EP0343596B1 - Container for freeze drying under sterile conditions - Google Patents

Container for freeze drying under sterile conditions Download PDF

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Publication number
EP0343596B1
EP0343596B1 EP89109246A EP89109246A EP0343596B1 EP 0343596 B1 EP0343596 B1 EP 0343596B1 EP 89109246 A EP89109246 A EP 89109246A EP 89109246 A EP89109246 A EP 89109246A EP 0343596 B1 EP0343596 B1 EP 0343596B1
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EP
European Patent Office
Prior art keywords
membrane
container according
container
drying
freeze
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EP89109246A
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German (de)
French (fr)
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EP0343596A2 (en
EP0343596A3 (en
Inventor
Thomas Dr. Rer. Nat. Bergmann
Herbert Brustmann
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Roche Diagnostics GmbH
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Roche Diagnostics GmbH
Boehringer Mannheim GmbH
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Priority to AT89109246T priority Critical patent/ATE73226T1/en
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Classifications

    • F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
    • F26—DRYING
    • F26B—DRYING SOLID MATERIALS OR OBJECTS BY REMOVING LIQUID THEREFROM
    • F26B5/00—Drying solid materials or objects by processes not involving the application of heat
    • F26B5/04—Drying solid materials or objects by processes not involving the application of heat by evaporation or sublimation of moisture under reduced pressure, e.g. in a vacuum
    • F26B5/06—Drying solid materials or objects by processes not involving the application of heat by evaporation or sublimation of moisture under reduced pressure, e.g. in a vacuum the process involving freezing

