Classifications

• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
  • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
  • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
  • C12Q1/22—Testing for sterility conditions
• • 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
  • A61L2/26—Accessories or devices or components used for biocidal treatment
  • A61L2/28—Devices for testing the effectiveness or completeness of sterilisation, e.g. indicators which change colour
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
  • B01J13/00—Colloid chemistry, e.g. the production of colloidal materials or their solutions, not otherwise provided for; Making microcapsules or microballoons
  • B01J13/0095—Preparation of aerosols
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
  • B01J19/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
  • B01J19/0006—Controlling or regulating processes
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
  • B01J19/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
  • B01J19/0006—Controlling or regulating processes
  • B01J19/0013—Controlling the temperature of the process
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
  • B01J19/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
  • B01J19/0053—Details of the reactor
  • B01J19/0073—Sealings
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
  • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
  • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
  • C12Q1/18—Testing for antimicrobial activity of a material
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
  • B01L1/00—Enclosures; Chambers
  • B01L1/02—Air-pressure chambers; Air-locks therefor
  • B01L1/025—Environmental chambers
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
  • G01N33/0078—Testing material properties on manufactured objects
  • G01N33/0081—Containers; Packages; Bottles

Definitions

• Aerosol product exposure system and method for evaluating the integrity of a container using such a system Aerosol product exposure system and method for evaluating the integrity of a container using such a system
• the invention relates to a system for exposure to a product in the form of an aerosol, such as an aerosol of microorganisms. Such a system is particularly useful for testing the integrity of containers, such as packages.
• the invention therefore also relates to a method for evaluating the integrity of a container by means of such an exposure system.
• Such a system can also be used to evaluate the antimicrobial properties of a composition.
• the subject of the invention is also a method for evaluating the antimicrobial activity of a product of interest by means of the exposure system according to the invention.
• Packaging integrity plays a key role in maintaining the sterility of packaged products.
• the integrity defects are not necessarily detectable to the naked eye. This is the case of microcracks or scratches of glass containers. Other defects may be masked by a subsequent crimping operation, or on the contrary be due to poor crimping.
• the sealing zones are frequently involved.
• the integrity of packaging must be guaranteed to maintain the sterility and quality of the products they contain throughout their shelf life. conservation, and more generally for the safety of the product and the consumer.
• CCIT Contassembly Integrity Testing
• numerous tests of integrity of the packaging (“Container Closure Integrity Testing” or “CCIT”) have been developed to make it possible to easily check the sealing zones of the packaging or containers, and to validate the sterility of the products. that they contain.
• CCIT method should evaluate the ability of a closure system of a container to maintain a sterile barrier against potential contaminants, such as microorganisms, reactive gases or other harmful substances. .
• Microbial Ingress Test is particularly known. This test is suitable for any container containment system that can withstand changes immersion and pressure.
• the test container is immersed in a broth containing one or more test microorganisms, such as Brevundimonas diminuta, Serratia marcescens, Escherichia coli.
• the test can be performed in static mode, where no pressure or vacuum is applied, or in dynamic mode, where pressure and vacuum are applied (to simulate air transport of the product).
• the main test factors are bacterial size and motility, differential pressure, challenge media, exposure time and viable number of microorganisms in the challenge media.
• the dye leak test (“Dye Ingress Test”) is also widespread.
• the test container is immersed in a dye solution (and in particular in a methylene blue solution) and a pressure and a vacuum are applied.
• the container is then inspected visually or spectrophotometrically for any traces of dye in the container.
• Key factors for this test include differential pressure, dye compatibility with product, liquid viscosity and surface tension, inspector training and experience (for visual inspection), and assay sensitivity (spectrophotometry). ).
• the Mass Extraction Assay can be used to detect leaks in non-porous, rigid or flexible packages.
• the test is performed by placing the test container inside a test chamber which is pneumatically connected to a mass extraction leak test system equipped with a vacuum generator. A predetermined vacuum level is maintained in the chamber for a predetermined time. A series of evacuation cycles is performed, each cycle being designed to identify the smallest leak rates. After each cycle, the test system is isolated from the vacuum source and the measurements of the absolute pressure, the rate of pressure decrease and / or the mass flow rate of the gas are recorded. Readings above predetermined limits established by negative controls indicate leakage from the container. At the end of these leakage vacuum cycles, a final vacuum is established. The mass flow rate is measured with all the flow of the test chamber directed through the mass flow sensor. A mass flow rate greater than a predetermined limit established by negative controls indicates leakage from the vessel. Such tests are however heavy to implement and require expensive equipment.
• the inventors have developed a device for projecting a product, such as a solution containing microorganisms, in the form of an aerosol inside an enclosure, in which the container has been previously placed or packaging to test.
• the aerosol spraying form makes it possible to diffuse very high concentrations of the product inside the enclosure and thus to reach an optimum concentration of said product on the surface of the container.
• the device according to the invention is particularly useful for performing microbial integrity tests.
• the device according to the invention makes it possible to project an aerosol of microorganism solution at concentrations such that it is possible to reach microorganism concentrations of up to 10 6 CLU / cm 2 on the surface of the container to be tested. .
• the device according to the invention makes it possible to test the integrity of all kinds of packaging, and in particular flexible packaging such as blood bags or other, rigid packaging such as vials, cartridges, pre-filled syringes, etc.
• Such a device can also be used to evaluate the antimicrobial properties of a composition. For this, it suffices to spray said aerosolized composition into the chamber in which a source of microorganisms was previously placed, and then to evaluate the rate of growth and / or survival of said microorganisms.
• the present application therefore proposes a system of exposure to a product in the form of an aerosol comprising
• a device for diffusing a product in the form of an aerosol comprising a source of spray product in the form of an aerosol, connected to an aerosol generator, the said aerosol generator cooperating with a desiccator so as to eliminate at least partially the moisture of the product in aerosol form;
• At least one supply duct intended to bring the product in the form of an aerosol of the diffusion device into the exposure chamber.
• the application also proposes a method for evaluating the integrity of a container by means of an aerosol exposure system comprising an exposure chamber, a device for diffusing a suspension of microorganisms in the form of aerosol comprising a source of said microorganism suspension connected to an aerosol generator, said aerosol generator cooperating with a desiccator so as to at least partially remove moisture from the aerosolized suspension of microorganisms; and at least one conduit feed device for supplying the suspension of microorganisms in the form of an aerosol into the exposure chamber, comprising the successive steps according to which:
• an aerosol is generated from the suspension of microorganisms, said aerosol being vaporized in the exposure chamber so that the microorganisms are in contact with the container;
• the application also proposes a method for evaluating the antimicrobial activity of a product of interest by means of an aerosol exposure system comprising an exposure chamber, a device for diffusing the product of interest under aerosol form comprising a source of the product of interest connected to an aerosol generator, said aerosol generator cooperating with a desiccator so as to at least partially remove the moisture of the product of interest in aerosol form; and at least one feed conduit for supplying the product of interest as an aerosol to the exposure chamber, the method comprising the steps of:
• the invention also relates to the use of the aerosol exposure system according to the invention for testing the integrity of a package and / or the antimicrobial activity of a product of interest. Brief description of the figure
• Figure 1 schematically shows an aerosol exposure system according to an exemplary embodiment of the invention.
• the inventors have succeeded in developing an aerosolized product exposure system in which product moisture is at least partially reduced prior to spraying into the exposure chamber.
• the aerosol product exposure system is mainly characterized by (i) an exposure chamber, intended to receive a product in the form of an aerosol and the test container or a source of microorganisms. (ii) an aerosol dispensing device comprising a source of aerosol sprays connected to an aerosol generator which cooperates with a desiccator and (iii) one or more supply ducts for to bring the aerosol into the exposure chamber.
• the term "aerosol” means a stable suspension of solid or liquid particles in a gas.
• the inventors have discovered that the at least partial elimination of the moisture of the aerosolized spray product before it is sprayed inside the exposure chamber, or chamber, makes it possible to concentrate the product. to spray.
• desiccation makes it possible to spray high concentrations of microorganisms and thus easily reach a concentration on the surface of a container placed inside the exposure chamber of 10. 6 CFU / m 2 .
• the product in aerosol form is a spore suspension.
• the aerosol product is a composition whose antimicrobial activity is to be tested.
• the aerosol generator comprises a container (containing the product to be sprayed) pressurized with a propellant.
• a container containing the product to be sprayed
• a propellant pressurized with a propellant.
• the skilled person is able to choose the most suitable aerosol generator, depending in particular on the product to be sprayed.
• any desiccator for removing at least partially the moisture of a composition or suspension can be used.
• the desiccant of Drier is dry air.
• the desiccant desiccant is a hydrophilic compound, such as silica gel. Those skilled in the art are able to choose the most suitable desiccator, depending in particular on the product to be sprayed.
• the desiccator is advantageously capable of removing at least 50% of the moisture initially contained in the product to be sprayed, more preferably at least 60%, 70%, 75%, 80%, 85% or more.
• the sprayed product in aerosol form has a moisture content of less than 50%.
• the aerosol product exposure system comprises one or more conduits for supplying the product aerosol form from the diffusion device into the exposure chamber.
• the exposure system comprises two ducts, opening on either side into the volume of the exposure chamber, in order to evenly distribute the aerosol throughout the exposure chamber.
• the supply duct or ducts open in the upper part of the exposure chamber, so as to bring the product aerosol form in the upper part of said chamber.
• the product sprayed inside the exposure chamber will fill the entire volume of said chamber by falling gradually by sedimentation.
• the "upper” part designates the part of the element considered furthest from the surface on which said element rests, as opposed to the "low” part.
• the system according to the invention also comprises evacuation means, able to evacuate the sprayed product from the exposure chamber at the end of the test.
• the evacuation means comprise an evacuation valve, arranged in the lower part of the exposure chamber, and at least one filter adapted to receive the product at the exit of the evacuation valve.
• the evacuation means comprise an air filter and / or an activated carbon filter. The filter advantageously makes it possible to retain the microorganisms in the case where the pulverized product is a suspension of microorganisms.
• the presence of at least one filter makes it possible in particular to maintain an atmospheric pressure in the exposure chamber while avoiding the exit of microorganisms outside the chamber, thus preventing contamination of the surrounding air.
• the evacuation means comprising at least one filter make it possible in particular to avoid the overpressure in the chamber while retaining the microorganisms, thus maintaining an optimum concentration of the particles sprayed in the exposure chamber.
• at least one filter is arranged in the lower part of the exposure chamber. This makes it possible in particular to retain in the exposure chamber the particles, in particular the microorganisms, having sedimented.
• the evacuation means comprising at least one filter capable of retaining the particles are arranged in the upper part and in the lower part of the exposure chamber.
• the filters may be arranged in the lower part and in the upper part of the exposure chamber, at the inlet and / or the outlet of the discharge valve.
• the exposure chamber is a chamber that can be sealed. Any means making it possible to guarantee the tightness of an enclosure may be used, and in particular joints, mechanical grasshoppers, etc.
• the system according to the invention comprises air supply means arranged in the upper part of the diffusion chamber.
• the aerosol exposure system also comprises means of control, and possibly adjustment, of at least one parameter among the pressure, the temperature and the hygrometry inside the chamber. exposure.
• control and possibly adjustment, of at least one parameter among the pressure, the temperature and the hygrometry inside the chamber. exposure.
• control means may be useful, if necessary, to control the temperature, the pressure, the humidity, etc., in order to possibly be able to adjust them.
• these control means comprise one or more sensors arranged inside the exposure chamber.
• Such control means are conventionally used by those skilled in the art, who will know how to choose the most appropriate according to the parameter or parameters to be controlled and possibly to adjust.
• the aerosol exposure system advantageously comprises means for maintaining a sample in the exposure chamber, in order to maintain the test container or the sample of test microorganisms during the implementation of a method of evaluating the integrity of a container or method of evaluating the antimicrobial activity of a product of interest.
• such holding means consist of a plate on which can be placed the container or the sample of microorganisms to be tested.
• the holding means are means for keeping the suspension in suspension. test container inside the exposure chamber.
• suspension holding means promote the deposition of the aerosol over the entire surface of said container. It is particularly possible to use a rod or cord at the end of which will be suspended the container to be tested.
• such a system can advantageously be used to implement a method for evaluating the integrity of a container, in which the source of product to be sprays in the form of an aerosol is a suspension of microorganisms, said process comprising successively the steps according to which:
• the container to be evaluated is introduced into the exposure chamber
• an aerosol is generated from the suspension of microorganisms, said aerosol being vaporized in the exposure chamber so that the microorganisms are in contact with the package;
• air is injected into the exposure chamber so as to remove the microorganisms from said exposure chamber;
• the container is recovered and incubated optionally under conditions allowing the growth of microorganisms of the suspension of microorganisms;
• the presence of microorganisms is systematically sought after recovery of the container on which the product in aerosol form, in particular the suspension of microorganisms, has been sprayed.
• the container designates any packaging, supposed to be hermetically closed, to contain a liquid product, solid or gaseous.
• a container may in particular be a flexible pouch containing a pharmaceutical product, with or without a connection, a flexible or rigid packaging, an overpouch, a film, a sachet, etc.
• the container and / or the exposure chamber preferably both, have been sterilized prior to the step of removing said container in said exposure chamber. Otherwise, it is possible to carry out a disinfection or thorough cleaning step.
• the vaporization time of the suspension of microorganisms, as well as the flow rate, may vary according to the container to be tested, the volume of the exposure chamber, the initial concentration of microorganisms, etc. In particular, the vaporization time may be between 5 minutes and 24 hours. Those skilled in the art are able to adapt the vaporization times and the flow rate according to these different parameters.
• the suspension of microorganisms is vaporized in the exposure chamber so as to obtain a concentration of microorganisms at the surface of the container of between 10 4 CFU / cm 2 and 10 8 CFU / cm 2 , preferably equal to 10 6 CFU / cm 2 , + / - 10 1 .
• the microorganism suspension used comprises spores.
• the suspension comprises from 10 1 spore / mL to 10 12 spores / mL.
• the suspension is sprayed with a pressure of between 3.5 and 5 bar, for 1.5 hours +/- 30 minutes, in an exposure chamber having a volume of 1 m 3 , so that the entire surface of the container suspended from the interior of said chamber is covered with microorganisms.
• test container Once the test container has been subjected to the aerosol of microorganisms for the desired time, said container is recovered in order to verify the possible presence of these microorganisms inside said container.
• any method for detecting microorganisms can be implemented. Those skilled in the art are able to choose both the culture conditions and the detection means to be used in particular according to the microorganisms of the suspension of microorganisms.
• the aerosol exposure system according to the invention can also be used in a method for evaluating the antimicrobial activity of a product of interest, in which the source of aerosol sprays is said product of interest, the process comprising the steps of:
• a source of microorganisms is arranged in the exposure chamber
• an aerosol is generated from the product of interest, the said aerosol being vaporized in the exposure chamber so that the microorganisms are in contact with the said product to be evaluated; optionally, air is injected into the exposure chamber so as to remove said product of interest from said exposure chamber;
• the rate of growth and / or survival of the microorganisms is evaluated.
• the source of microorganisms such as a petri dish on which bacteria are grown, is placed inside the exposure chamber, so as to be in contact with the product of interest to be tested, which will be sprayed in. bedroom.
• the source of microorganisms is recovered in order to evaluate the rate of growth and / or survival of said microorganisms.
• any evaluation method can be implemented. The skilled person is able to choose the evaluation method to be used in particular microorganisms of the source of microorganisms.
• the aerosol exposure system according to the invention can therefore be advantageously used to test the integrity of a container and / or the antimicrobial activity of a product of interest.
• the aerosol exposure system 1 comprises an enclosure 2, forming an exposure chamber, which can be sealed by means of mechanical locomotives 3 arranged all around the enclosure 2.
• the enclosure 2 is connected to a diffusion device an aerosol 4, by two ducts 5, 6, for bringing the aerosol from the diffusion device 4 to the upper part 7 of the chamber 2.
• Injection nozzles 9, 10 can inject aerosol inside the enclosure 2.
• the conduits 5, 6 extend from else of the chamber 2, so that they each open at a different end of the chamber 2, which allows a homogeneous distribution of the aerosol in the internal volume 8 of the chamber 2.
• a plate of support 16 is disposed in the lower part 15 of the chamber 2, on which a sample to be tested can be arranged.
• the sample may be a container, such as a package, or a culture of microorganisms.
• the diffusion device 4 is composed in this embodiment of an aerosol generator 11, connected to a source of product to be sprayed (not shown in the figure), a compressor 12 and an air dryer 13.
• a source of product to be sprayed not shown in the figure
• a compressor 12 and an air dryer 13.
• the product to be sprayed is partially dried when it is aerosolized, to be diffused through the conduits 5, 6 in the enclosure 2.
• the aerosol 14 spreads throughout the volume 8 of the chamber 2, and falls by sedimentation in the lower part 15 of the chamber 2.
• the aerosol exposure system as shown in FIG. 1 also comprises means for evacuating the aerosol 17, disposed at the bottom portion 15.
• the evacuation means 17 comprise a valve 18 through which the product in the form aerosol is evacuated, and a filter 19, such as a HEPA filter, connected to a blower 20.
• an air inlet / outlet 21 Such an air outlet, preferably situated in the upper part 7, makes it possible in particular to manage the pressure inside the enclosure 2. It can also be used at the end of the test, in order to help the evacuation of the microorganisms or the vaporized composition inside the enclosure 2.
• the bags were filled aseptically with sterile culture medium (caso broth). This step was carried out under a microbiological safety station (PSM). After this filling step, a fertility test of the culture medium is performed to ensure the quality of the medium.
• sterile culture medium caso broth
• PSM microbiological safety station
• Disks of the same material as the pouches are arranged in the chamber to control by counting the actual concentration of spores per cm 2 at the surface.
• the chamber air is rinsed with clean air (passed through a HEPA filter) for 45 minutes.

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• 2019
  • 2019-01-28 WO PCT/EP2019/051971 patent/WO2019145525A1/fr unknown
  • 2019-01-28 US US16/965,309 patent/US11466304B2/en active Active
  • 2019-01-28 JP JP2020562842A patent/JP2021512644A/ja active Pending
  • 2019-01-28 EP EP19701528.2A patent/EP3746563A1/fr active Pending

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