U.S. patent application number 14/686706 was filed with the patent office on 2016-02-04 for mixing device for creating an output mixture by mixing a first material and a second material.
The applicant listed for this patent is REVALESIO CORPORATION. Invention is credited to Gregory J. Archambeau, Richard L. Watson, Anthony B. Wood.
Application Number | 20160030901 14/686706 |
Document ID | / |
Family ID | 39325445 |
Filed Date | 2016-02-04 |
United States Patent
Application |
20160030901 |
Kind Code |
A1 |
Wood; Anthony B. ; et
al. |
February 4, 2016 |
MIXING DEVICE FOR CREATING AN OUTPUT MIXTURE BY MIXING A FIRST
MATERIAL AND A SECOND MATERIAL
Abstract
A mixing device for mixing a first and second material together
to create an output mixture. The device includes a first chamber
containing the first material coupled to a mixing chamber defined
between a rotor and a stator. The rotor is disposed inside the
stator and rotates therein about an axis of rotation. The first
chamber houses an internal pump configured to pump the first
material from the first chamber into the mixing chamber. The pump
may be configured to impart a circumferential velocity into the
first material before it enters the mixing chamber. At least one of
the rotor and stator have a plurality of through-holes through
which the second material is provided to the mixing chamber.
Optionally, a second chamber is coupled to the mixing chamber. The
second chamber may house an internal pump configured to pump the
output material from the mixing chamber into the second
chamber.
Inventors: |
Wood; Anthony B.; (Tacoma,
WA) ; Archambeau; Gregory J.; (Puyallup, WA) ;
Watson; Richard L.; (Ruston, WA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
REVALESIO CORPORATION |
TACOMA |
WA |
US |
|
|
Family ID: |
39325445 |
Appl. No.: |
14/686706 |
Filed: |
April 14, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13902663 |
May 24, 2013 |
9004743 |
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14686706 |
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|
12945703 |
Nov 12, 2010 |
8449172 |
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13902663 |
|
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|
11924589 |
Oct 25, 2007 |
7832920 |
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12945703 |
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60982387 |
Oct 24, 2007 |
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60862955 |
Oct 25, 2006 |
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60862904 |
Oct 25, 2006 |
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Current U.S.
Class: |
366/170.3 ;
261/34.1; 261/37; 261/84; 366/336; 435/289.1 |
Current CPC
Class: |
B01F 3/0807 20130101;
A61P 27/02 20180101; B01F 5/0665 20130101; A61J 3/00 20130101; B01F
3/04099 20130101; Y02W 10/10 20150501; C02F 1/727 20130101; C09K
3/00 20130101; A61K 9/08 20130101; B01F 7/008 20130101; B01F
7/00816 20130101; A23L 2/54 20130101; A61K 33/00 20130101; B01F
3/04531 20130101; B01F 3/0853 20130101; Y02W 10/15 20150501; C02F
3/02 20130101; A61K 9/10 20130101; C12M 27/00 20130101; A61P 17/02
20180101 |
International
Class: |
B01F 5/06 20060101
B01F005/06; B01F 7/00 20060101 B01F007/00; B01F 3/04 20060101
B01F003/04 |
Claims
1. A mixing device for creating an output mixture by mixing a first
material and a second material, the device comprising: a first
chamber configured to receive the first material from a source of
the first material; a stator; a rotor having an axis of rotation,
the rotor being disposed inside the stator and configured to rotate
about the axis of rotation therein, at least one of the rotor and
stator having a plurality of through-holes; a mixing chamber
defined between the rotor and the stator, the mixing chamber being
in fluid communication with the first chamber and configured to
receive the first material therefrom, and to receive the second
material into the mixing chamber via the plurality of through-holes
formed in the at least one of the rotor and stator; and a first
internal pump housed inside the first chamber, the first internal
pump being configured to pump the first material from the first
chamber into the mixing chamber, and wherein the first material
comprises a fluid and the second material comprises a gas.
2. The mixing device of claim 1, wherein the first internal pump is
configured to impart a circumferential velocity into the first
material before it enters the mixing chamber.
3. The mixing device of claim 2, wherein the rotor imparts a mixing
circumferential velocity into the first material and the second
material inside the mixing chamber, and the circumferential
velocity imparted into the first material by the first internal
pump approximates the mixing circumferential velocity imparted by
the rotor.
4. The mixing device of claim 1, further comprising a drive shaft
coupled to the rotor and extending though the first chamber along
the axis of rotation, the drive shaft being configured to rotate
the rotor about the axis of rotation and to power to the first
internal pump.
5. The mixing device of claim 4, wherein the rotor has a sidewall
defining a hollow portion into which the drive shaft extends, the
sidewall has a plurality of openings providing communication
between the hollow portion and the mixing chamber, and the drive
shaft comprises an internal channel having a first opening into the
hollow portion of the drive shaft and a second opening, the mixing
device further comprising a source of the second material, the
source being coupled to the second opening of the channel and the
source being configured to supply the second material into the
mixing chamber through the channel, the hollow portion of the
rotor, and the plurality of openings of the sidewall of the
rotor.
6. The mixing device of claim 1, wherein the stator comprises a
plurality of through-holes, the mixing device further comprising: a
housing comprising an input port, the stator being housed inside
the housing; a channel defined between the housing and the stator,
the input port being in communication with the channel, the
plurality of through-holes of the stator providing communication
between the mixing chamber and the channel; and a source of a third
material coupled to the input port and configured to supply the
third material to the mixing chamber through the input port, the
channel, and the plurality of through-holes of the stator.
7. The mixing device of claim 1, further comprising: a second
chamber in fluid communication with the mixing chamber and
configured to receive the output material therefrom; and a second
internal pump housed inside the second chamber, the second internal
pump being configured to pump the output material from the mixing
chamber into the second chamber.
8. The mixing device of claim 1, further comprising: a second
chamber in fluid communication with the mixing chamber and
configured to receive the output material therefrom; a drive shaft
coupled to the rotor and extending though the first chamber, the
rotor, and the second chamber along the axis of rotation; and a
second internal pump housed inside the second chamber, the second
internal pump being configured to pump the output material from the
mixing chamber into the second chamber, the drive shaft being
configured to rotate the rotor about the axis of rotation and to
power to the second internal pump.
9. The mixing device of claim 1, further comprising: a second
chamber in fluid communication with the mixing chamber and
configured to receive the output material therefrom; and a second
internal pump housed inside the second chamber, the second internal
pump being configured to pump the output material from the mixing
chamber into the second chamber, and to impart a circumferential
velocity into the output material after it enters the mixing
chamber.
10. The mixing device of claim 1, wherein both the rotor and the
stator have a substantially cylindrical shape with a longitudinal
axis aligned along the axis of rotation, and the mixing chamber has
a ring-shaped cross-sectional shape having a thickness of about
0.02 inches to about 0.08 inches.
11. The mixing device of claim 1, wherein the rotor rotates about
the axis of rotation in a rotation direction having an tangential
component, the first chamber comprises an input port configured to
receive the first material from a source of the first material, the
input port being configured to introduce the first material into
the first chamber traveling in a direction substantially equivalent
to the tangential component of the rotation direction.
12. The mixing device of claim 11, wherein the first chamber has an
internal shape configured to deflect the first material and direct
it to flow in the rotation direction.
13. The mixing device of claim 11, further comprising a second
chamber in fluid communication with the mixing chamber and
configured to receive the output material therefrom, the second
chamber comprising an output port through which the output material
may exit the mixing device, the input port being configured to
allow the output material to exit the second chamber traveling in a
direction substantially equivalent to the tangential component of
the rotation direction.
14. The mixing device of claim 13, wherein the second chamber has
an internal shape configured to deflect the output material and
direct it to flow in the rotation direction.
15. A mixing device for creating an output mixture by mixing a
first material and a second material, the device comprising: a
stator; a rotor having an axis of rotation, the rotor being
disposed inside the stator and configured to rotate about the axis
of rotation therein, at least one of the rotor and stator having a
plurality of through-holes; a mixing chamber defined between the
rotor and the stator, the mixing chamber having an open first end
through which the first material enters the mixing chamber and an
open second end through which the output material exits the mixing
chamber, the second material entering the mixing chamber through
the plurality of through-holes formed in the at least one of the
rotor and the stator; a first chamber in communication with at
least a majority portion of the open first end of the mixing
chamber; and a second chamber in communication with the open second
end of the mixing chamber, and wherein the first material comprises
a fluid and the second material comprises a gas.
16. (canceled)
17. The mixing device of claim 15, wherein the second chamber is in
communication with at least a majority portion of the open second
end of the mixing chamber.
18. The mixing device of claim 15, further comprising: a first
internal pump housed inside the first chamber and configured to
pump the first material from the first chamber into the open first
end of the mixing chamber and to impart a circumferential velocity
into the first material before it enters the open first end of the
mixing chamber.
19. The mixing device of claim 18, further comprising: a second
internal pump housed inside the second chamber and configured to
pump the output material from the open second end of the mixing
chamber into the second chamber and to impart a circumferential
velocity into the second material after it exits the mixing
chamber.
20. The mixing device of claim 15, wherein the first chamber
comprises an input port coupled to an external pump, the external
pump configured to pump the first fluid into the first chamber, the
input port being positioned to introduce the first material into
the first chamber traveling in a direction substantially tangential
to the axis of rotation, the first chamber having an internal shape
configured to deflect the first material traveling in a direction
substantially tangential to the axis of rotation into a
circumferential flow about the axis of rotation.
21. A bioreactor system, comprising a bioreactor in combination
with the mixing device of any one of claims 1 and 15, or with a
gas-enriched fluid derived using the mixing device of any one of
claims 1 and 15.
22. A method of mixing a first material and a second material in a
mixing chamber formed between two contoured surfaces to create an
output mixture, the arcuate mixing chamber having a first end
portion opposite a second end portion, the method comprising:
introducing the first material into the first end portion of the
arcuate mixing chamber in a flow direction having a first component
that is substantially tangent to the mixing chamber and a second
component that is directed toward the second end portion; and
introducing the second material into the mixing chamber though at
least one of the two contoured surfaces between the first end
portion of the arcuate mixing chamber and the second end portion of
the arcuate mixing chamber, and wherein the first material
comprises a fluid and the second material comprises a gas.
23. The method of claim 22, wherein the first end portion of the
mixing chamber is coupled to a first chamber, the method further
comprising: before introducing the first material into the first
end portion of the mixing chamber, introducing the first material
into the first chamber, and imparting a circumferential flow into
the first material in the first chamber.
24. The method of claim 22, wherein the first end portion of the
mixing chamber is coupled to a first chamber, the mixing chamber is
formed between an outer contoured surface of a rotating cylindrical
rotor and an inner contoured surface of a stationary cylindrical
stator, and the rotor rotates inside the stator about an axis of
rotation, the method further comprising: before introducing the
first material into the first end portion of the mixing chamber,
introducing the first material into the first chamber, and
imparting a circumferential flow substantially about an axis of
rotation into the first material in the first chamber; introducing
the second material into a hollow portion of a rotating rotor
having a plurality of through-holes, each through-hole of the
plurality extending from the hollow portion to the outer contoured
surface of the rotor; flowing the second material from the hollow
portion of the rotating rotor through the plurality of
through-holes into the mixing chamber; flowing the first material
from the first chamber into the mixing chamber; and rotating the
rotor relative to the stator thereby mixing the first material and
the second material together inside the mixing chamber.
25. The mixing device of claim 1, wherein the mixing chamber is
cylindrical or arcuate.
26. The mixing device of claim 15, wherein the mixing chamber is
arcuate or cylindrical.
27. The method of claim 22, wherein the mixing chamber is arcuate
or cylindrical.
28. The mixing device of claim 1, wherein the fluid is an aqueous
liquid, and the gas is oxygen gas.
29. The mixing device of claim 15, wherein the fluid is an aqueous
liquid, and the gas is oxygen gas.
30. The method of claim 22, wherein the fluid is an aqueous liquid,
and the gas is oxygen gas.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. patent
application Ser. No. 13/902,663, filed May 24, 2013, entitled
"MIXING DEVICE FOR CREATING AN OUTPUT MIXTURE BY MIXING A FIRST
MATERIAL AND A SECOND MATERIAL" (issuing on Apr. 14, 2015, as U.S.
Pat. No. 9,004,743), which is a continuation of U.S. patent
application Ser. No. 12/945,703, filed Nov. 12, 2010, entitled
"MIXING DEVICE FOR CREATING AN OUTPUT MIXTURE BY MIXING A FIRST
MATERIAL AND A SECOND MATERIAL" (now U.S. Pat. No. 8,449,172,
issued on May 28, 2013), which is a continuation of U.S. patent
application Ser. No. 11/924,589, filed Oct. 25, 2007, entitled
"MIXING DEVICE FOR CREATING AN OUTPUT MIXTURE BY MIXING A FIRST
MATERIAL AND A SECOND MATERIAL" (now U.S. Pat. No. 7,832,920,
issued on Nov. 16, 2010), which claims the benefit of U.S.
Provisional Patent Application No. 60/982,387, filed Oct. 24, 2007,
entitled "MIXING DEVICE," 60/862,955, filed Oct. 25, 2006, entitled
"OXYGENATED SALINE SOLUTION," and 60/862,904, filed Oct. 25, 2006,
entitled "DIFFUSER/EMULSIFIER." The disclosures of which are hereby
incorporated by reference herein in their entirety.
SEQUENCE LISTING
[0002] A Sequence Listing comprising SEQ ID NO:1, has been provided
in computer readable form (.txt) as part of this application, and
is incorporated by reference herein in its entirety as part of this
application.
BACKGROUND OF THE INVENTION
[0003] 1. Field of the Invention
[0004] The present invention is directed generally to mixing
devices and more particularly to mixing devices that mix two or
more materials between surfaces, including such as between a
rotating rotor and a stationary stator.
[0005] 2. Description of the Related Art
[0006] FIG. 1 provides a partial block diagram, partial
cross-sectional view of a prior art device 10 for diffusing or
emulsifying one or two gaseous or liquid materials ("infusion
materials") into another gaseous or liquid material ("host
material") reproduced from U.S. Pat. No. 6,386,751, incorporated
herein by reference in its entirety. The device 10 includes a
housing configured to house a stator 30 and a rotor 12. The stator
30 encompasses the rotor 12. A tubular channel 32 is defined
between the rotor 12 and the stator 30. The generally cylindrically
shaped rotor 12 has a diameter of about 7.500 inches and a length
of about 6.000 inches providing a length to diameter ratio of about
0.8.
[0007] The rotor 12 includes a hollow cylinder, generally closed at
both ends. A gap exists between each of the first and second ends
of the rotor 12 and a portion of the housing 34. A rotating shaft
14 driven by a motor 18 is coupled to the second end of the rotor
12. The first end of the rotor 12 is coupled to an inlet 16. A
first infusion material passes through the inlet 16 and into the
interior of the rotor 12. The first infusion material passes from
the interior of the rotor 12 and into the channel 32 through a
plurality of openings 22 formed in the rotor 12.
[0008] The stator 30 also has openings 22 formed about its
circumference. An inlet 36 passes a second infusion material to an
area 35 between the stator 30 and the housing 34. The second
infusion material passes out of the area 35 and into the channel 32
through openings 22.
[0009] An external pump (not shown) is used to pump the host
material into a single inlet port 37. The host material passes
through a single inlet port 37 and into the channel 32 where it
encounters the first and second infusion materials, which enter the
channel 32 through openings 22. The infusion materials may be
pressurized at their source to prevent the host material from
passing through openings 22.
[0010] The inlet port 37, is configured and positioned such that it
is located along only a relatively small portion (<about 5%) of
the annular inlet channel 32, and is substantially parallel to the
axis of rotation of the rotor 12 to impart an axial flow toward a
portion of the channel 32 into the host material.
[0011] Unfortunately, before entering the tubular channel 32, the
host material must travel in tortuous directions other than that of
the axial flow (e.g., including in directions substantially
orthogonal thereto) and down into and between the gap formed
between the first end of the rotor 12 and the housing 34 (i.e.,
down a portion of the first end of the rotor adjacent to the inlet
16 between the end of the rotor 12 and the housing 34). The
non-axial and orthogonal flow, and the presence of the host
material in the gap between the first end of the rotor 12 and the
housing 34 causes undesirable and unnecessary friction. Further, it
is possible for a portion of the host material to become trapped in
eddy currents swirling between the first end of the rotor and the
housing. Additionally, in the device 10, the host material must
negotiate at least two right angles to enter any aspect of the
annual of the annular inlet of the tubular channel 32.
[0012] A single outlet port 40 is formed in the housing 34. The
combined host material and infusion material(s) exit the channel 32
via the outlet 40. The outlet port 40, which is also located along
only a limited portion (<about 5%) of the annular outlet of
tubular channel 32, is substantially parallel to the axis of
rotation of the rotor 12 to impart or allow for an axial flow of
the combined materials away from the limited portion of the annular
outlet of tubular channel 32 into the outlet port 40. An external
pump 42 is used to pump the exiting fluid through the outlet port
40.
[0013] Unfortunately, before exiting the channel 32, a substantial
portion of the exiting material must travel in a tortuous direction
other than that of the axial flow (e.g., including in directions
substantially orthogonal thereto) and down into and between the gap
formed between the second end of the rotor 12 and the housing 34
(i.e., down a portion of the second end of the rotor adjacent to
the shaft 14 between the end of the rotor 12 and the housing 34).
As mentioned above, the non-axial and orthogonal flow, and the
presence of the host material in the other gap between the end (in
this case, the second end) of the rotor 12 and the housing 34
causes additional undesirable and unnecessary friction. Further, it
is possible for a portion of the host material to become trapped in
eddy currents swirling between the second end of the rotor and the
housing. Additionally, in the device 10, a substantial portion of
the exiting combined material must negotiate at least two right
angles as it exits form the annular exit of the tubular channel 32
into the outlet port 40.
[0014] As is apparent to those of ordinary skill in the art, the
inlet port 37 imparts only an axial flow to the host material. Only
the rotor 21 imparts a circumferential flow into the host material.
Further, the outlet port 40 imparts or provides for only an axial
flow into the exiting material. Additionally, the circumferential
flow velocity vector is imparted to the material only after it
enters the annular inlet 37 of the tubular channel 32, and
subsequently the circumferential flow vector must be degraded or
eliminated as the material enters the exit port 40. There is,
therefore, a need for a progressive circumferential acceleration of
the material as it passes in the axial direction through the
channel 32, and a circumferential deceleration upon exit of the
material from the channel 32. These aspects, in combination with
the tortuous path that the material takes from the inlet port 37 to
the outlet port 40, create a substantial friction and flow
resistance over the path that is accompanied by a substantial
pressure differential (26 psi, at 60 gallons/min flow rate) between
the inlet 37 and outlet 40 ports, and these factors, inter alia,
combine to reduce the overall efficiency of the system.
BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWING(S)
[0015] FIG. 1 is a partial cross-section, partial block diagram of
a prior art mixing device.
[0016] FIG. 2 is block diagram of an exemplary embodiment of a
mixing device.
[0017] FIG. 3 is an illustration of an exemplary system for
delivering a first material to the mixing device of FIG. 2.
[0018] FIG. 4 is a fragmentary partial cross-sectional view of a
top portion of the mixing device of FIG. 2.
[0019] FIG. 5 is a fragmentary cross-sectional view of a first side
portion of the mixing device of FIG. 2.
[0020] FIG. 6 is a fragmentary cross-sectional view of a second
side portion of the mixing device of FIG. 2.
[0021] FIG. 7 is a fragmentary cross-sectional view of a side
portion of the mixing device of FIG. 2 located between the first
side portion of FIG. 5 and the second side portion of FIG. 6.
[0022] FIG. 8 is a perspective view of a rotor and a stator of the
mixing device of FIG. 2.
[0023] FIG. 9 is a perspective view of an inside of a first chamber
of the mixing device of FIG. 2.
[0024] FIG. 10 is a fragmentary cross-sectional view of the inside
of a first chamber of the mixing device of FIG. 2 including an
alternate embodiment of the pump 410.
[0025] FIG. 11 is a perspective view of an inside of a second
chamber of the mixing device of FIG. 2.
[0026] FIG. 12 is a fragmentary cross-sectional view of a side
portion of an alternate embodiment of the mixing device.
[0027] FIG. 13 is a perspective view of an alternate embodiment of
a central section of the housing for use with an alternate
embodiment of the mixing device.
[0028] FIG. 14 is a fragmentary cross-sectional view of an
alternate embodiment of a bearing housing for use with an alternate
embodiment of the mixing device.
[0029] FIG. 15 is a cross-sectional view of the mixing chamber of
the mixing device of FIG. 2 taken through a plane orthogonal to the
axis of rotation depicting a rotary flow pattern caused by
cavitation bubbles when a through-hole of the rotor approaches (but
is not aligned with) an aperture of the stator.
[0030] FIG. 16 is a cross-sectional view of the mixing chamber of
the mixing device of FIG. 2 taken through a plane orthogonal to the
axis of rotation depicting a rotary flow pattern caused by
cavitation bubbles when the through-hole of the rotor is aligned
with the aperture of the stator.
[0031] FIG. 17 is a cross-sectional view of the mixing chamber of
the mixing device of FIG. 2 taken through a plane orthogonal to the
axis of rotation depicting a rotary flow pattern caused by
cavitation bubbles when a through-hole of the rotor that was
previously aligned with the aperture of the stator is no longer
aligned therewith.
[0032] FIG. 18 is a side view of an alternate embodiment of a
rotor.
[0033] FIG. 19 is an enlarged fragmentary cross-sectional view
taken through a plane orthogonal to an axis of rotation of the
rotor depicting an alternate configuration of through-holes formed
in the rotor and through-holes formed in the stator.
[0034] FIG. 20 is an enlarged fragmentary cross-sectional view
taken through a plane passing through and extending along the axis
of rotation of the rotor depicting a configuration of through-holes
formed in the rotor and through-holes formed in the stator.
[0035] FIG. 21 is an enlarged fragmentary cross-sectional view
taken through a plane passing through and extending along the axis
of rotation of the rotor depicting an alternate offset
configuration of through-holes formed in the rotor and
through-holes formed in the stator.
[0036] FIG. 22 is an illustration of a shape that may be used to
construct the through-holes of the rotor and/or the apertures of
the stator.
[0037] FIG. 23 is an illustration of a shape that may be used to
construct the through-holes of the rotor and/or the apertures of
the stator.
[0038] FIG. 24 is an illustration of a shape that may be used to
construct the through-holes of the rotor and/or the apertures of
the stator.
[0039] FIG. 25 is an illustration of a shape that may be used to
construct the through-holes of the rotor and/or the apertures of
the stator.
[0040] FIG. 26 is an illustration of an electrical double layer
("EDL") formed near a surface.
[0041] FIG. 27 is a perspective view of a model of the inside of
the mixing chamber.
[0042] FIG. 28 is a cross-sectional view of the model of FIG.
27.
[0043] FIG. 29 is an illustration of an experimental setup.
[0044] FIG. 30 illustrates dissolved oxygen levels in water
processed with oxygen in the mixing device of FIG. 2 and stored a
500 ml thin walled plastic bottle and a 1,000 ml glass bottle each
capped at 65.degree. Fahrenheit.
[0045] FIG. 31 illustrates dissolved oxygen levels in water
processed with oxygen in the mixing device of FIG. 2 and stored in
a 500 ml plastic thin walled bottle and a 1,000 ml glass bottle
both refrigerated at 39.degree. Fahrenheit.
[0046] FIG. 32 illustrates the dissolved oxygen levels in
GATORADE.RTM. processed with oxygen in the mixing device of FIG. 2
and stored in 32 oz. GATORADE.RTM. bottles having an average
temperature of 55.degree. Fahrenheit.
[0047] FIG. 33 illustrates the dissolved oxygen retention of a 500
ml braun balanced salt solution processed with oxygen in the mixing
device of FIG. 2.
[0048] FIG. 34 illustrates a further experiment wherein the mixing
device of FIG. 2 is used to sparge oxygen from water by processing
the water with nitrogen in the mixing device of FIG. 2.
[0049] FIG. 35 illustrates the sparging of oxygen from water by the
mixing device of FIG. 2 at standard temperature and pressure.
[0050] FIG. 36 is an illustration of a nanocage.
[0051] FIG. 37 illustrates the Rayleigh scattering effects produced
by a sample of the water processed with oxygen by the mixing device
of FIG. 2.
[0052] FIGS. 38-41 illustrate the inventive oxygen-enriched fluid
tested positive for reactivity with horseradish peroxidase by
pyrogallol, while the pressure pot and fine bubbled water samples
had far less reactivity.
[0053] FIG. 42 illustrates pyrogallol/HRP assays as described
herein, showing that oxygen is required for the reaction with
pyrogallol in the presence of horseradish peroxidase, as inventive
fluid enriched with other gases (argon and nitrogen) did not react
in the same manner.
[0054] FIG. 43 illustrates the hydrogen peroxide positive control
showed a strong reactivity, while none of the other fluids tested
reacted with the glutathione.
[0055] FIG. 44 illustrates T7 DNA shows a conformational change at
about 50 degrees Celsius in the control (deionized water), whereas
the DNA in the oxygen-enriched inventive fluid remains intact until
about 60 degrees Celsius.
[0056] FIGS. 45A and 45B illustrate a graphical representation of
an exemplary embodiment of a bioreactor system 3300a.
[0057] FIG. 46 shows detailed portions of exemplary embodiments of
the bioreactor system 3300a of FIGS. 45A and 45B.
DETAILED DESCRIPTION OF THE INVENTION
Overview
[0058] FIG. 2 provides a block diagram illustrating some of the
components of a mixing device 100 and the flow of material into,
within, and out of the device. The mixing device 100 combines two
or more input materials to form an output material 102, which may
be received therefrom into a storage vessel 104. The mixing device
100 agitates the two or more input materials in a novel manner to
produce an output material 102 having novel characteristics. The
output material 102 may include not only a suspension of at least
one of the input materials in at least one of the other input
materials (e.g., emulsions) but also a novel combination (e.g.,
electrostatic combinations) of the input materials, a chemical
compound resulting from chemical reactions between the input
materials, combinations having novel electrostatic characteristics,
and combinations thereof.
[0059] The input materials may include a first material 110
provided by a source 112 of the first material, a second material
120 provided by a source 122 of the second material, and optionally
a third material 130 provided by a source 132 of the third
material. The first material 110 may include a liquid, such as
water, saline solution, chemical suspensions, polar liquids,
non-polar liquids, colloidal suspensions, cell growing media, and
the like. In some embodiments, the first material 110 may include
the output material 102 cycled back into the mixing device 100. The
second material 120 may consist of or include a gas, such as
oxygen, nitrogen, carbon dioxide, carbon monoxide, ozone, sulfur
gas, nitrous oxide, nitric oxide, argon, helium, bromine, and
combinations thereof, and the like. In preferred embodiments, the
gas is or comprises oxygen. The optional third material 130 may
include either a liquid or a gas. In some embodiments, the third
material 130 may be or include the output material 102 cycled back
into the mixing device 100 (e.g., to one or more of the pumps 210,
220, or 230, and/or into the chamber 310, and/or 330).
[0060] Optionally, the first material 110, the second material 120,
and the optional third material 130 may be pumped into the mixing
device 100 by an external pump 210, an external pump 220, and an
external pump 230, respectively. Alternatively, one or more of the
first material 110, the second material 120, and the optional third
material 130 may be stored under pressure in the source 112, the
source 122, and the source 132, respectively, and may be forced
into the mixing device 100 by the pressure. The invention is not
limited by the method used to transfer the first material 110, the
second material 120, and optionally, the third material 130 into
the mixing device 100 from the source 112, the source 122, and the
source 132, respectively.
[0061] The mixing device 100 includes a first chamber 310 and a
second chamber 320 flanking a mixing chamber 330. The three
chambers 310, 320, and 330 are interconnected and form a continuous
volume.
[0062] The first material 110 is transferred into the first chamber
310 and flows therefrom into the mixing chamber 330. The first
material 110 in the first chamber 310 may be pumped into the first
chamber 310 by an internal pump 410. The second material 120 is
transferred into the mixing chamber 330. Optionally, the third
material 130 may be transferred into the mixing chamber 330. The
materials in the mixing chamber 330 are mixed therein to form the
output material 102. Then, the output material 102 flows into the
second chamber 320 from which the output material 102 exits the
mixing device 100. The output material 102 in the mixing chamber
330 may be pumped into the second chamber 320 by an internal pump
420. Optionally, the output material 102 in the second chamber 320
may be pumped therefrom into the storage vessel 104 by an external
pump 430 (e.g., alone or in combination with the internal pump 410
and/or 420).
[0063] In particular aspects, a common drive shaft 500 powers both
the internal pump 410 and the internal pump 420. The drive shaft
500 passes through the mixing chamber 330 and provides rotational
force therein that is used to mix the first material 110, the
second material 120, and optionally, the third material 130
together. The drive shaft 500 is powered by a motor 510 coupled
thereto.
[0064] FIG. 3 provides a system 512 for supplying the first
material 110 to the mixing device 100 and removing the output
material 102 from the mixing device 100. In the system 512, the
storage vessel 104 of the output material 102 and the source 112 of
the first material 110 are combined. The external pump 210 is
coupled to the combined storage vessel 104 and source 112 by a
fluid conduit 514 such as hose, pipe, and the like. The external
pump 210 pumps the combined first material 110 and output material
102 from the combined storage vessel 104 and source 112 through the
fluid conduit 514 and into a fluid conduit 516 connecting the
external pump 210 to the mixing device 100. The output material 102
exits the mixing device 100 through a fluid conduit 518. The fluid
conduit 518 is coupled to the combined storage vessel 104 and
source 112 and transports the output material 102 exiting the
mixing device 100 to the combined storage vessel 104 and source
112. The fluid conduit 518 includes a valve 519 that establishes an
operating pressure or back pressure within the mixing device
100.
[0065] Referring to FIGS. 2, 4-10, and 11, a more detailed
description of various components of an embodiment of the mixing
device 100 will be provided. The mixing device 100 is scalable.
Therefore, dimensions provided with respect to various components
may be used to construct an embodiment of the device or may be
scaled to construct a mixing device of a selected size.
[0066] Turning to FIG. 4, the mixing device 100 includes a housing
520 that houses each of the first chamber 310, the mixing chamber
330, and the second chamber 320. As mentioned above, the mixing
device 100 includes the drive shaft 500, which rotates during
operation of the device. Therefore, the mixing device 100 may
vibrate or otherwise move. Optionally, the mixing device 100 may be
coupled to a base 106, which may be affixed to a surface such as
the floor to maintain the mixing device 100 in a substantially
stationary position.
[0067] The housing 520 may be assembled from two or more housing
sections. By way of example, the housing 520 may include a central
section 522 flanked by a first mechanical seal housing 524 and a
second mechanical seal housing 526. A bearing housing 530 may be
coupled to the first mechanical seal housing 524 opposite the
central section 522. A bearing housing 532 may be coupled to the
second mechanical seal housing 526 opposite the central section
522. Optionally, a housing section 550 may be coupled to the
bearing housings 530.
[0068] Each of the bearing housings 530 and 532 may house a bearing
assembly 540 (see FIGS. 5 and 6). The bearing assembly 540 may
include any suitable bearing assembly known in the art including a
model number "202SZZST" manufactured by SKF USA Inc., of
Kulpsville, Pa., operating a website at www.skf.com.
[0069] Seals may be provided between adjacent housing sections. For
example, o-ring 560 (see FIG. 5) may be disposed between the
housing section 550 and the bearing housing 530, o-ring 562 (see
FIG. 5) may be disposed between the first mechanical seal housing
524 and the central section 522, and o-ring 564 (see FIG. 6) may be
disposed between the second mechanical seal housing 526 and the
central section 522.
Mixing Chamber 330
[0070] Turning now to FIG. 7, the mixing chamber 330 is disposed
inside the central section 522 of the housing 520 between the first
mechanical seal housing 524 and the second mechanical seal housing
526. The mixing chamber 330 is formed between two components of the
mixing device 100, a rotor 600 and a stator 700. The rotor 600 may
have a sidewall 604 with an inside surface 605 defining a generally
hollow inside portion 610 and an outside surface 606. The sidewall
604 may be about 0.20 inches to about 0.75 inches thick. In some
embodiments, the sidewall 604 is about 0.25 inches thick. However,
because the mixing device 100 may be scaled to suit a particular
application, embodiments of the device having a sidewall 604 that
is thicker or thinner than the values provided are within the scope
of the present teachings. The sidewall 604 includes a first end
portion 612 and a second end portion 614 and a plurality of
through-holes 608 formed between the first end portion 612 and the
second end portion 614. Optionally, the outside surface 606 of the
sidewall 604 may include other features such as apertures,
projections, textures, and the like. The first end portion 612 has
a relieved portion 616 configured to receive a collar 618 and the
second end portion 614 has a relieved portion 620 configured to
receive a collar 622.
[0071] The rotor 600 is disposed inside the stator 700. The stator
700 has a sidewall 704 with an inside surface 705 defining a
generally hollow inside portion 710 into which the rotor 600 is
disposed. The sidewall 704 may be about 0.1 inches to about 0.3
inches thick. In some embodiments, the sidewall 604 is about 1.5
inches thick. The stator 700 may be non-rotatably coupled to the
housing 520 in a substantially stationary position. Alternatively,
the stator 700 may integrally formed with the housing 520. The
sidewall 704 has a first end portion 712 and a second end portion
714. Optionally, a plurality of apertures 708 are formed in the
sidewall 704 of the stator 700 between the first end portion 712
and the second end portion 714. Optionally, the inside surface 705
of the sidewall 704 may include other features such as
through-holes, projections, textures, and the like.
[0072] The rotor 600 rotates with respect to the stationary stator
700 about an axis of rotation ".alpha." in a direction indicated by
arrow "C3" in FIG. 9. Each of the rotor 600 and the stator 700 may
be generally cylindrical in shape and have a longitudinal axis. The
rotor 600 has an outer diameter "D1" and the stator 700 may have an
inner diameter "D2." The diameter "D1" may range, for example, from
about 0.5 inches to about 24 inches. In some embodiments, the
diameter "D1" is about 3.04 inches. In some embodiments, the
diameter "D1" is about 1.7 inches. The diameter "D2," which is
larger than the diameter "D1," may range from about 0.56 inches to
about 24.25 inches. In some embodiments, the diameter "D2" is about
4 inches. Therefore, the mixing chamber 330 may have a ring-shaped
cross-sectional shape that is about 0.02 inches to about 0.125
inches thick (i.e., the difference between the diameter "D2" and
the diameter "D1"). In particular embodiments, the mixing chamber
330 is about 0.025 inches thick. The channel 32 between the rotor
12 and the stator 34 of prior art device 10 (see FIG. 1) has a
ring-shaped cross-sectional shape that is about 0.09 inches thick.
Therefore, in particular embodiments, the thickness of the mixing
chamber 330 is less than about one third of the channel 32 of the
prior art device 10.
[0073] The longitudinal axis of the rotor 600 may be aligned with
its axis of rotation ".alpha.." The longitudinal axis of the rotor
600 may be aligned with the longitudinal axis of the stator 700.
The rotor 600 may have a length of about 3 inches to about 6 inches
along the axis of rotation ".alpha.." In some embodiments, the
rotor 600 may have a length of about 5 inches along the axis of
rotation ".alpha.." The stator 700 may have a length of about 3
inches to about 6 inches along the axis of rotation ".alpha.." In
some embodiments, the stator 700 may have a length of about 5
inches along the axis of rotation ".alpha.."
[0074] While the rotor 600 and the stator 700 have been depicted as
having a generally cylindrical shape, those of ordinary skill in
the art appreciate that alternate shapes may be used. For example,
the rotor 600 and the stator 700 may be conically, spherically,
arbitrarily shaped, and the like. Further, the rotor 600 and the
stator 700 need not be identically shaped. For example, the rotor
600 may be cylindrically shaped and the stator 700 rectangular
shaped or vise versa.
[0075] The apertures 708 of the stator 700 and the through-holes
608 depicted in FIGS. 4-7 are generally cylindrically shaped. The
diameter of the through-holes 608 may range from about 0.1 inches
to about 0.625 inches. The diameter of the apertures 708 may range
from about 0.1 inches to about 0.625 inches. One or more of
apertures 708 of the stator 700 may have a diameter that differs
from the diameters of the other apertures 708. For example, the
apertures 708 may increase in diameter from the first end portion
712 of the stator 700 to the second end portion 714 of the stator
700, the apertures 708 may decrease in diameter from the first end
portion 712 of the stator 700 to the second end portion 714 of the
stator 700, or the diameters of the apertures 708 may vary in
another manner along the stator 700. One or more of through-holes
608 of the rotor 600 may have a diameter that differs from the
diameters of the other through-holes 608. For example, the
through-holes 608 may increase in diameter from the first end
portion 612 of the rotor 600 to the second end portion 614 of the
rotor 600, the through-holes 608 may decrease in diameter from the
first end portion 612 of the rotor 600 to the second end portion
614 of the rotor 600, or the diameters of the through-holes 608 may
vary in another manner along the rotor 600.
[0076] As described below with reference to alternate embodiments,
the apertures 708 and the through-holes 608 may have shapes other
than generally cylindrical and such embodiments are within the
scope of the present invention. For example, the through-holes 608
may include a narrower portion, an arcuate portion, a tapered
portion, and the like. Referring to FIGS. 7, each of the
through-holes 608 includes an outer portion 608A, a narrow portion
608B, and a tapered portion 608C providing a transition between the
outer portion 608A and the narrow portion 608B. Similarly, the
apertures 708 may include a narrower portion, an arcuate portion, a
tapered portion, and the like.
[0077] FIG. 8 provides a non-limiting example of a suitable
arrangement of the apertures 708 of the stator 700 and the
through-holes 608 of the rotor 600. The apertures 708 of the stator
700 may be arranged in substantially parallel lateral rows "SLAT-1"
through "SLAT-6" substantially orthogonal to the axis of rotation
".alpha.." The apertures 708 of the stator 700 may also be arranged
in substantially parallel longitudinal rows "SLONG-1" through
"SLONG-7" substantially parallel with the axis of rotation
".alpha.." In other words, the apertures 708 of the stator 700 may
be arranged in a grid-like pattern of orthogonal rows (i.e., the
lateral rows are orthogonal to the longitudinal rows) having the
longitudinal rows "SLONG-1" through "SLONG-7" substantially
parallel with the axis of rotation ".alpha.."
[0078] Like the apertures 708 of the stator 700, the through-holes
608 of the rotor 600 may be arranged in substantially parallel
lateral rows "RLAT-1" through "RLAT-6" substantially orthogonal to
the axis of rotation ".alpha.." However, instead of being arranged
in a grid-like pattern of orthogonal rows, the through-holes 608 of
the rotor 600 may also be arranged in substantially parallel rows
"RLONG-1" through "RLONG-7" that extend longitudinally along a
helically path. Alternatively, the through-holes 608 of the rotor
600 may also be arranged in substantially parallel rows "RLONG-1"
through "RLONG-7" that extend longitudinally at an angle other than
parallel with the axis of rotation ".alpha.."
[0079] The apertures 708 of the stator 700 and the through-holes
608 of the rotor 600 may be configured so that when the rotor 600
is disposed inside the stator 700 the lateral rows "SLAT-1" to
"SLAT-6" at least partially align with the lateral rows "RLAT-1" to
"RLAT-6," respectively. In this manner, as the rotor 600 rotates
inside the stator 700, the through-holes 608 pass by the apertures
708.
[0080] The through-holes 608 in each of the lateral rows "RLAT-1"
to "RLAT-6" may be spaced apart laterally such that all of the
through-holes 608 in the lateral row align, at least partially,
with the apertures 708 in a corresponding one of the lateral rows
"SLAT-1" to "SLAT-6" of the stator 700 at the same time. The
longitudinally extending rows "RLONG-1" through "RLONG-6" may be
configured such that the through-holes 608 in the first lateral row
"RLAT-1" in each of the longitudinally extending rows passes
completely by the apertures 708 of the corresponding lateral row
"SLAT-1" before the through-holes 608 in the last lateral row
"RLAT-6" begin to partially align with the apertures 708 of the
corresponding last lateral row "SLAT-6" of the stator 700.
[0081] While, in FIG. 8, six lateral rows and six longitudinally
extending rows have been illustrated with respect to the rotor 600
and six lateral rows and seven longitudinally extending rows have
been illustrated with respect stator 700, it is apparent to those
of ordinary skill in the art that alternate numbers of lateral rows
and/or longitudinal rows may be used with respect to the rotor 600
and/or stator 700 without departing from the present teachings.
[0082] To ensure that only one pair of openings between
corresponding lateral rows will be coincident at any one time, the
number of apertures 708 in each of the lateral rows "SLAT-1" to
"SLAT-6" on the stator 700 may differ by a predetermined number
(e.g., one, two, and the like) the number of through-holes 608 in
each of the corresponding lateral rows "RLAT-1" to "RLAT-6" on the
rotor 600. Thus, for example, if lateral row "RLAT-1" has twenty
through-holes 608 evenly spaced around the circumference of rotor
600, the lateral row "SLAT-1" may have twenty apertures 708 evenly
spaced around the circumference of stator 700.
[0083] Returning to FIG. 7, the mixing chamber 330 has an open
first end portion 332 and an open second end portion 334. The
through-holes 608 formed in the sidewall 604 of the rotor 600
connect the inside portion 610 of the rotor 600 with the mixing
chamber 330.
[0084] The rotor 600 is rotated inside the stator 700 by the drive
shaft 500 aligned with the axis of rotation ".alpha." of the rotor
600. The drive shaft 500 may be coupled to the first end portion
612 and the second end portion 614 of the rotor 600 and extend
through its hollow inside portion 610. In other words, a portion
720 of the drive shaft 500 is disposed in the hollow inside portion
610 of the rotor 600.
[0085] The collar 618 is configured to receive a portion 721 of the
drive shaft 500 disposed in the hollow inside portion 610 and the
collar 622 is configured to receive a portion 722 of the drive
shaft 500 disposed in the hollow inside portion 610.
[0086] The portion 721 has an outer diameter "D3" that may range
from about 0.5 inches to about 2.5 inches. In some embodiments, the
diameter "D3" is about 0.625 inches. The portion 722 has an outer
diameter "D4" that may be substantially similar to the diameter
"D3," although, this is not required. The diameter "D4" may range
from about 0.375 inches to about 2.5 inches.