Definitions

  • the invention relates to a container for freeze-drying under sterile conditions, in particular for drying biological and / or pharmaceutical material.
  • Freeze-drying of biological and pharmaceutical substances is generally known (see also Ullman's Encyclopedia of Industrial Chemistry, 3rd edition, Vol. I, p. 556 ff.), In order to avoid contamination of the dried goods with germs and other impurities , elaborate equipment and process engineering measures taken.
  • the procedure is such that vials containing the frozen good are provided with a bacterial filter and the good in the vial is dried in a first drying step until sublimation of the frozen solvent is complete is.
  • the so-called post-drying or residual drying the goods remaining moisture removed. Since this second drying step is usually carried out in a special apparatus, the ampoules or vials must be removed from the first drying apparatus in a further contamination-sensitive operation and introduced into the second drying apparatus.
  • the bacterial filter is removed and replaced by an aluminum cap with a rubber diaphragm and a hollow needle.
  • the drying room is filled with an inert gas and with a slight overpressure and the diaphragm opening is sealed as vapor-tight as possible by a sealing compound.
  • freeze-drying of biological and pharmaceutical material is also carried out on plates under sterile conditions.
  • a solution of the material to be dried is first sterilized, for example by filtration through a sterile filter, then poured onto plates under sterile conditions and freeze-dried using known methods.
  • this method presupposes that the entire freeze dryer can be sterilized. It is also necessary to keep the area around the drying plant germ-free.
  • the aim of the invention is now to overcome the disadvantages indicated above and to provide a simple container, with the aid of which a sterile, freeze-dried material can be obtained without the above-mentioned complex sterility requirements for the drying system and for the space surrounding it.
  • a container for freeze-drying in particular biological or pharmaceutical material under sterile conditions which is characterized in that the sides thereof at least partially consist of two walls which are tightly and pressure-tightly connected at their edges, one of which consists of a wall liquid-tight material and the other wall of a hydrophobic, porous, germ-tight, water vapor permeable membrane is formed.
  • the invention is based on the knowledge that the steam flow resulting from the sublimation of solvent molecules, in particular water molecules, which flows from the material to be dried to the condenser, is only hindered to a small extent by the membrane used in the container according to the invention. Freeze-drying of material enclosed by the membrane is therefore almost the same quickly like freeze-drying the same open, unpackaged material.
  • the membranes used according to the invention are hydrophobic membranes which contain pores which on the one hand are permeable to water vapor, but on the other hand are so small that they can no longer be passed through by microorganisms. Such pores preferably have a size of ⁇ 0.5 ⁇ m, in particular of ⁇ 0.2 ⁇ m.
  • Preferably membranes are used which are tear-resistant even under the respective process conditions even when wet.
  • the container according to the invention can also be used with less stable membranes, provided that these are reinforced with a carrier material or are not subjected to excessive mechanical stress.
  • the proportion of the membrane on the wall surface selected in the container according to the invention depends on the respectively selected conditions and the drying time and can easily be found out by a person skilled in the art by simply trying it out.
  • the entire wall surface consists of the membrane film, in a further preferred embodiment approximately half.
  • the container according to the invention can also be used advantageously if the wall surface consists of only 10% of the membrane film.
  • Semi-permeable papers made of cellulose and conventional cellulose derivatives, such as cellulose acetate, are particularly suitable.
  • Membranes made of films of polymer compounds, such as polytetrafluoroethylene or polypropylene, are also preferably used.
  • Films made of sterilization paper according to DIN 58 953 are also particularly suitable as water-vapor-permeable membranes.
  • any film membrane can be used regardless of its components, provided that it meets the requirements specified in DIN standard 58 953 with regard to germ density, air permeability and, in particular, strength.
  • the container according to the invention consists of a bag or hose, which consists of the two walls which are tightly and pressure-tightly connected at their edges, one wall of which consists of liquid-tight material and the other wall is formed by the membrane.
  • the membrane is preferably welded or glued to the vessel.
  • Troughs are particularly suitable as vessels according to the invention.
  • the trough is made of liquid-tight plastic and preferably has a wall thickness of 0.5 to 1 mm.
  • drying conditions such as pressure, temperature and quantity, depend on the material to be dried, the thickness of the membrane as well as the size and number of its pores and must be determined for the respective material and the packaging by simple and customary testing.
  • the germ density of a membrane was tested in accordance with DIN 58 953 in such a way that microorganisms in water drops were placed on the test pieces and after the water drops had dried on, it was examined whether microorganisms had passed through on the underside of the test pieces.
  • the membrane film to be tested was cut into squares with an edge length of approximately 50 mm.
  • the test pieces were sterilized and dried. Each test piece of the sterilized membrane was placed with the side that can be contaminated during use upwards on a sterilized surface and inoculated with 5 drops of 0.1 ml (corresponding to 107 to 107 germs).
  • the test pieces were stored at room temperature from 20 to 25 ° C under a relative humidity of 40 to 60%. The drops must be completely dry within 6 hours.
  • Each specimen was placed with the inoculated area up on the surface of a blood agar plate (1.5% agar) so that the entire film area came into contact with the agar. After 5 to 6 seconds the paper was removed.
  • the plates were incubated at 37 ° C for 16 to 25 h. If the agar plates treated with such film samples show no growth, the film is considered to be sufficiently germ-tight. Further information on the testing of the germ tightness of membranes, in particular the production of test germ suspensions, can be found in part 6 of DIN standard 58 953.
  • a nutrient solution was prepared which consisted of 10 g peptone, 5 g glucose, 5 g NaCl, 0.084 g KH2PO4, 0.187 g Na2-HPO4x2H2O and pyrogen-free water ad 1.0 l existed and which was adjusted to pH 7.0. It was then sterilized in a closed vial.
  • a transparent sterile bag consisting of a transparent film and a suitable paper was prepared to hold the sterile nutrient solution to be lyophilized.
  • the commercially available transparent sterilization pouch film from Wipak Medical, type Steri-King R 47, which is tubular, i.e. welded on both sides, otherwise open, located on a roll (width of the roll 400 mm), cut off a piece with a length of 800 mm. This tube was welded to a bag on the two open sides with a commercially available film device.
  • This bag was then sterilized in an autoclave with a filter program at 123 ° C and 2 bar steam pressure, the sterile bag with the transparent film facing downwards for better handling in an unsterile sheet metal tray (VA sheet, dimensions: length 800 mm, width 400 mm, height 30 mm) and opened in a laminar flow box under sterile conditions with disinfected scissors by cutting off a corner.
  • VA sheet unsterile sheet metal tray
  • 1.5 l of the sterile nutrient solution were introduced via a sterile tube pushed into the opening.
  • the bag filled in this way was sealed in the laminar flow box under sterile conditions by means of a commercially available film sealing device by corner sealing.
  • the entire arrangement (sheet tray, bag and sterile proximity solution) was placed on a plate of a commercially available, non-sterilizable freeze drying system from Edwards + Kniese pre-cooled to -45 ° C a total floor space of 1.5 m2 and the solution frozen. After completely freezing the solution under non-sterile conditions, freeze-drying was carried out at a pressure of 10 ⁇ 1 torr and a plate temperature of 22 ° C and the product was dried at 10 ⁇ 3 torr, also under non-sterile conditions. The total drying time was approx. 72 h.
  • the freeze-dried material thus obtained which was present as a light brown powder in the transparent sterilization bag, was placed in a laminar flow box including the bag and dissolved in 1.5 l of sterile water.
  • the intended puncture site on the paper side was disinfected with alcohol, a total of 1.5 l of sterile water was added to the bag using a sterile cannula and a suitable sterile syringe, the dried material was dissolved and the solution was transferred to a sterile bottle. This solution was incubated for 4 days at 37 ° C. and the germ count of the incubated solution was then determined using the membrane filter method.

