Note: Descriptions are shown in the official language in which they were submitted.
" 1056340
This invention relates to apparatus useful in
connection with biological liquids. More particularly,
this invention is concerned with novel apparatus for
storing and transporting anaerobic biological liquids of
the type which remain viable only when in a gaseous environ-
ment or atmosphere low in, or devoid of, oxygen.
Many diseases of man and lower animals are
bacterial in origin. The treatment of many bacterial
diseases requires that the infecting organism be identified.
A drug known to be effective against the infecting
organism can then be prescribed.
The identification of an infecting organism is
often by means of a liquid sample obtained from the ill
patient or animal. Cerebrospinal fluids, deep abscess
material, tissue biopsy, transtrachael aspirate, pleural
fluid, supra-pubic aspirated urine, bone marrow and aspirated
joint fluid are representative liquids obtained from a
patient which require transport to a testing laboratory.
The liquid sample is then tested as required, generally
including tests for identification of possible infecting
organisms. Such laboratories require highly trained
microbiologists and elaborate, expensive equipment.
Suitable testing laboratories, accordingly, are not always
readily available. It therefore becomes necessary for the
patient to visit, or animal be taken to, the laboratory
where the liquid sample can be obtained and put immediately
into the test procedures or for the liquid sample to be
taken at a location remote from the laboratory and then
transported to the laboratory for testing.
While the collecting of a liquid sample
generally presents no difficulties, the storage and/or
`- 1056340
transportation of the sample to a testing laboratory under
conditions which guarantee it will be viable and free of
contaimination upon arrival presents serious problems.
Although contamination from other organisms can generally
be avoided by suitable means, the maintenance of the
sample often requires that it be stored and transported in
a particular gaseous environment.
Since bacteria of the anaerobic type are known
to require an oxygen-deficient or oxygen-free gaseous
environmentj it is obvious that the transportation
and storage of an anaerobic biological liquid sample
should be effected in an environment having little or no
oxygen.
The Brewer U.S. patent 3,246,959 discloses a
gas-producing device for generating an atmosphere conducive
for maintaining and increasing the viability of organisms
which require a special non-toxic atmosphere. The patent ~ -
shows the chemical generation of hydrogen, carbon dioxide
and acetylene for the purpose of supplying a non-toxic
atmosphere to a culture in a container. A platinized
wire gauze in the container is heated by electricity for
the purpose of completely reacting oxygen in the container.
Anandam U.S. patent 3,616,263 discloses a culture
tube for anaerobic cultures. Oxygen is removed from the
tube by use of a divided capsule containing aqueous potas-
sium hydroxide and aqueous pyrogallic acid which when
combined form a strong reducing agent for the oxygen.
Although the prior art recognizes that various
cultures must be maintained under anaerobic conditions,
there has been a need for a low cost, reliable,
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1056340
disposable package for storage, transport and/or testing of an anaerobic
biological liquid sample.
According to the present invention there is provided an anaerobic
liquid storage and transport apparatus comprising a liquid receiver means for
receiving and holding an anaerobic liquid and a gas generator container means
containing a gas generator for generating a reducing gas. The gas generator
comprises a reducing gas generating solid material, an ampoule containing
a liquid which is reactive with the solid material to produce a reducing gas
catalytically reactive with oxygen at room temperature, and means in the
container means which prevents liquid from flowing from the container means
after the ampoule is opened but which permits flow of reducing gas out of the
container means. First means are provided for supplying reducing gas from the
gas generator container means to the liquid receiver means, and second means
other than said first means for exit of the reducing gas out of the liquid
receiver means without flow of liquid therefrom. A catalyst is positioned to
promote reaction between the reducing gas, when produced by the gas generator,
and oxygen to thereby produce an anaerobic envirorment in contact with a
liquid placed in the liquid receiver means.
The storage and transport apparatus thus provided by the subject
invention includes at le~st a receptacle for receiving an anaerobic biological
liquid ~md~mapparatus which is self-sufficient for producing at least a reduc-
ing gas atmosphere, and advisabl~ an atmosphere which isalso essentially oxy-
gen-free, in contact with the liquid. Also advisably included is a color
indicator appara~s which will show by color change the presence or absence
of oxygen in the atmosphere around the anaerobic liquid. The described ap-
paratus is c~mparatively easily manufactured, low in cost, light-weight and
easily utilized. The apparatus is intended to be employed only once and then
discarded, making it unnecessary to clean and re-use any parts of the
apparatus.
