DEVICES, SYSTEMS, AND METHODS FOR REMOVAL OF SOLUBLE GASES FROM FLUID SAMPLES

Abstract

Devices, systems and methods are disclosed which relate to using containers with a multitude of nucleation sites covering a major portion of the inside wall of the container to enable rapid and nearly complete removal of soluble gases from fluid samples, including carbonated beverages and other carbonated fluid samples. A fluid sample is rapidly poured into the described container initiating a catastrophic release of the soluble gas from the sample.

Claims

1. A system for degassing a fluid sample, the system comprising a container with a plurality of nucleation sites for exposing to the fluid sample, wherein a total area of the nucleation sites is at least 5% of a total surface area of contact of the fluid sample with the container.

2. The system of claim 1, wherein the fluid sample contains carbon dioxide, nitrogen, nitrous oxide, oxygen or other soluble gases or mixtures of soluble gases.

3. The system of claim 1, wherein the fluid sample is one of a carbonated beverage, beer, cider, sparkling wine, champagne, wine cooler, alcoholic beverage, juice, lemonade, coffee, soft drink, coke, fizzy drink, fizzy juice, cool drink, cold drink, lolly water, pop, seltzer, soda, soda pop, fountain drink, ginger ale, ginger beer, tonic water, mineral water, or other carbonated beverage.

4. The system of claim 1, wherein the container is made from one of glass, Styrofoam, ceramic, metal, plastic, thermoplastic, polytetrafluoroethylene or other material routinely used for producing bottles or sample tubes for laboratory use.

5. The system of claim 1, wherein the nucleation sites are produced during molding of the container, by using a mold containing a mirror image of the nucleation sites.

6. The system of claim 1, where in the nucleation sites are produced after molding of the container by sandblasting, laser etching, machining, acid etching or other chemical or mechanical means of creating nucleation sites on the surface.

7. The system of claim 6, wherein the sandblasting is performed using aluminum oxide, silicon carbide, crushed glass, glass beads, plastic abrasive, pumice, steel shot, steel grit, corn cob, walnut shell or garnet with a grit ranging from 8 to 1,000.

8. The system of claim 1, wherein a lid with open holes or holes covered with a membrane filter material is used to allow gas to escape during degassing.

9. A method for degassing a fluid, the method comprising: pouring a fluid sample into a container with a plurality of nucleation sites, wherein a total area of the nucleation sites is at least 5% of a total surface area of contact of the fluid sample with the container; and incubating the fluid in the container for a sufficient time to allow for degassing.

10. The method of claim 9, wherein the fluid sample is quickly poured into the container to agitate the fluid sample and increase the rate of degassing.

11. The method of claim 9, wherein the fluid sample is first heated to increase the rate of degassing.

12. The method of claim 9, wherein the fluid is cooled during or after degassing to increase the solubility of the gas in the fluid.

13. A device for degassing a fluid sample, the device comprising a container with a plurality of nucleation sites for exposure to the fluid sample, wherein a total area of the nucleation sites is at least 5% of a total surface area of contact of the fluid sample with the container.

14. The device of claim 13, wherein the container is made from one of glass, Styrofoam, ceramic, metal, plastic, thermoplastic, polytetrafluoroethylene or other material routinely used for producing bottles or sample tubes for laboratory use.

15. The device of claim 13, where in the nucleation sites are produced during molding of the container, by using a mold containing a mirror image of the nucleation sites.

16. The device of claim 13, where in the nucleation sites are produced after molding of the container by sandblasting, laser etching, machining or other mechanical means of creating nucleation sites on the surface.

17. The device of claim 16, wherein the sandblasting is performed using aluminum oxide, silicon carbide, crushed glass, glass beads, plastic abrasive, pumice, steel shot, steel grit, corn cob, walnut shell or garnet with a grit ranging from 8 to 1,000.

18. The device of claim 13, wherein the nucleation sites are on a device or plurality of devices which are placed in the container.

19. The device of claim 13, wherein the container is of sufficient size to allow the fluid sample to foam while not overflowing the container.

20. The device of claim 13, wherein a lid with open holes or holes covered with a membrane filter material is used to allow gas to escape during degassing.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0019] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments of the subject disclosure and technical data supporting those embodiments, and together with the written description, serve to explain certain principles of the subject disclosure.

[0020] FIG. 1 shows the solubility of carbo dioxide in water.

[0021] FIG. 2 shows a decarbonation container, according to an exemplary embodiment of the present subject disclosure.

