lungs, on the left-hand side of
the heart. In addition, it should also permit the
representation of the circulation of blood through
other organs, such as the myocardium, the liver, the
spleen and the kidneys.
The present invention provides a contrast agent
for use in the ultrasound diagnosis of the human or
animal body, which comprises micro particles of a solid
surface-active substance and micro-bubbles of a gas in
- a liquid carrier. If desired, the liquid carrier also
contains micro particles of a solid non-surface active
agent.
It will be understood that the constituents of
the contrast agents of the invention must be
physiologically tolerable, and this, of course, equally
applies to the liquid media and diagnostic kits
described below.
The ultrasound contrast agents of the present
kiwi
invention possess all the above-mentioned properties
that are expected of such a contrast agent.
The invention also provides a liquid medium for
use in making up the ultrasound contrast agent, which
comprises a suspension of micro particles of a solid
surface-active substance, and, if desired, micro-
particles of a solid non-surface active substance, in a
liquid carrier.
Surprisingly, we have found that, by suspending
micro particles of a solid surface-active substance,
optionally in conjunction with micro particles of a
solid non-surface-active substance, in a liquid
carrier, an ultrasound contrast agent is obtained
- which, after being injected into a peripheral vein,
permits reproducible ultrasound images even of blood in
the arterial left-hand side of the heart. Since the
left-hand side of the heart can be reached with the
ultrasound contrast agent of the invention after
intravenous administration, ultrasound contrasts of
other organs supplied with blood from the aorta, such
as the myocardium, the liver, the spleen, the kidneys,
inter alias are therefore also possible after venous
administration. The ultrasound contrast agent of the
invention is, of course, also suitable for contrasts on
the right-hand side of the heart and for all other uses
as an ultrasound image contrast medium.
All substances that are physiologically tolerable
--7--
in the quantities used, that is, that have a low
toxicity and/or are biologically degradable and the
melting point of which is higher than room temperature,
are suitable as the surface-active substance for the
production of micro particles. Especially suitable are
lecithins, lecithin fractions and their conversion
products, polyoxyethylene fatty acid esters,
polyoxyethylene fatty alcohol ethers, polyoxyethylated
sorbitan fatty acid esters, glycerine polyethylene
- 10 glycol oxystearate, glycerine polyethylene glycol
ricinoleate, ethoxylated soya strolls, ethoxylated
castor oils and their hydrogenated derivatives,
cholesterol, polyoxyethylene fatty acid struts
and polyoxyethylenepolyoxypropylene polymers
15 having a molecular weight of from 6800 to 8975, 13300
and 16250, succors esters, such, for example, as
sugar esters, for example succors dipalmitate and
succors monolaurate or succors glycerides and
xyloglycerides, saturated or unsaturated (C4-C20)-
fatty alcohols or (C4-C20)-fatty acids or their metal
salts, moo-, dip and triglycerides, sorbitan fatty
acid esters, fatty acid esters of succors or fatty
acid esters such, for example, as bottle Stewart and
acquirable palpitate; calcium Stewart, the succors
esters of Laurie acid Starkey acid and palmitic acid,
and acquirable palpitate are preferred.
The rate at which the micro particles of the
surface-active substance dissolve in the liquid carrier
should be slower than the rate at which these
micro particles dissolve in the blood. Advantageously,
the volubility of the micro particles of the surface-
active substance in the liquid carrier is such that
when they are introduced into it they do not start to
dissolve in it to a substantial extent for at least
10 minutes. It will be appreciated that upon
administration of the contrast agent the micro particles
- of the surface-active substance will start to dissolve
in the blood.
The solid surface-active substance of the contrast
agent is used in a concentration of from 0.01 to 5 %,
preferably from 0.Q4 to 0.5 %, by weight of the
contrast agent.
If desired, the micro particles of the surface-
active substance can be used in conjunction with
micro particles of a physiologically tolerable non-
surface-active crystalline solid. Organic and
inorganic substances can be used as the crystalline
solid, for example salts such, for example, as sodium
chloride, sodium citrate, sodium acetate or sodium
tart rate, monosaccharides such, for example, as
glucose, fructose or galactose; disaccharides such, for
example, as succors, lactose or maltose; pentoses
such, for example, as Arabians, Zulus or rubs; or
Jo kiwi
go
cyclodextrines such, for example, as a-, B- or-y-cyclo-
dextrine; galactose, lactose and -cyclodextrine are
preferred. They are contained in the contrast agent of
the invention in a concentration of from 5 to 50 %,
5 preferably from 9 to 40 %, by weight.