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  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Molecular Biology (AREA)
  • Mechanical Engineering (AREA)
  • General Engineering & Computer Science (AREA)
  • Medical Preparation Storing Or Oral Administration Devices (AREA)
  • Drying Of Solid Materials (AREA)
  • Freezing, Cooling And Drying Of Foods (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Separation Using Semi-Permeable Membranes (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)

Abstract

For the freeze-drying of, in particular, biological or pharmaceutical material under sterile conditions, the material to be dried is placed in a container whose sides are at least partly composed of a hydrophobic, porous, germproof membrane which is permeable to water vapour, the container is closed pressure-tight and then the material is freeze-dried in the closed container under usual conditions.

Description

Der Erfindung betriff einen Behälter zum Gefriertrocknen unter sterilen Bedingungen, insbesondere zum Trocknen von biologischem und/oder pharmazeutischem Material.The invention relates to a container for freeze-drying under sterile conditions, in particular for drying biological and / or pharmaceutical material.

Bei biologischem und pharmazeutischem Material ist es häufig notwendig, die Substanzen bis zu ihrer Verwendung völlig trocken zu lagern. Meist sind diese empfindlichen Substanzen nur durch Gefriertrocknung zugänglich. Zudem besteht in der Regel die Notwendigkeit, diese Substanzen völlig frei von mikrobiologischen Keimen zu halten, und zwar sowohl wegen der durch Mikroben verursachten Zersetzung von biologischen Substanzen als auch, um mögliche Infektionen bei ihrer Verwendung zu verhindern.With biological and pharmaceutical material, it is often necessary to store the substances completely dry until they are used. Most of these sensitive substances are only accessible by freeze drying. In addition, there is usually a need to keep these substances completely free of microbiological germs, both because of the decomposition of biological substances caused by microbes and in order to prevent possible infections when they are used.

Das Gefriertrocknen von biologischen und pharmazeutischen Substanzen ist allgemein bekannt (siehe auch Ullmans Enzyklopädie der Technischen Chemie, 3. Aufl., Bd. I, S. 556 ff.) Dabei werden, um eine Kontamination des getrockneten Gutes mit Keimen und anderen Verunreinigungen zu vermeiden, aufwendige apparative und verfahrenstechnische Maßnahmen getroffen.Freeze-drying of biological and pharmaceutical substances is generally known (see also Ullman's Encyclopedia of Industrial Chemistry, 3rd edition, Vol. I, p. 556 ff.), In order to avoid contamination of the dried goods with germs and other impurities , elaborate equipment and process engineering measures taken.

Beim Trocknen von pharmazeutischen Präparaten in Ampullen oder Fläschchen wird beispielsweise so vorgegangen, daß Fläschchen, die das gefrorene Gut enthalten, mit einem Bakterienfilter versehen werden und das Gut in den Fläschchen in einem ersten Trocknungsschritt so weit getrocknet wird, bis die Sublimation des gefrorenen Lösungsmittels abgeschlossen ist.When drying pharmaceutical preparations in ampoules or vials, for example, the procedure is such that vials containing the frozen good are provided with a bacterial filter and the good in the vial is dried in a first drying step until sublimation of the frozen solvent is complete is.

Anschließend wird in einem zweiten Trocknungsabschnitt, der sogenannten Nach- oder Resttrocknung, dem Gut die noch verbliebene Restfeuchtigkeit entzogen. Da dieser zweite Trocknungsschritt meist in einer besonderen Apparatur durchgeführt wird, müssen die Ampullen oder Phiolen in einem weiteren kontaminationsempfindlichen Arbeitsgang der ersten Trocknungsapparatur entnommen und in die zweite Trocknungsapparatur eingebracht werden. Dazu werden die Bakterienfilter entfernt und durch eine mit einem Gummidiaphragma und einer Hohlnadel versehenen Aluminiumkappe ersetzt. Nach einer je nach Art des zu trocknenden Gutes mehrtägigen Resttrocknung wird der Trockenraum mit einem Inertgas und mit leichtem Überdruck gefüllt und die Diaphragmaöffnung durch eine Vergußmasse möglichst dampfdicht verschlossen.Then, in a second drying section, the so-called post-drying or residual drying, the goods remaining moisture removed. Since this second drying step is usually carried out in a special apparatus, the ampoules or vials must be removed from the first drying apparatus in a further contamination-sensitive operation and introduced into the second drying apparatus. For this purpose, the bacterial filter is removed and replaced by an aluminum cap with a rubber diaphragm and a hollow needle. After a residual drying process lasting several days, depending on the type of material to be dried, the drying room is filled with an inert gas and with a slight overpressure and the diaphragm opening is sealed as vapor-tight as possible by a sealing compound.

Da die Sublimationsgeschwindigkeit bei dieser Art der Gefriertrocknung nur etwa halb so grob ist wie diejenige von offen ausgebreitetem Material, wird die Gefriertrocknung von biologischem und pharmazeutischem Material auch auf Platten unter sterilen Bedingungen durchgeführt. Dabei wird eine Lösung des zu trocknenden Gutes zuerst sterilisiert, beispielsweise durch Filtration über einen Sterilfilter, anschließend unter sterilen Bedingungen auf Platten gegossen und mittels bekannter Methoden gefriergetrocknet. Dieses Verfahren setzt jedoch voraus, daß die gesamte Gefriertrocknungsanlage sterilisierbar ist. Zudem ist es erforderlich, auch die Umgebung der Trocknungsanlage keimfrei zu halten.Since the rate of sublimation in this type of freeze-drying is only about half as large as that of openly spread material, the freeze-drying of biological and pharmaceutical material is also carried out on plates under sterile conditions. A solution of the material to be dried is first sterilized, for example by filtration through a sterile filter, then poured onto plates under sterile conditions and freeze-dried using known methods. However, this method presupposes that the entire freeze dryer can be sterilized. It is also necessary to keep the area around the drying plant germ-free.

Nach erfolgter Trocknung ist es notwendig, das Gut in der Trocknungsanlage selbst oder in ihrer Umgebung mit mechanischen Verfahren von den Platten unter sterilen Bedingungen zu entfernen und in ebenfalls sterile Aufbewahrungsbehältnisse zu füllen. Dieses Verfahren erfordert aufwendige Anlagen und sterile Räume sowie ein besonders sorgfältiges Arbeiten mit dem zu trocknenden bzw. dem bereits getrockneten Gut bis zu seiner gebrauchsfertigen Konfektionierung.After drying, it is necessary to remove the material from the plates in the drying system itself or in its surroundings by mechanical methods under sterile conditions and to fill it in sterile storage containers as well. This procedure requires complex facilities and sterile rooms as well particularly careful work with the goods to be dried or the goods that have already dried until they are ready for use.