[0022] FIG. 3 shows a vented lid for a decarbonation container, according to an exemplary embodiment of the present subject disclosure.

[0023] FIG. 4 shows a nucleated PYREX measuring cup, according to an exemplary embodiment of the present subject disclosure.

[0024] FIG. 5 shows beer poured into a nucleated container, according to an exemplary embodiment of the present subject disclosure.

[0025] FIG. 6 shows flattened beer being concentrated, according to an exemplary embodiment of the present subject disclosure.

[0026] FIG. 7 shows a 12 ounce can of flattened beer filtered, according to an exemplary embodiment of the present subject disclosure.

[0027] FIG. 8 shows nucleation sites being created inside a container, according to an exemplary embodiment of the present subject disclosure.

[0028] FIG. 9 shows an internally nucleated container, according to an exemplary embodiment of the present subject disclosure.

[0029] FIG. 10 shows a Concentrating Pipette Instrument, according to an exemplary embodiment of the present subject disclosure.

DETAILED DESCRIPTION OF THE SUBJECT DISCLOSURE

[0030] The following detailed description references specific embodiments of the subject disclosure and accompanying figures, including the respective best modes for carrying out each embodiment. It shall be understood that these illustrations are by way of example and not by way of limitation.

[0031] The present subject disclosure describes highly efficient and simple to use devices, systems, and methods for removing saturated gasses, such as carbon dioxide, from liquid samples. The technique uses a large surface area of nucleation sites along with methods for enhancing the nucleation of the gasses to quickly and efficiently remove these gases from samples prior to implementing liquid concentration or other laboratory methods. Specifically, the present technique may be used to decarbonate beer and other carbonated liquids prior to processing on a Concentrating Pipette Instrument, for example those described and patented by the Applicant.

[0032] FIG. 2 shows a decarbonation container 100, according to an exemplary embodiment of the present subject disclosure. A large zone of nucleation sites 101, on the inside surface of the container, extends from near the neck at the top end to the bottom edge of the inside walls and includes the entire bottom inside surface as well. Thus, any fluid which is poured into container 100 will come into direct contact with the nucleation sites 101, which extend to nearly the entire interior surface of the container 100. The higher the percentage of inside walls of container 100 is covered by nucleation sites 101, the more effective the decarbonation of a fluid deposited therein. Typically, the nucleated interior surface is 5%-100% of the interior surface. Further, a standard bottle thread 102 is provided to allow for use of a threaded lid.

[0033] FIG. 3 shows a screw top lid 200 including a hydrophobic membrane filter vent 201 for use on container 100. The user pours a carbonated sample into container 100 and carbon dioxide is nucleated by nucleation surface 101. After pouring the sample into the container 100, the user may place lid 200 onto container 100 to reduce the potential for contamination of the liquid sample. Vent 201 allows gas to escape during the decarbonation process so that pressure does not build up within container 100.

[0034] Applicant has demonstrated sufficient rapid degassing of carbonated beverage samples by pouring them into glass or plastic containers that have had a major portion of the inside walls etched or “frosted” and which contain enough volume to hold the sample as it catastrophically foams up, evolving the majority of the carbon dioxide gas, and collapsing as flat liquid in the bottom of the container.

[0035] An exemplary demonstration of the process was accomplished by grit blasting a 2 Liter Pyrex Measuring Cup (see FIG. 4). The blasting process “frosted” the interior of the container, forming nucleation sites over the internal surface. When a can or bottle of beer was poured into the container, it immediately foamed up to the top, then rapidly collapsed back to a flat liquid with very little residual carbonation (FIG. 5). As is shown in FIGS. 6-7, the entire bottle or can is then able to be rapidly processed using the Concentrating Pipette Instrument. In the example shown, the CP-150 Concentrating Pipette Instrument was used. This Instrument is shown again in FIG. 10. As an example, a 12 ounce can of Busch Light beer was filtered in 7 minutes and 43 seconds.

[0036] Various other glass and plastic containers may be treated by interior grit blasting and have demonstrated to behave in the same manner. Such a grit blasting process is shown in FIG. 8. Various beers and ales, including but not limited to Angry Orchard hard cider, Boulevard unfiltered wheat, single wide IPA, Tank 7 Farmhouse Ale, and Pale Ale; Budweiser, Carlsberg, Coors Banquet, Miller Lite, Guinness Draught, Heineken, Modelo Especial, Samuel Adams Boston Lager, Sapporo, Shiner Bock, Stella Artois, and TsingTao were shown to be quickly decarbonated using the nucleated containers.

[0037] Nucleation sites may be created on the container in a number of ways including, but not limited to: by use of a mold during manufacturing that contains a reverse image of the nucleation sites, by use of a container material that is naturally rough or naturally contains many nucleation sites, or by post treatment of the container after initial manufacturing using any number of well-known processes for creating a rough surface. These processes include: sandblasting, bead blasting, laser etching, acid etching, or other mechanical or chemical means for roughening the surface or creating nucleation sites.

[0038] Molding may also be used to create a rough surface, wherein the nucleation sites are produced during molding of the container, by using a mold containing a mirror image of the nucleation sites.

[0039] Sandblasting may be performed using aluminum oxide, silicon carbide, crushed glass, glass beads, plastic abrasive, pumice, steel shot, steel grit, corn cob, walnut shell or garnet with a grit ranging from 8 to 1,000. Laser etching may be performed with any number of commercially available laser cutter/etcher systems. Plastic containers, especially, may be manufactured with a multitude of nucleation sites by using a mold that has be roughened using sandblasting or other methods.

[0040] The nucleation sites may be created in any number of types of materials including, but not limited to: glass, ceramic, metal, plastic, thermoplastic, polytetrafluoroethylene or other material routinely used for producing bottles or sample tubes for laboratory use. The container may be autoclavable, such that it can be washed and then autoclaved between samples to ensure that no cross contamination occurs between samples. Alternatively, the container may be single use and may be packaged and treated, prior to sale, using e-beam irradiation, gamma irradiation, ethylene oxide, vaporous hydrogen peroxide, peracetic acid or other commonly used sterilization processes.

[0041] The container is sized such that entire sample may be vigorously poured into the container and not overflow due to the decarbonation process and associated foaming of the sample. In the case of beer, most sample 350 mL cans or bottles of beer can be rapidly poured into a container of 1 L nominal volume without overflowing during the decarbonation process. Other samples types and volumes may require larger containers or may be able to be processed in smaller containers.

[0042] Applicants have proven the effectiveness of the present technique with various volumes. For example, from 100 mL of beer to 750 mL of sparkling wine. Degassing of up to 12 ounces (335 mL) of beer has been performed in 32 ounce straight sided jars (like the jars shown in FIGS. 2 and 9). A 750 mL wine sample was degassed in a 60 ounce sand blasted beer pitcher and was run on the Concentrating Pipette with a 0.4 um Concentrating Pipette Tip in 12 minutes and 35 seconds. The present technique is effective for a wide range of volumes, from 5 mL to 10 L. Further, different shapes of containers may also be used to enhance the decarbonation process through increased turbulence associated with different container diameters and shapes.

[0043] A large surface area with nucleation sites may also be produced by using a multitude of devices such as glass beads or a single device or several devices with a large surface area of nucleation sites which are placed into the container prior to pouring the sample.

[0044] The decarbonation process causes significant quantities of gas to be released and as such the process to some extent helps maintain sterility due to an outflow of carbon dioxide from the container. Further this outflow requires that an open container or some type of vent be used during the decarbonation process. For example, a loose-fitting lid may be used to eliminate the chance of a user touching the container opening while also constricting the open area enough to ensure only an outflow of carbon dioxide takes place. Further, lids with an integral membrane filter of a small enough pore size to ensure sterility may also be used to allow for an outflow of gas while helping to reducing the chance of contamination.

[0045] In addition to the decarbonation action performed by use of a sample container with a large surface area of nucleation sites, other steps may be taken to enhance the rate and completeness of the decarbonation process. These include vigorous pouring of the beer into the container, heating of the sample prior to pouring into the container, stirring or shaking of the sample while in the container, as well as other methods for enhancing mixing and contact with the nucleation sites. Further, the ability to process the sample following decarbonation may also be enhanced by cooling the sample during or after the decarbonation process to increase the solubility of the remaining carbon dioxide in the sample and thus make it more difficult to come out of solution during subsequent filtration or concentration processes.

[0046] Vigorous pouring of beer samples or other carbonated or gassed samples may entail turning the sample container—in the case of beer the bottle or can—nearly or completely upside down from a height of a few inches to one foot or more in height above the nucleated container and letting the entire sample flow into the container.

[0047] Heating of the sample includes any temperature from room temperature to up to 37° C. or more to enhance the release of carbon dioxide. In the case of beer tests were performed by putting full, unopened cans or bottles of beer in a 37° C. incubator for 30 minutes before pouring into the container. This significantly enhanced the carbon dioxide removal and reduced the total time period required for decarbonation and sample processing on the Concentrating Pipette.

[0048] Further, in addition to beer, other sample types such as cider, sparkling wine, champagne, wine coolers, other alcoholic beverages, juice, lemonade, coffee, soft drink, coke, fizzy drink, fizzy juice, cool drink, cold drink, lolly water, pop, seltzer, soda, soda pop, fountain drink, ginger ale, ginger beer, tonic water, mineral water, or other carbonated beverages may also be decarbonated using this method. Additionally, other fluid samples containing carbon dioxide, nitrogen, nitrous oxide, oxygen or other soluble gases or mixtures of soluble gases may also be degassed using these methods.

[0049] The more complete the removal of carbon dioxide from the beer or carbonated beverage the more quickly and completely the sample may be processed on the Concentrating Pipette. Without decarbonation of beer samples, most beer fouls the Concentrating Pipette Tip and causes the instrument to shut down within seconds. Provided below in Table 1 through Error! Reference source not found. are data for processing of a number of brands and styles of beer following decarbonation. Table 1 through Error! Reference source not found. show varying volumes of beer processed and varying run times for different types of beers and different types of Concentrating Pipette Tips. Error! Reference source not found. and Error! Reference source not found. provide a comparison between beer samples at room temperature and beer samples heated at 37° C. prior to processing.

[0050] Membrane filtration and concentration of biological particles including beer spoilage organisms spiked into Coors beer was then demonstrated using the Concentrating Pipette equipped with 0.45 micron hollow fiber pipette tips, P/N CC 08018. The first three test runs (test runs 1-3) are shown with a prototype next generation instrument with 3.5 psi backpressure and demonstrate an average of 693× concentration of the organism in an average of 3.6 minutes (see Error! Reference source not found.). Test runs 4-6 used the current generation instrument (CP-150) and test runs 7-9 used the next generation instrument (CP SELECT) without backpressure, with the difference being that the next generation instrument has improved valves, foam control, software, etc.

[0051] Optimization of the grit blasting (nucleating) process (FIG. 8) was undertaken using a commercially available autoclavable culture vessel (32 oz wide mouth glass jar with HEPA-vent lid, P/N C607, Phyto Technology Laboratories, Mission, Kans. USA). The vented lid allows the gas to escape during decarbonation without building up pressure in the jar. Aluminum Oxide sandblasting media and other media were screened for the ability to create an optimum surface for beer decarbonation, including but not limited to 150 grit blasted for 8 minutes which was selected for production of decarbonation vessels for commercial sale. Other values and time ranges are also possible and within the purview of the present disclosure.

[0052] The blasting media size may range from 8-1,000 grit, and more specifically would be limited to 18-500 grit. Above 500 grit is the “micro fine” area and less conducive to creation of nucleation sites. Testing times have ranged from 1 minute (minimum to lightly blast the jar) to 32 minutes. The 32 minutes involved several instances of sandblasting with a fine grit blasting media. As such, the process was to blast, test, and repeat. Beyond 30 minutes is where effectiveness starts to drop. An exemplary preferable material used is 150 grit aluminum oxide for 6-8 minutes at −100 psi. An internally nucleated jar is shown in FIG. 9.

[0053] In another exemplary embodiment, Styrofoam cups and containers are used. Styrofoam cups and containers inherently provide a vast number of nucleation sites due to their method of manufacture. Observation of such cups under a microscope clearly shows nucleation sites where cells border each other and where edges are created on the cells in the interstices between cells by injection into mold cavities.

[0054] Three samples were degassed in Styrofoam cups and processed on the Concentrating Pipette Instrument (CP-150, FIG. 10). The appearance of the degassing process was similar to that in a frosted glass container. After pouring the beer in the cups, the standard lids with either the slit aperture for inserting a drinking straw, or the coffee drinking lid with the small sipping hole, were applied to the cups. Due to gas evolution from the cups, these small gas openings are sufficient to allow the evolving gas to escape without blowing the lids off. The evolving gas may also protect the sample from contamination during the settling and degassing step. In this case, the samples were chilled for an hour or less. See Table 9 for results of beer degassing using commercially available Styrofoam cups.

[0055] Following the above exemplary treatments, the Applicant Concentrating Pipette and other Applicant instruments based on the above patents and pending patent application Ser. No. 14/313,618, 15/456,981, 15/431,655, and 14/058,193 are able to concentrate the initial sample into a volume of from less than 100 to 1000 milliliters or more in a matter of minutes.

[0056] It will be appreciated that the foregoing instrumentalities teach by way of example, and not by limitation. Accordingly, those skilled in the art understand that the subject matter is not limited to what is strictly disclosed, but also pertains to what is understood by those skilled in the art on the basis of the teachings herein. The inventors hereby state their subject matter to rely, as may be needed, upon the Doctrine of Equivalents to protect the fullness of their rights in what is claimed.

TABLE-US-00001 TABLE 1 Concentrating Pipette Run Times for Decarbonated Beer - Best Results (with 4° C. chilling) Average Time at Volume time 4° C. Number Beer Tip processed (min) (min) Can/bottle of runs Coors 0.4 um all (355 mL) 13.86 20 (as low both 16 Banquet flat as 3) Heineken 0.4 um all (355 mL) 17.26 20 Bottle 7 flat Stella 0.4 um all (355 mL) 8.82 20 Bottle 3 Artoise flat TsingTao 0.4 um all (355 mL) 8.7 20 Bottle 3 hf-CPT

TABLE-US-00002 TABLE 2 Concentrating Pipette Run Times for Decarbonated Beer - 2.sup.nd Best Results (with 4° C. chilling) Average Average Time at volume time 4° C. Number Beer Tip processed (min) (min) Can/bottle of runs Budweiser 0.4 um 265.33 mL 22.74 20 Can 3 flat Shiner Bock 0.4 um  259.9 mL 34.13 20 Bottle 2 flat Modelo 0.4 um 167.67 mL 10.96 20 Can 3 Especial flat Guinness 0.45 um 170.11 16.74 20 Both 4 Draught hf-CPT Sapporo 0.45 um 238.99 17.5 20 Can 1 hf-CPT

TABLE-US-00003 TABLE 3 Concentrating Pipette Run Times for Decarbonated Beer - 3.sup.rd Best Results (with 4° C. chilling) Avg volume Average Time processed time at 4° C. Number Beer Tip (mL) (min) (min) Can/bottle of runs Boulevard 0.4 um 147.68 10.81 20 Bottle 4 Single Wide flat IPA

TABLE-US-00004 TABLE 4 Concentrating Pipette Run Times for Decarbonated Beer - 4.sup.th Best Results (with 4° C. chilling) Average volume Average Time at processed time 4° C. Number Beer Tip (mL) (min) (min) Can/bottle of runs Samuel 0.4 um 153.83 29.79 20 Can 2 Adams flat Boston Lager Angry 0.4 um 127.70 31.44 20 Bottle 2 Orchard had flat cider Sapporo 0.4 um 127.27 19.33 20 Can 1 flat Boulevard 0.45 147.68 11.1 20 bottle 2 Pale Ale um hf- CPT

TABLE-US-00005 TABLE 5 Concentrating Pipette Run Times for Decarbonated Beer - 5.sup.th Best Results (with 4° C. chilling) Average Average Time at volume time 4° C. Number Beer Tip processed (min) (min) Can/bottle of runs Carlsberg 0.4 um 32.54 31.83 20 Can 2 flat Boulevard 0.4 um 78.40 11.71 20 Bottle 2 Pale Ale flat Boulevard 0.4 um 32.64 27.43 20 Bottle 1 Tank 7 flat Boulevard 0.45 um 109.11 27.58 20 Bottle 1 Tank 7 hf-CPT Boulevard 0.45 um 63.72 11.72 20 Bottle 1 unfiltered hf-CPT wheat

TABLE-US-00006 TABLE 6 Concentrating Pipette Run Times for Decarbonated Beer - Comparison −25° C. and 37° C. prior to decarbonation) (with 4° C. chilling) Temp Average Average prior to volume time Number Beer pouring processed (min) Tip type Can/bottle of runs Coors 25 355 13.86 0.4 um flat Can 16 Banquet Coors 37 355 6.53 0.4 um flat Can 3 Banquet Budweiser 25 265.33 22.74 0.4 um flat Can 3 Budweiser 37 452.76 16.51 0.4 um flat Can 3 Heineken 25 354.1 17.26 0.4 um flat Bottle 7 Heineken 37 349.82 10.8 0.4 um flat Bottle 1