The micro particles may be produced by
recrystallizing the surface-active substances and, if
desired, non-surface-active substances under sterile
conditions. They are then commented under sterile
conditions, for example, by grinding in an air-jet
mill, until the desired particle size is obtained.
Preferably the micro particles should have a median
particle size of less than 10 em, advantageously less
than 8 em, more especially within the range of from 1
to 3 em. The particle size is determined in a suitable
measuring apparatus. The micro particles produced
comprise either the commented surface-active substance
alone or a mixture of the micro particles of the surface-
active substance and a solid non-surface-active
substance. In the latter case the ratio by weight of
solid surface-active substance to solid non-surface-
active substance is preferably from 0.01 to 5:100.
Both-the micro particle size achieved by the
comminution process and also the size of the micro-
bubbles containing a physiologically tolerable gas contained in the contrast agent of the invention ensure
safe passage through the capillary system and the
3'~)9.~
--10--
capillary bed of the lungs and preclude the occurrence
of embolism.
Some of the micro-bubbles required to produce
image contrast are transported by the suspended micro-
S particles, adsorbed at the surface of the micro-
particles and enclosed in the cavities between the
micro particles or enclosed in an inter crystalline
manner.
The volume of gas transported by the
micro particles in the form of gas micro-bubbles is from
0.02 to 0.6 ml per gram of micro particles.
Apart from its transporting function, the carrier
liquid also has the function of stabilizing the suspend
soon comprising micro particles and gas micro-bubbles,
for example of preventing the sedimentation of the
micro particles and the coalescing of the gas micro-
bubbles or of delaying the dissolving process of the
micro particles.
There may be used as the liquid carrier, for
example, water, aqueous solutions of one or more
inorganic salts such, for example, as physiological
sodium chloride solution and buffer solutions, aqueous
solutions of moo- or disaccharides such, for example,
as galactose, glucose or lactose, mandrake or
polyhydric alcohols, in so far as they are
physiologically tolerable such, for example, as
ethanol, propanol, isopropyl alcohol, polyethylene
kiwi
glycol, ethylene glycol, glycerine, propylene glycol,
propylene glycol methyl ester or their aqueous
solutions.
Water and physiological electrolyte solutions,
. 5 such, for example, as physiological sodium chloride
solution, and aqueous solutions of galactose and
lactose, are preferred. If solutions are used, the
concentration of the dissolved substance should be from
0.1 to on % by weight, preferably from 0.5 to 25 % by
weight, and, more especially there may be mentioned,
0.9 % aqueous sodium chloride solution or 20 % aqueous
galactose~
The invention also provides a process for the
preparation of the contrast agent of the invention,
wherein micro particles of a physiologically tolerable
solid surface-active substance and, if desired,
micro particles of a solid non-surface-active substance,
are mixed with a liquid carrier and agitated until a
homogeneous suspension is formed.
In order to prepare the ultrasound contrast agent
in a form ready for use, the sterile liquid carrier may
be added to the sterile solid surface-active substance,
which is in the form of micro particles and which is
optionally in conjunction with micro particles of a
sterile solid non-surface-active substance, and this
mixture with the carrier liquid is shaken until a
homogeneous suspension has formed, which takes
3 . d
-12-
approximately from 5 to 10 seconds. Immediately after
its preparation, and at the latest up to 5 minutes
thereafter, the resulting suspension is injected in the
form of a bonus into a peripheral vein or into a
catheter which is already present, from 0.01 ml to
1 ml/kg body weight being administered.
For reasons of expediency, the components
necessary for the preparation of the contrast agent of
the invention such, for example, as carrier liquid
and micro particles of the surface-active substance,
optionally in conjunction with micro particles of the
solid non-surface-active substance are stored under
sterile conditions in two separate vessels (A) and (B)
respectively in the quantity necessary to carry out an
examination. The size of vessel (B) should be such
that the contents of vessel (A) can be transferred to
(B) by means of an injection syringe and the resulting
mixture can be shaken.
The present invention also provides a diagnostic
kit for use in the ultrasound diagnosis of the human or
animal body, which comprises
(A) a container which contains a liquid carrier, and
(B) a second container which contains micro particles
of a solid surface-active substance and, if
desired, micro particles of a solid non-surface-
active substance.