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A catalyst may be placed in, or outside of, the liquid storage
and transport apparatus. A catalyst is generally located in the conduit
means for gas exiting from the liquid receiver means since any back flow of
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"` ~L056340
oxygen which may happen to take place will thereby be
caused to react with the reducing gas and thus oxygen kept
out of contact with a liquid sample in the liquid receiver
means. A catalyst can, however, be placed in the gas
generator container means, or in the color indicator when
joined to the transport apparatus, or a catalyst can be
placed in a bag of low permeability for the transport
apparatus. A catalyst can be placed in only one of the
; thus described places or in two or more of such places.
The anaerobic liquid storage and transport
apparatus provided by this invention is generally employed
in combination with a bag of flexible transparent sheet
material of low gas permeability. The liquid storage and
transport apparatus is advisably placed in such a bag, and
the bag is sealed before the gas generator and the color
indicator are activated. The bag serves to prevent oxygen
from entering the apparatus and also serves to provide an
oxygen-deficient, or essentially oxygen-free, environment
inside of the bag around the liquid storage and transport
apparatus. Such an environment is produced by the
flow of a reducing gas, such as hydrogen, from the liquid
storage and transport apparatus, together with any oxygen
flushed therefrom, into the bag where the reducing gas is
caused to react with oxygen in the bag at ambient temperature
by the inclusion of a-suitable catalyst in the bag. By
removing the oxygen from the bag the liquid storage and
transport apparatus permits a liquid sample to be held
under anaerobic conditions for a longer period of time,
such as for about 96 hours or more, than would otherwise
generally be the case.
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:1~56340
Both the gas generator and the color indicator
apparatus are constructed so that each may be activated
from outside, thus making it unnecessary to open either of
these elements constituting part of the anaerobic liquid
storage and transport apparatus.
The liquid receiver means is provided with a
port means for introducing an anaerobic liquid into the
liquid receiver means. The port means may constitute a
simple closure or cap or it may comprise a self-closing
elastomeric material which will reseal after penetration
by a hypodermic needle used to place an anaerobic liquid
in the liquid receiver means.
The gas generator will generally be one which
produces hydrogen as the reducing gas. However, other
reducing gases may be produced, such as acetylene. Further-
more, the gas generator may be so-formulated as to produce
simultaneously carbon dioxide and a reducing gas. The
production of carbon dioxide is often desirable since
increased amounts of this gas in the atmosphere surrounding
a sample anaerobic liquid are desirable to promote
viability of various bacteria. --
The gas generator forming part of the anaerobic
liquid storage and transport apparatus will usually contain
a reducing gas generating solid material, an amp~ule con-
taining a liquid which is reactive with the solid material
to produce a reducing gas catalytically reactive with
oxygen at room temperature, said ampoule being openable
from outside the container means to release the liquid to
contact the solid material, and means in the container
means which prevents liquid from flowing from the
container means after the ampoule is opened but which
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1~56340
permits flow of reducing gas generated in the container
means out of the opening. Furthermore, the gas generator
generally contains a solid desiccant material which absorbs
water which may enter the container before the ampoule is
opened, thereby preventing degradation or premature reaction
of the gas generating solid material.
The gas generator container means may be in the
- form of an elongated flexible tube closed at one end and
having an opening at the other end portion in communication
with conduit means leading to the liquid receiver
means. Furthermore, the ampoule may be placed snugly in
the tube and the gas generating solid material located
between the tube closed end and the ampoule. An absorbent
plug may be located in the tube between the ampoule and
the opening in the tube to keep liquid from flowing from
the tube after the ampoule is ruptured to release the
liquid therein for activation of the gas generating solid
material.
The color indicator apparatus used with, or as
part of, the anaerobic liquid storage and transport
apparatus may comprise an ampoule in the indicator container
means, said ampoule containing a redox color indicator
liquid, and an absorben* material, for the liquid in the
ampoule, in the indicator container means. The color
indicator container may be separate from the liquid storage
apparatus or it may communicate directly with the liquid
receiver means. Also, the color indicator container may
be a tube with the ampoule snugly fit in the tube so as to
be readily rupturable by squeezing the tube.