[0087] The rotor 600 may be non-rotationally affixed to the portion
721 and the portion 722 of the drive shaft 500 by the collar 618
and the collar 622, respectively. By way of example, each of the
collars 618 and 622 may be installed inside relieved portions 616
and 620, respectively. Then, the combined rotor 600 and collars 618
and 622 may be heated to expand them. Next, the drive shaft 500 is
inserted through the collars 618 and 622 and the assembly is
allowed to cool. As the collars 618 and 622 shrink during cooling,
they tighten around the portions 722A and 722B of the drive shaft
500, respectively, gripping it sufficiently tightly to prevent the
drive shaft 500 from rotating relative to the rotor 600. The collar
618, which does not rotate with respect to either the portion 721
or the relieved portion 616, translates the rotation of the drive
shaft 500 to the first end portion 612 the rotor 600. The collar
622, which does not rotate with respect to either the portion 722
or the relieved portion 620, translates the rotation of the drive
shaft 500 to the second end portion 614 of the rotor 600. The drive
shaft 500 and the rotor 600 rotate together as a single unit.
[0088] The drive shaft 500 may have a first end portion 724 (see
FIG. 5) and a second end portion 726 (see FIG. 6). The first end
portion 724 may have a diameter "D5" of about 0.5 inches to about
1.75 inches. In particular embodiments, the diameter "D5" may be
about 1.25 inches. The second end portion 726 may have a diameter
"D6" that may be substantially similar to diameter "D5."
[0089] The second material 120 may be transported into the mixing
chamber 330 through one of the first end portion 724 and the second
end portion 726 of the rotating drive shaft 500. The other of the
first end portion 724 and the second end portion 726 of the drive
shaft 500 may be coupled to the motor 510. In the embodiment
depicted in FIGS. 5 and 6, the second material 120 is transported
into the mixing chamber 330 through the first end portion 724 and
the second end portion 726 of the drive shaft 500 is coupled to the
motor 510.
[0090] Turning to FIG. 5, the drive shaft 500 may have a channel
728 formed therein that extends from first end portion 724 into the
portion 720 disposed in the inside portion 610 of the rotor 600.
The channel 728 has an opening 730 formed in the first end portion
724. When the mixing device 100 is operating, the second material
120 is introduced into the channel 728 through the opening 730.
[0091] A valve 732 may be disposed inside a portion of the channel
728 located in the first end portion 724 of the drive shaft 500.
The valve 732 may restrict or otherwise control the backward flow
of the second material 120 from inside the hollow inside portion
610 through the channel 728 and/or the forward flow of the second
material 120 into the channel 728. The valve 732 may include any
valve known in the art including a check valve. A suitable check
valve includes a part number "CKFA1876205A," free flow forward
check valve, manufactured by The Lee Company USA having an office
in Bothell, Wash. and operating a website at www.theleeco.com.
[0092] The drive shaft 500 may include an aperture 740 located in
the inside portion 610 of the rotor 600 that connects the channel
728 with the inside portion 610 of the rotor 600. While only a
single aperture 740 is illustrated in FIG. 5, it is apparent to
those of ordinary skill in the art that multiple apertures may be
used to connect the channel 728 with the inside portion 610 of the
rotor 600.
[0093] Referring to FIG. 2, optionally, the external pump 220 may
pump the second material 120 into the mixing device 100. The pump
220 may include any suitable pump known in the art. By way of
non-limiting example, the pump 220 may include any suitable pump
known in the art including a diaphragm pump, a chemical pump, a
peristaltic pump, a gravity fed pump, a piston pump, a gear pump, a
combination of any of the aforementioned pumps, and the like. If
the second material 120 is a gas, the gas may be pressurized and
forced into the opening 730 formed in the first end portion 724 of
the drive shaft 500 by releasing the gas from the source 122.
[0094] The pump 220 or the source 122 is coupled to the channel 728
by the valve 732. The second material 120 transported inside the
channel 728 exits the channel 728 into the inside portion 610 of
the rotor 600 through the aperture 740. The second material 120
subsequently exits the inside portion 610 of the rotor 600 through
the through-holes 608 formed in the sidewall 608 of the rotor
600.
[0095] Referring to FIG. 5, the mixing device 100 may include a
seal assembly 750 coupled to the first end portion 724 of the drive
shaft 500. The seal assembly 750 is maintained within a chamber 752
defined in the housing 520. The chamber 752 has a first end portion
754 spaced across the chamber from a second end portion 756. The
chamber 752 also includes an input port 758 and an output port 759
that provide access into the chamber 752. The chamber 752 may be
defined by housing section 550 and the bearing housing 530. The
first end portion 754 may be formed in the housing section 550 and
the second end portion 756 may be adjacent to the bearing housing
530. The input port 758 may be formed in the bearing housing 530
and the output port 759 may be formed in the housing section
550.
[0096] The seal assembly 750 includes a first stationary seal 760
installed in the first end portion 754 of the chamber 752 in the
housing section 550 and the bearing housing 530. The first
stationary seal 760 extends around a portion 762 of the first end
portion 724 of the drive shaft 500. The seal assembly 750 also
includes a second stationary seal 766 installed in the second end
portion 756 of the chamber 752 in the bearing housing 530. The
second stationary seal 766 extends around a portion 768 of the
first end portion 724 of the drive shaft 500.
[0097] The seal assembly 750 includes a rotating assembly 770 that
is non-rotatably coupled to the first end portion 724 of the drive
shaft 500 between the portion 762 and the portion 768. The rotating
assembly 770 rotates therewith as a unit. The rotating assembly 770
includes a first seal 772 opposite a second seal 774. A biasing
member 776 (e.g., a spring) is located between the first seal 772
and the second seal 774. The biasing member 776 biases the first
seal 772 against the first stationary seal 760 and biases the
second seal 774 against the second stationary seal 766.
[0098] A cooling lubricant is supplied to the chamber 752 and
around rotating assembly 770. The lubricant enters the chamber 752
through the input port 758 and exits the chamber 752 through output
port 759. The lubricant may lubricate the bearing assembly 540
housed by the bearing housing 530. A chamber 570 may be disposed
between the bearing housing 530 and the mechanical seal housing
524. The bearing housing 530 may also include a second input port
759 connected to the chamber 570 into which lubricant may be
pumped. Lubricant pumped into the chamber 570 may lubricate the
bearing assembly 540. The seal assembly 750 may significantly, if
not greatly, reduce frictional forces within this portion of the
device caused by the rotation of the rotor 600 and may increase the
active life of the seals 770. The seals may include surfaces
constructed using silicon carbide.
[0099] Referring to FIG. 9, as the rotor 600 rotates about the axis
of rotation ".alpha." in the direction indicated by arrow "C1," the
rotor expels the second material 120 into the mixing chamber 330.
The expelled bubbles, droplets, particles, and the like of the
second material 120 exit the rotor 600 and are imparted with a
circumferential velocity (in a direction indicated by arrow "C3")
by the rotor 600. The second material 120 may be forced from the
mixing chamber 330 by the pump 220 (see FIG. 2), the centrifugal
force of the rotating rotor 600, buoyancy of the second material
120 relative to the first material 110, and a combination
thereof.
Motor 510
[0100] Returning to FIG. 6, the second end portion 726 of the drive
shaft 500 may be coupled to a rotating spindle 780 of a motor 510
by a coupler 900. The spindle 780 may have a generally circular
cross-sectional shape with a diameter "D7" of about 0.25 inches to
about 2.5 inches. In particular embodiments, the diameter "D7" may
be about 0.25 inches to about 1.5 inches. While in the embodiment
depicted in FIG. 6, the diameter "D5" of the first end portion 724
of the drive shaft 500 is substantially equal to the diameter "D7"
and the spindle 780, embodiments in which one of the diameter "D5"
and the diameter "D7" is larger than the other are within the scope
of the present invention.
[0101] Referring also to FIG. 4, it may be desirable to cover or
shield the coupler 900. In the embodiment illustrated in FIGS. 4
and 6, a drive guard 910 covers the coupler 900. The drive guard
910 may be generally U-shaped having a curved portion 914 flanked
by a pair of substantially linear portions 915 and 916. The distal
end of each of the substantially linear portions 915 and 916 of the
drive guard 910 may have a flange 918 and 919, respectively. The
drive guard 910 may be fastened by each of its flanges 918 and 919
to the base 106.
[0102] The motor 510 may be supported on the base 106 by a support
member 920. The support member 920 may be coupled to the motor 510
near the spindle 780. In the embodiment depicted, the support
member 920 includes a through-hole through which the spindle 780
passes. The support member 920 may be coupled to the motor 510
using any method known in the art, including bolting the support
member 920 to the motor 510 with one or more bolts 940.
[0103] The coupler 900 may include any coupler suitable for
transmitting a sufficient amount of torque from the spindle 780 to
the drive shaft 500 to rotate the rotor 600 inside to the stator
700. In the embodiment illustrated in FIGS. 4 and 6, the coupler
900 is a bellows coupler. A bellows coupler may be beneficial if
the spindle 780 and the drive shaft 500 are misaligned. Further,
the bellows coupler may help absorb axial forces exerted on the
drive shaft 500 that would otherwise be translated to the spindle
780. A suitable bellows coupler includes a model "BC32-8-8-A,"
manufactured by Ruland Manufacturing Company, Inc. of Marlborough,
Mass., which operates a website at www.ruland.com.
[0104] The motor 510 may rotate the rotor 600 at about 0.1
revolutions per minute ("rpm") to about 7200 rpm. The motor 510 may
include any motor suitable for rotating the rotor 600 inside to the
stator 700 in accordance with the present teachings. By way of
non-limiting example, a suitable motor may include a one-half
horsepower electric motor, operating at 230/460 volts and 3450 per
minute ("rpm"). A suitable motor includes a model "C4T34NC4C"
manufactured by LEESON Electric Corporation of Grafton, Wis., which
operates a website at www.leeson.com.
First Chamber 310
[0105] Turning to FIGS. 4 and 7, the first chamber 320 is disposed
inside the central section 522 of the housing 520 between the first
mechanical seal housing 524 and the first end portions 612 and 712
of the rotor 600 and the stator 700, respectively. The first
chamber 310 may be annular and have a substantially circular
cross-sectional shape. The first chamber 310 and the mixing chamber
330 form a continuous volume. A portion 1020 of the drive shaft 500
extends through the first chamber 310.
[0106] As may best be viewed in FIG. 4, the first chamber 310 has
an input port 1010 through which the first material 110 enters the
mixing device 100. The first material 110 may be pumped inside the
first chamber 310 by the external pump 210 (see FIG. 2). The
external pump 210 may include any pump known in the art for pumping
the first material 110 at a sufficient rate to supply the first
chamber 310.
[0107] The input port 1010 is oriented substantially orthogonally
to the axis of rotation ".alpha.." Therefore, the first material
110 enters the first chamber 310 with a velocity tangential to the
portion 1020 of the drive shaft 500 extending through the first
chamber 310. The tangential direction of the flow of the first
material 110 entering the first chamber 310 is identified by arrow
"T1." In the embodiment depicted in FIGS. 4 and 7, the input port
1010 may be offset from the axis of rotation ".alpha.." As is
apparent to those of ordinary skill in the art, the direction of
the rotation of the drive shaft 500 (identified by arrow "C1" in
FIG. 9), has a tangential component. The input port 1010 is
positioned so that the first material 110 enters the first chamber
310 traveling in substantially the same direction as the tangential
component of the direction of rotation of the drive shaft 500.
[0108] The first material 110 enters the first chamber 310 and is
deflected by the inside of the first chamber 310 about the portion
1020 of the drive shaft 500. In embodiments wherein the first
chamber 310 has a substantially circular cross-sectional shape, the
inside of the first chamber 310 may deflect the first material 110
in a substantially circular path (identified by arrow "C2" in FIG.
9) about the portion 1020 of the drive shaft 500. In such an
embodiment, the tangential velocity of the first material 110 may
cause it to travel about the axis of rotation ".alpha." at a
circumferential velocity, determined at least in part by the
tangential velocity.
[0109] Once inside the first chamber 310, the first material 110
may be pumped from the first chamber 310 into the mixing chamber
330 by the pump 410 residing inside the first chamber 310. In
embodiments that include the external pump 210 (see FIG. 2), the
external pump 210 may be configured to pump the first material 110
into the first chamber 310 at a rate at least as high as a rate at
which the pump 410 pumps the first material 110 from the first
chamber 310.
[0110] The first chamber 310 is in communication with the open
first end portion 332 of the mixing chamber 330 and the first
material 110 inside the first chamber 310 may flow freely into the
open first end portion 332 of the mixing chamber 330. In this
manner, the first material 110 does not negotiate any corners or
bends between the mixing chamber 330 and the first chamber 310. In
the embodiment depicted, the first chamber 310 is in communication
with the entire open first end portion 332 of the mixing chamber
330. The first chamber 310 may be filled completely with the first
material 110.
[0111] The pump 410 is powered by the portion 1020 of the drive
shaft 500 extending through the first chamber 310. The pump 410 may
include any pump known in the art having a rotating pump member
2022 housed inside a chamber (i.e., the first chamber 310) defined
by a stationary housing (i.e., the housing 520). Non-limiting
examples of suitable pumps include rotary positive displacement
pumps such as progressive cavity pumps, single screw pumps (e.g.,
Archimedes screw pump), and the like.
[0112] The pump 410 depicted in FIGS. 7 and 9, is generally
referred to as a single screw pump. In this embodiment, the pump
member 2022 includes a collar portion 2030 disposed around the
portion 1020 of the drive shaft 500. The collar portion 2030
rotates with the portion 1020 of the drive shaft 500 as a unit. The
collar portion 2030 includes one or more fluid displacement members
2040. In the embodiment depicted in FIGS. 7 and 9, the collar
portion 2030 includes a single fluid displacement member 2040
having a helical shape that circumscribes the collar portion 2030
along a helical path.
[0113] Referring to FIG. 9, the inside of the first chamber 310 is
illustrated. The pump 410 imparts an axial flow (identified by
arrow "A1" and arrow "A2") in the first material 110 inside the
first chamber 310 toward the open first end portion 332 of the
mixing chamber 330. The axial flow of the first material 110
imparted by the pump 410 has a pressure that may exceed the
pressure obtainable by the external pump of the prior art device 10
(see FIG. 1).
[0114] The pump 410 may also be configured to impart a
circumferential flow (identified by arrow "C2") in the first
material 110 as it travels toward the open first end portion 332 of
the mixing chamber 330. The circumferential flow imparted in the
first material 110 before it enters the mixing chamber 330 causes
the first material 110 to enter the mixing chamber 330 already
traveling in the desired direction at an initial circumferential
velocity. In the prior art device 10 depicted in FIG. 1, the first
material 110 entered the channel 32 of the prior art device 10
without a circumferential velocity. Therefore, the rotor 12 of the
prior art device 10 alone had to impart a circumferential flow into
the first material 110. Because the first material 110 is moving
axially, in the prior art device 10, the first material 110
traversed at least a portion of the channel 32 formed between the
rotor 12 and the stator 30 at a slower circumferential velocity
than the first material 110 traverses the mixing chamber 330 of the
mixing device 100. In other words, if the axial velocity of the
first material 110 is the same in both the prior art device 10 and
the mixing device 100, the first material 110 may complete more
revolutions around the rotational axis ".alpha." before traversing
the axial length of the mixing chamber 330, than it would complete
before traversing the axial length of the channel 32. The
additional revolutions expose the first material 110 (and combined
first material 110 and second material 120) to a substantially
larger portion of the effective inside surface 706 (see FIG. 7) of
the stator 700.
[0115] In embodiments including the external pump 210 (see FIG. 2),
the circumferential velocity imparted by the external pump 210
combined with the input port 1010 being oriented according to the
present teachings, may alone sufficiently increase the revolutions
of the first material 110 (and combined first material 110 and
second material 120) about the rotational axis ".alpha.." Further,
in some embodiments, the circumferential velocity imparted by the
pump 210 and the circumferential velocity imparted by the pump 410
combine to achieve a sufficient number of revolutions of the first
material 110 (and combined first material 110 and second material
120) about the rotational axis ".alpha.." As is appreciated by
those of ordinary skill in the art, other structural elements such
as the cross-sectional shape of the first chamber 310 may
contribute to the circumferential velocity imparted by the pump
210, the pump 410, and a combination thereof.
[0116] In an alternate embodiment depicted in FIG. 10, the pump 410
may include one or more vanes 2042 configured to impart a
circumferential flow in the first material 110 as it travels toward
the open first end portion 332 of the mixing chamber 330.
Second Chamber 320
[0117] Turning now to FIGS. 4 and 7, the second chamber 320 is
disposed inside the central section 522 of the housing 520 between
the second mechanical seal housing 526 and the second end portions
614 and 714 of the rotor 600 and the stator 700, respectively. The
second chamber 320 may be substantially similar to the first
chamber 310. However, instead of the input port 1010, the second
chamber 320 may include an output port 3010. A portion 3020 of the
drive shaft 500 extends through the second chamber 320.
[0118] The second chamber 320 and the mixing chamber 330 form a
continuous volume. Further, the first chamber 310, the mixing
chamber 330, and the second chamber 320 form a continuous volume.
The first material 110 flows through the mixing device 100 from the
first chamber 310 to the mixing chamber 330 and finally to the
second chamber 320. While in the mixing chamber 330, the first
material 110 is mixed with the second material 120 to form the
output material 102. The output material 102 exits the mixing
device 100 through the output port 3010. Optionally, the output
material 102 may be returned to the input port 1010 and mixed with
an additional quantity of the second material 120, the third
material 130, or a combination thereof.
[0119] The output port 3010 is oriented substantially orthogonally
to the axis of rotation ".alpha." and may be located opposite the
input port 1010 formed in the first chamber 310. The output
material 102 enters the second chamber 320 from the mixing chamber
330 having a circumferential velocity (in the direction indicated
by arrow "C3" in FIG. 9) imparted thereto by the rotor 600. The
circumferential velocity is tangential to the portion 3020 of the
drive shaft 500 extending through the second chamber 320. In the
embodiment depicted in FIGS. 4, 6, and 7, the output port 3010 may
be offset from the axis of rotation ".alpha.." The output port 3010
is positioned so that the output material 102, which enters the
second chamber 320 traveling in substantially the same direction in
which the drive shaft 500 is rotating (identified in FIG. 9 by
arrow "C1"), is traveling toward the output port 3010.
[0120] The output material 102 enters the second chamber 320 and is
deflected by the inside of the second chamber 320 about the portion
3020 of the drive shaft 500. In embodiments wherein the second
chamber 320 has a substantially circular cross-sectional shape, the
inside of the second chamber 320 may deflect the output material
102 in a substantially circular path about the portion 3020 of the
drive shaft 500.
[0121] Referring to FIG. 2, optionally, the output material 102 may
be pumped from inside the second chamber 320 by the external pump
430. The external pump 430 may include any pump known in the art
for pumping the output material 102 at a sufficient rate to avoid
limiting throughput of the mixing device 100. In such an
embodiment, the external pump 430 may introduce a tangential
velocity (in a direction indicated by arrow "T2" in FIGS. 4 and 11)
to at least a portion of the output material 102 as the external
pump 430 pumps the output material 102 from the second chamber 320.
The tangential velocity of the portion of the output material 102
may cause it to travel about the axis of rotation ".alpha." at a
circumferential velocity, determined in part by the tangential
velocity.
Pump 420
[0122] Turning to FIGS. 6 and 7, the pump 420 residing inside the
second chamber 320 may pump the output material 102 from the second
chamber 320 into the output port 3010 and/or from the mixing
chamber 330 into the second chamber 320. In embodiments that
include the external pump 430, the external pump 430 may be
configured to pump the output material 102 from the second chamber
320 at a rate at least as high as a rate at which the pump 420
pumps the output material 102 into the output port 3010.
[0123] The second chamber 320 is in communication with the open
second end portion 334 of the mixing chamber 330 and the output
material 102 inside the mixing chamber 330 may flow freely from the
open second end portion 334 into the second chamber 320. In this
manner, the output material 102 does not negotiate any corners or
bends between the mixing chamber 330 and the second chamber 320. In
the embodiment depicted, the second chamber 320 is in communication
with the entire open second end portion 334 of the mixing chamber
330. The second chamber 320 may be filled completely with the
output material 102.
[0124] The pump 420 is powered by the portion 3020 of the drive
shaft 500 extending through the second chamber 320. The pump 420
may be substantially identical to the pump 410. Any pump described
above as suitable for use as the pump 410 may be used for the pump
420. While the pump 410 pumps the first material 110 into the
mixing chamber 330, the pump 420 pumps the output material 102 from
the mixing chamber 330. Therefore, both the pump 410 and the pump
420 may be oriented to pump in the same direction.
[0125] As is appreciated by those of ordinary skill in the art, the
first material 110 may differ from the output material 102. For
example, one of the first material 110 and the output material 102
may be more viscous than the other. Therefore, the pump 410 may
differ from the pump 420. The pump 410 may be configured to
accommodate the properties of the first material 110 and the pump
420 may be configured to accommodate the properties of the output
material 102.
[0126] The pump 420 depicted in FIGS. 6 and 7, is generally
referred to as a single screw pump. In this embodiment, the pump
member 4022 includes a collar portion 4030 disposed around the
portion 3020 of the drive shaft 500. The collar portion 4030
rotates with the portion 3020 of the drive shaft 500 as a unit. The
collar portion 4030 includes one or more fluid displacement members
4040. The collar portion 4030 includes a single fluid displacement
member 4040 having a helical shape that circumscribes the collar
portion 4030 along a helical path.
[0127] Referring to FIG. 11, the inside of the second chamber 320
is illustrated. The pump 420 imparts an axial flow (identified by
arrow "A3" and arrow "A4") in the output material 102 inside the
second chamber 320 away from the open second end portion 334 of the
mixing chamber 330.
[0128] The pump 420 may be configured to impart a circumferential
flow (identified by arrow "C4") in the output material 102 as it
travels away from the open second end portion 334 of the mixing
chamber 330. The circumferential flow imparted in the output
material 102 may help reduce an amount of work required by the
rotor 600. The circumferential flow also directs the output
material 102 toward the output port 3010.
[0129] In an alternate embodiment, the pump 420 may have
substantially the same configuration of the pump 410 depicted in
FIG. 10. In such an embodiment, the one or more vanes 2042 are
configured to impart a circumferential flow in the output material
102 as it travels away from the open second end portion 334 of the
mixing chamber 330.
[0130] As is apparent to those of ordinary skill, various
parameters of the mixing device 100 may be modified to obtain
different mixing characteristics. Exemplary parameters that may be
modified include the size of the through-holes 608, the shape of
the through-holes 608, the arrangement of the through-holes 608,
the number of through-holes 608, the size of the apertures 708, the
shape of the apertures 708, the arrangement of the apertures 708,
the number of apertures 708, the shape of the rotor 600, the shape
of the stator 700, the width of the mixing chamber 330, the length
of the mixing chamber 330, rotational speed of the drive shaft 500,
the axial velocity imparted by the internal pump 410, the
circumferential velocity imparted by the internal pump 410, the
axial velocity imparted by the internal pump 420, the
circumferential velocity imparted by the internal pump 420, the
configuration of disturbances (e.g., texture, projections,
recesses, apertures, and the like) formed on the outside surface
606 of the rotor 600, the configuration of disturbances (e.g.,
texture, projections, recesses, apertures, and the like) formed on
the inside surface 706 of the stator 700, and the like.
Alternate Embodiment
[0131] Referring to FIG. 12, a mixing device 5000 is depicted. The
mixing device 5000 is an alternate embodiment of the mixing device
100. Identical reference numerals have been used herein to identify
components of the mixing device 5000 that are substantially similar
corresponding components of the mixing device 100. Only components
of the mixing device 5000 that differ from the components of the
mixing device 100 will be described.
[0132] The mixing device 5000 includes a housing 5500 for housing
the rotor 600 and the stator 5700. The stator 5700 may be
non-rotatably couple by its first end portion 5712 and its second
end portion 5714 to the housing 5500. A chamber 5800 is defined
between the housing 5500 and a portion 5820 of the stator 5700
flanked by the first end portion 5712 and the second end portion
5714. The housing 5500 includes an input port 5830 which provides
access into the chamber 5800. The input port 5830 may be oriented
substantially orthogonally to the axis of rotation ".alpha.."
however, this is not a requirement.
[0133] The stator 5700 includes a plurality of through-holes 5708
that connect the chamber 5800 and the mixing chamber 330 (defined
between the rotor 600 and the stator 5700). An external pump 230
may be used to pump the third material 130 (which may be identical
to the second material 120) into the chamber 5800 via the input
port 5830. The third material 130 pumped into the chamber 5800 may
enter the mixing chamber 330 via the through-holes 5708 formed in
the stator 5700. The third material 130 may be forced from the
channel 5800 by the pump 230, buoyancy of the third material 130
relative to the first material 110, and a combination thereof. As
the rotor 600 rotates, it may also draw the third material 130 from
the channel 5800 into the mixing chamber 330. The third material
130 may enter the mixing chamber 330 as bubbles, droplets,
particles, and the like, which are imparted with a circumferential
velocity by the rotor 600.
Alternate Embodiment
[0134] An alternate embodiment of the mixing device 100 may be
constructed using a central section 5900 depicted in FIG. 13 and a
bearing housing 5920 depicted in FIG. 14. FIG. 13 depicts the
central section 5900 having in its interior the stator 700 (see
FIG. 7). Identical reference numerals have been used herein to
identify components associated with the central section 5900 that
are substantially similar corresponding components of the mixing
device 100. Only components of the central section 5900 that differ
from the components of the central section 522 will be described.
The central section 5900 and the stator 700 are both constructed
from a conductive material such as a metal (e.g., stainless steel).
The input port 1010 and the output port 3010 are both constructed
from a nonconductive material such as plastic (e.g., PET,
TEFLON.RTM., nylon, PVC, polycarbonate, ABS, DELRIN.RTM.,
polysulfone, etc.).
[0135] An electrical contact 5910 is coupled to the central section
5900 and configured to deliver a charge thereto. The central
section 5900 conducts an electrical charge applied to the
electrical contact 5910 to the stator 700. In further embodiments,
the central section 5900 may be constructed from a nonconductive
material. In such embodiments, the electrical contact 5910 may pass
through the central section 5900 and coupled to the stator 700. The
electric charge applied by the electrical contact 5910 to the
stator 700 may help facilitate redox or other chemical reactions
inside the mixing chamber 330.
[0136] Optionally, insulation (not shown) may be disposed around
the central section 5900 to electrically isolate it from the
environment. Further, insulation may be used between the central
section 5900 and the first and second mechanical seals 524 and 526
that flank it to isolate it electrically from the other components
of the mixing device.
[0137] Turning now to FIG. 14, the bearing housing 5920 will be
described. The bearing housing 5920 is disposed circumferentially
around the portion 726 of the drive shaft 500. An electrical
contact 5922 is coupled to the bearing housing 5920. A rotating
brush contact 5924 provides an electrical connection between the
drive shaft 500 and the electrical contact 5922.
[0138] In this embodiment, the drive shaft 500 and the rotor 600
are both constructed from a conductive material such as a metal
(e.g., stainless steel). The bearing housing 5920 may be
constructed from either a conductive or a nonconductive material.
An electrical charge is applied to the drive shaft 500 by the
electrical contact 5922 and the rotating brush contact 5924. The
electrical charge is conducted by the drive shaft 500 to the rotor
600.
[0139] The alternate embodiment of the mixing device 100
constructed using the central section 5900 depicted in FIG. 13 and
the bearing housing 5920 depicted in FIG. 14 may be operated in at
least two ways. First, the electrical contacts 5910 and 5922 may be
configured not to provide an electrical charge to the stator 700
and the rotor 600, respectively. In other words, neither of the
electrical contacts 5910 and 5922 are connected to a current
source, a voltage source, and the like.
[0140] Alternatively, the electrical contacts 5910 and 5922 may be
configured to provide an electrical charge to the stator 700 and
the rotor 600, respectively. For example, the electrical contacts
5910 and 5922 may be coupled to a DC voltage source (not shown)
supplying a steady or constant voltage across the electrical
contacts 5910 and 5922. The negative terminal of the DC voltage
source may be coupled to either of the electrical contacts 5910 and
5922 and the positive terminal of the DC voltage source may be
coupled to the other of the electrical contacts 5910 and 5922. The
voltage supplied across the electrical contacts 5910 and 5922 may
range from about 0.0001 volts to about 1000 volts. In particular
embodiments, the voltage may range from about 1.8 volts to about
2.7 volts. By way of another example, a pulsed DC voltage having a
duty cycle of between about 1% to about 99% may be used.
[0141] While the above examples of methods of operating the mixing
device apply a DC voltage across the electrical contacts 5910 and
5922, as is apparent to those of ordinary skill in the art, a
symmetrical AC voltage or non symmetrical AC voltage having various
shapes and magnitudes may be applied across the electrical contacts
5910 and 5922 and such embodiments are within the scope of the
present invention.
Mixing Inside the Mixing Chamber
[0142] As mentioned above, in the prior art device 10 (shown in
FIG. 1), the first material 110 entered the channel 32 between the
rotor 12 and the stator 30 via a single limited input port 37
located along only a portion of the open second end of the channel
32. Likewise, the output material 102 exited the channel 32 via a
single limited output port 40 located along only a portion of the
open first end of the channel 32. This arrangement caused
undesirable and unnecessary friction. By replacing the single
limited inlet port 37 and the single limited outlet port 40 with
the chambers 310 and 320, respectively, friction has been reduced.
Moreover, the first material 110 does not negotiate a corner before
entering the mixing chamber 330 and the output material 102 does
not negotiate a corner before exiting the mixing chamber 330.
Further, the chambers 310 and 320 provide for circumferential
velocity of the material prior to entering, and after exiting the
channel 32.
[0143] Accordingly, pressure drop across the mixing device 100 has
been substantially reduced. In the embodiments depicted in FIGS. 2,
4-9, and 11, the pressure drop between the input port 1010 and the
output port 3010 is only approximately 12 psi when the mixing
device 100 is configured to produce about 60 gallons of the output
material 102 per minute. This is an improvement over the prior art
device 10 depicted in FIG. 1, which when producing about 60 gallons
of output material per minute was at least 26 psi. In other words,
the pressure drop across the mixing device 100 is less than half
that experienced by the prior art device 10.
[0144] According to additional aspects, the inclusion of pumps 410
and 420, which are powered by the drive shaft 500, provides a
configuration that is substantially more efficient in mixing
materials and that requires less energy than the external pumps
used in the prior art.
Micro-Cavitation
[0145] During operation of the mixing device 100, the input
materials may include the first material 110 (e.g., a fluid) and
the second material 120 (e.g., a gas). The first material 110 and
the second material 120 are mixed inside the mixing chamber 330
formed between the rotor 600 and the stator 700. Rotation of the
rotor 600 inside the stator 700 agitates the first material 110 and
the second material 120 inside the mixing chamber 330. The
through-holes 608 formed in the rotor 600 and/or the apertures 708
formed in the stator 700 impart turbulence in the flow of the first
material 110 and the second material 120 inside the mixing chamber
330.
[0146] Without being limited by theory, the efficiency and
persistence of the diffusion of the second material 120 into the
first material 110 is believed to be caused in part by
micro-cavitation, which is described in connection with FIGS.
15-17. Whenever a material flows over a smooth surface, a rather
laminar flow is established with a thin boundary layer that is
stationary or moving very slowly because of the surface tension
between the moving fluid and the stationary surface. The
through-holes 608 and optionally, the apertures 708, disrupt the
laminar flow and can cause localized compression and decompression
of the first material 110. If the pressure during the decompression
cycle is low enough, voids (cavitation bubbles) will form in the
material. The cavitation bubbles generate a rotary flow pattern
5990, like a tornado, because the localized area of low pressure
draws the host material and the infusion material, as shown in FIG.
15. When the cavitation bubbles implode, extremely high pressures
result. As two aligned openings (e.g., one of the apertures 708 and
one of the through-holes 608) pass one another, a succussion (shock
wave) occurs, generating significant energy. The energy associated
with cavitation and succussion mixes the first material 110 and the
second material 120 together to an extremely high degree, perhaps
at the molecular level.
[0147] The tangential velocity of the rotor 600 and the number of
openings that pass each other per rotation may dictate the
frequency at which the mixing device 100. It has been determined
that operating the mixing device 100 within in the ultrasonic
frequency range can be beneficial in many applications. It is
believed that operating the mixing device 100 in the ultrasonic
region of frequencies provides the maximum succession shock energy
to shift the bonding angle of the fluid molecule, which enables it
to transport an additional quantity of the second material 120
which it would not normally be able to retain. When the mixing
device 100 is used as a diffuser, the frequency at which the mixing
device 100 operates appears to affect the degree of diffusion,
leading to much longer persistence of the second material 120
(infusion material) in the first material 110 (host material).
[0148] Referring now to FIG. 18, an alternate embodiment of the
rotor 600, rotor 6000 is provided. The cavitations created within
the first material 110 in the mixing chamber 330 may be configured
to occur at different frequencies along the length of the mixing
chamber 330. The frequencies of the cavitations may be altered by
altering the number and/or the placement of the through-holes 6608
along the length of the rotor 600. Each of the through-holes 6608
may be substantially similar to the through-holes 608 (discussed
above).
[0149] By way of non-limiting example, the rotor 6000 may be
subdivided into three separate exemplary sections 6100, 6200, and
6300. The through-holes 6608 increase in density from the section
6100 to the section 6200, the number of holes in the section 6100
being greater than the number of holes in the section 6200. The
through-holes 6608 also increase in density from the section 6200
to the section 6300, the number of holes in the section 6200 being
greater than the number of holes in the section 6300. Each of the
sections 6100, 6200, and 6300 create succussions within their
particular area at a different frequency due to the differing
numbers of through-holes 6608 formed therein.
[0150] By manufacturing the rotor 6000 with a desired number of
through-holes 6608 appropriately arranged in a particular area, the
desired frequency of the succussions within the mixing chamber 330
may be determined. Similarly, the desired frequency of the
cavitations may be determined by a desired number of apertures 708
appropriately arranged in a particular area upon the stator 700
within which the rotor 600 rotates. Further, the desired frequency
(or frequencies) of the succussions within the mixing chamber 330
may be achieved by selecting both a particular number and
arrangement of the apertures 708 formed in the stator 700 and a
particular number and arrangement of the through-holes 608 formed
in the rotor 600.
[0151] FIGS. 19-21, depict various alternative arrangements of the
apertures 708 formed in the stator 700 and the through-holes 608
formed in the rotor 600 configured to achieve different results
with respect to the cavitations created. FIG. 19 illustrates a
configuration in which the apertures 708 and the through-holes 608
are aligned along an axis 7000 that is not parallel with any line
(e.g., line 7010) drawn through the axis of rotation ".alpha." of
the rotor 600. In other words, if the rotor 600 has a cylindrical
shape, the axis 7000 does not pass through the center of the rotor
600. Thus, the first material 110 within the mixing chamber 330
will not be oriented perpendicularly to the compressions and
decompressions created by the apertures 708 and the through-holes
608. The compressions and decompressions will instead have a force
vector that has at least a component parallel to the
circumferential flow (in the direction of arrow "C3" of FIG. 9) of
first material 110 within the mixing chamber 330.
[0152] Relative alignment of the apertures 708 and the
through-holes 608 may also affect the creation of cavitations in
the mixing chamber 330. FIG. 20 illustrates an embodiment in which
the apertures 708 are in registration across the mixing chamber 330
with the through-holes 608. In this embodiment, rotation of the
rotor 600 brings the through-holes 608 of the rotor into direct
alignment with the apertures 708 of the stator 700. When in direct
alignment with each other, the compressive and decompressive forces
created by the apertures 708 and the through-holes 608 are directly
aligned with one another.
[0153] In the embodiment depicted in FIG. 21, the apertures 708 and
the through-holes 608 are offset by an offset amount "X" along the
axis of rotation ".alpha..". By way of non-limiting example, the
offset amount "X" may be determined as a function of the size of
the apertures 708. For example, the offset amount "X" may be
approximately equal to one half of the diameter of the apertures
708. Alternatively, the offset amount "X" may be determined as a
function of the size of the through-holes 608. For example, the
offset amount "X" may be approximately equal to one half of the
diameter of the through-holes 608. If features (e.g., recesses,
projections, etc.) other than or in addition to the through-holes
608 and the apertures 708 are included in either the rotor 600 or
the stator 700, the offset amount "X" may be determined as a
function of the size of such features. In this manner, the
compressive and decompressive forces caused by the apertures 708 of
the stator 700 and the through-holes 608 of the rotor 600 collide
at a slight offset causing additional rotational and torsional
forces within the mixing chamber 330. These additional forces
increase the mixing (e.g., diffusive action) of the second material
120 into the first material 110 within the mixing chamber 330.
[0154] Referring now to FIGS. 22-25, non-limiting examples of
suitable cross-sectional shapes for the apertures 708 and the
through-holes 608 are provided. The cross-sectional shape of the
apertures 708 and/or the through-holes 608 may be square as
illustrated in FIG. 22, circular as illustrated in FIG. 23, and the
like.
[0155] Various cross-sectional shapes of apertures 708 and/or the
through-holes 608 may be used to alter flow of the first material
110 as the rotor 600 rotates within the stator 700. For example,
FIG. 24 depicts a teardrop cross-sectional shape having a narrow
portion 7020 opposite a wide portion 7022. If the through-holes 608
have this teardrop shape, when the rotor 600 is rotated (in the
direction generally indicated by the arrow "F"), the forces exerted
on the first material 110, the second material 120, and optionally
the third material 130 within the mixing chamber 330 increase as
the materials pass from the wide portion 7022 of the teardrop to
the narrow portion 7020.
[0156] Additional rotational forces can be introduced into the
mixing chamber 330 by forming the apertures 708 and/or the
through-holes 608 with a spiral configuration as illustrated in
FIG. 25. Material that flows into and out of the apertures 708
and/or the through-holes 608 having the spiral configuration
experience a rotational force induced by the spiral configuration.
The examples illustrated in FIGS. 22-25 are provided as
non-limiting illustrations of alternate embodiments that may be
employed within the mixing device 100. By application of ordinary
skill in the art, the apertures 708 and/or the through-holes 608
may be configured in numerous ways to achieve various succussive
and agitative forces appropriate for mixing materials within the
mixing chamber 330.
Double Layer Effect
[0157] The mixing device 100 may be configured to create the output
material 102 by complex and non-linear fluid dynamic interaction of
the first material 110 and the second material 120 with complex,
dynamic turbulence providing complex mixing that further favors
electrokinetic effects (described below). The result of these
electrokinetic effects may be observed within the output material
102 as charge redistributions and redox reactions, including in the
form of solvated electrons that are stabilized within the output
material.
[0158] Ionization or dissociation of surface groups and/or
adsorption of ions from a liquid cause most solid surfaces in
contact with the liquid to become charged. Referring to FIG. 26, an
electrical double layer ("EDL") 7100 forms around exemplary surface
7110 in contact with a liquid 7120. In the EDL 7100, ions 7122 of
one charge (in this case, negatively charged ions) adsorb to the
surface 7120 and form a surface layer 7124 typically referred to as
a Stern layer. The surface layer 7124 attracts counterions 7126 (in
this case, positively charged ions) of the opposite charge and
equal magnitude, which form a counterion layer 7128 below the
surface layer 7124 typically referred to as a diffuse layer. The
counterion layer 7128 is more diffusely distributed than the
surface layer 7124 and sits upon a uniform and equal distribution
of both ions in the bulk material 7130 below. For OH- and H+ ions
in neutral water, the Gouy-Chapman model would suggest that the
diffuse counterion layer extends about one micron into the
water.
[0159] According to particular aspects, the electrokinetic effects
mentioned above are caused by the movement of the liquid 7120 next
to the charged surface 7110. Within the liquid 7120 (e.g., water,
saline solution, and the like), the adsorbed ions 7122 forming the
surface layer 7124 are fixed to the surface 7120 even when the
liquid 7120 is in motion (for example, flowing in the direction
indicated by arrow "G"); however, a shearing plane 7132 exists
within the diffuse counterion layer 7128 spaced from the surface
7120. Thus, as the liquid 7120 moves, some of the diffuse
counterions 7126 are transported away from the surface 7120, while
the absorbed ions 7122 remain at the surface 7120. This produces a
so-called `streaming current.`
[0160] Within the mixing chamber 330, the first material 110, the
second material 120, and optionally, the third material 130 are
subject to an electromagnetic field created by the inside surface
705 of the stator 700 and/or the outside surface 606 of the rotor
600, a voltage between the inside surface 705 and the outside
surface 606, and/or an electrokinetic effect (e.g., streaming
current) caused by at least one EDL formed in the first material
110. The at least one EDL may be introduced into the first material
110 by at least one of the inside surface 705 of the stator 700 and
the outside surface 606 of the rotor 600.
[0161] Movement of the first material 110 through the mixing
chamber 330 relative to surface disturbances (e.g., the
through-holes 608 and apertures 708) creates cavitations in the
first material 110 within the mixing chamber 330, which may diffuse
the second material 120 into the first material 110. These
cavitations may enhance contact between of the first material 110
and/or the second material 120 with the electric double layer
formed on the inside surface 705 of the stator 700 and/or the
electric double layer formed on the outside surface 606 of the
rotor 600. Larger surface to volume ratios of the mixing chamber,
an increased dwell time of the combined materials within the mixing
chamber, and further in combination with a smaller average bubble
size (and hence substantially greater bubble surface area) provide
for effectively imparting EDL-mediated effects to the inventive
output materials.
[0162] In embodiments in which the inside surface 705 and the
outside surface 606 are constructed from a metallic material, such
as stainless steel, the motion of the liquid 7120 and/or the
streaming current(s) facilitate redox reactions involving H.sub.2O,
OH-, H+, and O.sub.2 at the inside surface 705 and the outside
surface 606.
[0163] Referring to FIG. 27, without being limited by theory, it is
believed a section 7140 of the mixing chamber 330 between the
inside surface 705 and the outside surface 606 the may be modeled
as a pair of parallel plates 7142 and 7144. If the first material
110 is a liquid, the first material 110 enters the section 7140
through an inlet "IN" and exits the section 7140 through an outlet
"OUT." The inlet "IN" and the outlet "OUT" restrict the flow into
and out of the section 7140.
[0164] Referring to FIG. 28, the area between the parallel plates
7142 and 7144 has a high surface area to volume ratio. Hence, a
substantial portion of the counterion layer 7128 (and counterions
7126) may be in motion as the first material 110 moves between the
plates 7142 and 7144. The number of counterions 7126 in motion may
exceed the number allowed to enter the section 7140 by the inlet
"IN" and the number allowed to exit the section 7140 by the outlet
"OUT." The inlet "IN" and the outlet "OUT" feeding and removing the
first material 110 from the section 7140, respectively, have far
less surface area (and a lower surface area to volume ratio) than
the parallel plates 7142 and 7144 and thereby reduce the portion of
the counterions 7126 in motion in the first material 110 entering
and leaving the section 7140. Therefore, entry and exit from the
section 7140 increases the streaming current locally. While a
background streaming current (identified by arrow "BSC") caused by
the flowing first material 110 over any surface is always present
inside the mixing device 100, the plates 7142 and 7144 introduce an
increased "excess" streaming current (identified by arrow "ESC")
within the section 7140.
[0165] Without a conductive return current (identified by arrow
"RC") in the plates 7142 and 7144 in the opposite direction of the
flow of the first material 110, an excess charge 7146 having the
same sign as the adsorbing ions 7122 would accumulate near the
inlet "IN," and an excess charge 7148 having the same sign as the
counterion 7126 would accumulate near the at outlet "OUT." Because
such accumulated charges 7146 and 7148, being opposite and
therefore attracted to one another, cannot build up indefinitely
the accumulated charges seek to join together by conductive means.
If the plates 7142 and 7144 are perfectly electrically insulating,
the accumulated charges 7146 and 7148 can relocate only through the
first material 110 itself. When the conductive return current
(identified by arrow "RC") is substantially equivalent to the
excess streaming current (identified by arrow "ESC") in the section
7140, a steady-state is achieved having zero net excess streaming
current, and an electrostatic potential difference between the
excess charge 7146 near the inlet "IN," and the excess charge 7148
near the outlet "OUT" creating a steady-state charge separation
therebetween.
[0166] The amount of charge separation, and hence the electrostatic
potential difference between the excess charge 7146 near the inlet
"IN," and the excess charge 7148 near the outlet "OUT," depends on
additional energy per unit charge supplied by a pump (e.g., the
rotor 600, the internal pump 410, and/or the external pump 210) to
"push" charge against the opposing electric field (created by the
charge separation) to produce the a liquid flow rate approximating
a flow rate obtainable by a liquid without ions (i.e., ions 7122
and 7126). If the plates 7142 and 7144 are insulators, the
electrostatic potential difference is a direct measure of the EMF
the pump (e.g., the rotor 600, the internal pump 410 and/or the
external pump 210) can generate. In this case, one could measure
the electrostatic potential difference using a voltmeter having a
pair of leads by placing one of the leads in the first material 110
near the inlet "IN," and the other lead in the first material 110
near the outlet "OUT."
[0167] With insulating plates 7142 and 7144, any return current is
purely an ion current (or flow of ions), in that the return current
involves only the conduction of ions through the first material
110. If other conductive mechanisms through more conductive
pathways are present between the excess charge 7146 near the inlet
"IN," and the excess charge 7148 near the outlet "OUT," the return
current may use those more conductive pathways. For example,
conducting metal plates 7142 and 7144 may provide more conductive
pathways; however, these more conductive pathways transmit only an
electron current and not the ion current.
[0168] As is appreciated by those of ordinary skill, to transfer
the charge carried by an ion to one or more electrons in the metal,
and vise versa, one or more oxidation-reduction reactions must
occur at the surface of the metal, producing reaction products.
Assuming the first material 110 is water (H.sub.2O) and the second
material 120 is oxygen (O.sub.2), a non-limiting example of a redox
reaction, which would inject negative charge into the conducting
plates 7142 and 7144 includes the following known half-cell
reaction:
O.sub.2+H.sub.2O.fwdarw.O.sub.3+2H.sup.++2e.sup.-,
Again, assuming the first material 110 is water (H.sub.2O) and the
second material 120 is oxygen (O.sub.2), a non-limiting example of
a redox reaction includes the following known half-cell reaction,
which would remove negative charge from the conducting plates 7142
and 7144 includes the following known half-cell reaction:
2H.sup.++e.sup.-.fwdarw.H.sub.2,
[0169] With conducting metal plates 7142 and 7144, most of the
return current is believed to be an electron current, because the
conducting plates 7142 and 7144 are more conductive than the first
material 110 (provided the redox reactions are fast enough not to
be a limiting factor). For the conducting metal plates 7142 and
7144, a smaller charge separation accumulates between the inlet
"IN" and the outlet "OUT," and a much smaller electrostatic
potential exists therebetween. However, this does not mean that the
EMF is smaller.
[0170] As described above, the EMF is related to the energy per
unit charge the pump provides to facilitate the flow of the first
material 110 against the opposing electric field created by the
charge separation. Because the electrostatic potential is smaller,
the pump may supply less energy per unit charge to cause the first
material 110 to flow. However, the above example redox reactions do
not necessarily occur spontaneously, and thus may require a work
input, which may be provided by the pump. Therefore, a portion of
the EMF (that is not reflected in the smaller electrostatic
potential difference) may be used to provide the energy necessary
to drive the redox reactions.
[0171] In other words, the same pressure differentials provided by
the pump to push against the opposing electric field created by the
charge separation for the insulating plates 7142 and 7144, may be
used both to "push" the charge through the conducting plates 7142
and 7144 and drive the redox reactions.
[0172] Referring to FIG. 29, an experimental setup for an
experiment conducted by the inventors is provided. The experiment
included a pair of substantially identical spaced apart 500 ml
standard Erlenmeyer flasks 7150 and 7152, each containing a volume
of deionized water 7153. A rubber stopper 7154 was inserted in the
open end of each of the flasks 7150 and 7152. The stopper 7154
included three pathways, one each for a hollow tube 7156, a
positive electrode 7158, and a negative electrode 7160. With
respect to each of the flasks 7150 and 7152, each of the hollow
tube 7156, the positive electrode 7158, and the negative electrode
7160 all extended from outside the flask, through the stopper 7154,
and into the deionized water 7153 inside the flask. The positive
electrode 7158 and the negative electrode 7160 were constructed
from stainless steel. The hollow tubes 7156 in both of the flasks
7150 and 7152 had an open end portion 7162 coupled to a common
oxygen supply 7164. The positive electrode 7158 and the negative
electrode 7160 inserted into the flask 7152 where coupled to a
positive terminal and a negative terminal, respectively, of a DC
power supply 7168. Exactly the same sparger was used in each
flask.
[0173] Oxygen flowed through the hollow tubes 7156 into both of the
flasks 7150 and 7152 at a flow rate (Feed) of about 1 SCFH to about
1.3 SCFH (combined flow rate). The voltage applied across the
positive electrode 7158 and the negative electrode 7160 inserted
into the flask 7152 was about 2.55 volts. This value was chosen
because it is believed to be an electrochemical voltage value
sufficient to affect all oxygen species. This voltage was applied
continuously over three to four hours during which oxygen from the
supply 7164 was bubbled into the deionized water 7153 in each of
the flasks 7150 and 7152.
[0174] Testing of the deionized water 7153 in the flask 7150 with
HRP and pyrogallol gave an HRP-mediated pyrogallol reaction
activity, consistent with the properties of fluids produced with
the alternate rotor/stator embodiments described herein. The HRP
optical density was about 20% higher relative to pressure-pot or
fine-bubbled solutions of equivalent oxygen content. The results of
this experiment indicate that mixing inside the mixing chamber 330
involves a redox reaction. According to particular aspects, the
inventive mixing chambers provide for output materials comprising
added electrons that are stabilized by either oxygen-rich water
structure within the inventive output solutions, or by some form of
oxygen species present due to the electrical effects within the
process.
[0175] Additionally, the deionized water 7153 in both of the flasks
7150 and 7152 was tested for both ozone and hydrogen peroxide
employing industry standard colorimetric test ampoules with a
sensitivity of 0.1 ppm for hydrogen peroxide and 0.6 ppm for ozone.
There was no positive indication of either species up to the
detection limits of those ampoules.
Dwell Time
[0176] Dwell time is an amount of time the first material 110, the
second material 120, and optionally the third material 130 spend in
the mixing chamber 330. The ratio of the length of the mixing
chamber 330 to the diameter of the mixing chamber 330 may
significantly affect dwell time. The greater the ratio, the longer
the dwell time. As mentioned in the Background Section, the rotor
12 of the prior art device 10 (see FIG. 1) had a diameter of about
7.500 inches and a length of about 6.000 inches providing a length
to diameter ratio of about 0.8. In contrast, in particular
embodiments, the length of the mixing chamber 330 of the mixing
device 100 is about 5 inches and the diameter "D1" of the rotor 600
is about 1.69 inches yielding a length to diameter ratio of about
2.95.
[0177] Dwell time represents the amount of time that the first
material 110, the second material 120, and optionally the third
material 130 are able to interact with the electrokinetic phenomena
described herein. The prior art device 10 is configured to produce
about 60 gallons of the output material 102 per minute and the
mixing device 100 is configured to produce about 0.5 gallons of the
output material 102 per minute, the prior art device 10 (see FIG.
1) had a fluid dwell time of about 0.05 seconds, whereas
embodiments of the mixing device 100 have a substantially greater
(about 7-times greater) dwell time of about 0.35 seconds. This
longer dwell time allows the first material 110, the second
material 120, and optionally the third material 130 to interact
with each other and the surfaces 606 and 705 (see FIG. 7) inside
the mixing chamber 330 for about 7 times longer than was possible
in the prior art device 10.
[0178] With reference to Table I below, the above dwell times were
calculated by first determining the flow rate for each device in
gallons per second. In the case of the prior art device 10 was
configured to operate at about 60 gallons of output material per
minute, while the mixing device 100 is configured to operate over a
broader range of flow rate, including at an optimal range of about
0.5 gallons of output material per minute. The flow rate was then
converted to cubic inches per second by multiplying the flow rate
in gallons per second by the number of cubic inches in a gallon
(i.e., 231 cubic inches). Then, the volume (12.876 cubic inches) of
the channel 32 of the prior art device 10 was divided by the flow
rate of the device (231 cubic inches/second) to obtain the dwell
time (in seconds) and the volume (0.673 cubic inches) of the mixing
chamber 330 of the mixing device 100 was divided by the flow rate
(1.925 cubic inches/second) of the device (in cubic inches per
second) to obtain the dwell time (in seconds).
TABLE-US-00001 TABLE I Table 1. Inventive device can accommodate a
range of dwell times, including a substantially increased (e.g.,
7-times) dwell time relative to prior art devices. Volume Flow Rate
Mixing Flow Rate Flow Rate Cubic Chamber Dwell Gallons/ Gallons/
Inches/ (Cubic Time Device Minute Second Second Inches) (Seconds)
Prior art 60 1.000 231.000 12.876 0.056 device 10 Mixing 2 0.033
7.700 0.673 0.087 device 100 Mixing 0.5 0.008 1.925 0.673 0.350
device 100
Rate of Infusion
[0179] Particular aspects of the mixing device 100 provide an
improved oxygen infusion rate over the prior art, including over
prior art device 10 (see FIG. 1). When the first material 110 is
water and the second material 120 is oxygen, both of which are
processed by the mixing device 100 in a single pass (i.e., the
return block of FIG. 2 is set to "NO") at or near 20.degree.
Celsius, the output material 102 has a dissolved oxygen level of
about 43.8 parts per million. In certain aspects, an output
material having about 43.8 ppm dissolved oxygen is created in about
350 milliseconds via the inventive flow through the inventive non
pressurized (non-pressure pot) methods. In contrast, when the first
material 110 (water) and the second material 120 (oxygen) are both
processed in a single pass at or near 20.degree. Celsius by the
prior art device 10, the output material had dissolved oxygen level
of only 35 parts per million in a single pass of 56
milliseconds.
Output Material 102
[0180] When the first material 110 is a liquid (e.g., freshwater,
saline, GATORADE.RTM., and the like) and the second material 120 is
a gas (e.g., oxygen, nitrogen, and the like), the mixing device 100
may diffuse the second material 120 into the first material 110.
The following discusses results of analyses performed on the output
material 102 to characterize one or more properties of the output
material 102 derived from having been processed by the mixing
device 100.
[0181] When the first material 110 is saline solution and the
second material 120 is oxygen gas, experiments have indicated that
a vast majority of oxygen bubbles produced within the saline
solution are no greater than 0.1 micron in size.
Decay of Dissolved Oxygen Levels
[0182] Referring now to FIG. 30, there is illustrated the DO levels
in water enriched with oxygen in the mixing device 100 and stored
in a 500 ml thin-walled plastic bottle and a 1000 ml glass bottle
out to at least 365 days. Each of the bottles was capped and stored
at 65 degrees Fahrenheit. As can be seen in the Figure, the DO
levels of the oxygen-enriched fluid remained fairly constant out to
at least 365 days.
[0183] Referring to FIG. 31, there is illustrated the DO levels in
water enriched with oxygen in the mixing device 100 and stored in a
500 ml plastic thin-walled bottle and a 1000 ml glass bottle. Both
bottles were refrigerated at 39 degrees Fahrenheit. Again, DO
levels of the oxygen-enriched fluid remained steady and decreased
only slightly out to at least 365 days.
[0184] Referring now to FIG. 32, there is illustrated the dissolved
oxygen levels in GATORADE.RTM. enriched with oxygen in the mixing
device 100 and stored in 32 oz. GATORADE.RTM. bottles having an
average temperature of 55 degrees Fahrenheit at capping. The
GATORADE.RTM. bottles were subsequently refrigerated at 38 degrees
Fahrenheit between capping and opening. During the experiment, a
different bottle was opened at 20, 60, and 90 days, respectively,
to measure the DO levels of the GATORADE.RTM. stored therein.
[0185] The GATORADE.RTM. within a first group of GATORADE.RTM.
bottles was processed with oxygen in the mixing device 100 at
approximately 56 degrees Fahrenheit. The DO levels of the
GATORADE.RTM. at bottling were approximately 50 ppm as indicated by
point 8104. A first bottle was opened at approximately 20 days, and
the DO level of the GATORADE.RTM. was determined to be
approximately 47 ppm as indicated by point 8106. A second bottle
was then opened at 60 days, and the DO level of the GATORADE.RTM.
was measured to be approximately 44 ppm as indicated by point 8108.
Finally, a third bottle was opened at 90 days, and the DO level of
the GATORADE.RTM. was determined to be slightly below 40 ppm as
indicated by point 8110.
[0186] The GATORADE.RTM. within a second group of GATORADE.RTM.
bottles was processed with oxygen in the mixing device 100 at
approximately 52 degrees Fahrenheit. The initial DO level for
GATORADE.RTM. stored in this group of bottles was 45 ppm as
illustrated by point 8112. The GATORADE.RTM. in the bottle opened
at 20 days had a DO level of only slightly lower than 45 ppm as
indicated by point 8114. The second bottle of GATORADE.RTM. was
opened at 60 days and the GATORADE.RTM. therein had a DO level of
slightly more than 41 ppm. Finally, a third bottle of GATORADE.RTM.
was opened at 90 days and the GATORADE.RTM. therein had a DO level
of approximately 39 ppm as shown by point 8116. As before, with
respect to the water test in the plastic and glass bottles (see
FIG. 31), it can be seen that the DO levels remain at relatively
high levels over the 90 day period and substantially higher than
those levels present in normal (unprocessed) GATORADE.RTM. stored
in 32 oz. GATORADE.RTM. bottles. Point 8010 is the level
corresponding to inventive output fluid in a covered PET
bottle.
[0187] FIG. 33 illustrates the DO retention of 500 ml of braun
balanced salt solution processed with oxygen in the mixing device
100 and kept at standard temperature and pressure in an amber glass
bottle. The DO level of the solution before processing is 5 ppm.
After processing in the mixing device 100, the DO level was
increased to approximately 41 ppm (illustrated as point 8202). An
hour after processing, the DO level dropped to approximately 40 ppm
as indicated by point 8204. Two hours after processing, the DO
level dropped to approximately 36 ppm as indicated by point 8206.
The DO level dropped to approximately 34 ppm three hours after
processing as indicated by point 8208. At approximately four and a
half hours after processing, the DO level within the salt solution
dropped to slightly more than 30 ppm. The final measurement was
taken shortly before six hours after processing whereat the DO
level had dropped to approximately 28 ppm. Thus, each of the
experiments illustrated in FIGS. 30-33 illustrate that that the DO
levels remain at relatively high levels over extended periods.
[0188] Because the output material 102 may be consumed by human
beings, the materials used to construct the mixing device 100
should be suitable for food and/or pharmaceutical manufacture. By
way of non-limiting example, the housing 520, the housing 5520, the
rotor 600, the stator 700, and the stator 5700 may all be
constructed from stainless steel.
Bubble Size Measurements
[0189] Experimentation was performed to determine a size of the
bubbles of gas diffused within the fluid by the mixing device 100.
While experiments were not performed to measure directly the size
of the bubbles, experiments were performed that established that
the bubble size of the majority of the gas bubbles within the fluid
was smaller than 0.1 microns. In other words, the experiments
determined a size threshold value below which the sizes of the
majority of bubbles fall.
[0190] This size threshold value or size limit was established by
passing the output material 102 formed by processing a fluid and a
gas in the mixing device 100 through a 0.22 filter and a 0.1 micron
filter. In performing these tests, a volume of the first material
110, in this case, a fluid, and a volume of the second material
120, in this case, a gas, were passed through the mixing device 100
to generate a volume of the output material 102 (i.e., a fluid
having a gas diffused therein). Sixty milliliters of the output
material 102 was drained into a 60 ml syringe. The DO level of the
fluid was measured via the Winkler Titration. The fluid within the
syringe was injected through a 0.22 micron filter into a 50 ml
beaker. The filter comprised the Millipore Millex.RTM. GP50 filter.
The DO level of the material in the 50 ml beaker was then measured.
The experiment was performed three times to achieve the results
illustrated in Table II below.
TABLE-US-00002 TABLE II DO AFTER 0.22 MICRON DO IN SYRINGE FILTER
42.1 ppm 39.7 ppm 43.4 ppm 42.0 ppm 43.5 ppm 39.5 ppm
[0191] As can be seen, the DO levels measured within the syringe
and the DO levels measured within the 50 ml beaker were not changed
drastically by passing the output material 102 through the 0.22
micron filter. The implication of this experiment is that the
bubbles of dissolved gas within the output material 102 are not
larger than 0.22 microns otherwise there would be a significantly
greater reduction in the DO levels in the output material 102
passed through the 0.22 micron filter.
[0192] A second test was performed in which the 0.1 micron filter
was substituted for the 0.22 micron filter. In this experiment,
saline solution was processed with oxygen in the mixing device 100
and a sample of the output material 102 was collected in an
unfiltered state. The DO level of the unfiltered sample was 44.7
ppm. The output material 102 was filtered using the 0.1 micron
filter and two additional samples were collected. The DO level of
the first sample was 43.4 ppm. The DO level of the second sample
was 41.4 ppm. Then, the filter was removed and a final sample was
taken from the unfiltered output material 102. The final sample had
a DO level of 45.4 ppm. These results were consistent with those
seen using the Millipore 0.22 micron filter. These results lead to
the conclusion that there is a trivial reduction in the DO levels
of the output material 102 passed through the 0.1 micron filter
providing an indication that the majority of the bubbles in the
processed saline solution are no greater than 0.1 micron in
size.
[0193] As appreciated in the art, the double-layer (interfacial)
(DL) appears on the surface of an object when it is placed into a
liquid. This object, for example, might be that of a solid surface
(e.g., rotor and stator surfaces), solid particles, gas bubbles,
liquid droplets, or porous body. In the mixing device 100, bubble
surfaces represent a significant portion of the total surface area
present within the mixing chamber that may be available for
electrokinetic double-layer effects. Therefore, in addition to the
surface area and retention time aspects discussed elsewhere herein,
the relatively small bubble sizes generated within the mixer 100
compared to prior art devices 10, may also contribute, at least to
some extent, to the overall electrokinetic effects and output fluid
properties disclosed herein. Specifically, in preferred
embodiments, as illustrated by the mixer 100, all of the gas is
being introduced via apertures on the rotor (no gas is being
introduced through stator apertures. Because the rotor is rotating
at a high rate (e.g., 3,400 rpm) generating substantial shear
forces at and near the rotor surface, the bubble size of bubbles
introduced via, and adjacent to the spinning rotor surface
apertures would be expected to be substantially (e.g., 2 to 3-times
smaller) smaller than those introduced via and near the stationary
stator. The average bubble size of the prior art device 10 may,
therefore, be substantially larger because at least half of the gas
is introduced into the mixing chamber from the stationary stator
apertures. Because the surface area of a sphere surface varies with
r.sup.2, any such bubble component of the electrokinetic surface
area of the mixing device 100 may be substantially greater than
that of the prior art diffusion device 10.
[0194] Therefore, without being bound by theory, not only does the
mixing chamber of the mixing device 100 have (i) a substantially
higher surface to volume ratio than that of the prior art device 10
(the prior art device 10 has a ratio of surface to volume of 10.9,
whereas the present mixer 100 has a surface to volume ratio of
39.4), along with (ii) a 7-fold greater dwell-time, but (iii) the
unique properties of the current output solutions may additionally
reflect a contribution from the substantially larger bubble surface
area in the mixing device 100. These distinguishing aspects reflect
distinguishing features of the present mixing device 100, and
likely each contribute to the unique electrokinetic properties of
the inventive output materials/fluids.
Sparging Effects
[0195] FIGS. 34-35 illustrate the sparging effects of the mixing
device 100 on a fluid (e.g., the first material 110) passing
therethrough. Sparging refers to "bubbling" an inert gas through a
solution to remove a different dissolved gas(es) from the solution.
In each of the examples illustrated in FIGS. 34 and 35, the second
material 120 is nitrogen. The levels of dissolved oxygen in the
output material 102 are measured at various points in time. As can
be seen in the figures, the nitrogen gas sparges the oxygen from
the fluid passing through the mixing device 100 causing the DO
levels in the fluid to decay over a period of time.
[0196] The results of another experiment are illustrated in FIG. 34
wherein water is sparged with nitrogen using the mixing device 100.
Two sets of experiments were conducted, the first having a gas flow
rate of SCFH (Standard Cubic Feet per Hour) of 1 and the second
having a gas flow rate of SCFH of 0.6 The fluid flow rate was about
0.5 gal/min. As can be seen, when the process is begun, the DO
levels in each of the experiments was approximately 9 ppm. After
only one minute, the DO levels had dropped to slightly above 5 ppm.
At two minutes the DO levels had dropped to approximately 2.5 ppm.
The DO level appears to level out at a minimum level at
approximately 6 minutes wherein the DO level is slightly above zero
(0). Thus, the nitrogen sparges the oxygen from the water
relatively quickly.
[0197] FIG. 35 illustrates the sparging of oxygenated water in an 8
gallon tank at standard temperature and pressure. The decay rate of
the DO in the water is illustrated by line 8602. As can be seen,
initially the oxygenated water had a DO level of approximately 42
ppm. After 2 minutes of processing by the mixing device 100, the
nitrogen sparged the oxygenated water such that the DO level
dropped to slightly more than 20 ppm. At 6 minutes, the DO level
dropped from greater than 40 ppm to only 6 ppm. The DO level of the
oxygenated water reached a minimum value slightly greater than zero
(0) at approximately 14 minutes after the beginning of the process.
Thus, the above described sparging experiments illustrate that the
mixing device 100 is capable of quickly sparging oxygen from water
and replacing the oxygen with another gas such as nitrogen by
processing oxygenated water with mixing device 100 for a rather
short period of time. In other words, because total partial gas
pressure in the fluid remained at approximately the same level
despite the decrease in DO, the nitrogen gas replaced the oxygen in
the fluid.
[0198] These figures illustrate the manner in which nitrogen may be
diffused into water to sparge the oxygen from the water. However,
any gas could be used to sparge a selected gas from any selected
fluid and diffuse into the selected fluid the gas used to sparge
the selected gas from the selected fluid. For example, the
principals illustrated may also be applicable to sparging nitrogen
from water or another fluid using oxygen. Further, any gas
dissolved within a solution may be sparged therefrom using a
different gas to take the place of the gas sparged from the
solution. In other words, by processing a sparging gas and a
solution containing a dissolved gas through the mixing device 100
for a relatively short period of time, the dissolved gas could be
quickly and efficiently removed from the solution.
Molecular Interactions
[0199] A number of physicists have begun to describe the quantum
properties of water. Conventionally, quantum properties are thought
to belong to elementary particles of less than 10.sup.-10 meters,
while the macroscopic world of our everyday life is referred to as
classical, in that it behaves according to Newton's laws of motion.
Between the macroscopic classical world and the microscopic quantum
world is the mesoscopic domain, where the distinction between
macroscopic and microscopic is becoming increasingly blurred.
Indeed, physicists are discovering quantum properties in large
collections of atoms and molecules in the nanometer to micrometer
range, particularly when the molecules are packed closely together
in a liquid phase.
[0200] Recently, chemists have made a surprising discovery that
molecules form clusters that increase in size with dilution. These
clusters measure several micrometers in diameter. The increase in
size occurs non-linearly with dilution and depends on history,
flying in the face of classical chemistry. Indeed, there is yet no
explanation for this phenomena. It may well be yet another
reflection of the strangeness of water that depends on its quantum
properties.
[0201] In the mid 1990's, quantum physicist del Giudice and
Preparata and other colleagues at the University of Milan, in
Italy, argued that quantum coherent domains measuring 100
nanometers in diameter could arise in pure water. They show how the
collective vibrations of water molecules in the coherent domain
eventually become phase locked to the fluctuations of the global
electromagnetic field. In this way, long lasting, stable
oscillations could be maintained in water.
[0202] One way in which memory might be stored in water is through
the excitation of long lasting coherent oscillations specific to
one or more substances (such as a therapeutic agent) dissolved in
the water. Interactions between the water molecules and the
molecules of the substances dissolved in the water change the
collective structure of the water, which would in turn determine
the specific coherent oscillations that develop. If these
oscillations become stabilized and maintained by phase coupling
between the global field and the excited molecules, then, even when
the dissolved substances are diluted away, the water may still
carry the coherent oscillations that can seed other volumes of
water on dilution.
[0203] The discovery that dissolved substances form increasingly
large clusters is compatible with the existence of a coherent field
in water that can transmit attractive resonance between molecules
when the oscillations are in phase leading to clumping in dilute
solutions. As a cluster of molecules increases in size, its
electromagnetic signature is correspondingly amplified, reinforcing
the coherent oscillations carried by the water.
[0204] One should expect changes in some physical properties in
water that could be detectible. Unfortunately, all attempts to
detect such coherent oscillations by usual spectroscopic and
nuclear magnetic resonance methods have yielded ambiguous results.
This is not surprising in view of the finding that cluster size of
the dissolved molecules depends on the precise history of dilution
rather than concentration of the molecules.
[0205] It is possible that despite variations in the cluster size
of the dissolved molecules and detailed microscopic structure of
the water, a specificity of coherent oscillations may nonetheless
exist. Usual detection methods fail because they depend upon using
the microscopic particles of individual molecules, or of small
aggregates. Instead, what is needed is a method of detecting
collective global properties over many, many molecules. Some
obvious possibilities that suggest themselves are the measurements
of freezing points and boiling points, viscosity, density,
diffusivity, and magnet properties. One possibility for detecting
changes in collective global properties of water is by means of
crystallization. Crystals are formed from macroscopic collections
of molecules. Like other measurements that depend on global
properties, crystals simplify the subtle changes in the individual
molecules that would have been undetectable otherwise.
[0206] With reference to FIG. 36, a simplified protonated water
cluster forming a nanoscale cage 8700 is shown. A protonated water
cluster typically takes the form of H.sup.+(H.sub.20).sub.n. Some
protonated water clusters occur naturally, such as in the
ionosphere. Without being bound by any particular theory, and
according to particular aspects, other types of water clusters or
structures (clusters, nanocages, etc.) are possible, including
structures comprising oxygen and stabilized electrons imparted to
the inventive output materials. Oxygen atoms 8704 may be caught in
the resulting structures 8700. The chemistry of the semi-bound
nanocage allows the oxygen 8704 and/or stabilized electrons to
remain dissolved for extended periods of time. Other atoms or
molecules, such as medicinal compounds, can be caged for sustained
delivery purposes. The specific chemistry of the solution material
and dissolved compounds depend on the interactions of those
materials.
[0207] Fluids processed by the mixing device 100 have been shown
via experiments to exhibit different structural characteristics
that are consistent with an analysis of the fluid in the context of
a cluster structure.
Rayleigh Effects
[0208] If a strong beam of light is passed through a transparent
gaseous or liquid medium containing solid or liquid particles, or
even molecules of extremely high molecular weight, the light is
scattered away from the direction of its incident path. The
scattering is due to the interference effects that arise from the
density fluctuations in the scattering medium (i.e., the presence
of particles or very high molecular weight molecules.) There are
two types of light scattering. The first involves the wavelength of
the scattered light differing from that of the incident light and
is called Raman scattering. The other type scattering involves when
the scattered light has the same wavelength of the incident light
and is called Rayleigh scattering. In Rayleigh scattering, the
intensity of the scattered light is proportional to the product of
the intensity of the incident light and the attenuation constant, a
function of the refractive index and the Rayleigh constant. The
Rayleigh constant is a somewhat involved function of the molecular
weight of the scattering substance and thus a measurement of the
intensity of the scattered light can give a value for the molecular
weight. This scattering phenomenon is used in a number of liquid
chromatography detectors.
[0209] Water processed through the mixing device 100 has been
demonstrated to have detectible structural differences when
compared with normal unprocessed water. For example, processed
water has been shown to have more Rayleigh scattering than is
observed in unprocessed water. In the experiments that were
conducted, samples of processed and unprocessed water were prepared
(by sealing each in a separate bottle), coded (for later
identification of the processed sample and unprocessed sample), and
sent to an independent testing laboratory for analysis. Only after
the tests were completed were the codes interpreted to reveal which
sample had been processed by the mixing device 100.
[0210] At the laboratory, the two samples were placed in a laser
beam having a wavelength of 633 nanometers. The fluid had been
sealed in glass bottles for approximately one week before testing.
With respect to the processed sample, Sample B scattered light
regardless of its position relative to the laser source. However,
"Sample A" did not. After two to three hours following the opening
of the bottle, the scattering effect of Sample B disappeared. These
results imply the water exhibited a memory causing the water to
retain its properties and dissipate over time. These results also
imply the structure of the processed water is optically different
from the structure of the unprocessed fluid. Finally, these results
imply the optical effect is not directly related to DO levels
because the DO level at the start was 45 ppm and at the end of the
experiment was estimated to be approximately 32 ppm.
Generation of Solvated Electrons
[0211] Additional evidence indicates that the mixing occurring
inside the mixing device 100 generates solvated electrons within
the output material 102. This conclusion results from conditions
observed with respect to the dissolved oxygen probe effects used in
measuring the DO levels within various processed solutions. Due to
the experiences viewed with respect to the polarographic dissolved
oxygen probes, it is a belief that the processed fluid exhibits an
electron capture effect and thus the fluid includes solvated
electrons.
[0212] There are two fundamental techniques for measuring dissolved
oxygen ("DO") levels electrically: galvanic measuring techniques
and polarographic measurements. In both techniques, the DO level
sensor includes two electrodes, an anode and a cathode, which are
both immersed in electrolyte within the sensor body. An oxygen
permeable membrane separates the anode and cathode from the
solution being tested. The cathode is a hydrogen electrode and
carries negative potential with respect to the anode. The
electrolyte solution surrounds the electrode pair and is contained
by the membrane. With no oxygen, the cathode becomes polarized with
hydrogen and resists the flow of current. When oxygen passes
through the membrane, the cathode is depolarized and electrons are
consumed. In other words, oxygen diffuses across the membrane and
interacts with the internal components of the probe to produce an
electrical current. The cathode electrochemically reduces the
oxygen to hydroxyl ions according to the following equation:
O.sub.2+2H.sub.2O+4E.sup.-=4OH.sup.-
[0213] When attempting to measure DO levels in a solution processed
by the mixing device 100, an overflow condition has been repeatedly
experienced wherein the dissolved oxygen meter actually displays a
reading that is higher than the meter is capable of reading.
Independent means, a Winkler Titration, reveals a much lower DO
level for the solution than indicated by the probe. Typically, in a
device such as the Orion 862, having a maximum reading of 60 ppm,
the meter will overflow and have the high oxygen level indication
if left in bulk processed water for several minutes.
[0214] Because the overload is not caused by dissolved oxygen in
the fluid, it is believed solvated electrons must be causing the
overload. In other words, solvated electrons are accompanying the
processed water across the membrane. These electrons are attracted
to the anode and cause the current observed. It is a further belief
that these electrons are captured in a cage or cluster mechanism
within the solution.
[0215] Compositions comprising hydrated (solvated) electrons
imparted to the inventive compositions by the inventive
processes
[0216] In certain embodiments as described herein (see under
"Double-layer"), the gas-enriched fluid is generated by the
disclosed electromechanical processes in which molecular oxygen is
diffused into the fluid and may operate to stabilize charges (e.g.,
hydrated (solvated) electrons) imparted to the fluid. Without being
bound by theory or mechanism, certain embodiments of the present
invention relate to an oxygen-enriched fluid (output material)
comprising charges (e.g., hydrated (solvated) electrons) that are
added to the materials as the first material is mixed with oxygen
in the inventive mixer device to provide the combined output
material. According to particular aspects, these hydrated
(solvated) electrons (alternately referred to herein as `solvated
electrons`) are stabilized in the inventive solutions as evidenced
by the persistence of assayable effects mediated by these hydrated
(solvated) electrons. Certain embodiments may relate to hydrated
(solvated) electrons and/or water-electron structures, clusters,
etc. (See, for example, Lee and Lee, Bull. Kor. Chem. Soc. 2003, v.
24, 6; 802-804; 2003).
[0217] Novel HRP based assay. Horseradish peroxidase (HRP) is
isolated from horseradish roots (Amoracia rusticana) and belongs to
the ferroprotoporphyrin group (Heme group) of peroxidases. HRP
readily combines with hydrogen peroxide or other hydrogen donors to
oxidize the pyrogallol substrate. Additionally, as recognized in
the art, HRP facilitates autoxidative degradation of
indole-3-acietic acid in the absence of hydrogen peroxide (see,
e.g., Heme Peroxidases, H. Brian Dunford, Wiley-VCH, 1999, Chapter
6, pages 112-123, describing that autoxidation involves a highly
efficient branched-chain mechanism; incorporated herein by
reference in its entirety). The HRP reaction can be measured in
enzymatic activity units, in which Specific activity is expressed
in terms of pyrogallol units. One pyrogallol unit will form 1.0 mg
purpurogallin from pyrogallol in 20 sec at pH 6.0 at 20.degree. C.
This purpurogallin (20 sec) unit is equivalent to approx. 18 .mu.M
units per min at 25.degree. C.
##STR00001##
[0218] According to particular aspects of the present invention,
the oxygen-enriched inventive fluids (output materials) have been
described and disclosed herein to react with pyrogallol in the
presence of horseradish peroxidase. The reaction is most likely
based on an auto-oxidation of the pyrogallol, since no hydrogen
peroxide, superoxide, or other reactive oxygen species has been
detected in oxygen-enriched inventive fluid. The extent of this
reaction is greater than that of pressurized oxygen solutions
(pressure-pot oxygen solutions) and less than that of hydrogen
peroxide.
[0219] Specifically, the present applicants have determined that
while there is no hydrogen peroxide (none detected at a sensitivity
of 0.1 ppm), the inventive gas-enriched fluid may be consistently
characterized by its facilitation of the apparent autoxidation of
pyrogallol to purpurogallin in the presence of horseradish
peroxidase enzyme (HRP). That is, like the case of HRP facilitation
of the autoxidative degradation of indole-3-acietic acid in the
absence of hydrogen peroxide, applicants have discovered HRP
facilitation of the autoxidative degradation of pyrogallol in the
absence of hydrogen peroxide. According to particular aspects, the
presence and level of this activity are distinguishing features of
the inventive compositions in view of the prior art.
[0220] In certain embodiments, the inventive gas-enriched fluid
facilitates, in the presence of HRP and absence of hydrogen
peroxide, a pyrogallol autoxidation rate (under standard conditions
as defined herein under "Definitions") equivalent to approximately
0.5 ppm of hydrogen peroxide, approximately 0.8 ppm of hydrogen
peroxide, approximately 1 ppm of hydrogen peroxide, approximately 2
ppm of hydrogen peroxide, approximately 3 ppm of hydrogen peroxide,
approximately 4 ppm of hydrogen peroxide, approximately 5 ppm of
hydrogen peroxide, approximately 6 ppm of hydrogen peroxide,
approximately 7 ppm of hydrogen peroxide, approximately 8 ppm of
hydrogen peroxide, approximately 9 ppm of hydrogen peroxide,
approximately 10 ppm of hydrogen peroxide, approximately 11 ppm of
hydrogen peroxide, approximately 12 ppm of hydrogen peroxide,
approximately 20 ppm of hydrogen peroxide, approximately 40 ppm of
hydrogen peroxide, approximately 50 ppm of hydrogen peroxide or any
value therebetween or greater.
[0221] It is known that Horseradish peroxidase enzyme catalyzes the
auto-oxidation of pyrogallol by way of facilitating reaction with
the molecular oxygen in a fluid. (Khajehpour et al., PROTEINS:
Struct, Funct, Genet. 53: 656-666 (2003)). It is also known that
oxygen binds the heme pocket of horseradish peroxidase enzyme
through a hydrophobic pore region of the enzyme (between Phe68 and
Phe142), whose conformation likely determines the accessibility of
oxygen to the interior. Without being bound by mechanism, because
surface charges on proteins are known in the protein art to
influence protein structure, it is possible that the solvated
electrons present in the inventive gas-enriched fluid act to alter
the conformation of the horseradish peroxidase such that greater
oxygen accessibility results. The greater accessibility of oxygen
to the prosthetic heme pocket of the horseradish peroxidase enzyme
in turn would allow for increased reactivity with pyrogallol, when
compared with prior art oxygenated fluids (pressure-pot,
fine-bubbled). Alternatively, the added or solvated electrons of
the present output compositions may be acting in other ways to
enable facilitation of the apparent autoxidation of pyrogallol to
purpurogallin in the presence of horseradish peroxidase enzyme
(HRP).
[0222] In any event, according to particular aspects, production of
output material using the inventive methods and devices comprises a
process involving: an interfacial double layer that provides a
charge gradient; movement of the materials relative to surfaces
pulling charge (e.g., electrons) away from the surface by virtue of
a triboelectric effect, wherein the flow of material produces a
flow of solvated electrons. Moreover, according to additional
aspects, and without being bound by mechanism, the orbital
structure of diatomic oxygen creates charge imbalances (e.g., the
two unpaired electrons affecting the hydrogen bonding of the water)
in the hydrogen bonding arrangement within the fluid material
(water), wherein electrons are solvated and stabilized within the
imbalances.
[0223] The inventive combination of oxygen-enrichment and solvated
electrons imparted by the double-layer effects and configuration of
the presently claimed devices facilitates the auto-oxidation of
pyrogallol in the presence of HRP, which is a distinguishing
feature of the present inventive output material compositions that
can be readily monitored and quantified by way of optical density.
Typically, the inventive oxygen-enriched compositions are
characterized in that they provide for about a 20% higher optical
density read-out in the standard assay compared to either
pressurized (pressure pot) or fine-bubbled control fluid have
equivalent dissolved oxygen concentrations. The HRP is likely
providing added oxidative ability to the autoxidation.
Pyrogallol Reactivity Test
[0224] An aliquot of the inventive oxygen-enriched output material
was tested for peroxidase activity by using a commercially
available horseradish peroxidase and a pyrogallol assay (Sigma).
Briefly, pyrogallol stock solution was prepared with deionized
water. Pyrogallol measures peroxidase activity of the horseradish
peroxidase enzyme on the fluid as it reacts with a substrate (such
as hydrogen peroxide), to yield purpurogallin and water. Test fluid
with horseradish peroxidase, pyrogallol and the appropriate
potassium phosphate buffer were compared with other fluids.
Hydrogen peroxide served as the positive control. The other fluids
tested were water that was oxygenated and pressurized in a pressure
pot, up to 100 psi to reach the desired dissolved oxygen level
(Pressure Pot), while the other fluid was oxygenated with an air
stone in an open beaker (Fine Bubble). All fluids tested were
maintained at room temperature, and measured approximately 55 ppm
dissolved oxygen level (by FOXY probe). Water samples were tested
by adding the enzymatic reagents. Continuous spectrophotometric
rate determination was made at A.sub.420 nm, and room temperature
(25 degrees Celsius).
[0225] As indicated in FIGS. 38-41, the inventive oxygen-enriched
fluid tested positive for reactivity with horseradish peroxidase by
pyrogallol, while the pressure pot and fine bubbled water samples
had far less reactivity. As indicated in FIG. 42, oxygen is
required for the reaction with pyrogallol in the presence of
horseradish peroxidase, as inventive fluid enriched with other
gases did not react in the same manner.
[0226] Several chemical tests of the inventive oxygen-enriched
fluid for the presence of hydrogen peroxide were conducted, as
described herein, and none of these tests were positive
(sensitivity of 0.1 ppm hydrogen peroxide). Thus, the inventive
oxygen-enriched fluid of the instant application provides for
peroxidase facilitated auto-oxidation activity in the absence of
hydrogen peroxide.
[0227] In particular embodiments, Applicants have determined that
the horseradish peroxidase effect remains at least up to seven
hours after opening of the bottle in which it is stored. In other
embodiments, Applicants have determined that the horseradish
peroxidase effect remains after opening of closed container after
105 days of storage in the closed container. By contrast, in other
embodiments, Applicants have determined that when testing
equivalent dissolved oxygen levels made with just pressurizing
fluid (pressure pot fluids), the decline of a background HRP effect
takes place rapidly, declining precipitously in under 4 hours.
Glutathione Peroxidase Study
[0228] The inventive oxygen-enriched output fluid material was
tested for the presence of hydrogen peroxide by testing the
reactivity with glutathione peroxidase using a standard assay
(Sigma). Briefly, glutathione peroxidase enzyme cocktail was
constituted in deionized water and the appropriate buffers. Water
samples were tested by adding the enzymatic reagents. Continuous
spectrophotometric rate determination was made at A.sub.340 nm, and
room temperature (25 degrees Celsius). Samples tested were: 1.
deionized water (negative control), 2. inventive oxygen-enriched
fluid at low concentration, 3. inventive oxygen-enriched fluid at
high concentration, 4. hydrogen peroxide (positive control). As
illustrated in FIG. 43, the hydrogen peroxide positive control
showed a strong reactivity, while none of the other fluids tested
reacted with the glutathione peroxidase.
Differential Nucleic Acid Stability
[0229] Particular embodiments of the present invention provide
another distinguishing feature of the present inventive
compositions. Specifically, applicants have discovered that there
is a differential thermostability of nucleic acids associated with
the inventive output fluids compared to control fluids. For
example, the T7 promoter primer 5'-d(TAATACGACTCACTATAGGG)-3' (SEQ
ID NO:1) when measured in the inventive oxygen-enriched output
materials relative to non-enriched deionized water. As the
temperature of the water increases, the DNA oligomeric structure
performs a conformational change. As illustrated in FIG. 44,
consent with the art recognized melting temperature for this oligo
of about 48.degree. C., the T7 DNA begins to denature at about 50
degrees Celsius in the control (deionized water), whereas the DNA
in the oxygen-enriched inventive fluid remains intact until about
60 degrees Celsius. Thus, the inventive oxygen-enriched fluid
comprising solvated electrons imparts a higher thermostability for
DNA when compared to control fluid, and provides a further
distinguishing feature of the present inventive output material
compositions that can be readily monitored and quantified by way of
optical density measurements.
Bioreactor Systems Comprising the Inventive Mixing Devices
[0230] Producing significant quantities of target products, such as
proteins, polypeptides, nucleic acids, therapeutic agents, and
other products in host cell systems are possible due to advances in
molecular biology. For example, recombinant proteins are produced
in a host cell systems by transfecting the host cell with nucleic
acids (e.g., DNA) encoding a protein of interest. Next, the host
cell is grown under conditions which allow for expression of the
recombinant protein. Certain host cell systems can be used to
produce large quantities of recombinant proteins which would be too
impractical to produce by other means.
[0231] In addition, enzymatic and/or reaction fermentations, with
or without host cells, are utilized for example in producing
foodstuff and beverages, in treating wastewater, or in
environmental cleanup.
[0232] Cell culturing processes, or cellular fermentation,
typically use prokaryotic or eukaryotic host cells to produce
recombinant proteins. The fermentation is typically conducted in
physical containers (e.g., stirred tanks) called fermentors or tank
bioreactors. The fermentation process itself may comprise (1)
discontinuous operation (batch process), (2) continuous operation,
or (3) semi-continuous operations (such as the fed-batch process),
or any combination of these.
[0233] Since the aim of large scale production of pharmaceutical
drugs (e.g., biologicals) or other target products is to provide
improved manufacturing processes and reduced costs, there is a need
for improved bioreactor equipment, methods, and media for
preparation of these target products.
[0234] The present disclosure sets forth novel gas-enriched fluids,
including, but not limited to gas-enriched water, saline solutions
(e.g., standard aqueous saline solutions), cell culture media, as
well as novel methods and biological and chemical reactor systems
for use in these application processes, and others.
[0235] Certain embodiments disclosed herein relate to systems,
media, and methods for producing a target product, such as a
protein.
[0236] In certain embodiments, a target product may refer to a
protein, peptide, polypeptide, nucleic acid, carbohydrate, polymer,
micelle, and any mixture thereof.
[0237] The target product is typically produced by a vehicle, such
as a host cell, which is associated with the gas-enriched fluid in
a chemical or biological reactor. Reactors may include standard
reactors, such as continuous feed, discontinuous feed, and/or
semi-continuous feed. Reactors may also include a cell culture
vessel (such as a plate, flask, or tank), a plant, an animal, a
fungus, an alga, or other organism. For example, a plant that is
associated with the gas-enriched fluid of the present invention may
comprise plant cells acting as vehicles that aid in the production
of the target product (for example, naturally occurring plant
matter or genetically altered plant matter).
[0238] In certain embodiments, the vehicles utilized with the
gas-enriched fluids or solutions (including media) may include
prokaryotic cells or eukaryotic cells. More specifically, the
living cells may include bacterial (e.g., E. coli, Salmonella,
Streptococcus, Staphylococcus, Neisseria, Nocardia, Mycoplasma,
etc.), fungal (e.g. yeasts, molds, mushrooms, etc.), plant
(tobacco, maize, soybean, fruit or vegetable, etc.), animal
(mammalian, insect, etc.) archebacterial (blue green algae),
protist, human embryonic kidney (HEK) cells, HeLa cells, hybridoma
cells, Madin-Darby Canine Kidney (MDCK) cells, stem cells, cell
lines (including SP2/0 and NSO), African Green Monkey Kidney (Vero)
cells, Spodoptera frugiperda (army worm), Trichoplusia ni (cabbage
looper), and other cells. In addition, viruses (such as
bacteriophage, baculovirus, vaccinia, and other viruses) may be
employed in the bioreactors of the present invention.
[0239] The bioreactor may comprise an airlift reactor, a packed bed
reactor, a fibrous bed reactor, a membrane reactor, a two-chamber
reactor, a stirred-tank reactor, a hollow-fiber reactor, or other
reactor designed to support suspended or immobilized cell growth
.
[0240] In cases of recombinant or target protein production, a
balanced batch and/or feed medium must encourage optimal cell
growth and expression of the recombinant protein. The medium, or
media, is termed "minimal" if it only contains the nutrients
essential for growth. For prokaryotic host cells, the minimal media
typically includes a source of carbon, nitrogen, phosphorus,
magnesium, and trace amounts of iron and calcium. (Gunsalus and
Stanter, The Bacteria, V. 1, Ch. 1 Acad. Press Inc., N.Y. (1960)).
Most minimal media use glucose as a carbon source, ammonia as a
nitrogen source, and orthophosphate (e.g., PO.sub.4) as the
phosphorus source. The media components can be varied or
supplemented according to the specific prokaryotic organism(s)
grown, in order to encourage optimal growth without inhibiting
target protein production. (Thompson et al., Biotech. and Bioeng.
27: 818-824 (1985)). This allows for higher levels of production
with lower cost.
[0241] In addition to the chemical composition of the media, other
factors may affect cell growth and/or target protein production.
These factors include, but are not limited to pH, time, cultivation
temperature, amount of dissolved oxygen or other gas(es), and
partial pressure of those dissolved gasses. During the fermentation
process, the pH of the media is typically altered due to the
consumption of ammonia, or microorganism synthesis of certain
metabolic products, e.g., acetic acid and lactic acid. Since
altered pH may be unfavorable for optimal cell growth, it may be
necessary or desirable to maintain the medium at a certain pH (i.e.
by addition of acids or bases). The pH and other process parameters
can be monitored manually or by automatic devices.
Inventive Gas-Enriched Fluids
[0242] Enriching a fluid with another fluid may result in a
solution or suspension of the two fluids, depending on the physical
and chemical properties of the two fluids. In particular, enriching
a liquid with a gas (e.g., oxygen) may be beneficial for certain
applications, including therapeutic treatments. As utilized herein,
"fluid," may generally refer to a liquid, a gas, a vapor, a mixture
of liquids and/or gases, a liquid and/or gas solution, or any
combination thereof, for any particular disclosed embodiment.
Furthermore, in certain embodiments a "liquid" may generally refer
to a pure liquid or may refer to a gel, sol, emulsion, fluid,
colloid, dispersion, suspension, or mixture, as well as any
combination thereof; any of which may vary in viscosity.
[0243] In particular embodiments, the dissolved gas comprises
oxygen. In other particular embodiments, the dissolved gas
comprises nitrogen, carbon dioxide, carbon monoxide, ozone, sulfur
gas, nitrous oxide, nitric oxide, argon, liquefied petroleum gas,
helium, natural gas, or others.
[0244] One particular advantage of embodiments of the present
invention relates to the gas-enriched fluids' long-term diffused
gas (particularly oxygen) levels, which allows for long-term
bio-availability of the gas to cellular or chemical reactors. The
long-term bio-availability of gasses in the gas-enriched fluids of
the present invention allow for increased target product production
and/or improved enzymatic or other chemical reactions that benefit
from the gas-enriched fluids (including oxygenated media) of the
present invention.
[0245] In some instances, living cells may be grown in a bioreactor
or fermentation chamber in order to promote cell growth and/or
production of the intended target product. While some living cells
require a mixture of gasses in order to sustain or promote their
survival or propagation, cell growth may be hindered or ceased if a
particular gas, such as oxygen, is present at too high of a
concentration.
[0246] For example, mammalian cells, such as Chinese Hamster Ovary
(CHO) cells, require oxygen in order to proliferate. However, the
existing techniques in the art for diffusing gasses, such as
oxygen, into the bioreactor fluids have a detrimental effect on
mammalian cell cultures. For example, the cells may be destroyed or
rendered non-viable in instances where the diffused gas bubbles
rupture or coalesce within the culture media, which is particularly
common at a gas-to-liquid interface. Accordingly, the present
invention represents an advance that would not have occurred in the
ordinary course since the existing knowledge in the art teaches
that the levels of dissolved gas, particularly the levels of
dissolved oxygen, in the gas-enriched media disclosed herein is
predicted to be harmful or detrimental. However, the gas-enriched
fluid media as described herein result in imparting at least one
beneficial advantage to cell cultures selected from the group
consisting of: enhanced cell growth (e.g., rate and/or number)
increased target product yield (e.g., amount), increased rate of
target product production, improved vehicle cell viability,
increased efficiency of target product production, increased ease
in target product purification, and the like. In certain
embodiments, one or more of these beneficial advantages are
conveyed to cell cultures without proving injurious to the cells
themselves.
[0247] In other embodiments, an acellular reaction may utilize the
gas-enriched fluids and methods of the present invention, including
general chemical and/or enzymatic reactions. Examples of such
reactions include, but are not limited to, wastewater treatment,
purification of water (such as treating municipal water, home
drinking purifiers, cleaning swimming pools or aquariums, etc.),
homogenization of milk, hydrogenation of oils, gas-enriching fuels,
and others.
[0248] In further embodiments, the gas-enriched fluid maintains a
dissolved gas enrichment level of at least 10 ppm, at least 15 ppm,
at least 20 ppm, at least 25 ppm, at least 30 ppm, at least 35 ppm,
at least 40 ppm, at least 45 ppm, at least 50 ppm, at least 55 ppm,
at least 60 ppm, at least 65 ppm, at least 70 ppm, at least 75 ppm,
at least 80 ppm, at least 85 ppm, at least 90 ppm, at least 100
ppm, or any value greater or therebetween, at atmospheric pressure.
In certain instances, the gas-enriched fluid maintains its
dissolved gas enrichment level (i.e., the level of the gas enriched
in the fluid) for a period of at least 10 days, at least 20 days,
at least 30 days, at least 40 days, at least 50 days, at least 60
days, at least 70 days, at least 80 days, at least 90 days, at
least 100 days, at least 110 days, at least 120 days, at least 130
days, or greater or any value therebetween, within a sealed
container at atmospheric pressure.
[0249] In one particular embodiment, the host material comprises
water or water vapor. In another particular embodiment, the host
material comprises other fluids (i.e., gasses or liquids) such as
wastewater, toxic materials, potable water, milk, juice, yogurt,
soft drinks (particularly carbonated beverages), ethanol, methanol,
polymers (such as plastic or rubber compounds), oil (edible or
non-edible), emulsions, suspensions, aqueous carriers, non-aqueous
carriers, and the like.
[0250] In certain embodiments, multiple gasses may be used to
enrich or infuse a host fluid. In certain embodiments, ozone and/or
oxygen may be used to break down complex structures into smaller
substructures, particularly if used with sonochemistry techniques,
as described herein inter alia. In certain embodiments, the
gas-enriched fluid or other host material of the present invention
has characteristics that may be more similar to the gas that has
enriched the fluid or other host material, or it may have
characteristics that are more similar to the fluid (e.g., gas or
liquid) or other host material itself.
[0251] In certain embodiments, a gas-enriched fluid or solution
comprises gas-enriched culture media. In particular embodiments,
the gas-enriched media comprises oxygenated or oxygen-enriched
media. In certain embodiments, the gas-enriched fluid or
gas-enriched host material may include further processing, such as
by filtering, separating, modifying or altering various
constituents of the fluid or host material.
Packaged Gas-Enriched Fluids
[0252] Certain embodiments disclosed herein relate to gas-enriched
fluids that have high levels of dissolved or diffused gases
(particularly oxygen) that may be produced by various methods,
including those described herein. In certain embodiments, the
gas-enriched fluid may be produced in a biomass production facility
and applied directly to a bioreactor system. Alternatively, the
gas-enriched fluid may be packaged and distributed for use at other
locations. In the event that the gas-enriched fluid is packaged,
such packaging may include a sterile package such as a glass or
plastic container, flexible foil or plastic pouches, sealed boxes
(particularly waxed boxes), and the like. In the case of sealed
packages, the gas-enriched fluid may maintain a high level of
dissolved or diffused gas for several days, several weeks, or
several months. In certain embodiments, the sealed container (i.e.,
enclosed with a cap, cover or other enclosure that is at least
semi-impermeable to gas exchange) maintains the diffused nature of
the fluid at least 2 weeks, at least 4 weeks, at least 2 months, at
least 4 months, at least 6 months, at least 8 months, at least 10
months, at least 12 months, or any value greater or
therebetween.
Gas-Enriched Fluids in Biological or Chemical Reactors
[0253] As illustrated in FIGS. 45A and 45B, a biological or
chemical reactor system 3300a may be used for conventional
large-scale cell-culturing or chemical processing to achieve the
production of the target product 3318. The target product 3318 may
include, but not be limited to, proteins, antibodies, synthetic
peptides, active pharmaceutical agents, foodstuff or beverage
products (such as wine; beer; soft drinks; fruit or vegetable
juices); plant products (flowers, cotton, tobacco, wood, paper,
wood or paper pulp, etc.); ethanol, methanol, paints, fruit or
vegetables or fruit or vegetable products such as jellies, jams,
sauces, pastes, and the like; cheese or cheese products; nuts or
nut products (such as peanut butter, almond paste, etc.); meat or
meat products, grain flours or products including bread, cereal,
pasta, and the like; slurries or mixtures of any of these,
processed polymers (including plastics, and other polymers),
petroleum products, and others.
[0254] In certain embodiments, in the case of using a vessel
reactor, such as a tank reactor, the target product resides within
inclusion bodies, particularly when E.coli cells are utilized. The
target product may be obtained by processing the inclusion bodies,
for example by using high-pressure homogenizers or other
techniques.
[0255] In particular embodiments in which the reactor is a
biological reactor system, the system 3306 includes a source 3308
of culture cells 3310 to be cultured, a source 3302 of culture
media 3304, a biological reactor 3306, and a harvesting and
purification system 3316, for producing the target product 3318.
The culture cells 3310 are genetically predetermined to produce
proteins or the like that constitute the target product 3318, and
the culture medium 3304 may comprise a sterile medium of a type
that provides nourishment for the proliferation of culture cells
3310. In this particular exemplary embodiment, the sterile medium
3304 is introduced into the internal chamber (which may be referred
to as the "fermentation chamber") of the reactor 3306 from the
source 3302. From the source 3308, the culture cells 3310 are
provided such that the cells 3310 and medium 3304 are combined into
a broth 3312 in the fermentation chamber of the bioreactor
3306.
[0256] The appropriate base medium 3304 to be utilized in the
reactor system 3300a may be formulated to provide optimal
nourishment and growth to the cell culture 3310. Medium 3304 is
preferably a fluid (e.g., liquid or gas) medium, more preferably a
liquid medium or a solid-liquid medium that is selected based on
the certain variables, such as the characteristics and objectives
of the overall bioreactor system 3300a, the cost, the type of cells
being cultured from the cell culture 3310, the desired production
parameters, the type of culturing and media management process used
in the reactor3306, the type of downstream harvest and purification
processes 3316, and the target active pharmaceutical ingredient
3318. Various cell culture media presently used may be adapted for
use or gas-enrichment by the present invention.
[0257] In certain embodiments, a suitable base medium 3304 may
include but not be limited to a serum-supplemented medium, a
hydrolysate medium, chemically-synthesized medium,
chemically-defined medium, a serum-free medium, any combination of
these or other media.
[0258] In certain embodiments, the gas-enriched media may be
supplemented with transferrins, albumins, fetuins, protein
hydrolysates, or other additives, preservatives, nutrients,
fillers, shear protectants (such as Pluronic F68), or active or
inactive agents.
[0259] In addition, the medium may be formulated for cells that are
attached to some type of support within the bioreactor 3306, rather
than suspended in the broth 3312. In all embodiments that utilize a
medium 3304, the medium 3304 is formulated to meet the nutritional
requirements of the individual cell type in the cell culture 3310,
and typically comprise minerals, salts, and sugars.
[0260] In certain embodiments, medium 3304 and/or broth 3312 are
gas-enriched using the presently disclosed mixing devices 100, in
order to dissolve or diffuse gases (such as oxygen) into, for
example, the media, both or components thereof, in concentrations
of at least about 8 ppm, at least about 10 ppm, at least about 20
ppm, at least about 25 ppm, at least about 30 ppm, at least about
35 ppm, at least about 40 ppm, at least about 50 ppm, at least
about 60 ppm, at least about 70 ppm, at least about 80 ppm, at
least about 90 ppm, at least about 100 ppm, or any value greater or
therebetween. In certain embodiments, the gas-enriched medium
and/or broth contains less than about 160 ppm.
[0261] In certain embodiments, the typical biological or chemical
reactor is loaded with sterilized raw materials (nutrients,
reactants, etc.) along with air or specific gas, as well as cells
for a biological reactor. Other agents may be added to the mixture,
including anti-foaming chemicals or agents, pH controlling agents,
and/or other agents. The target product is typically recovered by
separating the cells, and/or disrupting the cells in order to
extract the product, concentrating the product, and purifying,
drying, or further processing the product.
[0262] Many different types of bioreactor systems are in use today,
any of which can be used with the gas-enriched media of the present
invention. For example, air-lift bioreactors are commonly used with
bacteria, yeast and other fungi; fluidized-bed bioreactors are
commonly used with immobilized bacteria, yeast and other fungi, as
well as activated sludge; microcarrier bioreactors are commonly
used with mammalian cells immobilized on solid particles; surface
tissue propagators are commonly used with mammalian cells, tissue
grown on solid surfaces, and tissue engineering; membrane
bioreactors, hollow fibers and roto-fermentors are typically used
with bacteria, yeast, mammalian cells, and plant cells; modified
stir-tank bioreactors are commonly used with immobilized bacteria,
yeast, and plant cells; modified packed-bed bioreactors are
commonly used with immobilized bacteria, yeast, and other fungi;
tower and loop bioreactors are commonly used with bacteria and
yeast; vacuum and cyclone bioreactors are commonly used with
bacteria, yeast, and fungi; and photochemical bioreactors are
commonly used with photosynthetic bacteria, algae, cyanobacteria,
plant cell culture, and/or DNA plant cells.
[0263] Since living cells, including bacteria, yeast, plant cells,
mammalian cells, and fungal cells require molecular oxygen as an
electron acceptor in the bioxidation of substrates (such as sugars,
fats, and proteins), cell culture media that is highly oxygenated
is beneficial to the living cells. In a standard
oxidation-reduction reaction, glucose is oxidized to make carbon
dioxide, while oxygen is reduced to make water. Molecular oxygen
accepts all of the electrons released from the substrates during
aerobic metabolism. Thus, in order to provide the maximum amount of
bio-available oxygen to the growing cells, it is necessary to
ensure that the oxygen transfer from the air bubbles (gas phase) to
the liquid phase occurs quickly. When no oxygen accumulates in the
liquid phase, the rate of the oxygen transfer is the same as the
rate of the oxygen uptake by the growing cells.
[0264] The oxygen requirements of microorganisms is defined as a
standard formula, that is in units of QO.sub.2. Where QO.sub.2 is
the mass of oxygen consumed divided by the unit weight of dry
biomass cells in the bioreactor multiplied by time. Conversely, the
rate of accumulation of oxygen is equal to the net rate of oxygen
supply from air bubbles minus the rate of oxygen consumption by
cells.
[0265] In addition to a multitude of bioreactor types, each
bioreactor may utilize a particular impeller type or types, such as
marine-type propellers, flat-blade turbines, disk flat-blade
turbines, curved-blade turbines, pitched-blade turbines, paddles,
and shrouded turbines. The impeller or turbine may create a vortex
pattern of mixing in the bioreactor, or a uniform mixing.
[0266] In certain embodiments, the gas-enriched fluid of the
present invention relates to a sustained bio-availability of the
gas such that a gradual release of the gas occurs over time. This
gradual or "time" release is beneficial to the vehicles, such as
cultured cells, particularly when the gas released from the
gas-enriched fluid comprises oxygen. Thus, fermentation, or the
biochemical synthesis of organic compounds by cells 3310, typically
involve a relatively fast growth phase, facilitated by the
concentrations of diffused or dissolved gas in the broth 3312, as
well as by temperature control and by mixing the medium 3304 and
the cell culture 3310 in the fermentation chamber of the bioreactor
3306. Particular exemplary embodiments are depicted in the figures,
but may include additional components or tanks. Mixing may be
enhanced by rotating vanes or the like within bioreactor 3306, and
by reintroduction of fresh and/or freshly re-diffused supplies of
medium 3304 from any of the lines 3332, 3338, or 3334, as described
herein inter alia.
[0267] In one particular exemplary embodiment depicted in FIG. 45A,
the enrichment processing of the medium and/or broth to introduce
the gas (e.g., oxygen) in a cell culture medium may occur at
various points in the system. For example, the medium may be
gas-enriched prior to introducing the medium 3304 into the system
3300a at source 3302, or after such introduction at one or more
locations "A," "B," "C," "D," "E," or combinations thereof.
Gas-enriched fluid that may be introduced at the source 3302,
whether enriched at the site of the bioreactor or at a separate
location. If the gas-enriched fluid is enriched at a separate
location, it may be transported to the source 3302 in appropriate
containers or plumbing.
[0268] In certain embodiments, each of the locations "A," "B," and
"C," of FIG. 45A represent alternative locations for introduction
of a gas-enrichment diffuser device 1--within the bioreactor system
3300A. In the event that the introduction occurs at point "A," the
flow of medium from tank 3304 through the upper section of 3332A of
3332, the medium may be directed through the gas-enrichment
mixer/diffuser device 100 located at position "A," and medium 3304
with dissolved gases therein proceeds from the mixer/diffuser
device 100 through 3332B and into the fermentation chamber of the
bioreactor 3306.
[0269] With reference to FIG. 45B, the medium 3304 from tank 3302
may be directed through line 3332A into a pump 3410 and,
subsequently into the host material input of the gas-enrichment
diffuser device 100. The pump 3410, is preferably a variable speed
pump, which may be controlled by a controller 3390, based, in part,
on pressure readings detected by pressure sensor 3415. While
certain embodiments will utilize manual gauges as pressure
detectors, from which an operator may manually adjust the speed of
the pump 3410, and other components of the system 3300a or 3300b,
controller 3390 preferably receives an electrical signal from
sensor 3415 such that controller 3390 will automatically adjust the
speed of pump 3410. As will be evident from further descriptions
herein, the speed of pump 3410 may also be based on algorithms
within the controller 3390 which depend, in part, on the state of
other components of the system 3400 (such as valves 3420, 3421, and
sensors 3425).
[0270] Alternatively, the gas-enrichment mixer/diffuser device 100
may be positioned at location "B" such that the medium 3304 is
processed together with medium 3310. In this particular case, cells
3310 and medium 3304 are mixed in flow using a conventional mixing
nozzle and subsequently introduced into the mixer/diffuser device
100, where beneficial gases are infused into the mixed liquid of
medium 3304 and cells 3310. The resulting gas-enriched medium is
then directed into the fermentation tank of the bioreactor
3306.
[0271] As shown in FIG. 45A, cells 3310 may be combined with medium
3304, and following fermentation and/or development of the target
product, the contents of the bioreactor 3306 may then be directed
through line 3336 to a harvesting and purification stage. Once
purified, the target product is directed through line 3339 to a
target production tank 3318.
[0272] With reference to FIG. 45B, in certain embodiments, the
gas-enrichment mixer/diffuser device 100 combines the flow of a
medium 3304 with a flow of a gas from line 3426. Preferably, the
gas to be combined with medium 3304 flows from an oxygen tank 3450
and is metered by a valve 3420, which is controlled by controller
3390.
[0273] In certain embodiments, the gas-enrichment mixer/diffuser
device 100 is directed through line 3332b directly into the
fermentation tank by a reactor 3306. Alternatively, the
gas-enriched fluid may be directed through line 3332b to another
blending.
[0274] With reference to FIG. 45A, a bioreactor system 3300a, may
include an additional system (such as a perfusion system) 3314 that
begins processing the broth from the bioreactor 3306. During the
perfusion process 3314, the medium 3304 is continuously added to
the broth 3312 to nourish the cell culture 3310, which is then
mixed throughout the broth 3312. Simultaneously, cell or other
waste may be continuously removed from the broth 3312, typically at
the same rate as new medium 3304 is added. As indicated herein
above, gas-enrichment may also occur at positions "D" or "E," or at
both positions "D" and "E."
[0275] The perfusion system can allow for removal of cellular waste
and debris, as well as the target product, while retaining the
cells in the bioreactor 3306. The perfusion system thus reduces
waste accumulation and nutrient fluctuations, thereby allowing for
higher cell density and productivity. Retention of the cells in the
bioreactor may be achieved through various methods, including
centrifugation, internal or external spin filters, hollow fiber
modules, cross-flow filtration, depth filtration, any combination
of these or other means. In other embodiments, the accumulation of
waste products may be regulated by use of a glutamine synthetase
expression system.
[0276] With reference to FIG. 46, particular exemplary embodiments
utilize multiple gas sources 3502 and 3504 as shown, such that the
nature of the gas being diffused into the broth 3312 can be changed
depending on the stage of fermentation within the bioreactor 3306.
Hence, in a preferred embodiment, the cell culture medium is
enriched with oxygen during the proliferative phase of
fermentation. Subsequently, carbon dioxide, nitrous oxide, or
another gas may be substituted to facilitate other stages of the
fermentation process, particularly with processes that vary from
aerobic to anaerobic.
[0277] The bioreactor may comprise an airlift reactor, a packed bed
reactor, a fibrous bed reactor, a membrane reactor, a two-chamber
reactor, a stirred-tank reactor, a hollow-fiber reactor, or other
reactor designed to support suspended or immobilized cell
growth.
[0278] In one particular embodiment, the bioreactor 3306 is a
continuous stirred-tank reactor, comprising heat exchange and
refrigeration capabilities, sensors, controllers, and/or a control
system to monitor and control the environmental conditions within
the fermentation chamber. Monitored and controlled conditions may
include gas (e.g. air, oxygen, nitrogen, carbon dioxide, nitrous
oxide, nitric oxide, sulfur gas, carbon monoxide, hydrogen, argon,
helium, flow rates, temperature, pH, dissolved oxygen levels,
agitation speed, circulation rate, and others. Additionally, the
bioreactor 3306 may further comprise Cleaning-in-Place (CIP) or
Sterilization-in-Place (SIP) systems, which may be cleaned and/or
sterilized without assembly or disassembly of the units.
[0279] In one particular embodiment, the bioreactor 3306 performs a
continuous fermentation cycle, continuously adding medium 3304 to
the fermentation system with a balancing withdrawal, or harvest, of
the broth 3312 for target product extraction.
[0280] In alternate embodiments, the bioreactor 3306 may perform
batch fermentation cycles, fed-batch fermentation cycles, or
fed-batch fermentation cycles with the gas-enriched fluids.
Typically, batch fermentation cycles--in which all of the reactants
are loaded simultaneously--are used for small scale operations or
for the manufacture of expensive products or for processes that may
be difficult to convert into continuous operations. In a typical
process, the broth is fermented for a defined period to completion,
without further additions of the medium. The concentration varies
with time, but is typically uniform at any one particular time
point. Agitation serves to mix separate feed lines as well as
enhance heat transfer.
[0281] For batch fermentation, typically the total mass of each
batch is fixed, each batch is a closed system, and the reaction or
residence time for all reactants of the medium is the same. After
discharging the batch, the fermentation chamber is cleaned and
re-started with the medium 3304 for another batch cycle. Separation
or purification of the desired product from the other constituents
in the harvest broth 3312, may include further processing,
including refolding, altering affinity, ion exchange purification,
alteration of hydrophobic interactions, gel filtration
chromatography, ultra filtration and/or diafiltration, depending on
the target product.
[0282] For fed-batch fermentation, typically an initial, partial
charge or aliquot of medium 3304 is added to the fermentation
chamber, and subsequently inoculated with cell culture 3304. The
medium 3304 may be added at measured rates during the remainder of
the fermentation cycle. The cell mass and the broth 3312 are
typically harvested only at the end of the cycle.
[0283] Following harvest and purification of the target product
(step 3316), (typically once the cell culture 3310 has attained a
peak cell growth density within the bioreactor 3306), the purified
product 3318 (in some cases, a pharmaceutical drug or Active
Pharmaceutical Ingredient, or API) is attained. The purified
product may then be processed as desired and optionally packaged in
appropriate containers during a sterile packaging process 3322 for
transfer to a pharmaceutical manufacturing plant, or other
facility. The purified product may then be used for any desired
purpose, including for prevention, treatment, and/or diagnosis of
disease.
Plants and Animals as Reactors
[0284] In addition, a reactor may include a plant or animal, which
is used to generate a plant or animal product, or recombinant
product. In certain embodiments, the plant or animal target product
may be a naturally occurring product (e.g., food bearing crops or
meat, or textile-related products such as cotton fibers, etc.), or
the target product may be a genetically altered product (for
example, therapeutic agents, such human growth hormone or insulin
or other biologically active proteins and polypeptides). A
genetically altered or recombinant product may be produced by a
transgenic or genetically altered plant, animal, or combination
thereof.
Fish Culture
[0285] Fish (e.g., Tilapia fish) may be grown in aquaculture for
food, or as a transgenic vehicle for production of a target
product. The preferred temperature range for optimum tilapia growth
is 82.degree. -86.degree. F. Growth diminishes significantly at
temperatures below 68.degree. F. and death will typically occur
below 50.degree. F. Also, at temperatures below about 54.degree.
F., the immune resistance of tilapia declines and the animals are
easily subjected to infection by bacteria, fungi, and
parasites.
[0286] Twenty years ago, aquaculture researchers in Nigeria
attempted to correlate dissolved oxygen concentrations in pond
waiter with Tilapia growth rates. UN FAO reports: The study was
conducted by examining growth rates of young Tilapia at high
dissolved oxygen levels (approximately 7.0 ppm); at mid-level DO
(approximately 3.5 ppm); and at low DO levels (less than 2 ppm).
The growth rates were determined by measuring the weight of the
fish. The final increase in weight at the end of the research was
19 grams for the high DO level fish; 5 grams for the mid-level DO
fish; and 1.5 g for the low DO level fish. This represents to a 74%
and 92% reduction in growth rates correlating to the DO levels.
Thus, as the DO levels decrease, the feeding and waste output also
decrease. It was observed that the Tilapia in the low DO level
water break the surface of the water in order to access ambient
oxygen required for survival.
[0287] The gas-enriched fluids of the present invention further
include oxygenated freshwater supplies in which the high dissolved
oxygen levels in the water are maintained for extended periods of
time. According to particular aspects, using the diffuser device of
the present invention in an aquaculture setting, dissolved oxygen
levels of over 35 ppm can be recorded in 103.degree. F. water
without significantly stressing the aquatic life.
Plant Growth
[0288] In addition to animal growth, the gas-enriched fluids of the
present invention may be utilized for plant growth and development.
Gases (such as oxygen) are required for plant root respiration,
which allows for the release of energy for growth, as well as water
and mineral uptake. Plant growth has been widely and unequivocally
proven to be boosted by maintaining high gas (e.g., oxygen and/or
nitrogen) levels within the root zone. In this regard, increasing
gas delivery to plant root systems represents a potential for crop
improvement through boosting root activity. Likewise, in
embodiments in which transgenic plants are grown, increasing gas
delivery to the plants may provide for increased production of the
target product (such as a therapeutic or biopharmaceutical
product).
[0289] Hydroponic crops represent one exemplary system for
production which may greatly benefit from the gas-enrichment
diffuser devices of the present invention through direct
gas-enrichment (e.g., oxygenation) of the nutrient solution bathing
the root zone. Hydroponic crops are typically produced in a limited
volume of growing media or root area and as such need constant
replacement of gases (e.g., oxygen) within the root zone.
Hydroponic crops such as lettuce, spinach, tomatoes, and cucumbers
have already demonstrated a direct and significant response to the
gas-enriched nutrient solution. Some of these responses include
increases in plant growth, increases in root volume, increases in
plant yield, and higher quality produce. Thus, hydroponic systems
may benefit from the gas-enriched fluids of the present
invention.
[0290] Other hydroponic crops have had similar responses to
gas-enrichment in the root zone. However, at warm temperatures,
crop production declines due to the increased requirement for gases
(such as oxygen) in the root zone. Thus, enrichment is effective
for preventing gas-starvation of root cells, as well as boosting
growth under less than favorable growing conditions.
[0291] Typically tropical crops that are able to be grown at high
densities due to high light levels and rapid rates of development
(and high root zone temperatures) have a gas requirement that is
many times greater than those grown in more temperate climates.
Thus, gas-enrichment will become necessary in many systems of
horticulture production. Highly populated countries, which rely
heavily on producing intensive horticultural crops for income and
sustenance from very limited areas of land, will benefit greatly
from this technology.
[0292] Soil-based cropping systems can also benefit from the
gas-enriched solutions of the present invention. Many crops are fed
via drip, trickle, or furrow irrigation and could potentially
benefit greatly from the use of gas-enriched irrigation water or
fertigation solutions. Such crops include, but are not limited to:
vegetables (tomatoes, salad crops such as lettuce, herbs,
cucurbits), cut flowers, ornamental flowers, turf, vineyards,
orchards, and long-term plantings. Gases, such as oxygen, can
directly impact the health and growth of the plant but can also act
indirectly by increasing the bio-availability of gases (e.g.,
oxygen) at the root zone, and can also improve the health of the
plant by promoting microbial life in the soil.
[0293] With regard to the microbial life in the soil, the microbial
populations are essential for mineral conversion in the soil and
organic systems and overall plant health through suppression of
plant diseases. While these microbes are beneficial and often
essential for crop production, the populations also require gases
(e.g., oxygen), which can compete with the gases for plant root
cells. Thus, supplying gases (e.g., oxygen) to the plant roots in
order to enable microbial life to flourish is vital to both
organically grown crops, as well as standard growing conditions.
High rates of gases supplied to the growing media/soil in organic
systems would potentially speed up the rate of organic fertilizer
conversion and mineralization of plant usable nutrients, thus
increasing the health and productivity of highly profitable organic
crops.
[0294] In addition, the available land for growing crops represents
a challenge in many countries with limited resources or unsuitable
soils.
[0295] In addition to hydroponic crops, the technology disclosed
herein may apply to seed germination, seed raising, cell transplant
production, propagation from cuttings, sprout production, animal
fodder production, soil based cropping, turf industries, ornamental
plants, and medicinal plants.
Systems for Making Gas-Enriched Fluids
[0296] As shown here, exemplary oxygenation systems comprises a
supply or reservoir of fluid which is drawn up and circulated
through tubing or other conduits by a pump which subsequently
delivers the fluid to the mixer/diffuser. The mixer/diffuser may be
of any number of various embodiments including those set forth and
described herein above. These diffusers significantly increase the
amount of dissolved gas (e.g., oxygen) present in a fluid by
introducing, for example, gaseous oxygen to the fluid using a
diffuser having coaxial cylindrical or frusto conical stator and
rotor components rotating discs or plates within a housing, Mazzie
diffusers and impellers to create the desired cavitation and
succussion desired for mixing of the fluid and the gas. It should
be noted that many of the fluids will be aqueous or water-based,
but that the present invention is not limited to these.
[0297] The diffuser is supplied with fluid by the pump and combines
this with, for example, gaseous oxygen from supply and returns the
oxygenated (or otherwise gas-enriched) fluid to the reservoir. The
diffuser may employ any number of possible embodiments for
achieving diffusion including, but not limited to, micro-membrane,
Mazzie injector, fine bubble, vortexing, electromolecular
activation, or other methods. The oxygen supply may be either a
cylinder of compressed oxygen gas or a system for generating oxygen
gas from the air or other chemical components. The oxygenated fluid
produced by the diffuser is returned to the reservoir and may be
recirculated through the pump and/or the diffuser again to further
increase the dissolved oxygen content. Alternatively, the fluid may
be drawn off using the oxygenated fluid outlet. Oxygenated fluids
which are drawn off through the outlet may be immediately put to
use in various applications or may be packaged for later use.
[0298] The packaging step may enclose gas-enriched (e.g.,
oxygenated) fluids in a variety of bottles, bags or other
containers formed of plastic, metal, glass, or other suitable
materials. Although the gas-enriched or oxygenated fluids produced
in accordance with the present invention have a relatively long
shelf life under atmospheric conditions, the shelf life may be
further extended by using packaging which hermetically seals the
gas-enriched fluid. In this manner, dissolved oxygen which works
its way out of the fluid during storage will form a pressure head
above the gas-enriched fluid and help to prevent the migration of
dissolved oxygen, or other gas, out of the fluid and back into the
atmosphere. In one preferred embodiment of the present invention
the gas-enriched fluid is packaged in an air tight container and
the void space is filled with the gas used for enrichment at a
pressure of greater than one atmosphere prior to sealing the
container. The packaging step may be used to produce bottles, bags,
pouches, or other suitable containers for holding oxygenated
solutions.
[0299] The presently disclosed systems and/or methods allow oxygen,
or other gases, to be dissolved stably at a high concentration with
minimal passive loss. These systems and/or methods can be
effectively used to dissolve a wide variety of gases at heightened
percentages into a wide variety of fluids. By way of example only,
a deionized water at room temperature that typically has levels of
about 7-9 ppm (parts per million) of dissolved oxygen can achieve
levels of dissolved oxygen ranging from about 8-70 ppm using the
disclosed systems and/or methods. In accordance with a particular
exemplary embodiment, an oxygenated water or saline solution may be
generated with levels of about 30-60 ppm of dissolved oxygen.
Culturing Chinese Hamster Ovary Cells
[0300] Chinese Hamster Ovary (CHO) cells are mammalian cells that
are frequently utilized in expression and production of recombinant
proteins, particularly for those that require post-translational
modification to express full biological function.
[0301] According to particular aspects, various characteristics of
CHO cells can be improved by integrating either a gas-enriching
diffuser device 100 or gas-enriched media produced by the device
100 and integrated into a CHO bioreactor.
[0302] According to particular aspects, in the cultivation of CHO
cells, it is possible to utilize the gas-enriched fluids or media
of the present invention including with a cell-line specific,
serum-free medium (for example from SAFC Biosciences, Inc.) for
long-term growth of transformed CHO cells. According to additional
aspects, CHO cells are not harmed by passing through the
gas-enrichment diffuser device in the process of gas-enriching
fluids (including media).
[0303] A test was conducted that measured the survival of CHO cells
in an inline bioreactor. Briefly, the inline bioreactor was used
with 2 L of CHO media, and CHO cells at a density of 10.sup.6 or
higher. The bioreactor was run for approximately 10 minutes
(including the gas-enriching diffuser), and a 25 mL sample was
removed. Cells were stained with 0.4% Trypan Blue, and cell
viability was assessed with a hemacytometer. According to this
measure, CHO cells were not significantly harmed by passing through
the gas-enrichment diffuser device in the process of gas-enriching
fluids (including media).
[0304] The foregoing described embodiments depict different
components contained within, or connected with, different other
components. It is to be understood that such depicted architectures
are merely exemplary, and that in fact many other architectures can
be implemented which achieve the same functionality. In a
conceptual sense, any arrangement of components to achieve the same
functionality is effectively "associated" such that the desired
functionality is achieved. Hence, any two components herein
combined to achieve a particular functionality can be seen as
"associated with" each other such that the desired functionality is
achieved, irrespective of architectures or intermedial components.
Likewise, any two components so associated can also be viewed as
being "operably connected," or "operably coupled," to each other to
achieve the desired functionality.
[0305] While particular embodiments of the present invention have
been shown and described, it will be obvious to those skilled in
the art that, based upon the teachings herein, changes and
modifications may be made without departing from this invention and
its broader aspects and, therefore, the appended claims are to
encompass within their scope all such changes and modifications as
are within the true spirit and scope of this invention.
Furthermore, it is to be understood that the invention is solely
defined by the appended claims. It will be understood by those
within the art that, in general, terms used herein, and especially
in the appended claims (e.g., bodies of the appended claims) are
generally intended as "open" terms (e.g., the term "including"
should be interpreted as "including but not limited to," the term
"having" should be interpreted as "having at least," the term
"includes" should be interpreted as "includes but is not limited
to," etc.). It will be further understood by those within the art
that if a specific number of an introduced claim recitation is
intended, such an intent will be explicitly recited in the claim,
and in the absence of such recitation no such intent is present.
For example, as an aid to understanding, the following appended
claims may contain usage of the introductory phrases "at least one"
and "one or more" to introduce claim recitations.
[0306] However, the use of such phrases should not be construed to
imply that the introduction of a claim recitation by the indefinite
articles "a" or "an" limits any particular claim containing such
introduced claim recitation to inventions containing only one such
recitation, even when the same claim includes the introductory
phrases "one or more" or "at least one" and indefinite articles
such as "a" or "an" (e.g., "a" and/or "an" should typically be
interpreted to mean "at least one" or "one or more"). The same
holds true for the use of definite articles used to introduce claim
recitations. In addition, even if a specific number of an
introduced claim recitation is explicitly recited, those skilled in
the art will recognize that such recitation should typically be
interpreted to mean at least the recited number (e.g., the bare
recitation of "two recitations," without other modifiers, typically
means at least two recitations, or two or more recitations).
[0307] Accordingly, the invention is not limited except as by the
appended claims.
Sequence CWU 1
1
1120DNAArtificial SequenceOligonucleotide 1taatacgact cactataggg
20
* * * * *
References