AL Ed
The contents of the containers are in a form ready
for mixing together immediately before use.
Preferably container (A) is provided with a
closure permitting the removal of the contents under --
s sterile conditions and container (B) is provided with a
closure permitting, under sterile conditions, the
addition of the contents of vessel (A) and the removal
of the resulting contrast agent.
Advantageously the containers A and B both have a
volume of from 5 to 10 ml. When the non-surface active
substance is present, the ratio by weight of the micro-
particles of the surface active substance to the micro-
particles of the non-surface-active substance is
preferably from 0.01 to 5:100.
The use of the contrast agent of the invention is
demonstrated by an echocardiographic examination of a
baboon weighing 10 kg. which will now be described.
5 ml of carrier liquid (prepared according to
Example 1 below) are removed from a Phil using an
2-0 injection syringe and are added to 2 g of micro-
particles (prepared according to Example 1 B below)
which are in a second Phil, and the mixture is shaken
for approximately from 5 to 10 seconds until a
homogeneous suspension has formed. 2 ml of this
suspension are injected into a peripheral vein (V.
jugulars, brachialis or siphon) via a three-
way tap having an infusion speed of at least 1 ml/sec.,
-14-
preferably 2-3 mljsec. Immediately after injecting the
contrast agent, 10 ml of physiological sodium chloride
solution are injected at the same speed so that the
contrast agent bonus is maintained as complete as
possible until the right-hand side of the heart is
reached. Before, during and after injection, a
commercially available transducer for echo cardiography
is held against the thorax of the experimental animal
so that a typical cross-section is obtained through the
right-hand side and the left-hand side of the heart.
- This test procedure is understood and well known to a
person skilled in the art.
If the ultrasound contrast agent reaches the right-
- hand side of the heart, it is possible to follow in a 2-
D echo image or an M-mode echo image how the blood
marked by the contrast agent first reaches the level of
the right-hand atrium and then the level of the right-
hand ventricle and the pulmonary artery, homogeneous
filling occurring for approximately 10 seconds. While
the cavities in the right-hand side of the heart in the
ultrasound image empty again, the blood which is
rendered visible with contrast agent, after passing
through the lungs, appears again in the pulmonary veins
and fills the left atrium, the left ventricle and the
aorta homogeneously, the contrast image lasting from 2
to 3 times longer than on the right-hand side of the
heart. In addition to the representation of the blood
I
-15-
flow through the cavities of the left-hand side of the
heart, there is also a representation of the myocardium
showing the circulation of the blood.
The use of the ultrasound contrast agent of the
invention is, however, not limited to rendering visible
the circulation of blood in the arterial part of the
heart after venous administration but is also used with
outstanding success as a contrast agent for examining
the right-hand side of the heart and other organs.
The invention still further provides a method of
ultrasound diagnosis of the human or animal body,
wherein a contrast agent of the invention containing
a dispersion of micro-bubbles is injected into a part
of the human or animal body, preferably
intravascularly, and an ultrasound image of the micro-
bubbles at a site in the body which it is desired to
investigate is obtained.
do
-16-
The following Examples illustrate the invention,
the parts and percentages being by weight unless
otherwise indicated.
Example 1
I) Preparation of the carrier liquid
80 g of galactose are dissolved in water for
injection purposes, made up to a volume of 400 ml and
forced through a 0.2 em filter; 5 ml phallus are each
filled with 4 ml of this filtrate and sterilized for 20
lo minutes at 120C.
B) Preparation of the micro particles:
Under sterile conditions, 198 g of galactose are
mixed intensively with 2 g of magnesium Stewart by
homoeopathic trituration, passed through a 0.8 mm
sieve, mixed loosely and ground in an air-jet mill
until the following distribution of the particle size
is obtained:
Median: 1.9 em
99 % < 6 em
< 3 em.
The particle size and the distribution thereof are
determined in a particle-measuring apparatus after
suspension in an hydrous isopropanol. 5 ml phallus are
each filled with 2 9 of the micro particles.
I
.
-17-
C) For the preparation of 5 ml of the ultrasound
contrast agent in a form ready for use, the contents
of a Phil containing carrier fluid (20 % galactose
solution in water, A) are introduced by means of an
injection syringe into a Phil containing micro-
particles (B) and shaken until a homogeneous suspension
is formed (from 5 to 10 seconds).
Example 2
A) Preparation of the carrier liquid-
Water for injection purposes is used and 5 ml
phallus are each filled with 4 ml of the water and
sterilized for 20 minutes at 120C.