The invention will be described further in
conjunction with the attached drawings, in which:
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1056340
Figure 1 is an isometric view, partially broken
away, of an anaerobic liquid storage and transport apparatus
provided by the invention positioned in a bag;
Figure 2 is a partial side view, partially in
section, of the liquid storage and transport apparatus
shown in Figure 1;
Figure 3 is a sectional view through the length
of the liquid storage and transport apparatus shown in
Figure l;
Figure 4 is a sectional view taken along the
line 4-4 of Figure 3;
Figure 5 is a sectional view taken along the
line 5-5 of Figure 2;
Figure 6 is a plan view of a second embodiment
of a liquid storage and transport apparatus provided by
the invention;
Figure 7 is a sectional view through the line 7-
7 of Figure 6;
Figure 8 is an exploded view of a further embodiment
of the invention;
Figure 9 is a partial sectional view of the
embodiment shown in Figure 8;
Figure 10 is a sectional view of the gas generator
apparatus used in the embodiment shown in Figures 8 and 9;
Figurs 11 is a sectional view of the color
indicator apparatus used in the embodiment shown in Figures
8 and 9;
Figure 12 is an exploded view of another embodiment
of the invention; and
Figure 13 is a partial sectional view of
the embodiment shown in Figure 12.
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So far as is practical, the same numbers will be
used to identify the same elements or parts which appear
in the various views of the drawings.
The anaerobic liquid storage and transport appa-
ratus 10 as shown in Figures 1 to 5 has a liquid receiver
means 11 for receiving and holding an anaerobic biological
liquid, a gas generator container means 12, and a color
indicator container means 13.
The liquid receiver means 11 has a cylindrical
10 body 14 which is closed at one end by wall 15. The
other end 16 of cylindrical body 14 is open initially but
is subsequently closed by a closure 17 of an elastomeric
material, such as rubber, which can be readily penetrated
by a hypodermic needle but which is self-sealing after
withdrawal of the needle. The space 18 inside of cylindrical
body 14 provides a receptacle for receiving a biological
liquid sample. A tubular stub 19 extends outwardly from
end wall 15 and also inwardly into the space 18 defined by
the cylindrical body 14. Opening 20 in tubular stub 19
20 provides gaseous communication between liquid receptacle
space 18 and the space 21 inside of tubular stub 19.
Fibrous plug 22 in tubular stub 19 permits flow of gas
therethrough but prevents flow of liquid. The end 31 of
tube 30 of the gas gen~rator container means 12 is slid
(, over the end of tubular stub 19 to securely join it thereto.
Z An additional elongated tube 25 is molded into
Z the side wall of cylindrical body 14. The tube 25 has a
tubular stub end 26 which extends outwardly from end wall
15 to receive the end 41 of tube 40 of the color indicator
30 container means 13. Opening 27 at the internal end
of tube 25 communicates with the liquid receptacle space
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-` ` 105634S)
18 so that there can be flow of gas from space 18 through
opening 27 and into the interior space 28 of tube 25.
Fibrous plug 29 is positioned inside of tube 25 adjacent
opening 27 to prevent liquid from flowing from space 18.
However, plug 29 permits ready flow of gas through it.
The complete liquid receiver means 10 consti~
tuting the cylindrical body 14, tubular stub 19 and tube
25 can be readily molded as a unitary object from a suitable
polymeric material, such as polypropylene.
The gas generator container means 12 comprises
elongated plastic tube 30 which is closed at end 32. The - -
tube 30 may be made of a flexible but self-supporting
polymeric material such as polyethylene, polypropylene or
a polypropylene copolymer such as the one available as
Avisun 6011.
One or more gas generating solid tablets or
pellets 33 is positioned in tube 30. The tablet 33 has a
composition which is suitable for generating a reducing
gas such as hydrogen or acetylene, or both a reducing gas
and carbon dioxide.
Ampoule 34 is positioned in tube 30 more or less
snugly so that it maintains its position. A liquid
35 is contained in ampoule 34. The composition of liquid
35 is selected so that, when released from ampoule 34, it
will react with tablet 33 to generate one or more gases. ~ -
The ampoule 34 can be made of glass or some other material
which is nonreactive with liquid 35 or the components of
gas generating tablet 33. The ampoule 34 is advisably
made so that it will rupture or break upon application of
finger pressure to the outside of tube 30 adjacent
the ampoule wall. In this way the ampoule may be opened
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1056340
and the liquid 35 freed to react with tablet 33, which
will drop into the freed liquid when the tube 30 is held
upright.
A liquid absorbent plug 36, such as of polyester
fibers, is positioned in tube 30 after the ampoule 34 is
placed in the tube. The absorbent plug 36 is thus located
between the ampoule 34 and the tube open end so that
liquid from the ampoule cannot flow from the tube.
One or more desiccant pellets 37 are advisably
positioned between liquid absorbent plug 36 and the
open end of the tube 30 as shown in Figure 3. Any suitable
desiccant or water dehydrating material can be used for
this purpose although it is preferred to use molecular
sieves. Nevertheless, magnesium sulfate or calcium chloride
are representative of other desiccant materials which may
be used satisfactorily.
Also positioned between liquid absorbent plug 36
and the open end of tube 30 is a catalyst pellet 38. The
catalyst pellet 38 is provided to induce catalytic reaction
20 between the reducing gas which is formed by the ---
combination of liquid 35 with tablet 33 and any oxygen
which may be in the entire apparatus 10. A 0.5% palladium-
on-alumina catalyst may be used when hydrogen is the
reducing gas although other catalysts which induce the
reaction at room temperature may be employed.
The gas generating tablet 33 may have the
following composition when it is desired to produce
simultaneously both carbon dioxide and hydrogen as the
reducing gas: -
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l()S6340
Potassium borohydride 13 mg.
Zinc 13 mg.
Sodium chloride 15 mg.
Sodium bicarbonate 52.5 mg.
Lactose DT 47 mg.
Microporous cellulose--Avicel PH 102 57.5 mg.
Tabletting lubricant--Calcium stearate 2.0 mg.
If it is desired to produce only hydrogen and no carbon
dioxide the sodium bicarbonate may be omitted from the
composition set forth above for tablet 33.
The ampoule 34 may contain as the liquid 35 one
ml. of 0.65 N hydrochloric acid in a glass ampoule 1-
13/16" long. It should be understood, however, that the
size of ampoule 34 and the composition and quantity of
liquid 35 in the ampoule are coordinated with the ingre-
dients of tablet 33 so as to result in the generation of a -
predetermined volume of one or more gases as for example
an amount which will fill bag 50 when used and closed
without developing a gas pressure which will cause it to
rupture.
Although the above example illustrates the pro-
duction of hydrogen as the reducing gas by the use of
specific chemicals, i.e. potassium borohydride, zinc,
sodium chloride and dilute hydrochloric acid, other solid
materials can be used in conjunction with other liquids to
produce hydrogen or some other reducing gas which will
react catalytically with oxygen to remove it from the
space around the biological liquid sample. Thus, water
alone can be placed in ampoule 34 and the solid pellet 33
can be formulated to contain a material which reacts
with water safely and reasonably quickly to produce hydrogen.
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1056340
Thus, sodium borohydride, lithium aluminum hydride, lithium
hydride, calcium hydride, aluminum hydride and lithium
borohydride can be used since they react with water as
well as aqueous acid to form hydrogen. Such hydrides also
react with other liquids such as alcohols to form hydrogen
so that sometimes it may be desirable to replace the water
or acid with an alcohol, provided it does not adversely
affect the biological liquid sample. Hydrogen can, of
course, be produced by the reaction of a metal such as
iron, zinc, aluminum and magnesium with a suitable
acid such as sulfuric acid and hydrochloric acid.
Instead of using hydrogen as the reducing gas to
remove oxygen it is feasible to form acetylene by the
reaction of calcium carbide in pellet 33 and water or
dilute acid in ampoule 34.
The chemical means suitable for generating the
gaseous carbon dioxide in the apparatus of this invention
is not to be limited to the specific embodiment set forth
herein. Other well known chemical means for gaseous
carbon dioxide generation may be used. Broadly, any
solid material which upon contact with a liquid releases
carbon dioxide in adequate amount in a reasonably short
time may be used. The least expensive method, of course,
is to contact a carbonate or bicarbonate salt with a
dilute acid which will not produce vapors having an ad-
verse effect on the biological liquid sample. Instead of
putting a dilute acid in the ampoule it can be filled with
water, and sodium bicarbonate and citric acid, or some
suitable acid salt, can be put in pellet 33 to generate
carbon dioxide. Other feasible systems will appear
readily to skilled chemists.
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` `` ~056340
The dehydrating agent or desiccant 37 is advis-
ably included in the gas generating apparatus to remove
water and water vapor therefrom which may enter the tube
30 during sterilization, such as by ethylene oxide gas
sterilization, in the manufacturing process, or to remove
water vapor which may penetrate the tube in one way or
another. Removal of water in this way is desirable to
preserve the stability of the gas generating tablet 33,
although it is understood that under some conditions the
desiccant may not be necessarily employed.
The color indicator means 13 has an elongated
flexible transparent tube 40 which is open at end 42.
Tube 40 may be made of any suitable material although a
flexible polymeric material such as polyethylene is par-
ticularly suitable. Ampoule 43 is snugly positioned
within tube 40. The ampoule 43 may be made of any suit-
able material but desirably is made of relatively thin
glass so that it can be easily opened by fracturing the
ampoule walls by applying finger pressure through tube 40.
The ampoule 43 contains a liquid redox
color indicator 44 which occupies most if not all of the
space in the ampoule. The ampoule 43 shown in Figure 3
contains about 0.3 to 0.6 ml. of liquid and has a top
space 45 filled with an inert gas such as nitrogen or
carbon dioxide.
A fibrous liquid absorbent plug 46 is snugly
positioned in tube 40 between ampoule 43 and open end 42
of the tube. The fibrous plug 46 is made of a material
which is nonreactive with the redox liquid such as cotton,
polyester fibers or some other such liquid absorbent
material.
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1~56340
It is considered advisable to cover end 42 of
tube 40 with a bacteriological filter 47 through which
micro-organisms will not pass. The filter 47 is gas
permeable. The filter 47 is held in place by a cap 48
having a hole 49 in the top portion. A catalyst pellet 38
is also advisably placed in tube 40 between plug 46 and
filter 47.
The redox color indicator liquid 44 may be se-
lected from any suitable material which will change color
when the atmosphere around it changes from one which
is oxygen-free to one where there is a significant or sub-
stantial amount of oxygen in the atmosphere. Thus, the
indicator may have one color in the presence of oxygen and
a different color in an atmosphere which is devoid of
oxygen. Also, the indicator may be colorless when no
oxygen is present and develop a color when oxygen is
present, or the indicator may be colorless when oxygen is
present and develop a color when no oxygen is present in
the surrounding atmosphere.
A particularly useful redox color indicator
is rQsazurin in water. This redox indicator is colorless
in an atmosphere devoid of oxygen but in an oxygen-containing
atmosphere it has a pink color. When this indicator is
used it is advisable to include a small amount of cysteine
hydrochloride with it since this ingredient facilitates
color change. Another specific redox color indicator
which may be used is methylene blue. This indicator is
colorless in the absence of oxygen but in oxygen, such as
in the presence of air, it has a blue color. It is further-
more desirable that the redox color indicator used be
one which is color reversible so that any change from an
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1~56340
oxygen-containing atmosphere to an atmosphere deficient of
oxygen, or from an atmosphere deficient of oxygen to one
containing oxygen, will be indicated by the color change.
The preferred redox color indicator for use in
the apparatus is aqueous resazurin containing cysteine. A
0.001~ by weight solution of resazurin in water is specifi-
cally useful. ~ -
When the liquid redox color indicator liquid 44
is released from the ampoule 43 it flows onto absorbent
fibrous plug 46. The liquid is absorbed in this way
and held in place so as to provide a relatively easily
seen mass which can be observed through the transparent
wall of the tube. By making the absorbent plug of a white
fibrous material the color of the redox liquid indicator
can be readily observed and the presence or absence of
oxygen thereby determined.
The described liquid storage and transport appa-
ratus constitutes a disposable throw-away unit which is
intended to be employed only once. It is used by placing
a liquid sample in space 18 by use of a hypodermic
needle which punctures self-sealing closure 17. Once the
liquid sample has been placed in the apparatus it is held
upright with closure 17 at the top. Ampoule 34 is then
broken to free the liquid 35 which reacts with pellet 33
to produce a reducing gas and perhaps also carbon dioxide.
Ampoule 43 is also broken to free the redox color indicator
liquid 44. Gas generated in tube 30 flows through opening
20, space 18, through opening 27 into tube 40 and out
filter 47. The reducing gas catalytically reacts with
oxygen in the apparatus to form water by means of
catalyst pellets 38 and thereby removes the oxygen,
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1056340
forming an oxygen-deficient atmosphere in the apparatus.
The described apparatus is advisably used in
conjunction with a bag 50 as shown in Figure l. The appa-
ratus 10 is preferably shipped in such a bag lO, which
contains a catalyst pellet 38. The bag 50 may be made of
transparent polymeric flexible film or sheet material of
low gas permeability. The bag 50 may be made of two
sheets of plastic film heat sealed 52 around three side
edges, thereby leaving an open mouth 53 through which the
apparatus 10 is inserted. Specifically, a laminate
identified as No. CL5040 (Clear Lam Products) may be used
for the bag. The mouth 53 may be left open or be sealed
shut in any suitable way, such as by means of heat seal
54. To use the apparatus the bag 50 is cut at heat seal
54 and the apparatus is removed. The liquid sample is
then placed in space 18 of the apparatus, the apparatus
replaced in bag 50 and the mouth of the bag sealed, such --
as by a heat seal or by folding the bag mouth over on
itself a few times and clipping it together. The ampoules
34 and 43 are then crushed to activate the gas
generator and the color indicator. The generated gas
flows through the apparatus and out filter 47 into bag 50
which balloons outwardly. The ballooning effect is evidence
that the gases have generated as expected. However,
immediately upon generation of hydrogen as the reducing
gas, the catalyst pellets 38 induce reaction of the hydrogen
with the oxygen to form water. The described catalytic
removal of oxygen from bag 50 does not significantly
affect the ballooning. However, about 48 hours or so
after the unit is activated any carbon dioxide which
is simultaneously generated may have penetrated the walls
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-` lOS6340
of bag 50 causing a vacuum to develop therein. The external
atmospheric pressure may then press or collapse the flexible
walls of the bag together. This condition may result even
without oxygen entering the bag because the gas permea-
bility of the wall material may permit preferential flow
of carbon dioxide out of the bag but not oxygen in.
The decrease in oxygen concentration in bag 50
can be evidenced by the pink color of a resazurin saturated
plug 46 changing to light pink and finally to white when
the plug is made of polyester fibers, indicating the
oxygen has been removed. If oxygen subsequently leaks
into bag 50 the plug 46 will redevelop a pink color since
the color change is reversible when resazurin is used as
the color indicator.
When the package as shown in Figure 1 and con-
taining a liquid sample reaches a laboratory for analysis,
the apparatus 10 is removed from bag 50. The liquid
sample is then taken out of space 18 and then tested.
Figures 6 and 7 of the drawings illustrate a
second embodiment of the invention which, it will be
seen, incorporates a gas generator and color indicator
essentially identical to those shown in Figures 1 to 5.
It should be noted, however, that fibrous plug 66 is
placed in tube 40 of the color indicator 13 shown in
Figures 6 and 7 to prevent flow of liquid from ampoule 43
into biological liquid sample receptacle means 60.
The biological liquid sample receptacle means 60
shown in Figures 6 and 7 is substantially T-shaped and
hollow. The tubular arms 61 and 62 of receptacle means 60
are axially aligned and in communication with lateral
tubular extension 63. The end of tube 30 of the gas gen-
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lOS6340
erator container fits tightly over the end of arm 61 andthe end of tube 40 of the color indicator container means
fits tightly over the end of arm 62. Closure 64 covers
the end of tubular extension 63. Closure 64 can be made
of an elastomeric material which is self-sealing after
puncturing by a hypodermic needle to place an anaerobic
biological liquid sample in space 65 for storage and
transport.
The embodiment of the invention shown in Figures
6 and 7 operates in the same way as the embodiment
shown in Figures 1 to 5. However, it should be clear that - -
gas generated from pellet 33 and liquid 35 flows through
the entire length of the apparatus and out filter 47. It
is advisable, furthermore, to use the apparatus in con-
junction with a bag 50 as previously described herein.
Figures 8 to 11 illustrate a further embodiment
of the invention. As shown in Figures 8 and 9, the anaerobic
liquid storage and transport apparatus 70 has a liquid
receiver means 71 for receiving and holding an anaerobic
biological liquid, a gas generator apparatus 72, and
a color indicator apparatus 73.
The liquid receiver means 71, which may be
molded of clear or translucent polymeric material, has a
body with an exterior surface appearance of two elongated
hollow cylindrical portions 75 and 76 fused together with
tubular extensions 77 and 78 extending from the two opposing
ends of the body. The cylindrical portion 75 has a closed
end 79 while the end 80, which is slanted, of cylindrical
portion 76 is provided with a circular opening in which -~-
snap plug stopper 81 is positioned. The needle of
hypodermic syringe 82 penetrates stopper 81 to put in or
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i 1056340
remove a liquid sample from the space 83 inside of liquidreceiver means 71.
The gas generating apparatus 72 shown in Figures
8, 9 and 10, comprises a container in the form of an
elongated plastic tube 86 which is closed at end 88 and is
open at end 87. The tube 86 may be made of a flexible but
self-supporting polymeric material.
One or more gas generating solid tablets or
pellets 89 is positioned in tube 86 above the ampoule 90.
The tablet 89 has a composition which is suitable for
generating a reducing gas, or both a reducing gas and
carbon dioxide, as previously described herein.
Ampoule 90 is positioned in tube 86 more or less
snugly so that it maintains its position. A liquid 91 is
contained in ampoule 90. The composition of liquid 91 is
selected so that it, when released from ampoule 90, will
react with tablet 89 to generate one or more gases. The
ampoule g0 is advisably made so that it will rupture or
break upon application of finger pressure to the outside
of tube 86 adjacent the ampoule wall. In this way
the ampoule may be opened and the liquid 91 freed to react
with tablet 89 which drops into the freed liquid.
A liquid absorbent plug 92, such as of polyester
fibers, is positioned i~ tube 86 after the ampoule 90 is
placed in the tube. The absorbent plug 92 is thus located
between the ampoule 90 and the tube open end 87 so that
liquid cannot flow from the tube. The plug 92 may be
covered with a polytetrafluoroethylene-polypropylene
laminate (Gore-Tex) which allows passage of gas at low
pressure (7 psig) but not liquid.
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1056340
One or more desiccant pellets 93 are positioned
between liquid absorbent plug 92 and the open end 87 of
the tube 86. Any suitable desiccant or water dehydrating
material can be used for this purpose.
Also positioned between liquid absorbent plug 92
and the open end 87 of tube 86 is one or more catalyst
pellets 94. The catalyst pellet 94 is provided to induce
catalytic reaction between the reducing gas which is
formed by the combination of liquid 91 with tablet 89 and
any oxygen which may be in the liquid receiver means
71. A 0.5~ palladium-on-alumina catalyst may be used
although other catalysts which induce the reaction at room
temperature may be employed.
A polymeric cap 95 having a central hole 96 and
a covering of a non-woven sheet material 97 is pressed
firmly over the open end of tube 86. The sheet material
97 is one which will permit gas generated in tube 86 to
flow from the tube readily but which will not permit
liquid from space 83 to pass through it. A commercial
material marketed as Gore-Tex may be used for the sheet
material 97. Specifically, the sheet material 97 may
be Gore-Tex L-10477 (W. L. Gore & Assoc., Inc.) which is
1.0 mm of polytetrafluoroethylene on non-woven polypropylene.
The gas generator apparatus 72 is secured to the
liquid receiver means 71 by firmly forcing the cap 95 into
the opening in tubular extension 78.
The color indicator apparatus 73 shown in Figures
8, 9 and 11 has an elongated flexible transparent tube 101
which is open at its ends 102 and 103. Tube 101 can be
made of any suitable material although a flexible
polymeric material such as polypropylene is particularly
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suitable. Ampoule 104 is snugly positioned within tube
101. The ampoule 104 may be made of any suitable material
but desirably is made of relatively thin glass so that it
can be easily opened by fracturing the ampoule walls by
applying finger pressure against the adjacent surface of
tube 101. The ampoule 104 contains a liquid redox color
indicator 105 which occupies most if not all of the space
in the ampoule. A catalyst tablet or pellet 112 is posi-
tioned in tube 101 just past ampoule 104 to further facili-
tate reaction of the reducing gas with oxygen. A
fibrous liquid absorbent plug 107 is snugly positioned in
tube 101 below ampoule 104. The fibrous plug 107 is made
of a material which is nonreactive with the redox liquid,
such as polyester fibers or some other such liquid absor-
bent material. Each end of tube 101 is covered with a
non-woven sheet material 108, such as Gore-Tex L-10477,
which permits gas to pass but not micro-organisms and
which is not wetted by a liquid sample in space 83. The
sheet material 108 at each end of the tube 101 is held in
20 place by caps 109 and lO9A. Each cap has a hole 110
in the top portion.
The redox color indicator liquid 105 may be
selected from any suitable material which will change
color when the atmosphere around it changes from one which
is oxygen-free to one where there is a significant or
substantial amount of oxygen in the atmosphere.
The color indicator apparatus 73 is secured to
the liquid receiver means 71 by firmly forcing the cap 109
into the opening in tubular extension 77.
The embodiment of the invention shown in
Figures 8 to 11 can be used in the same way as the two
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other embodiments shown in Figures 1 to 7. Also, this
embodiment advisably would be used in conjunction with a
bag 50 as previously described herein.
Figures 12 and 13 illustrate still another
embodiment of the invention. This embodiment is very
similar to the embodiment shown in Figures 8 and 9. The
main difference is that the embodiment shown in Figures 8
and 9 incorporates a color indicator with the liquid
carrier whereas the embodiment of Figures 12 and 13 does
not employ an integral, but a separated, color indicator.
The anaerobic liquid storage and transport
apparatus 170 shown in Figures 12 and 13 has a liquid
receiver means 71 for receiving and holding an anaerobic
biological liquid and a gas generator apparatus 72. These
parts of the apparatus 170 are essentially identical with
the corresponding parts shown in Figures 8 to 11 except
that the gas generating tablet 130 is placed below the
ampoule 90 rather than above it. Furthermore, inside of
the tubular extension 77 there is positioned a water
impermeable, gas permeable filter plug 120. Filter
plug 120 may contain a catalyst to induce reaction between
generated hydrogen and oxygen in the liquid carrier,
although the catalyst is not essential but advisably
included. The particular filter plug 120 shown in Figures
12 and 13 has tubular body 121 covered at one end by a cap
122 having an opening at the top. A covering of a non-
woven sheet material is pressed over the end of tube 121
before the cap 122 is placed thereon. The non-woven sheet
material is gas permeable but liquid impermeable. The
described cap and sheet material are identical with
previously described cap 95 and sneet material 97. The
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cap 122 fits snugly inside of tubular extension 77 forabout the full length of the cap. Catalyst pellets 123
are positioned inside of the tube 121 and a gas permeable
polyethylene plug 124 is put in the end of the tube to
hold the catalyst pellets in place. Plug 124 also prevents
bacteria from entering or leaving space 83.
The described liquid transport apparatus develops
a lower back pressure in use than the apparatus shown in
Figures 8 and 9. This is largely due to the absence of
the color indicator from tubular extension 77 in the
apparatus of Figures 12 and 13.
The liquid transport apparatus 170 is used in
the same way as are the other embodiments disclosed herein.
Furthermore, the liquid transport apparatus 170 is advisably
employed in conjunction with a bag 50 as previously described
herein. In addition, it is desirable to include in the
bag 50, a color indicator 130 and a catalyst holder 140.
The color indicator 130 has an elongated flexible
transparent tube 131. Rupturable ampoule 132 is snugly
20 positioned within tube 131. The ampoule 132 contains
a liquid redox color indicator 133 which occupies most if
not all of the space in the ampoule. A fibrous liquid
absorbent plug 134 is snugly positioned in tube 131. A
plug 135 of gas-permeable, liquid impermeable expanded
polyethylene is positioned inside of each end of tube 131
to complete the color indicator.
The catalyst holder 140 is composed of a flexible
tube 141, such as of polyethylene, catalyst pellets 142
and a gas-permeable plug 143 inside each end of the tube.
The catalyst pellets 142 are selected to induce
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reaction between hydrogen gas generated by gas generator
72 and oxygen in bag 50.
It is intended that the unit shown in Figure 13
comprising bag 50 containing liquid transport apparatus
170, color indicator 130 and catalyst holder 140, be
marketed in a second bag, not shown, to preserve the
sterility of all the described components. When the unit
is to be used, the second bag is opened and discarded.
Liquid transport apparatus 170 is then removed from bag
50, a liquid sample placed in space 83, the apparatus
then replaced in bag 50 and the bag closed against gas
passage. Ampoules 90 and 133 are then crushed to actuate
the gas generator and the color indicator. Bag 50 will
balloon as hydrogen is generated but subsequently may
partially collapse as the hydrogen and oxygen react to
form water to thereby lower the gas pressure. The atmos-
phere inside of bag 50, as well as the liquid transport
apparatus 170, becomes oxygen-free or at least very low in
oxygen, thus providing an anaerobic environment for the
liquid sample.
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