Aus der GB-A-995 930 war es schon bekannt, radioaktive wäßrige Lösungen in dampfdurchlässigen Beuteln gefrierzutrocknen um dadurch für die Sublimation eine größere Oberfläche als beim Trocknen in Flaschen zur Verfügung zu stellen. Das Problem einer mikrobiellen Kontamination des zu trocknenden Guts stellte sich dabei nicht.From GB-A-995 930 it was already known to freeze-dry radioactive aqueous solutions in vapor-permeable bags in order to provide a larger surface for sublimation than when drying in bottles. The problem of microbial contamination of the items to be dried did not arise.

Die Erfindung hat nun zum Ziel, die oben aufgezeigten Nachteile zu überwinden und einen einfachen Behälter bereitzustellen, mit dessen Hilfe ohne die oben angeführten aufwendigen Sterilitätsanforderungen an die Trocknungsanlage sowie an den diese umgebenden Raum ein steriles, gefriergetrocknetes Material gewonnen werden kann.The aim of the invention is now to overcome the disadvantages indicated above and to provide a simple container, with the aid of which a sterile, freeze-dried material can be obtained without the above-mentioned complex sterility requirements for the drying system and for the space surrounding it.

Dieses Ziel wird erfindungsgemäß erreicht durch einen Behälter zum Gefriertrocknen von insbesondere biologischem oder pharmazeutischem Material unter sterilen Bedingungen, der dadurch gekennzeichnet ist, daß dessen Seiten zumindest teilweise aus zwei an ihren Rändern dicht- und druckfest miteinander verbundenen Wänden bestehen, von denen die eine Wand aus flüssigkeitsdichtem Material und die andere Wand aus einer hydrophoben, porigen, keimdichten, wasserdampfdurchlässigen Membran gebildet wird.This object is achieved according to the invention by a container for freeze-drying in particular biological or pharmaceutical material under sterile conditions, which is characterized in that the sides thereof at least partially consist of two walls which are tightly and pressure-tightly connected at their edges, one of which consists of a wall liquid-tight material and the other wall of a hydrophobic, porous, germ-tight, water vapor permeable membrane is formed.

Die Erfindung beruht auf der Erkenntnis, daß der durch die Sublimation von Lösungsmittelmolekülen, insbesondere von Wassermolekülen, entstehende Dampfstrom, der vom zu trocknenden Gut hin zum Kondensator fließt, durch die im erfindungsgemäßen Behälter verwendete Membran nur in geringem Umfang behindert wird. Somit läuft das Gefriertrocknen von Material, das von der Membran umschlossen ist, nahezu gleich schnell ab wie das Gefriertrocknen desselben offenen, nicht verpackten Materials. Bei den erfindungsgemäß verwendeten Membranen handelt es sich um hydrophobe Membranen, die Poren enthalten, die einerseits für Wasserdampf durchlässig, andererseits jedoch so klein sind, daß sie von Mikroorganismen nicht mehr passiert werden können. Solche Poren haben vorzugsweise eine Größe von ≦ 0,5 µm, insbesondere von ≦ 0,2 µm. Vorzugsweise werden Membranen verwendet, die auch noch im nassen Zustand unter den jeweiligen Verfahrensbedingungen reißfest sind. Allerdings ist der erfindungsgemäße Behälter auch mit weniger stabilen Membranen brauchbar, sofern diese mit einem Trägermaterial verstärkt sind oder nicht übermäßig mechanisch beansprucht werden.The invention is based on the knowledge that the steam flow resulting from the sublimation of solvent molecules, in particular water molecules, which flows from the material to be dried to the condenser, is only hindered to a small extent by the membrane used in the container according to the invention. Freeze-drying of material enclosed by the membrane is therefore almost the same quickly like freeze-drying the same open, unpackaged material. The membranes used according to the invention are hydrophobic membranes which contain pores which on the one hand are permeable to water vapor, but on the other hand are so small that they can no longer be passed through by microorganisms. Such pores preferably have a size of ≦ 0.5 μm, in particular of ≦ 0.2 μm. Preferably membranes are used which are tear-resistant even under the respective process conditions even when wet. However, the container according to the invention can also be used with less stable membranes, provided that these are reinforced with a carrier material or are not subjected to excessive mechanical stress.

Der im erfindungsgemäßen Behälter jeweils gewählte Anteil der Membran an der Wandfläche hängt von den jeweils gewählten Bedingungen und der Trocknungsdauer ab und kann vom Fachmann mittels einfachem Ausprobieren leicht herausgefunden werden. In einer bevorzugten Ausführungsform besteht die gesamte Wandfläche aus der Membranfolie, in einer weiteren bevorzugten Ausführungsform etwa zu Hälfte. Der erfindungsgemäße Behälter ist auch noch dann vorteilhaft brauchbar, wenn die Wandfläche nur zu 10 % aus der Membranfolie besteht.The proportion of the membrane on the wall surface selected in the container according to the invention depends on the respectively selected conditions and the drying time and can easily be found out by a person skilled in the art by simply trying it out. In a preferred embodiment, the entire wall surface consists of the membrane film, in a further preferred embodiment approximately half. The container according to the invention can also be used advantageously if the wall surface consists of only 10% of the membrane film.

Im besonderen eignen sich halbdurchlässige Papiere aus Cellulose und üblichen Cellulosederivaten, wie Celluloseacetat. Vorzugsweise finden auch Membranen aus Folien von Polymerverbindungen, wie Polytetrafluorethylen oder Polypropylen, Verwendung. Ganz besonders eignen sich als wasserdampfdurchlässige Membranen auch Folien aus Sterilisationspapier nach DIN 58 953. In weiteren bevorzugten Ausführungsformen der Erfindung werden Goretex- und ähnliche Membranen oder auch handelsübliche Folienschläuche eingesetzt. Im Prinzip ist jede Folienmenbran unabhängig von ihren Bestandteilen verwendbar, sofern sie die in der DIN-Norm 58 953 angegebenen Anforderungen bezüglich Keimdichte, Luftdurchlässigkeit und insbesondere Festigkeit erfüllt.Semi-permeable papers made of cellulose and conventional cellulose derivatives, such as cellulose acetate, are particularly suitable. Membranes made of films of polymer compounds, such as polytetrafluoroethylene or polypropylene, are also preferably used. Films made of sterilization paper according to DIN 58 953 are also particularly suitable as water-vapor-permeable membranes. In further preferred embodiments of the invention, Goretex and similar membranes or commercially available membranes Foil tubes used. In principle, any film membrane can be used regardless of its components, provided that it meets the requirements specified in DIN standard 58 953 with regard to germ density, air permeability and, in particular, strength.

In einer bevorzugten Ausführungsform besteht der erfindungsgemäße Behälter aus einem Beutel oder Schlauch, der aus den zwei an ihren Rändern miteinander dicht und druckfest verbundenen Wänden besteht, wovon die eine Wand aus flüssigkeitsdichtem Material besteht und die andere Wand von der Membran gebildet wird.In a preferred embodiment, the container according to the invention consists of a bag or hose, which consists of the two walls which are tightly and pressure-tightly connected at their edges, one wall of which consists of liquid-tight material and the other wall is formed by the membrane.

Die Membran ist mit dem Gefäß vorzugsweise verschweißt oder verklebt. Als Gefäße eignen sich erfindungsgemäß besonders Wannen.The membrane is preferably welded or glued to the vessel. Troughs are particularly suitable as vessels according to the invention.

In einer weiteren bevorzugten Ausführungsform besteht die Wanne aus flüssigkeitsdichtem Kunststoff und hat vorzugsweise eine Wandstärke von 0,5 bis 1 mm.In a further preferred embodiment, the trough is made of liquid-tight plastic and preferably has a wall thickness of 0.5 to 1 mm.

Die günstigsten Trocknungsbedingungen, wie Druck, