Abstract:
Buffer solutions within a plurality of containers can be adjusted and controlled by exposing the containers while open to a controlled environment. In particular, an inventory of containers holding bicarbonate buffers can be adjusted to a precisely controlled pH by exposing the containers while open to a controlled carbon dioxide environment in a treatment chamber.

Description:
CROSS-REFERENCES TO RELATED APPLICATIONS 
       [0001]    The present application claims the benefit of prior provisional application Ser. Nos. 61/270,571, filed on Jul. 9, 2009, and 61/276,137, filed on Sep. 8, 2009, the full disclosures of which are incorporated herein by reference. 
     
    
     BACKGROUND OF THE INVENTION 
       [0002]    1. Field of the Invention 
         [0003]    The present invention relates generally to methods and systems for pH adjustment and control. More specifically, the present invention relates to methods and systems for adjusting the pH of buffers and other medical solutions to achieve uniformity among pluralities of individual containers. 
         [0004]    A buffer is a substance that minimizes the change in pH of a solution when an acid or base is added to the solution. Medical buffer solutions frequently contain bicarbonate ions and are used in numerous medical applications including antidotes, dialysates, body replacement fluids, body irrigating solutions, cardiac perfusates, and for many other purposes. One of the most commonly used medical bicarbonate buffer solutions consists of sodium bicarbonate (NaHCO 3 ) mixed with water, which among other things, can be used to buffer parenteral injections to a more physiologic pH prior to injection. Of particular interest to the present application, bicarbonate solutions may be used to buffer acidic local anesthetic injections in order to enhance anesthetic effectiveness, reduce injection pain, and limit tissue trauma. For use of sodium bicarbonate for buffering local anesthetic, as well as for other medical uses including but not limited to treatment of acidosis, it is desirable that sodium bicarbonate solutions be precisely maintained at or near a particular known pH. Using a buffer solution having a known pH allows a medical practitioner to mix a predetermined ratio of the buffer solution with the parenteral solution in order to achieve precise control over the resulting pH of parenteral solution, which has significant benefits over using a parenteral solution where the pH is not precisely known. 
         [0005]    As an example, when compounding 8.4% sodium bicarbonate buffer solution with commercially available cartridges of 2% lidocaine with epinephrine 1:100,000, the pH of the buffer solution will tend to drive the pH of the combination, almost exclusively of the pH of the commercially available anesthetic cartridges, such that a relatively small volume of buffer solution will have a disproportionately large impact on the pH of the combined solution. Thus, to achieve a parenteral solution with a predictable pH, it is important that the pH of the buffer solution be precisely known and controlled. 
         [0006]    Commercially produced sodium bicarbonate buffers do not provide buffer packages with precisely controlled pH. For example, commercially available buffering solutions list a very large pH range on their labels, with sodium bicarbonate solutions typically labeled from pH 7.0 to pH 8.5. An assay of the commercially available sodium bicarbonate solutions performed by the inventors showed a pH range from 7.62 to 8.26 in a number of commercially obtained bicarbonate buffer cartridges. Presumably the actual range of the product available in the marketplace is even wider than the range shown in this assay. 
         [0007]    It must be appreciated, in this context, that a medical buffer having an actual pH of 7.0 may perform significantly differently than a medical buffer having an actual pH of 8.5. This is true whether the medical buffer is designed to buffer the pH of the body&#39;s fluids, for instance in the treatment of acidosis, or the medical buffer is designed to buffer the pH of a parenteral solution prior to its use. In the example where a practitioner uses sodium bicarbonate solution to buffer anesthetic to achieve physiologic pH, the ratio of buffer solution to anesthetic solution will be quite different when the pH of the bicarbonate solution is 7.0 compared to when the pH is 8.5. Thus, prior art methods of combining buffering solution with parenteral solutions which rely on adding the same ratio of buffering solution to the parenteral solution (regardless of the actual pH of the buffer) will not consistently arrive at the desired pH for the buffered parenteral. 