TABLE-US-00007 TABLE 7 Concentrating Pipette Run Times for Decarbonated Beer - Comparison −25° C. and 37° C. prior to decarbonation) (with 4° C. chilling) Temp Average Average prior to volume time Number Beer pouring processed (min) Tip type Can/bottle of runs TsingTao 25 119.74 20.91 0.4 um flat Bottle 3 TsingTao 37 131.98 6.27 0.4 um flat Bottle 1 Stella Artois 25 328.53 8.81 0.4 um flat Bottle 3 Stellla Artois 37 324.96 6.22 0.4 um flat Bottle 1

TABLE-US-00008 TABLE 8 InnovaPrep Concentration of Beer Spoilage Organism from Decarbonated Beer L. brevis spiked Miller High Life Mar. 14, 2017 Runs 1 2 3 avg st dev 4 5 6 avg st dev time 3.00 4.00 3.82 3.6067 0.4352 6.70 4.82 4.17 5.2300 1.0728 Feed Titer (~1 CFU/mL) plated 100 196 196 uL/counts 187 187 average 164 164 182.3333 182.3333 dilution 1.0000 1.0000 total spike 182.3333 182.3333 feed/titer, CFU/mL 0.5136 0.5136 Concentrate tare 3.7157 3.7149 3.69 3.7033 3.7036 3.7017 net 3.9074 4.0485 4.0881 4.3726 4.4142 4.5058 volume 0.1917 0.3336 0.3981 0.3078 0.0862 0.6693 0.7106 0.8041 0.7280 0.0564 counts, CFUs 93 99 114 106 105 102 conctiter CFU/mL 485.1330 296.7626 286.3602 158.3744 147.7625 126.8499 % Efficiency 51.01% 54.30% 62.52% 55.94% 4.84% 58.14% 57.59% 55.94% 57.22% 0.93% concentration 944.5460 577.7919 557.5386 693.2922 177.8556 308.3524 287.6911 246.9747 281.0060 25.4993 factor for Extraction 2 tare 3.6966 3.7267 3.6951 3.6965 3.6936 3.716 net 3.9142 4.0673 3.9811 4.3905 4.4625 4.4615 volume 0.2176 0.3406 0.286 0.2814 0.0503 0.6940 0.7689 0.7455 0.7361 0.0313 counts, CFU 36 9 9 22 4 3 Efficiency 19.74% 4.94% 4.94% 9.87% 6.98% 12.07% 2.19% 1.65% 5.30% 4.79% total elution volume 0.4093 0.6742 0.6841 1.3633 1.4795 1.5496 total efficiency 70.75% 59.23% 67.46% 65.81% 4.84% 70.20% 59.78% 57.59% 62.52% 5.50% Runs 7 8 9 avg st dev time 4.13 3.75 3.73 3.8700 0.1840 Feed Titer (~1 CFU/mL) plated 100 196 uL/counts average 187 164 182.3333 dilution 1.0000 total spike 182.3333 feed/titer, CFU/mL 0.5136 Concentrate tare 3.7258 3.7249 3.688 net 4.1903 4.2205 4.0338 volume 0.4645 0.4956 0.3458 0.4353 0.0645 counts, CFUs 117 111 81 conctiter CFU/mL 251.8837 223.9709 234.2394 % Efficiency 64.17% 60.88% 44.42% 56.49% 8.64% concentration 490.4135 436.0677 456.0603 460.8472 22.4433 factor for Extraction 2 tare 3.6991 3.7065 3.6897 net 4.1750 4.1963 4.086 volume 0.4759 0.4898 0.3963 0.4540 0.0412 counts, CFU 10 6 2 Efficiency 5.48% 3.29% 1.10% 3.29% 1.79% total elution volume 0.9404 0.9854 0.7421 total efficiency 69.65% 64.17% 45.52% 59.78% 10.33%

TABLE-US-00009 TABLE 9 Beer Degassing Using Commercially Available Styrofoam Cups Chill 1st time Processing Processed Elution Sample Beer Degassing Vessel (min) Time (m:s) Sample (g) (uL) 1 Miller 32 Oz. Styrofoam Cup 40 7:03 277 380 High Life (Dart 32AJ20) 2 Coors 32 Oz. Styrofoam Cup 50 6:24 266 260 Banquet (Dart 32AJ20) 3 Miller 12 Oz. Styrofoam 60 3:38 118 350 High Life (Sweetheart X12 67240)