EP1494723A1 - Verfahren und gerät zur dekontamination von flüssigkeiten - Google Patents

Verfahren und gerät zur dekontamination von flüssigkeiten

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Publication number
EP1494723A1
EP1494723A1 EP02807229A EP02807229A EP1494723A1 EP 1494723 A1 EP1494723 A1 EP 1494723A1 EP 02807229 A EP02807229 A EP 02807229A EP 02807229 A EP02807229 A EP 02807229A EP 1494723 A1 EP1494723 A1 EP 1494723A1
Authority
EP
European Patent Office
Prior art keywords
fluid
ozone
chamber
ultrasonic energy
treatment
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP02807229A
Other languages
English (en)
French (fr)
Other versions
EP1494723A4 (de
Inventor
Howard E. Purdum
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Throwleigh Technologies LLC
Original Assignee
Throwleigh Technologies LLC
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Throwleigh Technologies LLC filed Critical Throwleigh Technologies LLC
Publication of EP1494723A1 publication Critical patent/EP1494723A1/de
Publication of EP1494723A4 publication Critical patent/EP1494723A4/de
Withdrawn legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
    • A61L2/0005—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor for pharmaceuticals, biologicals or living parts
    • A61L2/0082—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor for pharmaceuticals, biologicals or living parts using chemical substances
    • A61L2/0094—Gaseous substances
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
    • A61L2/0005—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor for pharmaceuticals, biologicals or living parts
    • A61L2/0011—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor for pharmaceuticals, biologicals or living parts using physical methods
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
    • A61L2/0005—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor for pharmaceuticals, biologicals or living parts
    • A61L2/0011—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor for pharmaceuticals, biologicals or living parts using physical methods
    • A61L2/0029—Radiation
    • A61L2/0035—Gamma radiation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
    • A61L2/0005—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor for pharmaceuticals, biologicals or living parts
    • A61L2/0011—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor for pharmaceuticals, biologicals or living parts using physical methods
    • A61L2/0029—Radiation
    • A61L2/0041—X-rays
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
    • A61L2/0005—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor for pharmaceuticals, biologicals or living parts
    • A61L2/0011—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor for pharmaceuticals, biologicals or living parts using physical methods
    • A61L2/0029—Radiation
    • A61L2/0058—Infrared radiation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
    • A61L2/02—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor using physical phenomena
    • A61L2/025—Ultrasonics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
    • A61L2/16—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor using chemical substances
    • A61L2/20—Gaseous substances, e.g. vapours
    • A61L2/202—Ozone

Definitions

  • the present invention relates to methods for decontaminating fluids, including protein-containing biological fluids, in particular blood products, other natural biologicals, and synthetic biotechnology products.
  • the present invention also relates to apparatus useful for decontaminating fluids, including protein-containing biological fluids, in particular blood products, other natural biologicals, and synthetic biotechnology products.
  • the present invention further relates to apparatus for contacting ozone with a liquid.
  • Protein-containing biological fluids are important for a number of reasons.
  • protein-containing fluids such as whole blood and blood products, such as red blood cells, platelets, and plasma, are important components of the health care system.
  • modern health care is also dependent on other important protein-containing biological fluids, including synthetic biotechnology products such as recombinant clotting factors, as well as natural biological products, such as antitoxins and vaccines.
  • synthetic biotechnology products such as recombinant clotting factors
  • natural biological products such as antitoxins and vaccines.
  • the source of these fluids and the fact that these fluids contain proteins make them susceptible to contamination by a variety of infectious agents, such as parasites, bacteria, fungi, and viruses.
  • infectious agents such as parasites, bacteria, fungi, and viruses.
  • the common factor in all of these contaminants is that they contain DNA and/or
  • RNA RNA decontamination of the protein-containing fluid thus does not necessarily require the removal of the contaminating agents, but only the disruption of the contaminating agents' DNA and/or RNA so that these agents cannot propagate and thus spread disease.
  • RNA graft versus host disease
  • plasma is the straw-colored material left after the cellular blood components have been removed. Rich in proteins and nutrients, plasma can harbor many contaminants, but the smallest of the above contaminants, and thus the most difficult to treat, are the viruses. Specifically, potentially lethal viruses, such as HIV and Hepatitis B, are of great concern. These contaminants pose a great hazard when contaminated units are inadvertently included in the large pools of plasma used for the manufacture of pharmaceuticals, thus possibly leading to large scale infection among the treated population.
  • erythrocytes and platelets also have proteins similar to those found in contaminant, DNA and RNA
  • problems of protein damage and incomplete decontamination also extend to these blood components.
  • similar problems also arise in the treatment of biologies other than blood products. Specifically, these other biologies, whether of synthetic or natural origin, should contain no untreated genetic material of their own, and should also not be contaminated with foreign DNA and/or RNA.
  • the proteins in these biologies are similar to the proteins in the contaminating DNA and/or RNA. The net result of any treatment is thus again at least some protein damage, along with limited decontamination.
  • a fluid such as a protein-containing biological fluid
  • a fluid such as a protein-containing biological fluid may be effectively decontaminated by a method involving:
  • a fluid may be effectively decontaminated by a method involving: (a') a step for the treatment of a fluid with ultrasonic energy to obtain a de- oxygenated fluid; and (b') a step for the irradiation of said de-oxygenated fluid.
  • the inventor has further discovered, in a ninth main embodiment, that such fluids may be effectively decontaminated by a method involving: (a) treating a fluid with ultrasonic energy to obtain a de-oxygenated fluid; and
  • a fluid such as a protein-containing biological fluid may be effectively decontaminated by a method involving:
  • a fluid such as protein-containing biological fluid
  • a fluid such as a protein-containing biological fluid
  • a fluid including a protein-containing biological fluid, may be effectively decontaminated by a method involving:
  • a fluid including a protein-containing biological fluid, may be effectively decontaminated by a method involving:
  • a' a step for the treatment of a fluid with ultrasonic energy to obtain a de- oxygenated fluid
  • a step for the irradiation of said de-oxygenated fluid to obtain an irradiated fluid
  • step for the treatment of said irradiated fluid to obtain an ozone-containing fluid
  • step for the treatment of said ozone-containing fluid with ultrasonic energy may be effectively decontaminated by means of an apparatus which contains:
  • a source of ultrasonic energy coupled to the chamber, wherein said chamber comprises (i) a flat panel, (ii) an inlet, and (iii) an outlet; and wherein said flat panel of said chamber and said inlet are dimensioned such that a fluid flowing through said inlet and across said flat panel to said outlet will form a thin film and travel in plug flow at least during some portion of its flow across said flat panel.
  • a means for introducing ultrasonic energy into said means for containing said fluid wherein said means for containing said fluid comprises (i) a means for the introduction of said fluid into said containing means, (ii) a means for said fluid to flow through said containing means, and (iii) a means for the removal of said fluid from said containing means; and wherein said containing means is dimensioned such that a protein-containing fluid flowing through said containing means will form a thin film and travel in plug flow at least during some portion of its flow through said containing means.
  • a fluid including a protein-containing biological fluid
  • an apparatus which contains:
  • a chamber for containing a fluid (2) a vacuum source coupled to the chamber;
  • a fluid including a protein-containing biological fluid
  • an apparatus which contains: (1 ') a means for containing said fluid;
  • (2') means for contacting said fluid with a vacuum
  • a fluid including a protein-containing biological fluid
  • an apparatus which contains:
  • a source of ozone wherein said chamber comprises: (i) an inlet for introducing ozone from the source of ozone; (ii) an inlet for introducing plasma; and (iii) a device for mixing ozone from the source of ozone with a fluid.
  • a fluid including a protein-containing biological fluid, may be effectively decontaminated by means of an apparatus which contains:
  • a means for containing said fluid comprises: (i) a means for the introduction of ozone from said means for generating ozone into said containing means; (ii) a means for the introduction of said fluid into said containing means; and (iii) a means for mixing said ozone from said means for generating ozone with said fluid in said containing means.
  • a source of ozone wherein said chamber comprises: (i) an inlet for introducing ozone from the source of ozone; (ii) an inlet for introducing plasma; and (iii) a device for mixing ozone from the source of ozone with a fluid.
  • a fluid including a protein-containing biological fluid
  • an apparatus which contains: (1 ') a means for containing said fluid;
  • ozone may be effectively contacted with a liquid with an apparatus which comprises:
  • a substrate which has a lower surface and an upper surface and which has a plurality of passage-ways connecting said lower surface with said upper surface; (2) a source of ultrasonic energy coupled to said substrate, such that said ultrasonic energy is introduced into the liquid by the vibration of said substrate; (3) a source of ozone connected to said lower surface of said substrate.
  • Figure 1 is a flow chart which depicts one embodiment of the method according to the present invention.
  • FIG. 2 is a schematic representation of an ultrasonic degassing chamber according to the present invention.
  • Figure 3 is a schematic representation of an ultrasound treatment apparatus according to the present invention
  • Figure 4 is a schematic representation of another embodiment of a combined ultrasound treatment and UV treatment apparatus according to the present invention
  • Figure 5 is a schematic representation of a combined ozone and ultrasound treatment apparatus according to the present invention.
  • Figure 6 is a cross-sectional view of an UV treatment chamber and components according to the present invention.
  • Figure 7 is a schematic representation of an ozone contactor according to the present invention.
  • Figure 8 is a schematic representation of an ozone contactor according to the present invention
  • Figure 9 is a schematic representation of a preferred embodiment of an ozone contactor
  • Figure 10 is a schematic representation of a preferred embodiment of another ozone contactor
  • FIG 11 is a schematic representation of another ozone contactor which is useful for platelets.
  • the present invention provides novel methods and apparatus for decontaminating a fluid.
  • the fluid is a liquid such as a protein-containing biological fluid.
  • suitable protein-containing biological fluids include body fluids, such as whole blood, saliva, semen, spinal fluid, etc.
  • the protein-containing biological fluid may be a blood product, such as plasma, sera, and the red blood cell (ery hrocyte) or platelet fractions of whole blood.
  • the protein-containing biological fluid may also be any natural or synthetic protein-containing fluid derived from various in vitro or in vivo processes, such as a fermentation broth.
  • the protein- containing biological fluid is plasma.
  • the present methods and apparatus are discussed in detail below primarily in the context of the decontamination of plasma. However, it is to be understood that the present methods and apparatus may also be used to decontaminate the protein-containing biological fluids discussed above, including foodstuffs (including eggs) and reaction mixtures containing fermentation products such as those obtained by recombinant DNA technology.
  • the present invention is applicable not only to protein- containing biological fluids, but other heat-sensitive materials as well.
  • the pulsed electric field (PEF) methods discussed below are effective for decontaminating apple juice.
  • the fluid may be any liquid which is desired in decontaminated form, and includes juices such as apple juice, orange juice, tomato juice, etc.
  • the term fluid also includes liquid-like materials which are not typically thought of as liquids.
  • the present methods and apparatus may also be applied to the decontamination of eggs, etc. for in vitro fertilization (IVF).
  • the present methods may also be applied to foodstuffs, which do not necessarily have to include proteins.
  • the present methods may be used anywhere there is a problem of contamination, particularly in regard to PEF.
  • the plasma to be treated may be that of any mammal, such as dog, cat, cow, horse, pig, chimpanzee, and human.
  • the plasma to be decontaminated is human plasma.
  • the plasma to be decontaminated may be collected by any conventional technique, such as from whole blood donation or apheresis, in which cells are returned to the donor.
  • Whole blood donation involves taking less volume from the donor (about 200 ml), but requires a fairly long time (on the order of months) between donations.
  • Apheresis involves taking a greater volume from the donor (about 600 ml) but, since cells are returned to the donor, requires a shorter time (on the order of weeks) between donations.
  • the collection of plasma is described in AABB (American Association of Blood Banks) Press Technical Manual, 13 th Edition, Baltimore MD, 1999, which is inco ⁇ orated herein by reference.
  • the plasma to be decontaminated may be an individual unit obtained from a single donor.
  • the plasma to be decontaminated may be obtained by pooling a large number of individual units taken from a correspondingly large number of donors.
  • a method of decontaminating plasma need not remove or even inactivate all infectious agents to be considered useful.
  • many methods of decontaminating plasma are specifically designed to address only certain types of infectious agents, e.g., enveloped viruses, and none guarantees removal or inactivation of 100 % of even the infectious agent for which it is designed.
  • the term "method of decontaminating plasma” refers to a method which is capable of removing and/or inactivating a significant portion of at least one infectious agent found in plasma.
  • the present methods for decontaminating plasma are capable of achieving a log reduction factor or log kill of at least 4, preferably at least 5, more preferably at least 6, for at least one infectious agent found in plasma.
  • the present methods are also capable of affecting the decontamination of plasma, while minimizing the damage to plasma proteins.
  • the amount of protein damage will depend on the particular protein in question, the particular embodiment of the decontamination method used, and, to some extent the source and prior handling of the plasma.
  • the present methods are capable of achieving the above-noted log kills of at least one infectious agent while causing protein damage of less than
  • Examples of the types of infectious agents which may be removed and/or inactivated by the present methods include parasites, bacteria, fungi, and viruses, and possibly prions. Of these agents, parasites pose significant threats mainly in tropical climates. The greatest such risk is malaria, which is spread by all four different species of Plasmodium, but principally by Plasmodium malariae. Another parasitic risk is Trypanosoma cruzi, which causes Chaga's Disease, a serious problem in Central and South America. In the United States, two species of Babesi protozoans, which cause Babesiosis, can be transmitted by transfusion, although the more common route is tick bite. Leishmania infantum is also a parasite which may be found in blood products.
  • Bacteria also present a continuous threat in transfusion. For this reason, the CDC and leading members of the blood community have recently launched the BaCon (Bacterial Contamination) study to determine the risks of transfusion related infections.
  • BaCon Bacterial Contamination
  • Yerisinia enterocolitica Escherichia coli
  • Citrobacter freundii as well as Bartonella and Brucella species.
  • Fungal infections are a continuous and escalating problem in medical care, particularly for those patients with compromised immune systems due to cancer therapy, HIV, etc.
  • mold refers to the typically woolly appearance of a growing fungus.
  • a yeast is then a particular minute fungus, especially of the Saccharomycetaceae family.
  • viruses of concern are the various strains of hepatitis (A through E, and G), HIV (human immunodeficiency virus), HTLV (human t-cell lymphotropic virus) types I and II, CMV (cytomegalovirus), EBV (Epstein-Barr virus) and parvovirus B19.
  • the unique equipment described in the following can also be used for pu ⁇ oses other than decontamination.
  • the ozone treatment unit is also useful for adding gasses to liquids in general.
  • the device can be used to oxygenate blood during cardiopulmonary bypass.
  • the ozone treatment unit can be used to add carbon dioxide to aqueous solutions; other similar applications are also possible.
  • the present invention provides a method for decontaminating plasma which comprises: (a) treating plasma with ultrasonic energy.
  • ultrasonic energy and “ultrasound” refer to sonic waves with frequencies in the range of 20 kHz, the upper limit of human hearing, to several hundred MHz.
  • ultrasonic vibrations may also be generated by other conventional means, in particular by electromagnetic, electrostrictive, or magnetostrictive devices. Such devices are described in published PCT Patent Application WO 92/20420, which is inco ⁇ orated herein by reference. Because of its relatively high frequency range, ultrasound has many applications in industry and medicine. In particular, ultrasound has many unique and beneficial applications in the treatment of liquids.
  • Cavitation is a localized vaporization that occurs when the low pressure part of the ultrasonic wave becomes less than the vapor pressure of the liquid. Under these conditions, the local temperatures and pressures become extremely high as the cavitation bubbles grow and then collapse.
  • the treated liquids thus include all non-cellular protein containing liquids.
  • the treated liquids are plasma or sera, but not red blood cells or platelets.
  • the overall arrangement for this first embodiment is therefore much like that used for cell disruption. Specifically, three separate components are required: a source of ultrasound, a target, and some means of coupling the source to the target.
  • the limiting factor here is that the high frequency waves of ultrasound do not propagate well in gasses such as air, and therefore require a carrier medium such as a rigid metal or a liquid.
  • waveguides are commonly made of aluminum. With careful shaping into geometries called horns, these waveguides can produce waves of high amplitudes and energies, resulting in efficient transfer of the ultrasonic energy into the target.
  • plasma is therefore treated with ultrasonic energy having a frequency sufficient to result in cavitation of the plasma.
  • the plasma is suitably treated with ultrasonic energy having a frequency of 20 kHz to 10 MHz, preferably 20 kHz to 1 MHz, more preferably 20 kHz to 500 kHz, even more preferably 20 kHz to 100 kHz.
  • the decontamination unit must carefully preserve the desired protein components, to a level well beyond the degree of protection required for cell disruption devices.
  • the first problem in the protection of these proteins is limiting the strong chemical reactions that are induced by ultrasonic cavitation.
  • One such reaction is the breaking of long chain organic compounds by the severe agitation due to bubble growth and collapse during cavitation. This breaking may be a significant concern for the large, relatively delicate proteins involved in the clotting process, as described by El'piner (I. E. El'piner, Ultrasound Physical, Chemical, and Biological Effects. Consultants Bureau, New York, p. 217, 1964).
  • El'piner I. E. El'piner, Ultrasound Physical, Chemical, and Biological Effects. Consultants Bureau, New York, p. 217, 1964.
  • damage of the plasma proteins may be managed by two techniques.
  • the treatment times are kept short, i.e., less than 5 minutes, preferably less than 2 minutes, more preferably less than 30 seconds.
  • the intensity levels are kept low, i.e., 0.1 to 50 W/cm 2 , preferably 0.5 to 10 W/cm 2 , and more preferably 1 to 6 W/cm 2 .
  • these techniques must be adjusted to specific protein solutions and contaminants.
  • the ultrasonic energy is applied after the gas above the plasma has been replaced with an inert atmosphere.
  • "inert” does not include the noble gasses, because such monatomic species have too few degrees of freedom to disperse the ultrasonic energy (S. Y. Wang, in Symposium on Biological Effects and Characterizations of Ultrasound Sources, edited by D.G. Hazzard, et al, US Dept. HEW (FDA) 78-8948, US Government Printing Office, p.196, 1977).
  • suitable inert gasses must be polyatomic, notably carbon dioxide.
  • forming a carbon dioxide gas layer simply amounts to dropping a pellet of dry ice into the solution to be treated (High Intensity Ultrasonic Processor User's Guide, Sonics & Materials, Inc. Newton, CT, 1999).
  • the evolved gas then displaces the oxygen, and without oxygen, no oxygen radicals can form.
  • some of the dissolved gasses are also displaced by carbon dioxide. Part of this displacement occurs by a concentration gradient in the immediate vicinity of the pellet, but most of this displacement is due to transport from the enriched carbon dioxide gas layer above the liquid surface.
  • the net result is preferential removal of oxygen, which again is beneficial because oxygen radicals are quite damaging to proteins. Note that the overall process is thus similar to the helium sparging technique commonly used in hplc, but helium cannot be used here because of the above described formation of noble gas radicals.
  • a preferred means of limiting the source heat is to apply a water flow to the ultrasound source and/or horn.