         [0008]    For these reasons, it would be desirable to provide packaged buffers with precisely controlled pH&#39;s so that the pH in each individual package is substantially identical and consistent. It would be further desirable to provide methods and systems for producing packaged buffer solutions which allow for the preparation of unlimited numbers of packages, such as cartridges, capsules, carpules, vials, and other containers, each of which contains a stable and precisely known pH. Such methods and systems should provide for the economical preparation of such buffer packages, allow for storage of such packages for prolonged periods while maintaining pH stability, and be compatible with sterilization protocols and systems. At least some of these objectives will be met by the inventions described below. 
         [0009]    2. Description of the Background Art 
         [0010]    Glass vials and cartridges for storing medical solutions are described in U.S. Pat. Nos. 1,757,809; 2,484,657; 4,259,956; 5,062,832; 5,137,528; 5,149,320; 5,226,901; 5,330,426; and 6,022,337. Injection pens which employ drug cartridges are described in U.S. Pat. No. 5,984,906. Exemplary disposable drug cartridge that could be loaded with buffer solution in accordance with the present invention are described in U.S. Pat. No. 5,603,695 and in commonly owned, copending application US2009/0292271 (U.S. Ser. No. 12/406,670), both of which are incorporated herein by reference. A device for delivering a buffering agent into an anesthetic cartridge using a transfer needle is described in U.S. Pat. No. 5,603,695. Devices for maintaining a dissolved gas in solution in a pouch are described in U.S. Pat. Nos. 5,690,215; 5,610,170; and 4,513,015, and U.S. Patent Publ. No. 2007/0265593. Other patents and applications of interest include U.S. Pat. Nos. 2,604,095; 3,993,791; 4,154,820; 4,630,727; 4,654,204; 4,756,838; 4,959,175; 5,296,242; 5,383,324; 5,603,695; 5,609,838; 5,779,357; and U.S. Patent Publ. No. 2004/0175437. Literature publications describing buffering anesthetics with widely different ratios of a sodium bicarbonate solutions include Ridenauer et al., Anesth Prog, vol. 48, p. 9-15 (2000); Palmon et al., Anesth Analg, vol. 86, pp. 379-81 (1998); Metzinger et al., Southern Med J, vol. 87, no. 2 (1994); Nelson, Contracept, vol. 55, p. 299 (1995); Samdal, Scand J Plast and Recons Surg and Hand Surg, vol. 28, p. 33-37 (1993); Master, Br. J. Plast Surg, vol. 51, p. 385 (1998); Difazio et al. Anesth Analg, vol. 65, p. 760 (1986); Fitton et al., Br. J. Plast Surg, vol. 49, pp. 404-08 (1996); Peterfreund et al., Region Anesth, vol. 14, no. 6, p. 265 (1989); Momsen et al., Ugeskr Laeger, vol. 162, no. 33, p. 4391 (2000); Schwab et al. Am J Emerg Med, vol. no. 3 (1996); McGlone et al. Arch Emerg Med, vol. 7, pp. 65-68 (1990); and Sapin P, et al. Catheterization and Cardio Diag, vol. 23, pp. 100-102 (1991). 
       BRIEF SUMMARY OF THE INVENTION 
       [0011]    The present invention provides methods and systems for preparing container inventories of bicarbonate medical buffer solutions (buffers) having a precisely controlled pH with minimum variability among the solutions in any individual container. By providing such inventories of buffer containers where the buffer in each container has an identical pH (within a very small tolerance, typically±−0.03), the buffers can be used to control the pH of other medical solutions, such as local anesthetics, to a predictable physiologic pH by combining a predetermined volume of the buffer with the medical solution. In this way, the buffered pH of the combined buffer and medical solution will also be precisely controlled and predictable, a result that would not be possible if the pH of the buffer solution were not itself precisely controlled and predictable. 
         [0012]    Medical bicarbonate buffer solutions are acidic buffers that rely on the combination of carbon dioxide (CO 2 ) with water to form carbonic acid (H 2 CO 3 ) which dissociates to a hydrogen ion (H + ) and a bicarbonate ion (HCO 3   − ). 
         [0013]    In buffering medical solutions, including local anesthetics such as lidocaine, articaine, prilocalne, and mepivacaine, both the amount of the buffering solution and the pH of the buffering solution can have a significant effect on the “buffered” pH of the combined solutions. Viewed differently, the amount of buffering solution needed to stabilize a medical solution at a particular target pH will depend on the pH of the buffering solution itself. Thus, a measured volume of buffering solution cannot be relied on to adjust the pH of a medical solution toward a target pH when the actual pH of the buffering solution vary significantly from its nominal pH, and the stabilized pH of the buffered medical solution can differ significantly from the target value. Conversely, by providing inventories of medical buffer solutions having precisely controlled and highly predictable pH values in accordance with the present invention, the user can adjust the pH of a local anesthetic or other medical solution in a highly predictable manner by combining a predetermined or calculated volume of the buffer with a given volume of the medical solution. 
         [0014]    In a first aspect of the present invention, a method for adjusting pH of a bicarbonate buffer in a plurality of buffer containers comprises positioning each of the containers in a chamber with the containers open to expose the buffer inside each container to a carbon dioxide containing atmosphere in the chamber. The pressure, temperature, carbon dioxide (CO 2 ) level, and usually relative humidity within the chamber are controlled to levels selected to provide a target equilibrium pH of the buffer. Maintaining a high relative humidity level will limit evaporation from the containers. By “equilibrium,” it is meant that an equilibrium exists between the partial pressure of the dissolved CO 2  in the buffer solution within the container and the gaseous CO 2  in the chamber&#39;s atmosphere. As described above, the pH of the bicarbonate buffer solution depends on the four-way equilibrium between CO 2 , carbonic acid, hydrogen ions, and bicarbonate ions. By providing a virtually unlimited source of CO 2  within the chamber at controlled pressure and temperature, the amounts of carbonic acid and bicarbonate ion within the buffer solution in each container will be driven to equilibrium values which are precisely equal within ±0.05, preferably within ±0.03, and more preferably within ±0.02. To achieve such precision, the concentration of CO 2  and temperature in the atmosphere within the chamber must be carefully controlled. The atmosphere will preferably be mixed to assure that it is substantially constant in all regions within the chamber. Such mixing can be achieved by introducing and/or recirculating fresh carbon dioxide gas through the chamber and by further mechanically mixing the gas within the chamber, as will be described in more detail below. Optionally, the containers may be vibrated, shaken or otherwise displaced during equilibration to mix the solution within the containers and to prevent pH gradients from forming in the containers. 
         [0015]    The containers are held in the controlled atmosphere of the chamber for a time sufficient for the buffer in each of the containers to equilibrate with the carbon dioxide atmosphere to reach the target pH. Typically, the carbon dioxide atmosphere will be at least 99% by volume; preferably 99.9% by volume, most preferably being essentially pure carbon dioxide (100% by volume). These CO 2  values are based on a “dry” analysis with water vapor removed. In use, however, the atmosphere will typically have a high relative humidity as discussed below. The time necessary to equilibrate and reach the target equilibrium value with all containers will usually be about 36 hours, more usually about 40 hours, and often being 42 hours or more. When using 100% carbon dioxide at a temperature of about 72.5° F. (22.5° C.), at atmospheric pressure, a pH of 7.68±0.05 can be achieved. After sealing, the steam sterilization process (autoclaving of the containers) can have a further effect on the pH, for example, leading to a small increase of approximately 0.03, but such change is predictable and can be calculated in determining the final pH of the buffer in the sealed container. 
         [0016]    The atmosphere in the chamber will usually be humidified to provide a humidity as close to 100% as possible, typically being at least 80%, preferably at least 90%, and usually 95% relative humidity or higher. As described in more detail below, the atmosphere can be humidified by passing the CO 2 -containing gas through water columns prior to introducing the gas into the chamber. The temperature in the chamber will typically be maintained in the range from 72° F. to 74° F., preferably from 72° F. to 73° F. Open containers of water may be placed in the chamber during the period of time when the chamber is being filled with CO 2  in order to more efficiently achieve high humidity. Alternatively, the container may include an amount of liquid water that is expected to evaporate during the equilibration process. 
         [0017]    In the exemplary embodiments, the carbon dioxide-containing gas is introduced to and exhausted from the chamber at a substantially constant flow rate. The actual flow rate will depend on the volume of the chamber, where the constant flow rate per hour will typically be in the range from about 25% of the volume to about 150% of the volume of the chamber, typically being about 40% to 75% of the chamber volume. Usually, the carbon dioxide-carrying gas will be circulated through the chamber prior to placing the buffer containers within the chamber so that the chamber is “primed” before receiving the containers. The carbon dioxide-containing gas will usually be introduced into the bottom of the chamber and exhausted from the top of the chamber. Because the humidified carbon dioxide gas is about twice as heavy as the air initially present in the chamber, the carbon dioxide will blanket the bottom of the chamber and drive the air upwardly and out of the exhaust port at the top of the chamber, eventually establishing a highly uniform gas composition within the chamber prior to beginning the treatment of the individual containers. 
         [0018]    After the buffer solutions in the containers have equilibrated with the carbon dioxide atmosphere in the chamber, the containers may be sealed to inhibit changes in the pH during transfer and storage prior to use. The containers are preferably sealed with no head space so that there is no gas in the container which might allow CO 2  to evolve out of solution and enter the head space causing the pH of the solution in the container to change. Further preferably, a pressure is maintained on the buffer solution within the container to prevent the evolution of CO 2  out of solution, which could similarly cause the pH to change. Apparatus and methods for storing the buffers while applying pressure are described in commonly-owned application US 2009/029271, the full disclosure of which has previously been incorporated herein by reference. 
         [0019]    In a further preferred aspect of the present invention, the containers may have a reduced diameter neck, where the neck provides the opening through which the gas equilibrates with the buffer solution. In such instances, the reduced area of the neck will limit the surface area of the buffer available for the molecular exchange necessary for equilibration. In such cases, the level of the buffer can be lowered to a larger diameter region of the container which affords a larger area for the equilibration reactions to occur. The larger area, in turn, will reduce the time needed to achieve equilibrium. After equilibrium has been achieved, the level of the buffer within the container can be raised to eliminate the head space, usually by advancing a plug or other support member within the container to push the buffer solution upwardly to fill the neck. Other means for raising the surface of the solution can also be used, such as reducing the width of the container, introducing an inert volume (e.g., beads) into the interior of the chamber to displace fluid, or the like. 
         [0020]    In this way, inventories of buffer containers where the buffer in each container has a nearly identical pH can be prepared. Usually the inventories will comprise containers which are substantially identical in shape and volume, and the containers will then be packaged into larger boxes or other storage and/or shipping enclosures. Usually, such inventories will include at least four containers, but sometimes hundreds, thousands, or even tens of thousands of containers. 
         [0021]    The containers may be sealed while they remain within the controlled environment of the chamber. Usually, however, the open containers will be removed from the chamber and sealed outside of the chamber. While it is possible to maintain the controlled environment even outside of the chamber for sealing, it will usually not be necessary. Exposure of the open containers to a normal atmosphere (without high levels of carbon dioxide) will have little effect on the containers so long as the air-liquid interface is small in relation to the fluid volume of the container, and the exposure is of short duration, typically less than 10 minutes. 
         [0022]    In a further aspect of the present invention, a system for adjusting the pH of a plurality of buffer containers comprises a chamber having a perimeter wall and an interior. The system further comprises a supply (source) of CO 2 , such as one or more tanks of compressed CO 2  connected to one or more regulators, to deliver a CO 2  gas flow into the interior of the chamber, where the gas flow comprises at least about 99% by volume carbon dioxide, usually being substantially pure carbon dioxide, except that water vapor may be present in an amount sufficient to raise the relative humidity enough to limit or prevent evaporation during the equilibration process, as discussed elsewhere herein. A plurality of spaced-apart supports within the interior of the chamber are adapted to removably receive the plurality of buffer containers or carriers that may hold a small number of containers, typically being shelves on a revolving carousel or other moving frame. A multiplicity of pneumostatic passages are formed through the perimeter wall and located to allow placement of the buffer containers onto and removal of the buffer containers from each of the supports through the passages. The pneumostatic passages allow physical entry and exit of the buffer containers, typically on carriers, through the wall while inhibiting the exchange of gases from within the chamber with the exterior environment. Exemplary pneumostatic passages include valve structures, curtain structures, and port holes with sealing doors. Because the CO 2  source will maintain a constant flow of CO 2  into the chamber and the pneumatic passages are located below the level of the exhaust port any air which enters goes to the top of the chamber and is driven out by the continued flow of CO 2  gas into the chamber, thus re-establishing and maintaining the elevated carbon dioxide levels within the chamber. 
         [0023]    Usually, the chamber comprises a free-standing cabinet, but the chamber could comprise a room or other structure built within a larger structure. The gas flow will typically flow freely into the bottom of chamber and out the exhaust port at the top of the chamber, where exhaust port is large enough to prevent any buildup of pressure within the chamber. Optionally, the supply lines could be adapted to maintain the temperature of the gas being introduced to the chamber, but typically the temperature within the chamber will be allowed to reach the temperature of the room or other surrounding environment in which the chamber is being maintained. Thus, temperature control within the chamber will be affected by temperature control within the room or surrounding containing the chamber. Still another option would be to build one or more of the walls out of a double layer of material, with the space in between being heated or chilled to create the desired temperature. Usually, the system will further comprise a water bath or water column, most typically a series of water columns, through which the inlet carbon dioxide gas flow would pass before the gas enters the chamber. Optionally, the last water column can be maintained at a temperature slightly above the chamber temperature so that the relative humidity of the gas does not drop when entering the chamber. In the exemplary embodiments, the supports comprise a rotatable carousel having a plurality of stacked, circular shelves, where the shelves rotate adjacent to the multiplicity of pneumostatic passages so that different portions of the shelves will be accessible from each of the pneumostatic passages to facilitate loading and unloading of the supports with the buffer containers. Optionally, the carousel may vibrate, causing the solution in the containers to mix over time, inhibiting the formation of pH gradients that might limit uniform equilibration. 
         [0024]    In a still further aspect of the present invention, an inventory of buffer containers comprises a plurality of containers filled with an aqueous solution of a bicarbonate buffer solution, wherein each container in the inventory has a volume of bicarbonate solution in the range from 2.5 ml to 3.1 ml and a pH variation below ±0.05 when compared to the pH of other containers in the inventory. The pH containers may comprise any conventional medical solution container, including bottles, syringes, tubes, and the like, but usually will comprise glass cartridges having a neck and a cap with a needle-penetrable septum. Such cartridges, sometimes referred to as carpules, will preferably further include a bottom support which can be advanced within the cartridge in order to dispense the aqueous buffer through the neck under positive pressure. Such containers are illustrated, for example, in commonly-owned application US2009/0292271, the full disclosure of which has previously been incorporated herein by reference. In preferred aspects, the bicarbonate solution in each container will be maintained at a pressure above atmospheric to inhibit the evolution of carbon dioxide during storage. The containers will usually be maintained in storage receptacles, such as boxes, trays, or the like, for storage, shipping, and the like. 
         [0025]    The fluid in the containers may be held under pressure by a spring or other structure that provides enough force to prevent CO 2  from evolving out of solution during transportation or storage. If the cartridge is steam sterilized, as an alternative to aseptic filling, then the force applied by the spring must be sufficient to prevent the sodium bicarbonate from boiling during the autoclave process. However the spring force must not be so great that the fluid cannot undergo thermal expansion, in which case the container may fail. Finally, the spring force must not allow the pressure in the vessel to rise sufficiently to cause the bicarbonate salt to leave solution. 
     
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         [0026]      FIG. 1  illustrates a medical solution container useful in the practice of the present invention. 
           [0027]      FIG. 2  illustrates a plurality of the containers of  FIG. 1  held in a carrier used for placing them within a chamber for treatment in accordance with the principles of the present invention. 
           [0028]      FIG. 3  illustrates an exemplary treatment chamber in accordance with the principles of the present invention. 
           [0029]      FIG. 4  illustrates a humidifying system useful with the treatment chamber of  FIG. 3 . 
           [0030]      FIGS. 5A-5C  illustrate placement of the container carriers into the treatment chamber through pneumostatic ports. 
           [0031]      FIG. 6  is a schematic illustration of the pH adjustment protocols of the present invention. 
       
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
       [0032]    A buffer container  10  useful in the systems and methods of the present invention is illustrated in  FIG. 1 . The buffer container  10  comprises a cartridge  12  including a glass cylindrical body having an open bottom  14 . A reduced diameter neck  16  is at the top with an opening  18  which can be covered and sealed with a cap  20  with a needle-penetrable septum  22 . The bottom of the cartridge  12  is defined by a movable plunger  24  which can be pushed in an upward direction (indicated by the arrow) to apply pressure on the aqueous buffer held within the chamber  12  between the plunger and the opening  18  in neck  16 . The construction of buffer container  10  as just described is conventional within the art. 
         [0033]    Buffer containers  10  may be held in a carrier  30  which includes a plurality of receptacles  32 , each of which receives an individual container  10  and holds that container in an upright orientation with the opening  18  in the neck  16  being upwardly exposed. The carriers may have a variety of configurations, but will usually be elongate structures which allow for convenient insertion and removal of the trays through pneumostatic ports in the equilibration chamber, as described in more detail below. For example, the carriers  30  may comprise a number of receptacles  32 , typically from 1 to 32, which are arranged in a linear fashion with a minimum spacing between adjacent receptacles, typically from 3 mm to 3.5 mm. Typical buffer containers  10  will have a diameter in the range from 11 mm to 12 mm. 
         [0034]    Referring now to  FIG. 3 , a treatment chamber  40  comprises an enclosure with perimeter walls  42 , a top  44 , and a bottom  46 . The perimeter walls may include one or more large doors (not shown) to permit access to the interior of the chamber. The walls, top, and bottom will provide a contained interior in which the atmosphere can be controlled. The chamber need not be hermetically sealed, and small leakage from the container will be acceptable, provided that the flow rate of gas into the chamber is greater than the amount of leakage. The gas flow which is being introduced (as described below) will continuously replenish the atmosphere and maintain substantially constant conditions within the atmosphere to achieve the desired equilibration of the buffer containers  10 , as described below. The exhaust port  58  is sized large enough that gas will escape freely, preventing any buildup of pressure within the chamber. 
         [0035]    A plurality of horizontal supports  50  will be provided within the interior of the chamber  40 , typically being part of rotatable carousels  52  which will be adapted to rotate at a rate in the range from 2 rpm to 5 rpm. A plurality of pneumostatic ports  54  will be provided in one or more of the perimeter walls  42  to provide access to each of the supports  50  for introducing and removing the buffer containers  10 , typically by passing the carriers  30  through the ports. The ports will be designed to minimize the loss of gas from within the chamber during the pass-through process. 
         [0036]    The carbon dioxide treatment gas will be introduced through an inlet port  56  near the bottom of the chamber and will be exhausted through an outlet port  58  at or near the top of the chamber. The gas humidifying and supply assembly  60  ( FIG. 4 ) may be provided, typically being mounted on a side of the chamber as shown in  FIG. 3 . The gas humidifying and supply assembly includes a gas supply  62  which provides a carbon dioxide gas to a series of water columns  64 . Typically, the gas will pass through a pressure regulator  65  which maintains sufficient pressure that the gas flows through the columns and the chamber to achieve the flow rate previously described. A valve  66  is provided to allow shut off, and water fill lines  68  on each of the columns  64  to allow filling to the proper level. Optionally, the last column may include a heating element, such as an electric incandescent light bulb, to assure that the gas entering the treatment chamber  40  is slightly above the control temperature for the chamber. By maintaining a slight temperature elevation, typically 1-2° C., the relative humidity of the gas will rise as its temperature lowers to the chamber temperature, thus assuring that the relative humidity remains above the target level. In another configuration, one of the first columns may include a heating element, such as an electric incandescent light bulb, to raise the temperature of this column slightly so that the typically colder gas entering the water column does not reduce the temperature of the other water columns below the temperature of the room. In this manner, the water in the last columns is very close to or equal the temperature of the room, as is the chamber, so that there is little or no change in relative humidity of the gas as it leaves the last water column and enters the chamber. Usually, a flow meter  70  will be provided in the line leading into the treatment chamber  40  and a gas filter  72  will be provided to remove any contaminants and help maintain a clean sterile environment within the chamber. 
         [0037]    Referring now to  FIGS. 5A-5C , passage of the carriers  30  with the containers  10  into the treatment chamber  40  through the hemostatic ports  54  will be described. Each of the hemostatic ports  54  present in the perimeter wall  42  has a gas barrier  76  in the form of a large flap valve, port hole door, curtain or the like. When nothing is passing through the port  54 , the gas barrier  76  is closed, as shown in  FIG. 5A , to inhibit the loss of gas from the interior of the treatment chamber. The gas barrier  76 , however, can be opened by simply passing the carrier  34  therethrough, as shown in  FIG. 5B . This allows the carrier to be introduced quickly and placed on the support  50  with only a minimum loss of gas from within the treatment chamber  40 . After the carrier  30  is in place on the support  50 , as shown in  FIG. 5C , the gas barrier  76  will self-close to restore the barrier to gas loss. Even though some gas loss will occur, as the flow of gas is maintained above the volume of losses due to any leakage, the gas will be lost from the interior chamber to the exterior, thus having minimum or no effect on the composition, pressure, and temperature of the gas within the chamber. 
         [0038]    Referring to  FIG. 6 , the methods for treatment according to the present invention can now be described. Initially, the individual buffer containers  10  are filled with the aqueous bicarbonate buffer B to appoint near meniscus M. Usually, the volume of buffer B introduced will be selected so that the upper surface or meniscus M of the buffer lies below the reduced diameter neck  16  so that the meniscus M has a larger area than it would have if it were present within the neck. This larger area allows the carbonic acid within the buffer to reach equilibrium with the carbon dioxide in the atmosphere within the chamber  40  more rapidly than if the area were reduced. The containers  10  are then introduced to the treatment chamber  40 , typically after placement on carriers  30  as described with reference to  FIGS. 5A-5C  above. After the containers  10  have been loaded onto the supports  50 , they are left to equilibrate with the carbon dioxide environment which is continuously being replenished with the carbon dioxide gas for a number of hours under the conditions described above. After a sufficient time has passed for the buffer within the containers to equilibrate with the carbon dioxide within the treatment chamber  40 , the containers are removed and the plungers  24  advanced to raise the level of buffer to the opening  18  in the neck  16 . The caps  20  are then placed over the necks  16 , leaving little or no head space, and the individual sealed containers  10  then placed in a storage tray  80  where springs  82  can be engaged against the plungers  24  to pressurize the buffers to inhibit the evolution of carbon dioxide gas from the buffer solution. By limiting such evolution, the pH of the buffer will be more stably maintained. The treatment trays  80  are also suitable for autoclaving the containers to assure sterility. The containers may then be stored and distributed within the trays  80  or may be stored and distributed in separate containers. 
         [0039]    While the above is a complete description of the preferred embodiments of the invention, various alternatives, modifications, and equivalents may be used. Therefore, the above description should not be taken as limiting the scope of the invention which is defined by the appended claims.