B) Preparation of the micro particles:
Under sterile conditions, 198 g of galactose are
mixed intensively with 2 g of acquirable palpitate by
homoeopathic trituration and further processed as
described in Example 1 under B), the following
distribution of the particle size being obtained:
Median: 1.9 em
100 % 6 em
90 % < 3 em.
The particle size is determined as described in
Example 1 under B).
5 ml phallus are each filled with 1200 my of the
micro particles.
I
-18-
C) For the preparation of 4.5 ml of the ultrasound
contrast agent in a form ready for use, the contents
of a Phil containing carrier liquid (water, A) are
introduced by means of an injection syringe into a
Phil containing micro particles (B) and shaken until a
homogeneous suspension is formed (from 5 to 10
seconds).
Example 3
A) Preparation of the carrier liquid:
4.5 g of sodium chloride are dissolved in water to
a volume of 500 ml and the solution is forced through a
0.2 em filter; 5 ml phallus are each filled with 4 ml of
- this solution and sterilized for 20 minutes at 120C.
B) Preparation of the micro particles:
Under sterile conditions, 198 g of an hydrous
lactose ( < 0.3 mm) are mixed intensively with 2 g of
acquirable palpitate by homoeopathic trituration and the
mixture is further processed as described in Example 1
under B), the following distribution of the particle
size being obtained:
Median: . 2.8 em
100 % < 48 em
99 % < 12 em.
The particle size is determined as described in
Example 1 under B).
C~)6~
--19--
5 ml phallus are each filled with 1.6 9 of the
micro particles.
C) For the preparation of 5 ml of the ultrasound
contrast agent in a form ready for use, the contents
of a Phil (0.9 % sodium chloride solution in water, A)
are introduced by means of an injection syringe into a
Phil containing micro particles (B) and shaken until a
homogeneous suspension is formed (from 5 to 10 seconds).
Example 4
A) Preparation of the carrier liquid:
In the same manner as described in Example 3 under
A), Ox % aqueous sodium chloride solution is prepared,
introduced in 4 ml portions into 5 ml phallus and
sterilized for 20 minutes at 120C.
B) Preparation of the micro particles:
Under sterile conditions, 199 g of â-cyclodextrine
are mixed intensively with 1 g of acquirable palpitate by
homoeopathic trituration and the mixture is further
processed as described in Example 1 under B), micro-
particles having the following size distribution being obtained:
Median: 2 em.
99 < 6 em
- 90 % < 4 em.
I
-20-
The particle size is determined as described in
Example 1 under 8).
5 ml phallus are each filled with 400 my of the
micro particles.
C) For the preparation of 4 ml of the ultrasound
contrast agent in a form ready for use, the contents
of a Phil (0.9 % aqueous sodium chloride solution, A)
are introduced by means of an injection syringe into a
Phil containing micro particles (B) and shaken until a
homogeneous suspension is formed (from 5 to 10
seconds).
Example 5
A) Preparation of the carrier liquid:
50 g of lactose are dissolved in water for
injection purposes, made up to a volume of 500 ml and
forced through a 0.2 em filter; 5 ml phallus are each
filled with 4 ml of the solution and sterilized for 20
minutes at 120C.-
B) Preparation of the micro particles:
Acquirable palpitate is dissolved in methanol,
filtered under sterile conditions through a 0.2 em
filter, recrystallized under sterile conditions, dried
and passed through a 0.8 mm sieve. The sterile
acquirable palpitate is then ground under sterile
clue
-21-
conditions in an air-jet mill until the following size
distribution of the particles is obtained:
Median value: 1.9 em
99 < 6 em
90 % < 3 em.
The particle size and the distribution thereof are
determined in a particle-measuring apparatus after
(a my
suspension in cold aqueous 0.1 Pluronic F68 solution.
Sterile 5 ml phallus are each filled with 40 my of
the micro particles.
C) For the preparation of 4 ml of the ultrasound
contrast agent in a form ready for use, the contents
of a Phil containing carrier liquid (10 lactose
solution, A) are introduced by means of an injection
syringe into a Phil containing the micro particles and
shaken until a homogeneous suspension is formed.
- Example 6
A) Preparation of the carrier liquid:
4.5 g of sodium chloride are dissolved in water to
a volume of 500 ml and the solution is forced through a
0.2 em filter; 5 ml phallus are each filled with 4 ml of
this solution and sterilized for 20 minutes at 120C.
B) Preparation of the micro particles:
Under sterile conditions, a solution, filtered
~3C~
I
under sterile conditions, of OHS g of acquirable
palpitate in 40 g of isopropanol is absorbed on 199.5 g
of sterile galactose particles, the isopropanol is
removed by drying at 40 and 200 torn and comminution
is carried out in an air-jet mill until the following
size distribution of the particles is obtained:
Median value: 1.9 em
99 % < 6 em
90 % < 3 em.
The particle size and the distribution thereof are
determined in a particle-measuring apparatus, for
example after suspension in isopropanol. 2 g portions
of the micro particles are each packed into 5 ml phallus.
C) For the preparation of 5 ml of the ultrasound
contrast agent in a form ready for use, the contents
of a Phil containing carrier liquid (0.9 % sodium
chloride solution in water, A) are introduced by means
of an injection syringe into a Phil containing micro-
particles (B) and-shaken until a homogeneous suspension
is formed (from 5 to 10 seconds).
Example 7
A) Preparation of the carrier liquid:
4.5 g of sodium chloride are dissolved in water
and made up to a volume of 500 ml; the solution is
forced through a 0.2 em filter and 5 ml falser each
cut
-23-
filled with 4 ml of this solution and sterilized for 20
minutes at 120C.
B) Preparation of the micro particles: -
Under sterile conditions, 199.5 g of galactose are
triturated with 0.5 g of acquirable palpitate, mixed
intensively, passed through a 0.8 mm sieve and
commented in an air-jet mill until the following size
distribution of the particles is obtained:
Median value: 1.9 em
99 % 6 em
90 % 3 em.
.
The particle size and the distribution thereof are
determined in a particle-measuring apparatus, for
example after suspension in isopropanol. 2 g portions
of the micro particles are each packed into 5 ml phallus.
- C) For the preparation of 5 ml of the ultrasound
contrast agent in a form ready for use, the contents
of a Phil containing carrier liquid (0.9 % sodium
chloride solution in water, A) are introduced by means
of an injection syringe into a Phil containing micro-
particles (B) and shaken until a homogeneous suspension
is formed (from 5 to 10 seconds).
Jo
~24-
Example 8
A) Preparation of the carrier liquid:
Water for injection purposes is used and 5 ml
phallus are each filled with 4 ml of the water and
sterilized for 20 minutes at 120C.
B) Preparation of the micro particles:
Under sterile conditions, 0.5 g of succors
monopalmitate is triturated with 199.5 g of galactose,
mixed intensively, passed through a 0.8 mm sieve and
ground in an air-jet mill until the following size
distribution of the particles is obtained:
Median value: 1.9 em
at least 99 % 6 em
at least 90 % < 3 em.
The particle size and the distribution thereof are
determined in a particle-measuring apparatus, for
example after suspension in isopropanol. 2 g portions
of the micro particles are each packed into 5 ml phallus.
C) For the preparation of 5 ml of the ultrasound
contrast agent in a form ready for use, the contents
of a Phil containing carrier liquid (sterile water
for injection purposes, A) are introduced by means
of an injection syringe into a Phil containing micro-
particles (B) and shaken until a homogeneous suspension
is formed (from 5 to 10 seconds).
I
-25-
Example 9
A) Preparation of the carrier liquid:
5 ml phallus are each filled with 4 ml of water
used for injection purposes and sterilized for 20
minutes at 120C.
B) Preparation of the micro particles:
Under sterile conditions, a solution, filtered
under sterile conditions, of 0.5 g of succors
monopalmitate in 40 g of isopropanol is absorbed on
199.5 g of sterile galactose particles, the isopropanol
is removed by drying at 40C and 200 torn and
- grinding is carried out in an air-jet mill until the
following size distribution of the particles is
obtained:
Median value: 1.9 em
at least 99 % 6 em
at least 90 % < 3 em.
- The particle size and the distribution thereof are
- determined in a particle-measuring apparatus after
suspension in isopropanol. 2 g portions of the
micro particles are each packed into 5 ml phallus.
C) For the preparation of 5 ml of the ultrasound
contrast agent in a form ready for use, the contents
- of a Phil containing carrier liquid (water for
injection purposes, A) are introduced by means of an
do to
-26-
injection syringe into a Phil containing micro-
particles (B) and shaken until a homogeneous suspension
is formed (from 5 to 10 seconds).
Example 10
A) Preparation of the carrier liquid:
5 ml phallus are each filled with 4 ml of water
used for injection purposes and sterilized for 20
minutes at 120C.
B) Preparation of the micro particles:
Under sterile conditions, a solution, filtered
under sterile conditions, of 0.5 g of succors
- menstruate in 40 g of isopropanol is absorbed on
199.5 g of sterile galactose particles, the isopropanol
is removed by drying at 40C and 200 torn and
grinding is carried out in an air-jet mill until the
following size distribution of the particle is
obtained:
Median value: 1.9 em
at least 99 % < 6 em
at least 90 % < 3 em.
The particle size and the distribution thereof are
determined in a particle-measuring apparatus after
suspension in isopropanol. 2 g portions of the
micro particles are each packed into 5 ml phallus.
I
-27-
C) For the preparation of 5 ml of the ultrasound
contrast agent in a form ready for use, the contents
of a Phil containing carrier liquid (water for
injection purposes, A) are introduced by means of an
injection syringe into a Phil containing micro-
particles (B) and shaken until a homogeneous suspension
is formed (from 5 to 10 seconds).
Example 11
A) Preparation of the carrier liquid:
Water for injection purposes is used and 5 ml
phallus are each filled with 4 ml of the water and
sterilized for 20 minutes at 120C.
B) Preparation of the_microparticles:
under sterile conditions, 0.5 g of succors
menstruate is triturated with 199.5 g of galactose,
mixed intensively, passed through a 0.8 mm sieve and
ground in an air-jet mill until the following size
distribution of the particles is obtained:
Median value: 1.9 em
at least 99 % < 6 em
at least 90 % < 3 em.
The particle size and the distribution thereof are
determined in a particle-measurinq apparatus after
suspension in isopropanol. 2 g portions of the
micro particles are each packed into 5 ml phallus.
-28-
C) For the preparation of 5 ml of the ultrasound
contrast agent in a form ready for use, the contents
of a Phil containing carrier liquid (sterile water
for injection purposes, A) are introduced by means
of an injection syringe into a Phil containing micro-
particles (B) and shaken until a homogeneous suspension
is formed (from 5 to 10 seconds).
Example 12
A) Preparation of the carrier liquid:
5 ml phallus are each filled with 4 ml of water
for injection purposes and sterilized for 20 minutes at
120C.
B) Preparation of the-microparticles:
Under sterile conditions, a solution, filtered
under sterile conditions, of 0.5 9 of succors
distrait in 40 9 of isopropanol is absorbed on
199.5 9 of sterile galactose particles, the isopropanol
is removed by drying at 40C and 200 torn and
grinding is carried out in an air-jet mill until the
following size distribution of the particles is
obtained:
Median value: 1.9 em
at least 99 % 6 em
at least 90 % < 3 em.
The particle size and the distribution thereof are
~L~3'~)9
--29--
determined in a particle-measuring apparatus after
suspension in isopropanol. 2 g portions of the
micro particles are each packed into 5 ml phallus.
C) For the preparation of 5 ml of the ultrasound
contrast agent in a form ready for use, the contents
of a Phil containing carrier liquid (water for
injection purposes, A) are introduced by means of an
injection syringe into a Phil containing micro-
particles (B) and shaken until a homogeneous suspension
is formed (from 5 to 10 seconds).
Example 13
A) Preparation of the carrier liquid:
Water for injection purposes is used and 5 ml
phallus are each filled with 4 ml of the water and
sterilized for 20 minutes at 120C.
B) Preparation of the micro particles:
Under sterile conditions, 0.5 g of succors
distrait is triturated with 199.5 g of galactose,
mixed intensively, passed through a 0.8 mm sieve and
ground in an air-jet mill until the following size
distribution of the particles is obtained:
Median value: 1.9 em
at least 99 % < 6 em
at least 90 < 3 em.
-30-
The particle size and the distribution thereof are
determined in a particle-measuring apparatus after
suspension in isopropanol. 2 g portions of the
micro particles are each packed into 5 ml phallus. --
C) For the preparation of 5 ml of the ultrasound
contrast agent in a form ready for use, the contents
of a Phil containing carrier liquid (sterile water
for injection purposes, A) are introduced by means
of an injection syringe into a Phil containing micro-
- 10 particles (B) and shaken until a homogeneous suspension
is formed (from 5 to 10 seconds).