  • a preferred means to cool the target is immersion in a water bath, which also yields strong acoustic coupling to the ultrasound horn and source. With these techniques, it is thus possible to maintain the target at any selected temperature. This temperature, however, depends on concerns that are often conflicting. Specifically, proteins are typically heat sensitive, particularly clotting factors such as Factor VIII. For maximum protection, the temperature should therefore be kept relatively low, within the FDA specified range of 2 to 10 °C. On the other hand, cavitation in water or dilute aqueous systems is most effective at about 50°C (J. Blitz, Ultrasonics: Methods and
  • the unit should be operated at about 50°C, if the target can withstand such elevated temperatures.
  • use of the highest possible range provides thermal inactivation of pathogens, which is an additional safety measure.
  • temperatures slightly greater than 50°C may result in some decrease in cavitation efficiency, but this is more than compensated by the resulting improvement in thermal inactivation.
  • even higher decontamination temperatures can be used as a separate step, with cavitation done in the lower, 50°C range.
  • the liquid should be kept cold until immediately before treatment, at which time rapid heating techniques should be applied to small samples to raise the operating temperature to no more than 50°C.
  • the ultrasound should be applied for as short a duration and intensity as possible, with the sample then rapidly cooled back to storage temperatures.
  • the sonic treatment should be performed at the highest allowable temperature, followed by rapid cooling to remove any residual heat from the ultrasound source or from the cavitation process.
  • the volume to be treated is first broken into smaller units, or into a continuous, low volume per time flow. These smaller units or flows are then passed through a separate bag with a large surface area where they are subjected to heat transfer from any convenient source, such as from a water bath, microwave, air blast, etc.
  • suitable cooling mechanisms are water bath immersion, or contact with plates that are cooled by gas expansion or Peltier effects.
  • the method of the first main embodiment may be carried out in either a batch-wise, semi-continuous, or continuous fashion.
  • single bag units may be individually treated with the ultrasonic energy.
  • single units or individual small volumes of plasma may flow through one or more stations or stages in which they are treated with ultrasonic energy.
  • each individual unit or volume is held at each station or stage for processing, and is then passed in bulk to the next station or stage.
  • a continuous operation in which the plasma may flow without interruption through the stations or stages. Both the semi-continuous and continuous modes are applicable to fractionation or to other processes involving very large pools of material to be treated.
  • the first condition on this chamber is that the inlet tube or tubes must not harbor any pathogens.
  • the underlying problem can be seen in conventional blood bags, in which the inlet tube passes through a port in the top seam of such containers. As such, any untreated fluid that remains in this tube can subsequently contaminate the treated fluid. This is a particular problem for ultrasound and the other treatment technologies described below because after such treatment processes are terminated, no residual material remains in the fluid to prevent any recurrence of the pathogens. To prevent this problem, the inlet tube may be heat sealed close to the bag, but this approach still leaves the port, which is relatively hard to seal.
  • a modified compression approach is preferred.
  • the treatment chamber is clamped tightly just below the tube orifice, as described above, but in this case, no heat seal is used. Instead, the fluid in the chamber is treated and then drained before the next batch of fluid is allowed to enter by releasing the clamp.
  • the face of the clamp jaws are made of aluminum or stainless steel, thereby preventing any dampening of the waves at the clamp.
  • solid metal clamp jaws would allow the sound waves to propagate through the clamp and into the input reservoir, thus possibly causing excessive sound treatment.
  • the backs of the clamp jaws are coated with rubber or other sound insulation.
  • the second condition on the treatment chamber is that the liquid layer must be relatively thin.
  • a thin layer is necessary to ensure uniform temperatures and uniform cooling.
  • thin layers also allow the evolved gas bubbles in the liquid to rise quickly to the surface. This is important because rapid bubble rising reduces the time that the ultrasound can induce strong surface oscillations of the bubbles or strong slip-streaming around the bubbles, thereby reducing protein damage.
  • rapid bubble rising also prevents agglomeration of the bubbles to excessive size. Compared to such large bubbles, smaller bubbles are preferable because they provide more uniform treatment, and have less surface oscillation and slip-streaming.
  • the plasma is therefore preferably formed into a thin film having a thickness of 2 to 20 mm, preferably 2 to 10 mm, and more preferably then 2 to 4 mm, at least during some part of the application of the ultrasonic energy.
  • the plasma is formed into such a thin film prior to the commencement of the application of the ultrasonic energy and maintained in such a thin film during the entire application of the ultrasonic energy.
  • these thin layers must be relatively broad.
  • the broad surface can be of any desired shape, such as a circle, square, etc., as long as the total resulting volume can be treated uniformly by the ultrasound.
  • the limiting factor here is that ultrasonic waves typically do not produce uniform exposures in containers of water.
  • the ultrasonic cleaning industry has developed a number of ways to avoid "hot” and "cold” spots, primarily by using a mixture of frequencies over a narrow bandwidth and by building treatment tanks to dimensions that avoid resonant standing waves. These approaches can be used directly for batch and semi-continuous decontamination units, although the liquid layer in decontamination devices is much more shallow than that commonly used for ultrasonic cleaners.
  • the alternative is to use a plug flow, in which all of the fluid moves in bulk through the processor.
  • the fluid entering through an inlet tube is first spread out through an expanding section called a diffuser.
  • a rectangular geometry is used to provide the plug flow region.
  • this region can be 30 cm wide, 60 cm long, with a fluid depth of 0.4 cm.
  • a converging section which is essentially a reversed diffuser, is then used to guide the flow into an exit tube.
  • This simple geometry is used in ultraviolet flow cells and similar, common laboratory equipment. For decontamination applications, the ultrasonic sources are placed directly beneath the rectangular section.
  • the ultrasound can not only cause decontamination, but can also reduce the effective viscosity of the fluid.
  • This reduction in viscosity is important because lower viscosities reduce the tendency of the plug flow to become laminar, which would otherwise occur over several chamber flow diameters. The net result of this geometry is therefore quite uniform ultrasonic treatment of the fluid.
  • step (a') a step for the treatment of plasma with ultrasonic energy.
  • the step (a') "for the treatment of plasma with ultrasonic energy” may be carried out in the same way as the step “(a) treating plasma with ultrasonic energy” is carried out in the context of the first main embodiment.
  • the present invention provides a method for decontaminating a fluid, which comprises: (a) treating a fluid with ultrasonic energy, while degassing the fluid.
  • a vacuum is applied to the fluid during the application of the ultrasonic energy.
  • the ultrasonic energy may be applied to the fluid using the same equipment described above in the context of the first and second embodiments, except for the means of controlling the free radicals.
  • the ultrasonic energy is applied after the gas above the plasma has been replaced with an inert atmosphere. While quite effective, this approach unfortunately suffers from the material costs for sterile consumables, and the problems of introducing these materials without also allowing contaminants into the system.
  • An alternative approach is to apply a vacuum to remove the gasses above the liquid being treated with ultrasound (see, High Intensity Ultrasonic Processor User's Guide. Sonics & Materials, Inc. Newton, CT, 1999).
  • the liquid being decontaminated may be any of fluids discussed above.
  • the fluid is a protein-containing biological fluid, such as plasma.
  • plasma a protein-containing biological fluid
  • the gas above the fluid is effectively removed by applying a vacuum of about 2 to 100 mbar, preferably about 10 to 80 mbar, more preferably 20 to 60 mbar, to the gas above the fluid.
  • a vacuum of about 2 to 100 mbar, preferably about 10 to 80 mbar, more preferably 20 to 60 mbar, to the gas above the fluid.
  • the limiting factor here is the evaporation of the solvent: at low enough pressures, the liquid will boil uncontrollably. Since different liquids may require different levels of vacuum, it is preferred that the apparatus be configured such that the level of vacuum can be varied.
  • the vacuum may be applied by means of a vacuum pump.
  • the vacuum pump should use a scroll, or other dry, evacuation method.
  • the vacuum be applied to the gas above the fluid, e.g., plasma, at least at the time of commencement of the application of ultrasonic energy to the plasma.
  • the vacuum is applied to the gas above the plasma prior to the commencement of the application of ultrasonic energy to the plasma. Even more preferably, the vacuum is applied to the gas above the plasma: (1) prior to the commencement of the application of ultrasonic energy to the plasma; and (2) and during the application of at least a portion of the ultrasonic energy to the plasma. Of course, ultrasonic energy may be continued to be applied after cessation of exposure to the vacuum.
  • this vacuum technique also has additional benefits in the decontamination of protein solutions.
  • the main such benefit is that the applied vacuum reduces the vapor pressure above the liquid and thereby reduces the energy required to induce cavitation. With less applied energy, there is less undesirable protein damage, as would otherwise occur as described above in the first and second embodiments.
  • degassing is used in industries ranging from soda and beer production (Marks' Standard Handbook for Mechanical Engineers. Tenth Edition; McGraw- Hill, New York, 12_121-12_123, 1996) to hplc oil cleaning.
  • the goal is to remove at least some of the gasses dissolved in a liquid product. It is important to note that there are actually two sources of gas involved in the sonification of a liquid. A general discussion is provided in U.S. Patent No. 4,597,876.
  • the first gasses to be evolved are the dissolved gasses, in a process referred to as "rectified diffusion” or “gaseous cavitation.”
  • the dissolved gasses are simply trapped in progressively large bubbles because the gasses are forced out of solution more rapidly than they can diffuse back into the liquid.
  • the process more often called “cavitation” or “vaporous cavitation” in the sonochemistry literature refers to the formation of bubbles from the sonified liquid phase itself.
  • rectified diffusion requires only strong vibrations, as readily demonstrated by shaking a soda can. Much greater energy, however, is required to vaporize a liquid to yield what is conventionally called cavitation.
  • sonochemical systems and various ultrasonic cleaners are first "degassed” by extended operation and/or the use of various additives (soaps) before actual processing; otherwise, the dissolved gasses "soften” the sound waves and thus decrease the performance of the ultrasonic device.
  • soaps are acceptable for decontamination work.
  • applying sound waves of sufficient energy to plasma yields a combination of rectified diffusion and cavitation of water.
  • rectified diffusion can occur from the dissolved gasses towards the bubbles formed by cavitation.
  • An enhancement of this process is to use low intensity ultrasound and vacuum to degas the liquid before the higher intensity, cavitation inducing ultrasound is applied.
  • the advantage of this approach is that the dissolved oxygen is thus largely removed before any cavitation occurs, thus minimizing the protein damage.
  • hydrophone monitoring is used to separate the different steps in this sequence. Specifically, hydrophones record a slight hissing or “frying” sound as the liquid degasses under sonification, followed by a sharp "popping" sound as vapor formation and collapse occurs (A.A. Atchley and L.A. Crum, “Acoustic Cavitation and Bubble Dynamics," in Ultrasound: Its Chemical Physical, and Biological Effects, edited by K.S. Suslick, VCH Publishers, Inc., NY, pp. 19-20, 1988). The difference in these two signals is so distinct that it can be recognized by automated equipment, thus providing the basis for activating the different levels of ultrasound described above.
  • a final additional benefit of vacuum operation is improved feeding of the liquid into the system.
  • protein solutions such as blood products are easily damaged by pumping, whether by piston or peristaltic arrangements.
  • a vacuum system avoids this problem by drawing in the fluids under a body force.
  • a "body force” refers to an action on each component of the entire fluid stream, much like gravity. Gravity feeding, however, requires a sufficient height to provide adequate flows, and this height can sometimes be difficult to arrange in a laboratory setting.
  • valves are placed between the source bag or container and the vacuum chamber. Upon activation, these valves thus allow the fluid to enter the processor with minimum transfer damage. To prevent excessive fluid velocities, a flow restrictor is placed in the fluid path.
  • valves can be arranged to match or control the flow through the heat transfer devices described in embodiments one and two.
  • the heat transfer unit described earlier is placed directly beneath the source bag or container. This arrangement provides supplemental gravity feeding, as well as complete draining of the input bag for maximum yield.
  • the output of the warming unit is placed directly over the inlet of the ultrasonic processing chamber. Two valves are thus used for operation, one valve on each side of the warming bag. Opening the valve between the source bag and the warming bag allows the warming bag to fill to capacity. This valve is then closed, and the fluid is warmed. Next, the valve between the warming bag and the ultrasound chamber is opened, allowing the fluid to enter this chamber under the influence of both gravity and/or the applied vacuum.
  • vacuum operation also imposes several special requirements.
  • effective decontamination requires that the vacuum operations must be performed under sterile conditions.
  • One possible solution to this problem is extensive cleaning and decontamination of conventional vacuum hardware.
  • a preferred approach is to use a special disposable that can be easily changed between applications.
  • One possible approach is to use a disposable chamber that can withstand a 1 atmosphere vacuum. Such a device, however, would require a great deal of material and would therefore be quite expensive.
  • a preferred alternative is to use a thin disposable plastic liner inside a conventional vacuum chamber made of metal, preferably stainless steel.
  • this disposable amounts to a tent arrangement, with an inlet and outlet at opposite sides of the base.
  • a connecting tube provides vacuum access, which thus equalizes the inner and outer pressures.
  • an FDA approved filter prevents entry or exit of any pathogens through this tube.
  • a plastic cover is attached to this filter; this cover is opened before use of the unit and closed afterward.
  • a connecting hose with a sterile coupling device can also be used for direct vacuum connection. SCDs are also used for the inlet and exits.
  • the entire disposable can thus be sterilized by gamma radiation, autoclave, gas treatment, or any other conventional sterilization technique.
  • the tent can be made of either rigid or flexible plastic. Rigid plastic tents would be clamped to the ultrasound driver for highly effective sound transfer and great structural integrity. Unfortunately, such an arrangement would be relatively expensive and would require a significant amount of storage space.
  • An alternative is a flexible tent with mounting hooks at the corners to maintain the proper shape. This cheaper arrangement is preferable. If desired, the tent can be immersed in a liquid bath to improve acoustic coupling and heat transfer.
  • any contaminated liquid aspirated from the plasma container is thus captured in a separate chamber for disposal.
  • this trap is placed beyond the sterile filter so that the trap can be used for multiple cycles.
  • the trap is located externally. Under this approach, the connection to the plasma chamber is made through a molded insert placed in a recess of the vacuum chamber door seal. Routing the vacuum hose, and any feed lines, through this insert thus allows the disposable to be mounted and dismounted quickly and easily.
  • the vacuum chamber trap in this approach thus captures any immersion liquid, and also provides an additional safety measure should the plasma container fail.
  • the vacuum chamber trap also provides a convenient mounting location for the vacuum gauge that is required to monitor the process. Specifically, this gauge must be mounted on the pump side of the trap and sterile filters for protection from any aspirated material.
  • the system can be operated in batch, semi-continuous, or continuous mode, as described in the previous embodiments.
  • the system In batch mode, the system is evacuated simply by closing the vacuum chamber and turning on the vacuum pump. If the system is not pre-filled, the vacuum then draws the fluid to be treated into the processing chamber. Next, a vacuum sensor and relay activate the ultrasound at the selected vacuum level. Degassing and cavitation are then performed as described above, with the process continuing for a preset time. At the conclusion of this process, the vacuum is released, and the processed material is removed. After fitting a new disposable unit, the system is then ready for another operation.
  • Semi-continuous operation is performed in a like fashion, except that the filling is done in steps from a large reservoir, and the disposable is not changed at each cycle. In this case, the processed material is collected in a second reservoir at the end of each cycle. Finally, after draining the input reservoir, there will be some residual material left in the processor. To achieve maximum yield, with minimal residual material to pose a biohazard waste product, the treatment unit is then tilted slightly to drain the residual product. This action can be driven by a pneumatic piston, an electric motor, a solenoid, etc.
  • the first problem is that a continuous flow precludes the use of variable intensity and time pulses of ultrasound on a single batch, as described above. Instead, the ultrasound treatment may be achieved by passing the plasma through sequential pools, each pool having its own ultrasound sources acting at successively higher powers and longer times. Plug flow, as described above in embodiment I, is used in each pool. Because water is an excellent conductor of ultrasound, however, the pools must be acoustically isolated. This is achieved by having the flow cascade from one pool to the next, with sufficient space so that the fluid thins out to sheets. Next, these sheets flow over a saw tooth pattern of streamers cut from the outlet plate, thus forming a series of ligaments. Under the action of gravity and ultrasound, these ligaments then break up into multiple droplets. The space between these droplets cannot propagate ultrasound, thus providing the necessary acoustic isolation between treatment pools.
  • peristaltic pump which is quite useful for those fluids that can withstand the action of such pumps.
  • the limiting factor here is that. conventional peristaltic pumps do not work well in vacuum due to heat buildup and lubricant degassing. It is therefore necessary to use either an external driver with an access port through the vacuum chamber wall, or a sealed peristaltic pump.
  • Another means of removal is to collect the treated fluid in a vessel that is alternately exposed to vacuum and atmospheric environments by a valve and vacuum pump arrangement.
  • opening the valve at the exit of the treatment chamber allows the treated fluid to flow into a collection bag that is also in an evacuated chamber.
  • the treatment chamber valve is closed, and the vacuum in the collection chamber is released.
  • the outlet valve is then released.
  • the collection bag is completely drained, the exit valve is closed, and the collection chamber is pumped back to vacuum conditions. The process is then repeated as necessary.
  • This approach thus provides minimum protein damage during fluid transfer.
  • the present invention provides a method for decontaminating a fluid, which comprises: (a') a step for the treatment of a fluid with ultrasonic energy, while degassing the fluid.
  • the step (a') "for the treatment of a fluid with ultrasonic energy, while degassing the fluid” may be carried out in the same way as step "(a) treating a fluid with ultrasonic energy, while degassing the fluid" is carried out in the context of the third main embodiment.
  • the present invention provides a method for decontaminating a fluid, which comprises:
  • the limiting factor here is the interaction of the ultrasound with the bubbles in the liquid, which arise from either dissolved gasses or from vaporous cavitation of the liquid itself, as described above. From wave mechanics, the effectiveness of the interaction between these bubbles and the sound waves depends principally on the intensity and the frequency of the ultrasound. As discussed above, higher intensities promote cavitation of the liquid, while lower intensities promote degassification. Depending on the size of the bubble, however, it is also possible for the sound waves to cause the gas bubbles to be absorbed entirely back into the liquid. In this case, the growing bubbles are said to be unstable.
  • Another approach is to use two different frequencies simultaneously (see, CP Zhu, R Feng, YY Zhao, "Sonochemical effect of a bifrequency radiation," Chinese Science Bulletin, vol. 25, No. 2 (Jan), pp. 142-145 (2000)).
  • the advantage of this approach is that the combined exposure of two separate sources of greatly different frequencies is significantly greater than the sum of the two sources acting alone.
  • the importance of this result in the context of the present invention is that the use of multiple frequencies provides a means of achieving the same level of cavitation, but with less power applied to the system. With less input power, there is less sample heating, and less intense shear and oscillation around the bubbles. As a result, the material suffers less damage during sonification, while also providing the option of increased total power to improve decontamination and/or to shorten the processing times.
  • a second consideration in multiple source arrangements is that the frequencies must also be sufficiently great to prevent beating or even prolonged superposition of extrema. For this reason, Zhu et al used frequencies in the low tens of kHz range to the MHz range.
  • the general relationship is that the waves must be separated by at least an order of magnitude in frequency, and preferably also separated in addition by a small constant scale factor. As such, suitable separations can be on the order of 15 or 20 or so.
  • the lowest frequency band is on the order of 20 to 100 kHz
  • the next band is on the order of 500 kHz to 1.5 MHz
  • the highest band is on the order of 10 MHz.
  • the frequency separation is at least an order of magnitude, these ranges are not extremely critical; the main limitation in this procedure is simply the availability of commercial generating equipment.
  • a useful enhancement is to use cavitation power for the highest frequency, with or without pulsing.
  • the evolved gas pockets are quite small, and thus provide nucleation points for subsequent growth by rectified diffusion under the influence of the lower frequency sources.
  • the present invention provides a method for decontaminating a fluid, which comprises:
  • step (a') a step for the simultaneous treatment of a fluid with at least two different frequencies of ultrasonic energy.
  • the step (a') "for the simultaneous treatment of a fluid with at least two different frequencies of ultrasonic energy” may be carried out in the same ways as step "(a) simultaneously treating a fluid with at least two different frequencies of ultrasonic energy” is carried out in the context of the fifth main embodiment.
  • the present invention provides a method for decontaminating a fluid, which comprises:
  • the ultrasonic energy may be applied to the fluid in the same way and by using the same apparatus discussed above in the context of the first, second, third, fourth, fifth, and sixth main embodiments.
  • the main role of the ultrasonic energy is to affect degassing of the fluid prior to exposure to the radiation.
  • the major limitation in applying radiation, such as UV, gamma, or x-rays, for decontamination is the formation of free radicals of oxygen.
  • radiation such as UV, gamma, or x-rays
  • one possible solution to this problem is to use some type of scavenging agent, but these agents are expensive and often toxic, and they must therefore be removed before the product can be used.
  • the present invention uses a degassing technology before irradiating the liquid. In particular, this technology is directed towards the removal of oxygen. Without dissolved oxygen, no oxygen radicals can form under subsequent irradiation, thereby sparing the proteins being decontaminated.
  • degassing liquids in general industrial practice.
  • embodiment III describes the protein damage expected during degassing prior to ultrasound decontamination.
  • quenching agents may have little or no inactivating effects on such species. The net result is that quenching agents do not eliminate all oxygen effects, leaving some radical damage and some damage due to other reactive oxygen species. Because quenching agents are known to be effective even with these limitations, it is therefore not necessary that the new degassing technology remove all of the dissolved oxygen to be effective in protecting the proteins during irradiation. In practice, the actual amount of residual dissolved oxygen must be determined on an individual basis. One factor to be considered in this determination is the damage that the degassing process itself inflicts upon the proteins.
  • the dissolved oxygen concentration can easily be decreased to about 1 to 2 ppm in 5 minutes or less, depending on the starting temperature and oxygen concentration. To achieve concentrations in the hundreds of ppb range, however, the processing time increases to about 30 minutes.
  • the target dissolved oxygen range for blood products is preferably 10 to 3000 ppb, more preferably 100 to 2500 ppb, and most preferably 500 to 2000 ppb.
  • conventional electric resistance-based dissolved oxygen meters are not accurate because the samples are too small and there is inadequate flow to ensure representative reactions at the electrodes.
  • This flow limitation is particularly important in the ppb range, where a significant amount of liquid must be tested in order to obtain enough oxygen for an accurate result; otherwise, false low readings will occur as the local environment is depleted of the few oxygen molecules present.
  • an optical abso ⁇ tion meter Model VVR, CHEMetrics Calverton, VA
  • VA optical abso ⁇ tion meter
  • the remaining concern is the radiation to be used for the subsequent decontamination. Specifically, it is necessary to select the type of radiation, the required dosage of this radiation, and the means of applying this radiation to the material to be treated.
  • gamma radiation is commonly used in the decontamination industry, typically from Cobalt-60 or Cesium- 137 sources.
  • the required dosages are known for most pathogens, but for general decontamination work, a dosage must be selected that will treat most pathogens, without causing excessive protein damage.
  • An appropriate test virus must therefore be selected; the conditions that inactivate this virus are then considered to be adequate for other pathogens as well.
  • parvovirus A particularly useful test subject is parvovirus.
  • This small, non-enveloped virus is quite difficult to inactivate, thus assuring destruction of less robust viruses such as HIV.
  • parvovirus In its porcine form, parvovirus is harmless to humans, and is therefore easy to handle in the laboratory. Furthermore, because of its potential damage to a developing human fetus, and because of its ease of transmission by transfusion, the human form of this virus is of clinical significance.
  • parvovirus is therefore commonly used as a benchmark for inactivation technologies (SI Miekka et al, "New methods for inactivation of lipid-enveloped and non-enveloped viruses," Haemophilia 1998 Jul;4(4):402-8).
  • the appropriate dosage range for gamma radiation of blood products is between 1 to 100 kGy (kiloGray), more preferably 2 to 60 kGy, and even more preferably 4 to 40 kGy.
  • the radiation source is placed outside the sterile tent arrangement described above. As such, the source can be placed inside the vacuum chamber, or outside the vacuum chamber with access through a thin, low-absorption window.
  • Another option is to irradiate the liquid after the chamber vacuum has been released.
  • a valve is placed on the tube leading to the processing tent. Closing this valve after the tent and vacuum chamber have been evacuated and sonified allows the chamber to be returned to atmospheric conditions, while still maintaining a vacuum on the processed material. As such, the flexible bag then simply collapses upon the essentially incompressible fluid being treated.
  • the advantages of this approach include very simple shielding and control arrangements. The only constraint is that the treatment bag must be relatively impermeable to oxygen so that the irradiation is completed before the dissolved oxygen concentration rises to unacceptable levels by diffusion from the surrounding air.
  • the irradiation must be completed before significant amounts of oxygen pass through less impermeable bags.
  • batch and semi-continuous operations can be readily achieved by simply irradiating the target for a specified time, with continuous monitoring of the applied radiation by conventional detectors and recorders.
  • the previously described plug flow must be constrained in a closed channel arranged so that the fluid is continuously rising in height. This constraint ensures that all of the fluid has sufficient residence time in the exposure chamber to receive full treatment; otherwise, the fluid could flow out of the channel too rapidly under the force of gravity.
  • the driving force for the upward flow can be a pump, or a gravity head from the higher-placed vacuum chamber. The type of force is particularly important for the cellular components of blood that are readily damaged by most mechanical pumps, as noted above. In this regard, gravity feeding thus provides little or no such damage.
  • the present invention is well suited for the leukoreduction of cellular blood components.
  • leukoreduction by gamma irradiation avoids the problems of filter clogging, long processing times, high expense, and the related difficulties that complicate existing, filter-based technologies.
  • an additional advantage in the present invention is that degassing prior to leukoreduction prevents oxygen radical attack on the walls of the leukocytes, and thus prevents the leakage of the cellular contents that are quite detrimental to erythrocytes. Note in this application that although cellular blood components require oxygen, these cells can survive in an oxygen-depleted environment long enough (several minutes) for decontamination. Oxygen can then be supplied to the fluid after the irradiation is completed.
  • the above advantages of gamma exposure are particularly significant in terms of the recent recommendation for universal leukoreduction (Wall Street Journal, "Federal Panel Hears Debate On Filtering All Donated Blood,” 1/25/2001).
  • gamma radiation has sufficient energy to dissociate and/or excite water molecules.
  • protein concentrates can be used.
  • PCT Application number WO 0016872 describes a procedure for generating such concentrates. Because concentrates contain relatively little water by definition, the number of reactive water species is reduced proportionately, compared to the original protein solutions.
  • x-rays can also achieve all of the above benefits. Unlike the nuclear decay sources used for gamma irradiation, x-rays are instead generated by high voltage, electron acceleration sources. Because of the energy requirements and maintenance costs of such accelerators, however, gamma radiation is currently the preferred of these two radiations for decontamination work. Unfortunately, both gamma and x-ray sources are not only relatively expensive, but they also require strict shielding, licensure and other constraints.
  • UV sources are more commonly used for decontamination.
  • There are four types of UV radiation, which in terms of increasing energy are UVA, UVB, UVC, and VUV.
  • UVA and UVB are the relatively weak radiations associated with sun exposure. Being too weak to induce radical formation, these radiations unfortunately require some type of light-activated chemical to induce decontamination. As described above, however, the intent of this invention is to avoid the costs of any such chemicals, so these radiations are not useful here by themselves.
  • VUV or vacuum ultraviolet, is much more energetic, being on the border of soft x-rays in terms of energy. VUV, however, is difficult to generate and is absorbed so readily that it is not practical for decontamination work.
  • UVC is therefore the preferred form of ultraviolet radiation in the present invention, particularly the UVC produced by mercury sources.
  • These sources are useful because they emit light mostly at about 254 nm. Because these sources thus have relatively low emissions at wavelengths less than the 185 nm threshold required for ozone generation, they produce little of this undesirable gas. More importantly, however, this wavelength is in the middle of a local minimum of absorption that exists in the range of 250 to 260 nm for most proteins, but both DNA and RNA have a local absorption maximum in this range. For this reason, mercury lamp emissions thus selectively attack the DNA and RNA found in most contaminants, while sparing the other proteins in the material being treated. In particular, this selectivity is most useful for treating blood components because, as discussed earlier, most blood components lack the genetic materials DNA and RNA.
  • UVC has all of the clinical advantages described above for gamma rays, as well as the advantage of selective protection of the surrounding proteins.
  • UVC dosages in terms of water purification and food processing, but protein solutions have unique requirements, notably regarding the types of contaminants.
  • H. Sugawara has recently summarized the current state of the art of UVC inactivation of this virus in regard to plasma products (H. Sugawara et al, "Inactivation of parvovirus B19 in coagulation factor concentrates by UVC radiation: assessment by an in vitro infectivity assay using CFU-E derived from peripheral CD34+ cells," Transfusion 2001 Apr;41(4):456-61).
  • the net result of this and similar studies is that the UVC irradiation should preferably be on the order of 1 to 10,000 J/m 2 , more preferably 10 to 5,000 J/m 2 , and even more preferably 100 to 3,000 J/m 2 .
  • the treatment chamber must be of uniform thickness, as discussed above for the ultrasound system. Also as discussed for ultrasound systems, moving systems must have plug flow as described in first main embodiment. Unlike ultrasound systems, however, the depth of the treatment chamber is critical for
  • UVC systems The underlying problem is that the optical absorption in the treated liquid follows an exponential curve according to Beer's Law. As such, the exposure on one side of the treatment chamber can be much less than the exposure on the other side of the chamber, even for relatively thin units.
  • the UVC irradiation is therefore preferably applied from both sides of the treatment chamber. Under this arrangement, the exposure sum from the two opposing sources is thus nearly constant for moderate target widths.
  • the walls of the treatment chamber can also absorb UVC. For this reason, these walls must also be of uniform thickness, and of UVC transparent material, preferably fused quartz.
  • an integrating monitor (Model RX 003, UVItec, Cambridge, UK) to ensure that the specified exposures are actually maintained over the lifetime of the lamp. Furthermore, placement of such a monitor beside each lamp thus ensures that a bank of lamps functions as intended. In this case, at least one monitor should also be placed opposite at least one lamp, beyond the liquid being treated, to ensure that the appropriate absorption levels are maintained; this is a particularly important consideration for blood products, in which the optical properties often differ greatly from sample to sample.
  • UVC lamps produce significant amounts of heat, and while grid lamps produce relatively little heat compared to their quite high UVC intensity, their heat is still a problem. Part of this heat can be controlled by fan-assisted convection over the lamps. This approach, however, does not limit the infrared component from the lamps, which is absorbed directly within the target. For this reason, a flow or spray of water is therefore used to cool the target, since water is a very poor absorber of UVC light. To prevent this cooling water or spray, or any leaking contaminated fluid, from falling into the lamps and causing electrical problems or breakage, the treatment module and lamps should be oriented vertically. This geometry also aids the maintenance of controlled flow for continuous systems, as discussed earlier.
  • UVC treatment The final problem with UVC treatment is that after the sources have been turned off, there can be some recovery of the pathogens, apparently by an inherent repair mechanism.
  • Recent work with discharge lamps, however, (see, WH Cover, "Effect of Broad Spectrum Pulsed Light (BSPL) on Platelet Function," Cambridge Healthtech Eighth Annual Symposium on Blood Product Safety, February 4-7, 2002) indicates that other wavelengths of light can inhibit such recovery.
  • discharge lamps also produce a great deal of heat. Since mixtures of UVB and UVA are known to be more effective than either acting alone, and since the UV wavelengths are the most energetic in the emission of discharge lamps, a preferred approach is to include UVA and/or UVB sources in the treatment protocol to suppress pathogen repair.
  • any polymer that is transparent to UVC is also transparent to UVA and UVB, so the design considerations reduce to only building a system capable of UVC treatment.
  • UVC systems can be built for a variety of operating conditions and contaminated liquids.
  • the overall approach is similar to that described above for gamma irradiation systems, and can thus be used for batch, semi-continuous, and continuous operation.
  • the major difference is the limited penetration capability of UVC versus gamma irradiation, which leads to several concerns in regard to bag design.
  • the access ports and connecting tubes are typically not UVC transparent, and can thus shield the pathogens from UVC exposure.
  • the solution to this problem is similar to that described in embodiment I for ultrasound feed ports, except in this case any clamping must be done by UVC transparent materials to prevent shading effects.
  • the clamps can be made of fused quartz, but for greater mechanical strength, polymers such as Teflon® AF are preferred.
  • the clamping should be done in a section of the bag that is fully illuminated, and also elevated to drain the fluid away from the clamping location. Under this arrangement, the available light will be minimally attenuated, thereby providing maximum treatment at the clamp location.
  • the final bag problem is that for relatively large treatment volumes, the thickness must remain small, and thus the surface area must be large. Beyond the matter of costs, this restriction also causes substantial hydrostatic pressures, particularly for the vertically oriented systems described earlier.
  • One alternative is to use very heavy quartz plates to contain the treatment bags, but this approach is expensive and causes undesirable attenuation of the UVC light. It is therefore preferred that the center sections of the bags be joined front to back, thus reducing the loading on the fused quartz plates.
  • the boundary seal of the treatment bags include holes to match registration pins. These pins can be either permanently mounted in the periphery of the treatment chamber, or mounted in a plastic frame that allows for rapid insertion and removal of the bag assembly.
  • ultrasound is used to provide extremely effective vibration.
  • the underlying principle is that ultrasound can agglomerate or disperse solids in a liquid, as noted earlier in the discussion of aggregate treatment.
  • the vibrating ultrasound is provided by a horn in direct contact with the treatment bag. Specifically, this horn should be in contact with the bottom of the treatment bag, thus ensuring adequate vibration for even partially filled bags. This approach thereby avoids vibrating the entire treatment chamber, which would be difficult due to problems in impedance matching, as well as possible chipping of the expensive fused quartz plates.
  • the present invention provides a method for decontaminating a fluid, which comprises:
  • the step (a') "for the treatment of a fluid with ultrasonic energy to obtain a de-oxygenated fluid” may be carried out in the same ways that step "(a) treating a fluid with ultrasonic energy to obtain a de-oxygenated fluid” is carried out in the context of the seventh main embodiment.
  • the step (b') "for the irradiation of said de-oxygenated fluid” may be carried out in the same ways that step "(b) irradiating said de- oxygenated fluid” is carried out in the context of the seventh main embodiment.
  • PEF Planar electrostatic electrostatic field
  • tens of kV very high voltage
  • PEF very high voltage
  • the limiting factor is that the target pathogen must be sufficiently large to establish a voltage gradient. While this limitation effectively excludes pathogens on the size of viruses or smaller, many pathogens can nevertheless be treated by PEF.
  • a major limitation of PEF is that the sample may break down during treatment. In particular, dissolved gasses readily cause breakdown. For this reason, Q.H.
  • This embodiment relies on a synergistic effect between the application of ultrasonic energy and PEF.
  • the underlying principle of PEF is that the various fields can act on charged species, or even upon polar molecules.
  • Constant electric and magnetic fields are the simplest to analyze and implement. Beginning with electric fields, there is a long history of applying a current through a contaminated liquid to affect some kind of cleaning treatment. The overall approach is simply to put two electrodes on opposite sides of a pool of liquid and then apply electricity. The essential problem of this approach is that electrochemical reactions, primarily at the electrodes, can contaminate the product. This is a critical concern for biological materials, such as plasma, that will be used for medical treatment.
  • a new means of avoiding this problem is to use a salt bridge across a sterile filter to couple the electrodes to the fluid to be treated.
  • a thin tube leads from the filter and salt bridge, extending to the sides of the treatment bag.
  • a flow restriction is placed at the juncture of the tube and bag. After treatment, this tube is then heat sealed at the flow restriction, and the filter and salt bridge may then be discarded. Under this arrangement, undesired compounds are first trapped at the leading edge of the salt bridge. Any residual compounds that escape this trap are then caught in the connecting tubes before they reach the bulk of the fluid.
  • PEF pulsed electric field
  • PEF typically involves very high voltages, on the order of 20kV, but very short duration.
  • PEF and ozone are known to have a synergistic effect (R Unal, JG Kim, and AE Yousef, "Inactivation of Escherichia coli 01 57:H7, Listeria monocytogenes, and Latobacillus leichmannii by combinations of ozone and pulsed electric field," J. Food Prof, Jun;64(6), pp. 777-782 (2001)).
  • PEF is therefore combined with the above salt bridge and tube arrangement.
  • the preferred location to apply PEF is thus immediately after the degassing step.
  • PEF can be done before, during, or after UVC or gamma irradiation (as described below).
  • UVC or gamma irradiation as described below.
  • the improved speed of ultrasonic vacuum degassing is of immense use in the PEF treatment of foodstuffs.
  • a tenth main embodiment such a fluid may be effectively decontaminated by a method involving: (a') a step for the treatment of a fluid with ultrasonic energy to obtain a de- oxygenated fluid; and (b') a step for contacting said de-oxygenated fluid with a pulsed electric field.
  • the step (a') "for the treatment of a fluid with ultrasonic energy to obtain a de-oxygenated fluid" may be carried out in the same ways that step "(a) treating a fluid with ultrasonic energy to obtain a de-oxygenated fluid" is carried out in the context of the ninth main embodiment.
  • step (b') "for contacting said de- oxygenated fluid with a pulsed electric field” may be carried out in the same ways that step “(b) contacting said de-oxygenated fluid with a pulsed electric field” is carried out in the context of the ninth main embodiment.
  • the present invention provides a method for decontaminating a fluid, which comprises:
  • decontamination is best achieved by applying multiple, independent processes.
  • the pathogens that escape one decontamination technique may not escape a second, or third, technique etc.
  • any given technique can destroy only a limited number of pathogens before it also causes a significant amount of protein damage, so it is preferable to use multiple techniques at partial power instead of one technique carried to extreme limits. It is therefore desirable to integrate the above technologies with yet another independent technique.
  • a particularly useful such technique is ozone exposure.
  • Ozone is a triatomic molecule of oxygen, while the common form of oxygen is diatomic.
  • ozone is an unstable molecule and is thus extremely reactive.
  • this high reactivity makes ozone an extremely strong decontamination agent.
  • the underlying mechanism is that ozone rapidly attacks the complicated protein structures that pathogens require to propagate, thus causing rapid inactivation.
  • An additional benefit is that after it has reacted, ozone then reverts to non-toxic molecules that are naturally present. As such, ozone and its products do not have to be removed from the treated material, thus saving a separate, expensive, and time-consuming step that is typically required for other decontaminating agents.
  • ozone has been used for many years in a variety of decontamination devices. Some of these applications involve treatment of a given area or volume by gas exposure, such as a room or a device inside an enclosure. While these applications are quite numerous, the main concern of the present invention is liquid treatment, whereby the ozone is absorbed into an aqueous solution. Such aqueous solutions, in turn, have many particular applications. In this regard, the most common use of ozone decontamination is for processing water, including both potable water treatment and pollution control. While some aspects of the present invention are applicable to such processes, however, the main application here is the decontamination of biological products, particularly blood products. But even in this somewhat restricted discipline, several such devices have already been disclosed.
  • U.S. Patent No. 4,632,980 discloses an ozone blood treatment device, in particular a technique for controlling damage to blood products while preferentially attacking enveloped viruses.
  • enveloped viruses such as HIV were of primary concern in the early 1980's when this patent was under development, the subsequent development of advanced viral testing and the emergence of more non-enveloped viruses have greatly changed the needs of the blood industry.
  • Another problem is that although this patent mentions pH, the pH of blood and blood products depends strongly on the choice of anticoagulant, and as discussed later, the pH strongly affects the behavior of dissolved ozone.
  • ozone can also be used for XCT, or extraco ⁇ oreal treatments, as disclosed in U.S. Patent No. 6,027,688.
  • blood is withdrawn from the patient, treated, and then re-infused, with the intent of reducing the HIV burden.
  • This device is quite complicated and would thus be expensive to buy and operate.
  • this device also has a glass treatment tube, which, as discussed above, could cause severe clotting, and thus lead to pulmonary embolism and death.
  • the disclosed 99% (or log 2) viral reduction is quite small compared to the log 6 or 7 that is desired.
  • the concentration of the input ozone gas is the concentration of the input ozone gas: higher concentrations are preferable because they yield more collisions among the reacting species, and thus yield more product in less time.
  • the limiting factor here is that gaseous ozone concentrations of greater than 20% are explosive. At lesser concentrations, a number of practical concerns limit the effective concentrations that can be achieved. The main such concern is that ozone is so reactive that it cannot be stored for prolonged time periods. Instead, ozone is typically generated at the site where it is to be used.
  • Ultraviolet light sources function by first splitting diatomic oxygen molecules into singlet oxygen, which then reacts with other diatomic molecules to form triatomic ozone. Although this process is the origin of the earth's ozone layer, ultraviolet exposure is inefficient. The underlying problem is that while certain frequencies of light are quite effective in generating ozone, other frequencies are almost as effective at dissociating the formed ozone. As a result of these competing processes, UV units are limited to concentrations of less than 1%. Although UV sources are quite clean and easy to control, this low ozone concentration limits their use in decontamination work.
  • An alternative ozone generation technique is corona discharge.
  • oxygen is passed through a channel bound by high voltage electrodes.
  • the resulting discharge ruptures the diatomic molecules, and some of the resulting, high energy single oxygen molecules react with some of the neighboring oxygen molecules to form ozone.
  • Typical yields are in the range of 1 to 15% by volume ozone.
  • corona discharge systems there are several problems with corona discharge systems.
  • One such problem is that the feed gas may also contain nitrogen, water vapor, or other gasses. If so, there is a possibility that molecules other than oxygen and ozone may contaminate the product. Medical applications of discharge systems therefore typically use high grade oxygen as a feedstock, but this entails additional costs.
  • Medical applications of discharge systems therefore typically use high grade oxygen as a feedstock, but this entails additional costs.
  • degradation of the electrodes may introduce solid contaminants into the product; therefore, expensive filters are required.
  • the eroding electrodes also produce electromagnetic noise, which is undesirable in a medical environment.
  • Yet another problem with discharge systems is that the resulting gas is so hot and dry that it can damage the proteins being treated.
  • electric discharges are difficult to control, particularly at partial load operation. For these reasons, corona discharge units can be used for decontamination work, but a great deal of conditioning is necessary.
  • U.S. Patent No. 5,709,992 discloses one such technique for adding activated ceramic particles directly into the pool to be treated.
  • U.S. Patent No. 5,989,407 by Lynntech, Inc. discloses a device that produces ozone at concentrations of 10 to 15%. This device works on the principles of electrolysis of water, thereby avoiding the cost of expensive medical grade oxygen as a feedstock while also avoiding the problem of electromagnetic noise.
  • the ozone produced by this device is self-pressurizing, relatively cool, and fully humidified.
  • electrochemical units are currently the preferred ozone sources for the present invention.
  • electrochemical ozone generators are described in detail, along with the modifications and extensions that are necessary that are necessary to exploit these features in practice.
  • the ozone must be either generated at elevated pressure or generated at lower pressure and then compressed. Because compressing ozone is difficult and expensive, the above-noted ability to generate ozone at pressure is a major advantage of electrochemical ozone units.
  • the available electrochemical units are not yet capable of generating ozone at pressures beyond about 50 psi. It is therefore necessary to compress the gas to reach higher pressures.
  • the preferred means of compressing ozone is a diaphragm pump (BA series, Fluitron, Ivyland, PA). Diaphragm pumps are useful because they have no seals that can be destroyed by ozone contact, and the entire flow path can be constructed of easy to clean, chemically inert materials.
  • the limiting factor in the design of such compressors is that ozone decomposes at higher temperatures. It is therefore necessary to start with the ozone as cold as possible, and then compress the ozone through multiple stages. At each stage, the temperature should not exceed about 40 °C.
  • the maximum compression ratio in each stage can be calculated by the standard adiabatic ideal gas laws. Water cooling of the compression heads thus ensures that the peak temperatures are kept well below the theoretical upper limit. While effective, such compressors are quite expensive, and will be discarded as advances are made in electrochemical generation. In the meantime, the output from the existing generators provides a cool, partially compressed feedstock.
  • the 50 psi output from an electrochemical ozone generator can be fed directly into a two stage diaphragm pump, with each stage operating at a pressure ratio of 1.7: 1. The resulting gas is therefore at a pressure of about 150 psi, with a peak temperature of less than 35 °C.
  • the ozone must be applied to the material to be treated.
  • one option is simply to add the pressurized ozone into a sterile blood bag system.
  • the difficulty here is that conventional bags are not designed to handle such pressures. Although new bags could be built, they would be much more expensive than conventional units. Even then, should a bag rupture during treatment, it would then spray potentially contaminated plasma throughout the laboratory. Finally, from a practical standpoint, generating sufficient ozone to pressurize the bag would be expensive and time- consuming, and would waste much of the valuable ozone that could otherwise be used for attacking the contaminants.
  • This system consists of a pressure cell, which is driven by a standard air compressor. By pressurizing this cell with air to the same pressure as the ozone, the pressure on both sides of the treatment bags is equalized.
  • Cheap bags can thus be used, there is no risk of rupture, and the ozone requirements are greatly reduced.
  • pressures up to about 10 atmospheres (about 150 psig) can be achieved easily, and if desired even higher pressures can be generated by 220 VAC equipment.
  • ozone During the time that ozone is not required, such as during bag changes or ultrasonic vacuum degassing, it is desirable to maintain the ozone source at pressure so that processing can be continued immediately when necessary.
  • the pressure is typically maintained in a simple storage tank. Ozone, however, degrades so rapidly that this is not an option.
  • electrolytic units such as the Lynntech device, must be operated continuously for best output. For these reasons, the use of a bypass circuit is preferred.
  • the first part of this circuit is a solenoid valve placed at the outlet of the ozone generator. When activated, this valve diverts the ozone around the treatment vessel and through a check valve that maintains the desired pressure.
  • electrolytic units it is necessary to balance the pressure loading on the generator cell. Specifically, electrolytic units produce ozone and oxygen on one side of this cell, and hydrogen on the opposite side. In practice, the hydrogen back pressure can be maintained simply by a check valve. Downstream processing can then be done at approximately atmospheric pressure, using a simple drain trap for water and an optional hydrogen destruct unit. This equipment can thus be conveniently arranged parallel to the ozone bypass circuit.
  • the next concerns are temperature and humidity, which are interdependent.
  • the potential temperature problem here is that the gas may be so hot or cold that it damages the proteins.
  • the humidity can be so low that the proteins could be excessively dried, or so high that the proteins could be diluted with excess moisture.
  • a Peltier system For precise control of the ozone temperature, a Peltier system is desirable (Model TLC-1400, TEC A, Inc., Chicago, IL). Alternatively, conventional refrigeration and heating devices can also be used if proper controls are provided (Model RTE, Neslab, Portsmouth, NH). Both systems provide a source of either heated or cooled water. Connecting a heat exchanger to these devices therefore provides a simple means of regulating the ozone temperature.
  • Teflon® or similar plastic tube to connect the ozone source to the treatment unit.
  • Teflon® is desirable in these applications because it is quite resistant to attack by ozone. Although Teflon® is somewhat permeable to gasses, these losses are not excessive. If desired, however, lower permeability forms of Teflon®, notably Teflon® PFA, can also be used.
  • Teflon® laminate with a low permeability plastic can also be used.
  • Placing loops of the selected tubing in the controlled temperature bath thus provides the desired ozone heating or cooling.
  • metal heat exchangers could also be used, but in this case it is necessary to protect the metal surface from attack by the highly reactive ozone.
  • An effective protective layer is Teflon®.
  • stainless steel tubing can be used. In particular, tubing that has been treated with nitric acid rapidly forms an inert layer that resists further corrosion, without greatly reducing the heat flow (Stainless steel tubing: Nitric acid treated, Upchurch Scientific, Oak Harbor, WA).
  • a water trap is used to collect and remove any condensates in high humidity systems. The problem here is that the condensed water must be removed without losing the system pressure.
  • the first part of the water trap is a small pressure vessel.
  • This vessel is connected to a scale, float, optical or electrical sensor to determine the level of the water in the trap.
  • a solenoid valve is then actuated to release the pressurized liquid into a drain.
  • a "flip-flop" circuit is used to keep the valve actuated during the entire draining process, with the state of the circuit reversed by a switch placed at the closing location.
  • This valve should have Teflon® flow surfaces to resist attack by the ozone. Also, for minimal energy consumption, this valve should be "normally closed.” Downstream of this valve, a flow restriction must be placed in the outlet tube to prevent excessive spraying at elevated pressures.
  • An alternative approach is to use a peristaltic pump, if it is necessary to transport the liquid to a higher level than the ozone pressure can support. In either case, the drain must be closed before the trap is completely empty; otherwise, there will be some leakage of the ozone gas.
  • a device to increase the humidity to the required level replaces the water trap. In this case, a source of high purity water is required, as well as some means to vaporize this water at low temperatures. Sonic humidifiers are well suited for this application.
  • the ozone is thus delivered to the treatment chamber at the proper concentration, pressure, temperature and humidity.
  • these conditions are quite effective in decontamination, it is possible to accelerate the process even more by inco ⁇ orating the previously described degassing technology.
  • each gas has its own characteristic solubility in a given liquid; furthermore, Fick's first law describes the diffusion of this gas in the liquid, while Henry's law describes the concentration of this gas in the liquid, relative to the partial pressure of this gas above the liquid.
  • water or a dilute aqueous solution thus has an oxygen concentration of about 35%, and a nitrogen concentration of about 63%; these values differ from the respective 21 ) and 78% values in air because oxygen is more soluble than nitrogen.
  • the nitrogen concentration When subsequently exposed to a saturated mixture of 15% ozone and 85% oxygen, the nitrogen concentration then decreases as the liquid takes up ozone and oxygen.
  • Ozone is about 13 times more soluble than oxygen, so the uptake of oxygen is more rapid.
  • ozone reacts with the liquid in which it is dissolved.
  • some of the oxygen in the ozone combines with some of the other components in the solution, and some of the remaining oxygen reverts to the normal, diatomic form. In either case, the incoming ozone eventually completely reacts to a lower energy form, leaving a decontaminated liquid that is enriched with oxygen, and depleted from other gasses.
  • ozone reacts with the liquid in which it is dissolved.
  • ozone has a half-life in once-distilled water of about 20 minutes, but a half-life of 80 minutes or more in water that has had multiple distillations.
  • acids or neutral salts increase the solubility of ozone and extend the half-life of the solution.
  • alkalis decrease the solubility of ozone (see, Atherton Seidell, Solubilities of Inorganic and Organic Compounds. Van Nostrand, New York, p. 473, 1919).
  • gaseous concentrations can be on the order of hundreds or even thousands of parts per million.
  • Conventional liquid concentrations are on the order of 0.3 to 10 mg/L. This range is also the base for the present invention, but the peak transient concentrations are on the order of 100 to 200 mg/L.
  • the present invention utilizes the previously described degassing equipment to remove the excess ozone as soon as the decontamination is completed.
  • contactors To be effective, the contactor must be designed to match the properties of the fluid being treated. For example, in the preparation of drinking water or in the treatment of toxic wastes, high levels of turbulence and shear can be tolerated without concern for damaging the liquid being processed; furthermore, the contactors can be fabricated from any reasonably strong construction material. As noted above, however, protein systems, notably those involving blood products, must be handled much more carefully, and plastics that will not induce the clotting sequence must be used instead of materials such as glass.
  • FIG. 11 A particularly preferred chamber for the contacting of platelets is shown in Figure 11; the treatment chamber consists of a rectangular or similarly shaped block 1101 with staggered, opposed shelves in the shape of sha ⁇ wedges 1102. With the chamber in the horizontal position, the liquid enters a trough or inlet port 1103 along one side. After filling this trough, the chamber is then rotated upwards to about 80 degrees, at which point the fluid flows over the first shelf 1102a towards the opposing wall. Because the shelf does not actually touch the opposing wall, however, the fluid drops down to the next shelf 1102b and the flow then reverses. Meanwhile, ozone is introduced through ports 1104. Note that this arrangement is unlike the above cited patents because the flow reversal thoroughly mixes the material at each step, with the top layer becoming largely the bottom layer and vice versa.
  • Additional enhancements of this device include an ultrasonic driver to improve the rate of fluid flow and to aid in the mixing of the ozone; a pressurized treatment cell; and an ultrasonic degassing option with vacuum assist.
  • FIG. 5 This device employs an ultrasonic spray nozzle at the top of an enclosed chamber. Unlike the previously described spray system (US Patent No. 5,882,591), this device has no electrostatic fields, but it does inco ⁇ orate elevated pressures, direct ultrasonic processing of the sprayed fluid, and other enhancements described more fully below.
  • this contactor The key feature of this contactor is that the fluid to be treated flows through an enclosed channel. Ozone is then fed into the liquid through a series of small holes in the channel wall. While this arrangement thus has some similarities to conventional gas/liquid bubbling devices, there are however significant differences. Specifically, the ability to rotate the chamber aids in the bulk mixing of the fluid, thereby providing more uniform treatment. This mixing is further aided by the ultrasound, but ultrasound also has other important effects in the present application. Historically, the use of ultrasound to aid ozone contacting has been discussed by W.S.
  • ultrasound will drive small bubbles into the liquid, but larger bubbles will grow to the point that they can be removed.
  • ultrasound is coupled with a unique contactor to force as much ozone into the solution as possible.
  • the contactor in the present invention is driven directly by ultrasound.
  • this ultrasound is delivered by a high amplitude horn, so that the oscillations are large in displacement.
  • this displacement is larger than the diameter of the holes through which the ozone flows. The net result is that the ultrasound shears off extremely small bubbles into the surrounding liquid. Being much smaller than resonance, or even stable, size these small bubbles are then forced into the liquid rapidly under the action of ultrasound.
  • UV light is a particularly useful measurement technique (Ocean Optics Model 2000, Dunedin, FL).
  • Teflon® is ideal, but expensive; lower quality plastics may be used if a very bright UV source is available.
  • the major limitation in this technology is the presence of gas bubbles. Although present as a result of decontamination, these bubbles are a significant problem in the measuring process because they are optically quite different from the concentrated ozone solution. To minimize this problem, the measurement cell may be made with a wide top and a narrow bottom. Under this geometry, the gas bubbles rise to the surface, leaving only liquid in the path ofthe measuring UV light beam.
  • the present invention provides a method for decontaminating plasma by:
  • step (b') a step for the treatment of said de-oxygenated fluid with ozone.
  • the step (a') "for the treatment of a fluid with ultrasonic energy to obtain a de-oxygenated fluid” may be carried out in the same ways that step "(a) treating a fluid with ultrasonic energy to obtain a de-oxygenated fluid” is carried out in the context of the eleventh main embodiment.
  • the step (b') "for the treatment of said de-oxygenated fluid with ozone” may be carried out in the same ways that step "(b) contacting said de-oxygenated fluid with ozone” is carried out in the context ofthe eleventh main embodiment.
  • the present invention provides a method for decontaminating a fluid by:
  • the treatment ofthe fluid with the ultrasonic energy may be carried out in the same ways and by using the same apparatus as described above in the context ofthe first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, and twelfth main embodiments.
  • ultrasonic energy is used to enhance the decontamination effect of ozone.
  • ozone treatment is a standard decontamination technique. While quite effective, however, ozonation unfortunately suffers from relatively long treatment times.
  • This thirteenth main embodiment ofthe present invention therefore uses ultrasonic energy to accelerate the ozone decontamination process, and to improve the overall effectiveness ofthe combined system. It is already known that ultrasound is effective in enhancing the speed of bacterial decontamination (see, W.S. Masschelein, "Handbook of Ozone Technology and Applications. Volume One.” R. G. Rice and A.
  • the unique aspects of applying ultrasound to a liquid containing dissolved ozone is that the above conditions and techniques are applied to protect the proteins in solution during the decontamination process.
  • this embodiment will be described in the context of plasma, and amounts to first mixing the ozone with the plasma. Next, the ozone-containing plasma is then treated with ultrasonic energy. In this embodiment, the plasma is treated with ultrasonic energy as described above.
  • the term "treating said ozone-containing fluid with ultrasonic energy” does not require that the application of ultrasonic energy to the ozone-containing fluid commence after the introduction of ozone into the fluid has ceased.
  • this term means that the application of ultrasonic energy to the ozone- containing fluid may commence: (1) prior to the commencement ofthe introduction of ozone into the fluid; (2) at the time the introduction of ozone into the fluid is commenced; (3) after the introduction of ozone into the fluid has commenced; or (4) after the introduction of ozone into the fluid has ceased.
  • the ultrasonic energy is applied to the fluid during the entire time that the ozone is introduced into the fluid.
  • the present invention provides a method for decontaminating a fluid by:
  • step (a') a step for mixing a fluid with ozone, to obtain an ozone-containing fluid; and (b') a step for the treatment of said ozone-containing fluid with ultrasonic energy.
  • the step (a') "for mixing a fluid with ozone, to obtain an ozone-containing fluid" may be carried out in the same ways that step "(a) mixing a fluid with ozone, to obtain an ozone-containing fluid" is carried out in the context ofthe thirteenth main embodiment.
  • step (b') "for the treatment of said ozone-containing fluid with ultrasonic energy" may be carried out in the same ways that step “(b) treating said ozone-containing fluid with ultrasonic energy” is carried out in the context ofthe thirteenth main embodiment.
  • the present invention provides a method for decontaminating a fluid by:
  • the fifteenth main embodiment is essentially a combination ofthe eleventh and thirteenth main embodiments.
  • the fluid is first degassed using ultrasonic energy as discussed above.
  • the degassed fluid is then contacted with ozone, and the ozone-containing fluid is treated with ultrasonic energy to enhance the reactivity ofthe ozone, as described in the ninth main embodiment.
  • the present invention provides a method for decontaminating fluid by:
  • step (b') a step for the treatment of said de-oxygenated fluid, to obtain an ozone- containing fluid; and (c') a step for the treatment of said ozone-containing fluid with ultrasonic energy.
  • step (a') "for the treatment of a fluid with ultrasonic energy to obtain a de-oxygenated fluid" may be carried out in the same ways that step "(a) treating a fluid with ultrasonic energy to obtain a de-oxygenated fluid" is carried out in the context ofthe fifteenth main embodiment.
  • step (b') "for the treatment of said de-oxygenated fluid, to obtain an ozone-containing fluid" may be carried out in the same ways that step "(b) contacting said de-oxygenated fluid with ozone, to obtain an ozone- containing fluid” is carried out in the context ofthe fifteenth main embodiment.
  • step (c') "for the treatment of said ozone-containing fluid with ultrasonic energy” may be carried out in the same ways that step "(c) treating said ozone-containing fluid with ultrasonic energy” is carried out in the context ofthe fifteenth main embodiment.
  • the present invention provides a method for decontaminating a fluid by: (a) treating a fluid with ultrasonic energy to obtain a de-oxygenated fluid;
  • This seventeenth main embodiment represents a combination ofthe irradiation and ozone-treatment embodiments described above.
  • this main embodiment represents a combination ofthe fifth and seventh main embodiment, and the steps (a), (b), and (c) may be carried out in the same ways and with the same apparatus described above.
  • combined decontamination processes are quite attractive because they produce very high log reduction rates.
  • the ozone-treatment decontamination may either precede or follow the irradiation decontamination.
  • ultrasonic degassing is very effective, it is generally not desired to add extra dissolved oxygen species in the ozone process before subsequently removing them in the irradiation process.
  • performing the ozone-treatment decontamination after the irradiation decontamination would allow any residual ozone additional time to react with the pathogens, which would thus improve the overall kill effectiveness.
  • this method comprises: (a") treating plasma with ultrasonic energy to obtain deoxygenated plasma;
  • the present invention provides a method for decontaminating a fluid by:
  • step (a') a step for the treatment of a fluid with ultrasonic energy to obtain a deoxygenated fluid;
  • step (b') a step for the irradiation of said de-oxygenated fluid, to obtain an irradiated fluid; and
  • step (c') a step for the treatment of said irradiated fluid, to obtain an ozone-containing fluid.
  • the step (a') "for the treatment of a fluid with ultrasonic energy to obtain a de-oxygenated fluid" may be carried out in the same ways that step "(a) treating a fluid with ultrasonic energy to obtain a de-oxygenated fluid" is carried out in the context ofthe seventeenth main embodiment.
  • step (b') "for the irradiation of said de-oxygenated fluid, to obtain an irradiated fluid” may be carried out in the same ways that step “(b) irradiating said de-oxygenated fluid, to obtain an irradiated fluid” is carried out in the context ofthe seventeenth main embodiment.
  • step (c') "for the treatment of said irradiated fluid, to obtain an ozone-containing fluid” may be carried out in the same ways that step "(c) contacting said irradiated fluid with ozone, to obtain an ozone-containing fluid” is carried out in the context ofthe seventeenth main embodiment.
  • the present invention provides a method for decontaminating a fluid by:
  • This embodiment represents another combination ofthe irradiation and ozone- treatment embodiments described above.
  • this nineteenth main embodiment represents a combination ofthe fifth and thirteenth main embodiments, and the steps (a), (b), (c), and (d) may be carried out in the same ways and with the same apparatus described above.
  • the ozone-treatment decontamination may either precede or follow the irradiation decontamination.
  • ultrasonic degassing is very effective, it is generally not desired to add extra dissolved oxygen species in the ozone process before subsequently removing them in the irradiation process.
  • performing the ozone-treatment decontamination after the irradiation decontamination would allow any residual ozone additional time to react with the pathogens, which would thus improve the overall kill effectiveness.
  • the present invention provides a method for decontaminating a fluid by: (a') a step for the treatment of a fluid with ultrasonic energy to obtain a deoxygenated fluid; (b') a step for the irradiation of said de-oxygenated fluid, to obtain an irradiated fluid; (c') a step for the treatment of said irradiated fluid, to obtain an ozone-containing fluid; and
  • step (d') a step for the treatment of said ozone-containing fluid with ultrasonic energy.
  • step (a') ' for the treatment of a fluid with ultrasonic energy to obtain a de-oxygenated fluid may be carried out in the same ways that step "(a) treating fluid with ultrasonic energy to obtain a de-oxygenated fluid” is carried out in the context ofthe nineteenth main embodiment.
  • step (b') "for the irradiation of said de-oxygenated fluid, to obtain an irradiated fluid” may be carried out in the same ways that step "(b) irradiating said de-oxygenated fluid, to obtain an irradiated fluid” is carried out in the context ofthe nineteenth main embodiment.
  • step (c') "for the treatment of said irradiated fluid, to obtain an ozone-containing fluid" may be carried out in the same ways that step "(c) contacting said irradiated fluid with ozone, to obtain an ozone-containing fluid” is carried out in the context ofthe nineteenth main embodiment.
  • step (d') “for the treatment of said ozone-containing fluid with ultrasonic energy” may be carried out in the same ways that step "(d) treating said ozone-containing fluid with ultrasonic energy” is carried out in the context ofthe nineteenth main embodiment.
  • the present invention provides an apparatus for decontaminating a fluid, comprising:
  • a source of ultrasonic energy coupled to the chamber, wherein said chamber comprises (i) a flat panel, (ii) an inlet, and (iii) an outlet; and wherein said flat panel of said chamber and said inlet are dimensioned such that a fluid flowing through said inlet and across said flat panel to said outlet will form a thin film and travel in plug flow at least during some portion of its flow across said flat panel.
  • any surface ofthe chamber which comes into contact with the plasma should be constructed of materials that will have no have no deleterious effect on the fluid, especially when the fluid is plasma. Suitable materials for those portions ofthe chamber which come into contact with the plasma are as specified by FDA for contact with blood. Although not an absolute requirement, it is preferred that at least a portion ofthe chamber be constructed of a transparent material to permit visual inspection ofthe decontamination process.
  • PVC polyolefin bags under development. The main concern with these new materials is that the plasticizer may leach out over time. For the present methods, however, the contact time is quite short. On the other hand, the sonification may accelerate the leaching process. However, because tests to date show no measurable degradation, there appears to be no unique restrictions for the present method and apparatus.
  • the chamber is configured to contain a flat panel or plane at the bottom.
  • a flat panel or plane at the bottom.
  • the flat panel would be approximately 25 by 25 cm.
  • continuous, large scale units for pool processing would have planar sections on the order of several meters.
  • the chamber also contains an inlet and an outlet.
  • the inlet is preferably located near the bottom ofthe chamber and extends along the width of one end ofthe flat panel.
  • the inlet is preferably a divergent spreader to assist in forming the plasma into a thin film as it flows across the flat panel at the bottom ofthe chamber.
  • the height ofthe inlet is preferably dimensioned such that plasma forms a thin film.
  • the exact thickness ofthe film is not by itself critical. All that is required is that the gas bubbles reach the surface relatively quickly. In the case of very durable proteins, this is not even a consideration. For less durable proteins and cells, a thickness of 2 to 20 mm, preferably 2 to 10 mm, and more preferably then 2 to 4 mm may be used. This is by no means precise, and it is possible the thickness may be varied by simply changing the vacuum settings, power, etc. and then tuning to a different range.
  • the creation of plug flow is well known (John A. Roberson, Clayton T. Crowe,
  • the dimensions of inlet and the flat panel are preferably adjusted such that the plasma flows across the flat panel in plug flow.
  • the ratio ofthe length ofthe flat panel to the width ofthe inlet is less than twenty, preferably less than fifteen, more preferably less than about ten.
  • the inlet is connected to a device for controlling the flow rate of plasma across the flat panel.
  • the flow ofthe fluid may be controlled as follows.
  • the treatment range includes plasma, as well as platelets and erythrocytes (red blood cells).
  • all blood applications should include a means to remove the white blood cells
  • leukocytes While leukocytes are obviously useful in the donor, transfusion of these cells can result in a number of adverse immune reactions. Even worse, these cells also present an opportunity to transmit diseases, notably nvCJD (new variant Creutzfeldt- Jacob Disease). For this reason, these cells should either be destroyed, or preferably, removed.
  • nvCJD new variant Creutzfeldt- Jacob Disease
  • plasma should be heated to about 53 °C for one hour. This procedure alone kills many viruses. Another advantage of this heating is that the dissolved oxygen drops rapidly at such elevated temperatures. Another advantage is that cavitation is much easier at elevated temperatures. The actual heating method is not critical, as long as it is reasonably fast. There are several blood, plasma, and IV solution warmers on the market capable of providing the necessary heating.
  • the fluid to be treated is in a bag, which may or may not be heated.
  • the next task is to get this fluid into the degassing unit.
  • a peristaltic pump While quite effective for robust materials such as plasma, rbc's and platelets would however suffer severe degradation because only finely spaced rollers on a very thin tube can achieve the required low, steady flow rate. This arrangement, unfortunately, would cause excessive pumping damage to the entrained cells. Furthermore, the vacuum on the discharge side would exacerbate the pumping damage. For these reasons, cellular systems will use a body force system for fluid flow.
  • the suction from the vacuum system will provide the overall driving force.
  • the flow will be retarded by several techniques.
  • One option is to use a very narrow tube, thus causing frictional losses.
  • Another option is a flow restriction, such as a pin hole in an occluding membrane.
  • a third option is to place the inlet bag below the degassing unit, so that the suction must overcome gravity.
  • a fourth option is to include the bag inside a partial vacuum system, so that the pressure difference between the degassing and feed side can be controlled.
  • a fifth option is a variable screw arrangement, which can be tightened or loosened as necessary to control the flow through the connecting tube. All of these approaches, as well as other standard metering techniques, can be applied.
  • the only remaining concern is how to control the process in practice.
  • the problem here is that the vacuum must be established, the ultrasound made ready, the UV lamps warmed, etc., before the liquid is drawn into the system.
  • the necessary control can be achieved by placing a shut off valve on the feed tube. For complete automation, this valve may be controlled electronically.
  • the inlet is configured to be connected to the outlet of an individual apheresis donation unit.
  • the device for controlling the flow rate ofthe plasma may be contained within the individual apheresis donation unit itself or located between the inlet and the individual apheresis donation unit.
  • the inlet may be configured to be easily connected and disconnected to any plasma container, such as a plasma bag.
  • the outlet is also preferably located near the bottom ofthe chamber at the end ofthe flat panel opposite that ofthe inlet. In any event, the outlet is positioned such that the plasma flowing across the flat panel will exit the chamber through the outlet after having traversed the flat panel.
  • the outlet is configured so as to be easily connected and disconnected to a container for receiving the decontaminated plasma.
  • a container may range in size from many hundreds or even thousands of liters for apparatus used for the continuous decontamination of large pools of plasma units to as small as a few hundreds or even tens of ml for apparatus used to decontaminate individual units.
  • the chamber includes a second outlet which is in communication with a vacuum source, such as a vacuum pump.
  • the second outlet is preferably located near the top ofthe chamber or at least above the top ofthe plasma layer, such that plasma is not sucked into the second outlet when a vacuum is applied to the chamber through the second outlet.
  • the vacuum source can provide a vacuum to the space above the thin film of plasma in the chamber of 2 to 100 mbar, preferably about 10 to 80 mbar, more preferably 20 to 60 mbar.
  • the apparatus comprises a liquid trap with a sterile filter located between the second inlet and the vacuum source.
  • the source of ultrasonic energy may be any which is capable of generating ultrasonic energy having the desired frequency and intensity.
  • ultrasound generators include those described above.
  • the source ofthe ultrasonic energy is coupled to the chamber such that the desired intensity and frequency of ultrasonic energy may be applied to the thin film of plasma flowing across the flat panel.
  • the apparatus comprises an ultrasound driver located beneath the flat panel.
  • the apparatus comprises a water jacket located between the ultrasound driver and the flat panel.
  • the apparatus comprises a resonator plate located between the ultrasound driver and the water jacket.
  • the water jacket is preferably connected to a cooling and circulation system such that cooled water circulates through the water jacket when the ultrasonic energy is being applied to the plasma.
  • the present apparatus may further comprise additional sensors and data loggers to ensure regulatory compliance.
  • additional sensors may include a hydrophone to ensure adequate cavitation or degassing, thermocouples to ensure adequate temperature maintenance, digital scales on the input and output bags to ensure proper flow rates as functions of time, and bar code readers and data printers to maintain a traceable path. Direct radical detection and recording are also possible.
  • the hydrophone and thermocouples should be located in the chamber such that they are in communication or contact with the thin film of plasma as it flows across the flat panel.
  • the apparatus may be constructed such that all ofthe components are permanent or semi-permanent, i.e., such that all or most ofthe components are intended to be used repeatedly for the processing of large amounts of plasma.
  • the apparatus may be divided into a permanent or semi-permanent subunit and a disposable subunit.
  • the permanent or semi-permanent subunit is constructed such that all or most of the components are intended to be used repeatedly for the processing of large amounts of plasma.
  • the permanent or semi-permanent subunit may comprise:
  • the permanent or semi-permanent subunit may further comprise other fixed hardware, including a peristaltic pump, a water jacket, and a vacuum pump.
  • the peristaltic pump is positioned such that it can be used to control the flow rate of plasma through the disposable unit.
  • the water jacket is positioned such that it will be between the ultrasound driver and the chamber when the chamber is placed in the region designed to accept it.
  • the vacuum pump is placed such that it can supply a vacuum to the gas above a thin film of plasma flowing through the chamber when the chamber is placed in the region designed to accept it.
  • the permanent or semi-permanent subunit may further optionally comprise a resonator plate which is positioned such that it will be located between the water jacket and the ultrasound driver.
  • the disposable subunit may comprise: (1) a chamber, wherein said chamber has a flat panel, an inlet, and an outlet, and wherein said flat panel of said chamber and said inlet are dimensioned such that plasma flowing through said inlet and across said flat panel to said outlet will form a thin film and travel in plug flow.
  • the disposable unit may further comprise a second outlet which may be connected to the vacuum pump ofthe permanent or semi -permanent subunit for supplying a vacuum to the gas above the plasma.
  • Figure 3 shows a decontamination system 30 designed for use in a method in which the plasma is decontaminated by the application of ultrasonic energy without application of UVC radiation or subsequent ozone treatment.
  • the plasma enters the system from a plasma bag 31 or other source on the left, with the flow rate ofthe plasma controlled by a peristaltic pump 32.
  • the plasma flow then crosses a divergent spreader 33, thus yielding a uniform plug flow of a thin film of plasma across the flat panel at the bottom of chamber 34, toward the collection bag 311.
  • the plasma enters the unit from the left, the flow controlled by a peristaltic pump 42.
  • Plug flow is achieved by keeping the planar section short relative to the entrance zone, which guarantees continuous plug flow in this design, using the general fluid mechanics rule that a flow section of about 20 times the inlet width is required to develop laminar flow under non-turbulent, low Reynolds numbers.
  • the flow rate ofthe plasma across the flat panel is as described above.
  • the ultrasonic energy is applied to the plasma by means ofthe ultrasound driver 35, which is coupled to the flat panel at the bottom of chamber 34 via a resonator plate 36.
  • the ultrasound driver 35 which is coupled to the flat panel at the bottom of chamber 34 via a resonator plate 36.
  • the sonification is driven by an ultrasonic driver 35 acting on a metal plate 36 which is resonance coupled for efficient energy transfer.
  • the temperature ofthe plasma flowing across the flat panel is controlled by the water jacket 37.
  • the water jacket 37 between resonator plate 36 and the flat panel prevents excess heat from the ultrasonic driver 35 from reaching the plasma; water is an excellent sound transmission medium, and any losses of ultrasonic energy are thus insignificant.
  • the decontaminated plasma After flowing across the flat panel, the decontaminated plasma then exits the chamber via outlet 310 and enters the collection bag 311.
  • the gas above the plasma in chamber 34 and gas evolved from the plasma during application ofthe sonic energy to the plasma are removed from the chamber by the vacuum pump 38.
  • Biological materials such as infectious agents are captured by the filter trap 39 to prevent contamination ofthe vacuum pump 38.
  • the entire process may be carried out under refrigeration, and the entire apparatus 30 or at least one or more ofthe starting plasma bag 31, chamber 34, and collection bag 311 may be contained in one or more refrigeration units.
  • Apparatus 30 in Figure 3 may be constructed as a complete permanent or semipermanent unit, with only the starting plasma and collecting plasma bags being disposable or consumable subunits.
  • apparatus 30 in Figure 3 is constructed as a permanent or semi-permanent subunit and a disposable or consumable subunit.
  • pump 32, ultrasound driver 35, resonator plate 36, water jacket 37, and vacuum pump 38 may be part ofthe permanent or semi-permanent subunit, while starting plasma bag 31, inlet 33, chamber 34, outlet 310, and collection bag 311 may be part of one or more disposable or consumable units.
  • the vacuum line including the filter trap 39 may be part of either the permanent or semi-permanent subunit or a disposable or consumable subunit.
  • the disposable or consumable units may be preferably blow molded from inexpensive plastics.
  • the collection bag preferably should meet these standards. Accordingly, it is preferred to use a conventional plasma bag.
  • any disposable parts ofthe present apparatus should have no metal parts so the disposables or consumables can be incinerated.
  • the walls ofthe chamber can be made of quite thin and/or flexible material.
  • the disposable chamber may be a bag or liner for the region which is designed to accept it.
  • the chamber is a bag or flexible liner, it may be made to hold a desired shape or to conform to the shape ofthe region designed to accept it, by applying a slight vacuum to draw the liner out to the required dimensions under differential pressure, thus allowing the use of a very cheap treatment bag as the chamber.
  • the disposable bag used as the chamber further comprises a virus tight filter at one end ofthe bag to equilibrate the pressures inside and outside ofthe bag during vacuum processing. This FDA approved component also allows for easier mounting ofthe bag inside the region designed to accept the chamber.
  • the disposable bag used as the chamber further comprises grommets at the inlet and outlet tubes to prevent them from collapsing during the application ofthe vacuum.
  • the chamber has a roughened inner surface.
  • a roughened inner surface allows the evolved gas bubbles to travel up to local spikes on the bag liner. From these points, the ultrasonic vibrations can dislodge the bubbles relatively easily. For comparison, bubbles flattened along one side of a smooth bag surface are more difficult to remove, even with agitation.
  • the apparatus may further comprise certain safety features, including electrical shielding, splash guards, and particularly a commercial ultrasound shielding enclosure.
  • the present apparatus may further comprise a device or means for detecting when a particular amount of fluid has been processed.
  • a device or means for detecting when a particular amount of fluid has been processed For example, when individual units are being processed into storage containers, it may be preferred to include a scale to detect when the storage container is full. Alternatively, an optical device which measures the level of fluid in the container may also be used.
  • a scale to measure the amount of fluid in the input container or bag.
  • mounting the input bag on a scale provides a means to measure the flow rate, given the time from the digital controller.
  • This flow rate provides information which may be used to control the opening or closing ofthe valve system.
  • the present invention provides an apparatus for decontaminating a fluid, comprising: (V) a means for containing said fluid; (2') a means for contacting said fluid with a vacuum; and
  • a means for introducing ultrasonic energy into said means for containing said fluid wherein said means for containing said fluid comprises (i) a means for the introduction of said fluid into said containing means, (ii) a means for said fluid to flow through said containing means, and (iii) a means for the removal of said fluid from said containing means; and wherein said containing means is dimensioned such that a fluid flowing through said containing means will form a thin film and travel in plug flow at least during some portion of its flow through said containing means.
  • the "means for containing said fluid” may be the same as described for the "chamber for containing a fluid” described above in the context ofthe twenty-first main embodiment; the “means for contacting said fluid with a vacuum” may be the same as the “vacuum source coupled to the chamber” described above in the context ofthe twenty-first main embodiment; and the “means for introducing ultrasonic energy into said means for containing said fluid” may be the same as the "source of ultrasonic energy coupled to the chamber” described above in the context ofthe twenty-first main embodiment.
  • all the optional and preferred components described above in the context ofthe twenty-first main embodiment may also be present in this twenty-second main embodiment.
  • the present invention provides an apparatus for decontaminating a fluid, comprising:
  • the "chamber for containing a fluid” may be the same as the "chamber for containing a fluid” described above in the context ofthe twenty-first main embodiment;
  • the "vacuum source coupled to the chamber” may be the same as the “vacuum source coupled to the chamber” described above in the context ofthe twenty-first main embodiment;
  • the “source of ultrasonic energy coupled to such chamber” may be the same as the “source of ultrasonic energy coupled to the chamber” described above in the context of the twenty-first main embodiment.
  • the chamber and source of ultrasonic energy in this embodiment may be the same as described above in the context ofthe twenty-first main embodiment.
  • the source of UV, gamma, or x-ray radiation is placed such that the UV, gamma, or x-ray radiation must pass through a portion ofthe chamber wall to reach the plasma, then at least that portion ofthe chamber wall must be sufficiently transparent to the radiation so that the desired degree of decontamination is achieved.
  • the principle difference between the apparatus of this embodiment and that ofthe embodiment described above is the presence ofthe UV, gamma, or x-ray radiation source.
  • the source of UV, gamma or x-ray radiation may be any that is capable of generating radiation ofthe desired frequency and intensity. Suitable sources of UV include those described above.
  • gamma radiation Cobalt-60 and Cesium- 137 are the most common medical application sources.
  • X-rays may be generated by standard, high voltage, electron accelerating sources.
  • the apparatus contains a dissolved-oxygen meter inside chamber.
  • the dissolved-oxygen meter is located such that it can detect the oxygen content in a thin film of plasma flowing across the flat panel.
  • This apparatus may also be constructed such that all ofthe components are permanent or semi-permanent, i.e., such that all or most ofthe components are intended to be used repeatedly for the processing of large amounts of plasma.
  • the apparatus may be divided into a permanent or semi-permanent subunit and a disposable subunit.
  • the permanent or semi-permanent subunit is constructed such that all or most of the components are intended to be used repeatedly for the processing of large amounts of a fluid, such as plasma.
  • the permanent or semi-permanent subunit comprises:
  • the permanent or semi-permanent subunit may further comprise other fixed hardware, including a peristaltic pump, a water jacket, and a vacuum pump.
  • the peristaltic pump is positioned such that it can be used to control the flow rate of plasma through the disposable unit.
  • the water jacket is positioned such that it will be between the ultrasound driver and the chamber when the chamber is placed in the region designed to accept it.
  • the vacuum pump is placed such that it can supply a vacuum to the gas above a thin film of plasma flowing through the chamber when the chamber is placed in the region designed to accept it.
  • the permanent or semi-permanent subunit may further optionally comprise a resonator plate which is positioned such that it will be located between the water jacket and the ultrasound driver.
  • the disposable subunit of this embodiment is essentially the same as that described above, with the proviso that at least one portion ofthe chamber wall must be constructed of material which is sufficiently UV-, gamma-, and/or x-ray-transparent, so that the plasma can be effectively decontaminated by the UV, gamma, and/or x-ray- radiation.
  • the present invention provides an apparatus for decontaminating a fluid, comprising:
  • the "means for containing said fluid” may be the same as the "chamber for containing a fluid” described above in the context ofthe twenty- first and twenty-third main embodiments; the “means for contacting said fluid with a vacuum” may be the same as the "vacuum source coupled to the chamber” described above in the context ofthe twenty-first and twenty-third main embodiments; the “means for introducing ultrasonic energy into said means for containing said fluid” may be the same as the "source of ultrasonic energy coupled to such chamber” described in the context of twenty- first and twenty-third main embodiments; and the “means for the treatment of said fluid with UV, gamma, or x-ray radiation” may be the same as the "source of UV, gamma, or x-ray radiation” described above in the context ofthe twenty-third main embodiment.
  • all the optional and preferred components described above may be the same as the "chamber for containing a fluid” described above in the context ofthe twenty- first and twenty-third main embodiments; the “
  • the present invention provides an apparatus for decontaminating a fluid, comprising:
  • a source of ozone wherein said chamber comprises: (i) an inlet for introducing ozone from the source of ozone; (ii) an inlet for introducing plasma; and (iii) a device for mixing ozone from the source of ozone with a fluid.
  • the ozone may be generated as described above, in the context ofthe thirteenth through twentieth main embodiments. Having thus generated the ozone, the next concern is how to apply it to the fluid.
  • two alternative methods or contactors may be used.
  • the ozone is mixed with the fluid with a contactor which comprises: (1) a substrate which has a lower surface and an upper surface and which has a plurality of passage-ways connecting said lower surface with said upper surface; (2) a source of ultrasonic energy coupled to said substrate, such that said ultrasonic energy is introduced into the fluid by the vibration of said substrate; (3) a source of ozone connected to said lower surface of said substrate.
  • the ozone is introduced into the fluid by passing through the same substrate which couples the source of ultrasonic energy to the fluid.
  • the ozone passes through the passage-ways in the substrate and is introduced into the fluid in the form of bubbles.
  • the size ofthe bubbles may be controlled, at least in part, by controlling the size of the openings ofthe passage-ways to the fluid.
  • the openings are circular in shape with the diameters ofthe openings ofthe passage-ways having a size of 25 to 1000 microns, preferably 50 to 500 microns, depending on the ultrasonic frequency range.
  • the size ofthe ozone bubbles introduced into the fluid is also influenced, in part, by the frequency and amplitude ofthe vibration ofthe substrate.
  • the substrate will vibrate at a frequency of 20 to 250 kHz, preferably 20 to 100 kHz, with an amplitude greater than the diameter of the openings.
  • the first part ofthe ozone treatment system is the plasma input reservoir, which is in direct contact with heat transfer plates for cooling. As noted in a previous section, cooled liquids are much more receptive to gasses.
  • the overall geometry ofthe reservoir is a cylinder, decreasing in size towards the base. At the bottom of this cylinder, the reservoir becomes rectangular in cross section. This rectangular cross section matches the inlet ofthe ozone nozzle. This nozzle is shaped like a "V" with small (several micron) holes on both sides of each planar section. These holes connect to an ozone source. Ultrasound is applied normal to the plane ofthe "V" along the direction ofthe bottom channel.
  • a similar reservoir is placed to collect the treated fluid.
  • the height of this second reservoir is less than the height ofthe first so that the liquid flows under gravity; alternatively, the fluid can be pumped.
  • the ozone enters the liquid already divided into "ligaments.”
  • the direct action ofthe ultrasound on these gaseous ligaments is immediate disruption into bubbles.
  • the motion ofthe ultrasonic horn is on the order of a mm, which is much greater than the ozone orifice diameters.
  • the fine gas bubbles are typically spread over a wide area.
  • this motion allows many orifices to be spaced close together, with subsequent rows staggered, to yield a quite uniform distribution.
  • fewer orifices are placed on the inlet side because the incoming downward flow tends to force the rising bubbles together, leading to undesirable larger sizes.
  • buoyancy on the exit side has the opposite effect, so more gas can be introduced here.
  • the fluid in the reservoir is immediately exposed to some gas, thus improving the overall treatment time.
  • the requirement that all ofthe liquid must pass through the nozzle ensures uniform treatment.
  • the continued ozone treatment on the exit side also extends the total treatment time, under good mixing conditions.
  • the small, micron-sized bubbles are much less than the optimum for resonance for a typical 20 kHz source, and are therefore rapidly driven into solution by the applied ultrasound, as discussed earlier.
  • the low amplitude source improves mixing and diffusion, without excessive bubble growth or protein damage due to cavitation.
  • Allowing the narrow sides ofthe "V” to flex slightly under ultrasonic motion can further enhance this mixing.
  • the flexing allows the essentially incompressible fluid to move more readily relative to the orifices ofthe nozzle.
  • a single driver at the base ofthe "N" is more cost effective than a pair of drivers on each side.
  • the liquid is then collected into a second reservoir, as described above. From here, the liquid is then pumped by a peristaltic unit through a heater into a vacuum/ultrasound degasser as described earlier. As described earlier, the fluid is then partially degassed, preferentially removing the oxygen while leaving the ozone. After degassing at this slightly elevated temperature, the fluid is then recycled into the starting reservoir.
  • the entire process can be repeated as many times as desired.
  • the overall intent is to achieve a high concentration of ozone rapidly.
  • part ofthe fluid can be in the degassing component while the remainder ofthe fluid is in the ozone nozzle component. Some ofthe material is thus continuously being processed, thereby decreasing the overall system time requirements.
  • the ozone flow rate into the fluid depends on the pressure applied to the lower surface ofthe substrate and on the size and density ofthe passage-ways.
  • the size ofthe passage-ways determines the size ofthe bubbles introduced into the fluid.
  • the size ofthe bubbles is important because bubbles greater than a critical size are stable and grow so large that they escape the liquid, while bubbles smaller than this critical size are unstable and are driven back into the solution by the ultrasound. Because the critical size limit depends on the frequency ofthe ultrasound, all bubbles less than the critical size are suitable.
  • the amount of ozone introduced into the fluid is typically controlled by varying the pressure of ozone applied to the lower surface ofthe substrate and by careful selection of the size and the density of passage-ways in the substrate.
  • the ozone is applied to the lower surface ofthe substrate at a flow velocity of 1 to 10 mm/sec, preferably 1 to 5 mm/sec.
  • the critical factor limiting the flow velocity on the ozone is the exit pressure, after the ozone leaves the passage-ways.
  • the gas be moving slowly, on the order of less than 1 cm/sec, along with negligible residual pressure, to prevent damage to the delicate proteins and/or any cells.
  • passing the output ofthe above described ozone generator through 400 holes each of 75 micron diameter yields a maximum velocity of about 0.6 cm/sec. In actual practice, the flow velocity is much slower due to pressure losses, as desired. Using 100 holes per square cm, distributed as described above, yields a total surface area of 4 cm 2 .
  • the substrate is part of a v-shaped trough, with one "leg" ofthe “v” taller than the other.
  • the inside surface ofthe tall “leg” corresponds to the upper surface ofthe substrate described above, and the outside surface of the tall “leg” corresponds to the lower surface ofthe substrate described above.
  • the fluid flows down the inside surface ofthe tall “leg” (the upper surface ofthe substrate), where it is effectively contacted with the ozone, to the bottom and then up and over the short "leg.”
  • the substrate forms part of a hollow apparatus which has an approximate U shape.
  • the fluid flows from an inlet (preferably after degassing and even more preferably after exposure to UV, gamma, and/or x-ray radiation).
  • the outside member ofthe hollow "U" corresponds to the substrate, and its inside surface corresponds to the upper surface of the substrate, while its outside surface corresponds to the lower surface ofthe substrate.
  • both the inside and outside members ofthe hollow "U” correspond to the substrate, with both inside surfaces corresponding to the upper surface ofthe substrate and both outside surfaces corresponding to the lower surface ofthe substrate.
  • the present invention provides an apparatus for decontaminating a fluid, comprising:
  • the "means for containing said fluid” may be the same as the "chamber for containing a fluid” described above in the context ofthe twenty- fifth main embodiment; the “means for contacting said fluid with a vacuum” may be the same as the "vacuum source coupled to the chamber” described above in the context ofthe twenty- fifth main embodiment; the “means for introducing ultrasonic energy into said means for containing said fluid” may be the same as the "source of ultrasonic energy coupled to such chamber” described above in the context ofthe twenty-fifth main embodiment; and the “means for generating ozone” may be the same as the "source of ozone” described above in the context ofthe twenty-fifth main embodiment.
  • the "(i) a means for the introduction of ozone from said means for generating ozone into said containing means" and the "(iii) a means for mixing said ozone from said means for generating ozone with said fluid in said containing means” may together form any ofthe ozone contactors described above in the context ofthe twenty-fifth main embodiment.
  • the present invention provides an apparatus for decontaminating a fluid, comprising: (1) a chamber for containing a fluid;
  • a source of ozone wherein said chamber comprises: (i) an inlet for introducing ozone from the source of ozone; (ii) an inlet for introducing a fluid; and (iii) a device for mixing ozone from the source of ozone with a fluid.
  • the "chamber for containing a fluid;” (2) the “vacuum source coupled to the chamber;” (3) the “source of UV, gamma, or x-ray radiation;” (4) the “source of ultrasonic energy coupled to such chamber;” and (5) the “source of ozone” may be any ofthe corresponding elements described above in the twenty-first, twenty-third, and twenty-fifth main embodiments.
  • the "device for mixing ozone from the source of ozone with a fluid” may be any ofthe ozone contactors described above in the context ofthe twenty-fifth main embodiment.
  • the apparatus ofthe twenty-seventh main embodiment is designed for implementation of a process in which the fluid is first degassed, then exposed to UV, gamma, or x-ray radiation, and then treated with ozone, i.e., the methods of main embodiments seventeen through twenty.
  • the present invention provides an apparatus for decontaminating a fluid, comprising: (1 ') a means for containing said fluid; (2') a means for contacting said fluid with a vacuum;
  • the "means for containing said fluid” may be the same as the "chamber for containing a fluid" described above in the context ofthe twenty- first, twenty-third, twenty-fifth, and twenty-seventh main embodiments; the "means for contacting said fluid with a vacuum” may be the same as the "vacuum source coupled to the chamber” described above in the context ofthe twenty-first, twenty-third, twenty-fifth, and twenty-seventh main embodiments; the “means for introducing ultrasonic energy into said means for containing said fluid” may be the same as the "source of ultrasonic energy coupled to such chamber” described above in the context ofthe twenty-first, twenty-third, twenty- fifth, and twenty-seventh main embodiments; the "means for the treatment of said fluid with UV, gamma, or x-ray radiation” may be the same as the "source of UV, gamma, or x-ray radiation” described above in the context ofthe seventh main embodiment; and
  • a means for the introduction of ozone from said means for generating ozone into said means for containing and "(iii) a means for mixing said ozone from said means for generating ozone with said fluid in said means for containing” may together form any ofthe contactors described above.
  • the present invention provides an apparatus for contacting ozone with a liquid, which comprises:
  • a substrate which has a lower surface and an upper surface and which has a plurality of passage-ways connecting said lower surface with said upper surface;
  • This twenty-ninth main embodiment corresponds substantially to the contactor shown in Figure 8, which is described in detail below.
  • the basic principles behind the ozone contactor could also be applied to adding other gasses to liquids. Specifically, the underlying principle is to degas the liquid first, and then add the desired gasses immediately afterward, using a sonic assist contactor. Finally, partial degassing to remove reacted products and/or undesired species should then be done.
  • the most common application liquid is water, but this could be extended to include aqueous solutions, or even other liquids.
  • the gasses could include everything from ozone to carbon monoxide or dioxide to various nitrogen compounds, etc.
  • the end product would not necessarily be something for sterilization, but instead could include various feedstocks for the chemical industry.
  • the present invention provides an apparatus for contacting a gas, e.g., ozone, with a fluid, said apparatus comprising:
  • a source of ultrasonic energy coupled to said chamber wherein said chamber comprises an a fluid inlet; wherein said chamber comprises a first sidewall and a second sidewall and said first and second sidewalls are positioned opposite to each other; wherein said chamber further comprises a plurality of partitions, and said partitions are attached to said first and second sidewalls in an alternating arrangement, and each partition attached to said first sidewall projects toward said second sidewall, and each partition attached to said second sidewall projects toward said first sidewall, such that said plurality of partitions forms a plurality of shelves; wherein said inlet is positioned in said chamber such that a fluid entering said chamber through said inlet occupies a first shelf; wherein said chamber is capable of rotating such that on rotation of 90 to -90 ° of said chamber fluid which occupies said first shelf will flow to a second shelf; wherein said source of gas is connected to said chamber to permit mixing of a gas with a fluid in said chamber; and wherein said source of ultrasonic energy is coupled to at least one of said partition, to
  • the apparatus of this thirtieth main embodiment corresponds to that depicted in Figure 11 and which is described below.
  • the contactor of this thirtieth main embodiment is especially useful for contacting ozone with fluids which contain platelets, and its use and operation and described in detail in connection with the description of Figure 11.
  • the discussions above which relate to the materials used for the chamber, the source of ozone, and the source of ultrasonic energy apply to this thirtieth main embodiment.
  • any nd all ofthe steps and/or components > described in the above-disclosed methods and apparatus may be carried out or operated by means of computer control. Such computer control substantially reduces the possibility and risk of problems and/or malfunctions resulting from human error.
  • Figure 1 is a schematic flow chart ofthe method ofthe fifteenth through eighteenth main embodiments, which are especially preferred.
  • the fluid is subjected to a temperature preparation step.
  • the fluid is degassed by application of ultrasonic energy.
  • the degassed fluid is irradiated.
  • the irradiated fluid is treated with ozone.
  • the first step in the process may require an hour or more.
  • degassing and UV exposure may take a half-hour or so, while the ozone exposure may take a similar time, or more if multiple cycles are used.
  • a clinical unit would have two or more bags being warmed simultaneously. As one of these bags reaches the completion of its heat treatment, it is degassed, etc., while the other bag continues its heat processing.
  • several different bags can be undergoing ozone exposure, while other units are being heated, degassed, etc. As a result, the unit is kept processing at all times for maximum return on investment.
  • FIG. 2 shows a preferred apparatus which is useful for ultrasonic degassing as carried out in the third and fourth main embodiments.
  • the fluid enters the chamber, which is a flexible, disposable bag, 21, through an inlet, 22, and eventually exits through a drain port, 23.
  • the disposable bag is received inside a vacuum chamber, 24, which is equipped with water cooling and ultrasonic drivers, 25, such that ultrasonic energy is introduced into the fluid.
  • the flexible, disposable bag, 21, is maintained in an expanded shape by means of a vacuum applied to the outside ofthe bag via a chamber vacuum port, 26 or by fasteners mated to the fixed vacuum chamber walls.
  • a vacuum is applied to the fluid inside the bag via bag vacuum port, 27.
  • the vacuum chamber, 24, is also equipped with temperature and mass sensors, 28 and 29, so that the flow rate and heating due to the introduction of ultrasonic energy may be monitored and controlled.
  • Figure 3 shows a preferred apparatus ofthe present invention which corresponds to the twenty-first and twenty-second main embodiments.
  • the fluid is introduced into the main chamber 34 through an inlet 33 from a starting plasma bag 31.
  • the feed rate ofthe fluid may be controlled by pump 32.
  • the fluid is sonified from below, as it flows across the planar section ofthe main chamber 34.
  • the ultrasonic energy is provided by the ultrasound driver 35, which is coupled to the fluid via the resonator plate 36.
  • the temperature ofthe fluid may be controlled by a water jacket 37.
  • the dissolved gasses, including oxygen may be released from the fluid and then trapped in the plastic housing. This housing and the planar section are a sealed unit, thus preventing external air from being drawn into the system.
  • FIG. 4 shows another preferred apparatus ofthe present invention which corresponds to the twenty-first and twenty-second main embodiments.
  • Figure 4 shows a decontamination system 40 designed for use in a method in which the fluid, in particular plasma, is decontaminated by the application of ultrasonic energy without application of UVC radiation or subsequent ozone treatment.
  • the plasma enters the system from a plasma bag 41 or other source on the left, with the flow rate ofthe plasma controlled by a peristaltic pump 42.
  • the plasma flow then crosses a divergent spreader 43, thus yielding a uniform plug flow of a thin film of plasma across the flat panel at the bottom of chamber 44.
  • the ultrasonic energy is applied to the plasma by means ofthe ultrasound driver 45, which is coupled to the flat panel at the bottom of chamber 44 via a resonator plate 46.
  • the ultrasound driver 45 which is coupled to the flat panel at the bottom of chamber 44 via a resonator plate 46.
  • the sonification is driven by an ultrasonic driver 45 acting on a metal plate 46 which is resonance coupled for efficient energy transfer.
  • the temperature ofthe plasma flowing across the flat panel is controlled by the water jacket 47.
  • the water jacket 47 between resonator plate 46 and the flat panel prevents heat from the ultrasonic driver 45 from reaching the plasma; water is an excellent sound transmission medium, and any losses of ultrasonic energy are thus insignificant.
  • Chamber 44 is a sealed unit, thus preventing external air from being drawn into the system.
  • the vacuum line inco ⁇ orates a sterile coupling and a filter trap 49 to prevent any pathogens from entering into and contaminating the vacuum pump.
  • the plasma After the plasma has been deoxygenated, the plasma then passes under irradiation source (in this case, UV lights) 410 for decontamination.
  • irradiation source in this case, UV lights
  • the water jacket 47 extends under this section to prevent excess heating ofthe plasma by the UV lights 410.
  • the ultrasound driver 45 also extends under the section in which the plasma passes under the UV lights 410. Extension ofthe ultrasound driver 45 under this section provides enhanced decontamination due to improved mixing ofthe plasma during UV exposure, as well as the dispersal of any aggregates.
  • the ultrasound generator does not extend under the region where the plasma passes under the UV lights 410. However, even when the ultrasound generator does not extend under the region where the plasma passes under the UV lights 410, it is preferred that the water jacket 47 extends under the region where the plasma passes under the UV lights 410.
  • the flow then enters a converging zone (outlet) 411, which leads to a tube connected to a collection vessel or bag 412 for the decontaminated product.
  • a converging zone which leads to a tube that passes through an optional peristaltic pump and then into collection bag 412.
  • the entire process may be carried out under refrigeration, and the entire apparatus 40 or at least one or more ofthe starting plasma bag 41, chamber 44, and collection bag 412 may be contained in one or more refrigeration units.
  • Apparatus 40 in Figure 4 may be constructed as a complete permanent or semi- permanent unit, with only the starting plasma and collecting plasma bags being disposable or consumable subunits.
  • apparatus 40 in Figure 4 is constructed as a permanent or semi-permanent subunit and a disposable or consumable subunit.
  • pump 42, ultrasound driver 45, resonator plate 46, water jacket 47, and vacuum pump 48 may be part ofthe permanent or semi -permanent subunit, while starting plasma bag 41, inlet 43, chamber 44, outlet 411, and collection bag 412 may be part of one or more disposable or consumable units.
  • the vacuum line including the filter trap 49 may be part of either the permanent or semi-permanent subunit or a disposable or consumable subunit.
  • the disposable or consumable units with the exception ofthe collection bag, may be preferably blow molded from inexpensive plastics, while it is preferred to use a conventional plasma bag and the disposable parts ofthe present apparatus should have no metal parts so they can be incinerated.
  • the chamber walls can be made of quite thin and/or flexible material, with only a small window for UV transmission directly under the lamps.
  • the disposable chamber may be a bag or liner for the region which is designed to accept it.
  • the chamber is a bag or flexible liner, it may be made to hold a desired shape or to conform to the shape ofthe region designed to accept it, by applying a slight vacuum to draw the liner out to the required dimensions under differential pressure, thus allowing the use of a very cheap treatment bag as the chamber.
  • An additional enhancement is the presence of another window in the bottom ofthe bag to allow exposure from both sides ofthe fluid layer by a second source of UV, gamma, or x-ray radiation.
  • the disposable bag used as the chamber further comprises a virus tight filter at one end ofthe bag to equilibrate the pressures inside and outside ofthe bag during vacuum processing.
  • This FDA approved component also allows for easier mounting ofthe bag inside the region designed to accept the chamber.
  • the disposable bag used as the chamber further comprises grommets at the inlet and outlet tubes to prevent them from collapsing during the application ofthe vacuum.
  • the chamber has a roughened inner surface.
  • a roughened inner surface allows the evolved gas bubbles to travel up to local spikes on the bag liner. From these points, the ultrasonic vibrations can dislodge the bubbles relatively easily.
  • This apparatus may further comprise certain safety features, including electrical shielding, splash guards, and particularly a commercial ultrasound shielding enclosure.
  • the plasma is sonified from below, as it flows across the planar section at the bottom ofthe main chamber 44.
  • the dissolved gasses, including oxygen are thus released from the plasma and are then trapped in the plastic housing.
  • This housing and the planar section are a sealed unit, thus preventing external air from being drawn into the system.
  • the evolved gasses are then captured by the vacuum pump.
  • the vacuum line inco ⁇ orates a sterile coupling and a filter trap to prevent any pathogens from contaminating the vacuum pump.
  • Input reservoir 41.
  • the first of these components is simply a bag to receive the output from the degassing unit.
  • this reservoir inco ⁇ orates a heat exchanger.
  • this reservoir is preceded by a heating/cooling pack as described for the inlet to the degassing unit.
  • the main function ofthe reservoir is simply to provide a uniform pressure head for the gravity flow throw the rest ofthe system. As such, the reservoir is broad and shallow, so that there is relatively little pressure difference between a full reservoir and a nearly empty reservoir.
  • UVC lamps 410. To achieve rapid and thorough processing, high intensity UV lamps are necessary. Several such UVC sources have recently been marketed (Spectronics
  • the flow cell is modified to include a thin layer of cooling water or other heat exchange liquid around the material to be treated, but constrained to separate channels.
  • the lamps must be separated from the flow cell by a thin, UV transparent shield.
  • the connecting hose may lead to an ozone exposure unit, rather than collection bag 412.
  • the essential problem is that the ozone unit typically operates at much greater pressures than the atmospheric pressure that exists in the UV unit. Thus, some provision must be made to handle this pressure difference.
  • the two alternative approaches are a pressure lock chamber or a peristaltic pump. Again, each has the previously described advantages and disadvantages. 4. Monitoring equipment.
  • UV systems tend to degrade over time in a process called "solarization.”
  • UV manufacturers for example, the Spectroline part of Spectronics Co ⁇ oration, Westbury, NY
  • Flow regulation The remaining concern is to control the flow through the system. The most important consideration here is that the liquid must be exposed to the light long enough for effective treatment, but not so long as to lead to excessive protein damage.
  • the approach here is to control the flow as described for the inlet to the degassing unit, using flow restrictions, pulsed flow, mass measurements, etc.
  • the essential consideration here is to keep the UV exposure unit working quickly enough to process continuously all ofthe output from the degassing unit.
  • the reservoir has to maintain sufficient head to pass all liquids rapidly, even though there may be substantial variations in viscosity from one unit to the next. While this holds in most cases, a control loop is also inco ⁇ orated to terminate flow into the degassing unit if necessary.
  • the first step in the startup process is to evacuate the fluid path with the vacuum pump. This is necessary to ensure that there is no oxygen in the system that might be absorbed by the fluid, and thus form oxygen radicals during UV exposure.
  • the exposure chamber should be tilted as described for the degassing unit so that only a minimum amount of fluid is left behind.
  • the reason for this effort is that the fluid is very valuable and thus must be collected as well as possible; furthermore, any residual material is simply a biohazard, thus presenting a disposal problem.
  • the UVC illumination be done from both sides. This provides for much more uniform exposure. In particular, this uniformity makes red blood cell treatment possible; otherwise, the strong abso ⁇ tion by hemoglobin prevents adequate treatment.
  • using double side exposure allows the use of a flow layer on the order of 10 to 40 microns, more preferably in the range of 30-40 microns.
  • the variation in intensity is less than 10% even for high hematocrit samples.
  • these dimensions are based on the size of erythrocytes, which are about 10 microns in length.
  • the required level of machining is available from specialized companies, such as Mindrum Precision, Inc., Collinso Cucamonga, CA. This firm specifies a flatness tolerance of about 0.5 microns for its UV flowcells.
  • the illuminator must be sonified at low intensity to promote uniform mixing and ensure plug flow. It should be noted that illuminator sonification was previously mentioned to prevent aggregation of plasma proteins.
  • the flexible bags be mounted on a rigid frame that matches the processing equipment.
  • This frame can be either reused or discarded.
  • the bags should be manufactured with registration holes in their borders.
  • the frame and/or the processor must have matching pins for these holes. This arrangement thus provides an easy way of aligning the bags in the processor, and also helps to prevent accidental misalignment by the operators.
  • the frame and/or bag assembly must be mounted in a recess inside the quartz flow cell.
  • the quartz thus provides rigid support after the fluid enters the treatment zone.
  • the quartz surfaces thus are in direct contact at the boundary, thereby ensuring tight tolerances.
  • the opposing panels are mounted on rubber supports, which compress on contact. Note that the sonification must therefore be applied directly to the panels under this arrangement, which will otherwise excessively damp the sound waves.
  • the last problem is to control the flow ofthe system.
  • all ofthe donation sample is illuminated in one step (for example, plasma or platelets).
  • all ofthe degassed liquid is poured into the top ofthe exposure chamber in one operation.
  • the lamps are then turned on.
  • all ofthe liquid is then drained.
  • the only problem here is that the clamp at the exit ofthe tube must not shadow the treatment volume. This can be avoided by designing a protrusion onto the clamp to extend sha ⁇ ly beyond the clamp body. Constructing this protrusion from UVC transparent materials, such as Teflon® AF or quartz, eliminates any remaining shadow effects.
  • Figure 5 shows another preferred apparatus, which corresponds to the apparatus ofthe twenty-fifth and twenty-sixth main embodiments and is useful for carrying out the method of the thirteenth through sixteenth main embodiments.
  • the fluid, plasma enters the ozonation unit from the plasma bag, 51, via a pump, 52, where it is mixed with ozone in a mixing tip/spray nozzle, 55.
  • the ozone enters the mixing tip/spray nozzle, 55, from an ozone generator, 53, passing through a filter trap, 56.
  • the fluid and the ozone are mixed in the mixing tip/spray nozzle, 55, and enter the reaction vessel, 57, as a spray or mist.
  • the fluid collects at the bottom ofthe reaction vessel, 57, to the fill line, 511.
  • Ultrasonic energy is applied to the fluid at the bottom ofthe reaction vessel, 57, via an ultrasound driver, 58, and a water jacket, 510, is placed between the bottom ofthe reaction vessel, 57, and the ultrasound driver, 58, to control the degree of heating.
  • the fluid is drained from the reaction vessel, 57, via a line to a collection bag, 512.
  • FIG. 5 shows a decontamination system 50 in which plasma enters the system from a plasma bag 51 or other source on the right, the flow controlled by a peristaltic pump 52.
  • Ozone from a conventional generator 53 is then passed through a connecting tube 54 to the mixing tip/spray nozzle assembly 55.
  • the ozone feed tube is passed through a filter 56 and trapped across a sterile coupling to prevent inadvertent contamination ofthe ozone generator 53.
  • the product is then collected in the reaction vessel 57.
  • this vessel sits on an ultrasonic driver 58 coupled to a resonator plate 59 and separated from the reaction vessel 57 by a water-driven cooling jacket 510. After sonification, the product is then drained into a collection vessel.
  • Apparatus 50 in Figure 5 may be constructed as a complete permanent or semi- permanent unit, with only the starting plasma and collecting plasma bags being disposable or consumable subunits.
  • apparatus 50 in Figure 5 is constructed as a permanent or semi-permanent subunit and a disposable or consumable subunit.
  • pump, 52, ozone generator, 53, ultrasound driver, 58, resonator plate, 59, and water jacket, 510 may be part ofthe permanent or semi-permanent subunit, while starting plasma bag, 51 , reaction vessel, 57, and collection bag, 512, may be part of one or more disposable or consumable units.
  • the ozone line, 54, including the filter trap, 56, and the mixing tip/spray nozzle, 55 may each be part of either the permanent or semi-permanent subunit or a disposable or consumable subunit.
  • the disposable bag used as the chamber further comprises a virus tight filter at one end ofthe bag to equilibrate the pressures inside and outside ofthe bag during vacuum processing.
  • This FDA approved component also allows for easier mounting ofthe bag inside the region designed to accept the chamber.
  • the disposable bag used as the chamber further comprises grommets at the inlet and outlet tubes to prevent them from collapsing during the application ofthe vacuum.
  • the chamber has a roughened inner surface. A roughened inner surface allows the evolved gas bubbles to travel up to local spikes on the bag liner. From these points, the ultrasonic vibrations can dislodge the bubbles relatively easily.
  • Ultrasound provides several alternatives to this problem, the main benefits being in the mixing process itself.
  • the basis for these effects is the ability of ultrasound to modify the properties of a liquid.
  • One such effect is the tendency for ultrasound to mix gasses into the surface of a liquid if the applicator horn is not deeply immersed into the liquid. Because the resulting poor coupling causes reduced cavitation, such operation of conventional ultrasound equipment is to be avoided (High Intensity Ultrasonic Processor User's Guide. Sonics &
  • the next step is conceptually similar to the spraying systems, or "nebulizers" of existing ozone technologies.
  • the problem with these conventional systems, however, is that they cause too much shear for plasma proteins.
  • the alternative is to use ultrasound to spray the already partially mixed plasma and ozone mixture, thus yielding even better mixing.
  • this process is not unique to this application; various ultrasonic nozzles are available commercially to produce a fine, soft spray.
  • the only modification for this process is to use an extended length of plastic tubing as a nozzle, the end of which is free at an antinode to whip under sonification. While this simple arrangement is not as effective as commercial nozzles, it is quite cheap.
  • the first step in this process is to use a peristaltic pump to control the flow rate ofthe plasma.
  • Ozone from a conventional generator is then passed through a connecting tube to the mixing tip/spray nozzle assembly.
  • the ozone feed tube is filtered and trapped across a sterile coupling to prevent inadvertent contamination ofthe ozone generator.
  • the product is then collected in the reaction vessel.
  • this vessel sits on an ultrasonic driver separated by a water cooling pack. After sonification, the product is then drained into a collection vessel.
  • Figure 6 depicts a portion of another preferred apparatus of the present invention, which corresponds to the twenty-fifth and twenty-sixth main embodiments and is useful for carrying out the method ofthe thirteenth through sixteenth main embodiments, in continuous, as opposed to batch-wise manner.
  • the fluid enters from an earlier ultrasonic degassing unit, which is not shown, and may pass through an optional cooler, 61.
  • the fluid then is formed into a thin film in the treatment assembly, 62, where it passes between one, preferably two, light sources, 63.
  • the fluid then passes on to ozone treatment or packaging, 64.
  • Figure 7 shows a preferred embodiment ofthe apparatus ofthe twenty-fifth and twenty-sixth main embodiments, which is useful for carrying out the methods ofthe thirteenth through sixteenth main embodiments.
  • a portion of Figure 7 also corresponds to a preferred embodiment ofthe contactor ofthe twenty-eighth main embodiment.
  • the fluid passes from bag 1, 71, through ozone contactor, 72, to bag 2, 73.
  • Bag 1, 71 is equipped with a fill and drain port, 74, through which the fluid may be introduced, and an equalizer port, 75, through gas may be introduced to fill the void created by exiting ofthe fluid from the bag 1, 71.
  • Bag 1, 71 is maintained within a vacuum chamber, 76, to allow for partial degassing, so that the spent ozone, which becomes oxygen, can be replaced by fresh ozone.
  • An alternative approach to the vacuum chamber is to use very high treatment pressures, in excess of 150 psi. In this case, simply releasing the pressure to ambient causes the excess gas to evolve rapidly.
  • a further enhancement for high pressure operation is to surround the fluid bags to be treated by solid blocks inside a pressure chamber. Under this approach, the air compressor is not necessary, because the blocks fill the available residual space in the chamber, thereby preventing the disposable bags from over expanding and subsequent rupture. As the fluid passes through the contactor, 72, it is simultaneously exposed to ozone and ultrasonic energy.
  • the ozone is introduced into the contactor, 72, via an ozone inlet, 77, and into the fluid via a plurality of passage-ways in the inner surfaces (not shown) ofthe contactor, 72, while the ultrasonic energy is introduced into the fluid via the vibration ofthe inner surfaces ofthe contactor, 72, driven by an ultrasonic driver, 78.
  • the fluid After passing through the contactor, 72, the fluid enters bag 2, 73, which is equipped with a vent port, 79, to vent the gas displaced by the entering fluid, and a drain port, 710, for draining the fluid.
  • Both bag 1 and bag 2 may be equipped with a mounting ring, 711. For repeated treatment, such as indicated above for degassing, there are two options.
  • One such option is to progress from one disposable bag to another. This is preferred wherever possible.
  • the bags could be reused.
  • such reuse raises the question of residual contamination.
  • all parts ofthe system are subject to direct ozone gas exposure, and thus are continually being cleaned.
  • the surfaces are cleaner than the liquid passing through the system in bulk because when no bulk fluid is present, the surfaces are coated by at most a thin fluid layer, which is readily treated by ozone exposure.
  • FIG. 8 is a detailed cross-sectional view ofthe contactor portion ofthe apparatus shown in Figure 7.
  • the contactor of Figure 8 is made up of 4 distinct layers, which form three different flow fields. First there are upper and lower outer layers, 81 and 82, respectively.
  • upper and lower inner layers, 83 and 84 are perforated by a plurality of channels, 85.
  • the upper and lower inner layers correspond to the substrate(s) described above in the context ofthe twenty- seventh and twenty-eighth main embodiments, and the channels correspond to the passageways described above in the context ofthe twenty-seventh and twenty-eighth main embodiments.
  • the lumen formed by the upper outer and inner layers, 81 and 83, and by the lower outer and inner layers, 82 and 84, are connected to the ozone inlet, 77, depicted in Figure 7, and permit the flow of ozone through the contactor, and are referred to as ozone flow fields, 86.
  • the lumen formed by the upper and lower inner layers, 83 and 84, is connected to a source ofthe fluid such as bag 1, 71, depicted in Figure 7, and is referred to as the liquid flow field, 87.
  • a source ofthe fluid such as bag 1, 71, depicted in Figure 7, and is referred to as the liquid flow field, 87.
  • ozone is introduced into the fluid through the channels, 85.
  • ultrasonic energy is introduced into the fluid by the vibration ofthe upper and lower inner surfaces, 83 and 84 by means of an ultrasound generator (not shown) which is coupled to the upper and lower inner surfaces, 83 and 84.
  • Figure 9 shows another preferred embodiment ofthe apparatus ofthe twenty-fifth and twenty-sixth main embodiments, which is useful for carrying out the methods ofthe thirteenth through sixteenth main embodiments.
  • a portion of Figure 9 also corresponds to a preferred embodiment ofthe contactor ofthe twenty-ninth main embodiment.
  • the fluid passes from bag 1, 91, through ozone contactor, 92, to bag 2, 93.
  • Bag 1, 91 is equipped with a fill and drain port, 94, through which the fluid may be introduced, and an equalizer port, 95, through gas may be introduced to fill the void created by exiting ofthe fluid from the bag 1, 91.
  • Bag 1, 91 is maintained within a vacuum chamber, 96, to allow for partial degassing, so that the spent ozone, which becomes oxygen, can be replaced by fresh ozone.
  • the contactor, 92 As the fluid passes through the contactor, 92, it is simultaneously exposed to ozone and ultrasonic energy.
  • the ozone is introduced into the contactor, 92, via an ozone inlet (not shown) and into the fluid via a plurality of passage-ways in the inner surfaces (not shown) of the connector, 92, while the ultrasonic energy is introduced into the fluid via the vibration of the inner surfaces ofthe connector, 92, driven by an ultrasonic driver (not shown).
  • FIG. 10 shows a cross-sectional view of a preferred embodiment of an ozone contactor according to the twenty-ninth main embodiment ofthe present invention.
  • the fluid flows from the inlet, 1001, through the fluid flow field, 1002, to the outlet, 1003.
  • the ozone enters the ozone inlet, 1004, flows through the ozone flow field, 1005, and is introduced into the fluid flow field, 1002, through a plurality of channels, 1006, in the walls forming the fluid flow field, 1007. Any excess ozone exits the ozone flow field, 1005, through an exit, 1008.
  • the walls which form the fluid flow field, 1007, are made to vibrate by being coupled to one or more ultrasonic drivers, 1009.
  • Figure 1 1 shows a cross-sectional view of another ozone contactor which is particularly useful for platelets.
  • the treatment chamber consists of a rectangular or similarly shaped block 1 101 with staggered, opposed shelves in the shape of sha ⁇ wedges 1102.
  • the chamber With the chamber in the horizontal position, the liquid enters a trough or inlet port 1 103 along one side. After filling this trough, the chamber is then rotated upwards to about 80 degrees, at which point the fluid flows over the first shelf 1102a towards the opposing wall. Because the shelf does not actually touch the opposing wall, however, the fluid drops down to the next shelf 1102b and the flow then reverses. Meanwhile, ozone is introduced through ports 1104. Note that this arrangement is unlike the above cited patents because the flow reversal thoroughly mixes the material at each step, with the top layer becoming largely the bottom layer and vice versa.
  • the last step is to shut down the system and store the product.
  • small solenoids can be used to tilt the components to drain the quite valuable product as completely as possible.
  • This concept can also be extended to drop the exit side ofthe "v" ofthe spray nozzle.
  • One approach is to collect the product after the last ozone injection pass. This would leave a substantial amount of ozone in the liquid. As it reacts and decays to oxygen, this residual ozone would provide a slight increase in decontamination effectiveness. Also, the resulting oxygen would be quite beneficial to red blood cells and platelets.
  • the collection should be taken after the product is degassed as thoroughly as possible. This step reduces the formation of gas bubbles (commonly observed as small pockets and streaks in ice cubes) in the frozen product, and thus leads to less product damage during the freezing process.
  • the exact conditions and/or parameters ofthe present methods may be needed to achieve optimum results for certain types of pathogens and infectious agents and fluids.
  • some ofthe conditions and/or parameters ofthe present methods may need to be varied to achieve optimum results for certain types of pathogens and infectious agents, while minimizing plasma protein damage.
  • Such conditions and/or parameters which might need to be varied include the precise intensity and/or frequency ofthe ultrasonic energy; the precise intensity and/or frequency ofthe UV, gamma, and/or x-ray radiation; the precise amount of ozone to be mixed with the plasma; the pressure ofthe ozone; and the precise temperature and/or time of any step.
  • any ofthe present decontamination methods may be optimized by assessing the efficacy ofthe method against the pathogen in a first test, then again assessing the efficacy ofthe method after varying one or more parameters and/or conditions ofthe method, and then comparing the results ofthe two tests.
  • the amount of damage to a particular fluid (e.g., plasma) protein and the optimization of any ofthe present methods in regard to the minimization of plasma protein damage may be carried out in the same way, with the exception of determining protein concentration or activity rather than pathogen concentration or activity.
  • any such testing on plasma itself requires plasma.
  • the concentrations of plasma proteins vary widely from donor to donor. This is a significant problem because these variations are usually greater than the fractional protein damage caused by the decontamination methods themselves, thus making direct comparisons difficult.
  • the standard reference range for fibrinogen is 200 to 400 mg/dl, but even a relatively poor (in terms of harshness to the plasma proteins) decontamination technique would destroy less than 25% of this protein. As a result, the variation in the plasma proteins from individual unit to individual unit would thus mask the entire range of protein damage.
  • a reference plasma obtained by pooling several donations and then extracting multiple units ofthe same volume.
  • the use of such a reference plasma establishes a common basis for comparison. Pooling also eliminates much testing expense: with a pool, the protein levels can be tested once to establish starting conditions, but for individual units, each starting condition must be tested separately. This pooling technique has proven to be quite successful in earlier work done by the present inventor, in which it was found that the inherently large error in blood testing equipment could be better offset by multiple tests of a single pool, versus repeated tests of single units.
  • this pooling is for assessment and optimization pu ⁇ oses only, i.e., it does not restrict in any way the ability ofthe present methods to process individual plasma units.
  • pu ⁇ oses of initial assessment and optimization for a particular pathogen it may be preferred to lower costs by using non-human plasma.
  • bovine plasma provides a useful starting point without the cost or handling problems associated with human products.
  • human plasma may then be used.
  • the plasma as obtained from the donor(s) might not contain any detectable amounts ofthe pathogen or infectious agent of interest. In such cases, a known amount ofthe pathogen or infectious agent may be added to the plasma.
  • fibrinogen is the most appropriate: it is clinically significant, it is commercially valuable, it is easy to test, it is readily damaged by existing decontamination techniques and its widespread use by other investigators provides a means for direct comparisons.
  • model viruses which simulate actual human viruses, may be used for experimental decontamination testing. Specifically, their low cost, low risk, and direct applicability to actual pathogens have led to complete industry and regulatory acceptance. Because of these benefits, many test viruses have been isolated and are now in common use. Typical examples of such model viruses include Sindbis and BVDV (Bovine Viral Diarrhea Virus) for human HCV, and duck HBV for human HBV (B. Horowitz, "Virus Inactivation by Solvent Detergent Treatment and the Sindbis and BVDV (Bovine Viral Diarrhea Virus) for human HCV, and duck HBV for human HBV (B. Horowitz, "Virus Inactivation by Solvent Detergent Treatment and the
  • parvovirus B19 erythema infectiosum or "fifth disease" from parvovirus is mainly a concern during pregnancy, and thus poses much less risk to the general population than hepatitis or AIDS.
  • parvovirus infection is actually so common that Plas+SD is sold with the claim that the pooled donors effectively contribute antibodies.
  • parvovirus is extremely difficult to eradicate by conventional techniques. The net result is that there is some question within the blood industry about the cost-effectiveness of attacking this particular virus.
  • parvovirus is of great interest as a test standard because any technique that is effective against such a robust virus would also be extremely effective against lesser pathogens. Accordingly, it may be preferred to use parvovirus, in particular porcine parvovirus (PPV), as the standard test virus for the optimization ofthe conditions and/or parameters of any one ofthe present methods. Having thus selected the plasma and a test virus, the next concern is how to measure the effectiveness ofthe proposed technology. Fortunately, such measurements are actually quite simple using standard procedures. Specifically, the plasma may be first spiked with the test virus and then split into two fractions. One fraction is then maintained as a control, while the other fraction is subjected to the decontamination methods being assessed or optimized. For statistical analysis, six samples may be taken at each test point. The test results for a given pathogen can be reported in terms of a logarithmic reduction factor.
  • PSV porcine parvovirus
  • the logarithmic reduction factor is a quantitative measurement. As such, it can be inserted directly into a standard test matrix. Because ultrasonic effects are nonlinear, this matrix is in turn nonlinear. This matrix can, however, be reduced by standard steepest descent techniques. The result is an optimized system, within the limits of resolution ofthe test points. For example, given Beer's exponential law of optical abso ⁇ tion, it is anticipated that the logarithmic reduction factor will drop substantially beyond a critical fluid depth. Having thus established this critical value, the residence time can then be optimized by changing the pump rate and/or the dimensions ofthe treatment chamber.
  • the present methods may not be effective by themselves for the complete removal of all contaminants. Accordingly, it may be desired in certain circumstance to utilize a small amount of quencher in conjunction with those embodiments which involve irradiation.
  • the advantages in the context ofthe present invention are lower quench concentrations, reduced chemical cost, reduced removal cost, and less bioburden.
  • the present methods may also be used to prepare anaerobic synthesis systems for biotechnology applications. The advantage here is that the processed systems are then be ready for immediate UV and/or ozone treatment.
  • parvovirus is so widespread, many veterinary facilities and schools can analyze it.
  • a particularly well known group is the American BioResearch Laboratories, located in Sevierville, Tennessee.
  • plasma is particularly useful because it has both heat-tolerant components, notably fibrinogen, and heat-sensitive components, notably Factor VIII.
  • heat-tolerant components notably fibrinogen
  • heat-sensitive components notably Factor VIII.
  • bovine plasma can be used instead of human plasma; in any case, porcine plasma should not be used due to the high likelihood of existing antibodies.
  • bovine plasma is readily available from slaughterhouses, higher quality material is available from facilities dedicated to the maintenance of healthy donor animals, such as Quad Five, Ryegate,
  • erythrocytes can also be obtained from bovine sources; alternatively, out of date human cells can also be used, following the convention that scarce transfusable materials should not be used for basic experimental pu ⁇ oses.
  • human platelets which have a shelf life of only 5 days, are also available on an out of date basis. All blood banks have such materials, but the largest supplier is the American Red Cross, which has offices nationwide.
  • testing must then be arranged.
  • bovine blood products are routinely analyzed at the University of Georgia Veterinary School in Athens,
  • the first such test is a heat-tolerant plasma protein, fibrinogen.
  • This protein is particularly interesting because it is a crucial part of surgical glues, such as Tisseel by Baxter
  • Example 1 Heat-tolerant plasma protein This test is designed for a heat tolerant protein, fibrinogen. This test is also designed for a small quantity, on the order of several ml, because this is the volume of material that can be extracted from a single unit of donated plasma. The test procedure is described below.
  • the required system equipment includes a warming component, a small vacuum module, a UVC exposure module, and an ozone treatment module.
  • the disposable is a small, single unit device, consisting of a liner for the heater, a Teflon® bag to serve in both the vacuum chamber and UVC irradiator, a pair of bags for the ozone exposure unit, an ultrasonic ozone contactor, two virus tight filters, and sterile connecting tubing. Load the disposable items and apply vacuum to evacuate the system to 50 mbar. Next, dilute 1 ml of porcine parvovirus by 10:1 in bovine plasma. For statistical pu ⁇ oses, prepare 6 sets of six samples each. Retain one set as a control. Heat the remainder
  • results Compared to the controls, the heating and degassing alone show no significant differences in viral loading or protein damage. Conversely, the UVC irradiation alone shows a Log 6 reduction in active parvovirus loading; likewise the ozone treatment alone shows a Log 6 reduction in parvovirus. The combined UVC and ozone shows an apparent Log 12 reduction, although at such levels, detection is much more difficult. In all cases, the fibrinogen loss is less than 5%, which is essentially within the limit of error ofthe measuring equipment.
  • Example 3 Batch plasma This test is designed to evaluate the system when processing sequential batches of material.
  • the specific material is a single donor unit of plasma. As such, it is larger than the volumes tested above, but smaller than the volumes that would be treated by the continuous flows discussed below in Example 4. To accommodate this volume, the equipment described in the previous example must be changed as follows.
  • results The individual UVC and ozone cases each yield Log 6 virus reductions, with Log 12 for the combined process. For the repeated ozone exposure, the viral reduction is Log 9. In all cases, the Factor VIII damage is less than 5 %.
  • This test is designed to evaluate the system when processing a continuous flow of material.
  • the specific material is again plasma, but in this case the large flow corresponds to the treatment of a pool of material for subsequent fractionation.
  • the equipment described in the previous example must be changed as follows.
  • the major modification is to change the batch flow equipment to continuous flow equipment. Specifically, the major changes are made to the degassing and ozone treatment modules, along with their associated modules; the batch flow UVC module can also be used for continuous flows with only the addition of a flow controller on the feed pump.
  • UVC exposure follows immediately, and then the fluid is exposed to ozone.
  • the residence time in the UVC and ozone modules is about 15 minutes each, with the actual treatment time being continuously varied slightly about this value according to the respective dosage monitors.
  • This test is designed to determine the effect of decontamination on a living cellular structure.
  • the equipment and conditions are essentially as described for Example 1, except
  • the first concern is that a much larger illumination chamber (30 cm by 10 cm) is used to treat a larger surface area than required for the relatively transparent fluid used in Example 1 ; the volumes are nevertheless similar because the erythrocyte chamber is much thinner at about 40 microns.
  • the second difference is that the heating is done only to 45 °C, instead of 52 °C, because erythrocytes are known to withstand this lower temperature well 0 during hyperthermia treatments, but the higher (52 °C) temperature could compromise their cell membranes.
  • the third concern is that the erythrocytes are exposed to oxygen, or oxygen ozone, immediately after irradiation because these cells require oxygen to survive. Note that nerve cells in the brain are known to suffer irreversible damage after only 6 minutes without oxygen, but erythrocytes are much more durable. Furthermore, decreasing the first concern is that a much larger illumination chamber (30 cm by 10 cm) is used to treat a larger surface area than required for the relatively transparent fluid used in Example 1 ; the volumes are nevertheless similar because the ery
  • the viral test results are Log 6 reduction for both UVC and ozone, while the combined process is about Log 12.
  • Cell damage in such testing is commonly measured by hemolysis. Observing the number of damaged cells immediately after the process indicates gross mechanical damage, while
  • This test is designed for the platelet ozone contactor.
  • the equipment and conditions are essentially as described in Example 1 for heated, small volumes, except for three factors.
  • the first difference is the use of a special ozone contactor designed solely for platelets, and
  • the results of this test are Log 6 reduction of parvovirus during UVC and ozone, and Log 12 for the combined processes.
  • the first test is a simple optical procedure widely used in the platelet industry. This test amounts to simply observing the flow ofthe platelets in their special oxygen-permeable bag. Normal platelets should sparkle, while damaged platelets often agglomerate, and thus do not sparkle when illuminated. In this experiment, the treated platelets exhibit the same sparkle as shown by the controls.
  • the second test is to determine clotting effectiveness, which is done by forming a clot from the gel and then rupturing it, thereby indicating whether or not the platelets function as necessary. In this experiment, the clots formed from the control are indistinguishable from the clots formed from the treated material, thereby indicating no appreciable damage to the platelets during decontamination.

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US20070102858A1 (en) 2005-11-07 2007-05-10 Navigant Biotechnologies, Inc. Clamps and methods for displacing fluid from portions of fluid containers
DE102005062410A1 (de) 2005-12-23 2007-08-09 Forschungsgemeinschaft Der Drk-Blutspendedienste E.V. Verfahren zur Bestrahlung von Thrombozytenkonzentraten in flexiblen Behältnissen mit ultraviolettem Licht
DE102005062634A1 (de) 2005-12-23 2007-06-28 Blutspendedienst der Landesverbände des Deutschen Roten Kreuzes Niedersachsen, Sachsen-Anhalt, Thüringen, Oldenburg und Bremen gGmbH Verfahren zur Inaktivierung von Pathogenen in Spenderblut, Blutplasma oder Erythrozytenkonzentraten in flexiblen Behältnissen unter Bewegung
DE102006027227A1 (de) 2006-06-12 2008-01-03 DRK - Blutspendedienst Baden-Württemberg-Hessen GmbH Verfahren und Vorrichtung zum Inaktivieren von Viren und/oder Bakterien in flüssigen Medien, insbesondere in Blutplasmen und Serumkonserven
US7522702B2 (en) * 2006-08-29 2009-04-21 Harris Corporation Soft x-ray radiation for biological pathogen decontamination and medical sterilization applications
EP1902740A1 (de) * 2006-09-19 2008-03-26 Maco Pharma S.A. Blutbeutelsystem und Verfahren, um Pathogene in Blutplättchenkonzentraten mit Hilfe des Blutbeutelsystems zu neutralisieren
US7776260B2 (en) * 2006-12-11 2010-08-17 Ethicon, Inc. Apparatus and method for the irradiation of radiation sensitive materials
RU2419467C2 (ru) 2006-12-20 2011-05-27 Хару МИЯКЕ Устройство для ультразвуковой терапии
EP2008669A1 (de) 2007-06-22 2008-12-31 Maco Pharma S.A. Bestrahlungsvorrichtung zur Deaktivierung von Krankheitserregern und/oder Leukozyten in einer biologischen Flüssigkeit und entsprechendes Verfahren
ITMI20111439A1 (it) * 2011-07-29 2013-01-30 Paolo Benatti Apparato per l'ozonizzazione di fluidi biologici, particolarmente per sangue.
US8940228B2 (en) 2012-01-11 2015-01-27 Terumo Bct, Inc. Slidable clamp for port isolation
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KR102252218B1 (ko) * 2019-11-01 2021-05-14 김진왕 친환경 스마트 혈액 모듈레이션 장치
CN114225067B (zh) * 2021-12-22 2024-01-26 中国医学科学院输血研究所 一种血液病原体灭活方法

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AU2002306757B2 (en) 2009-07-02
AU2002306757A1 (en) 2003-10-27

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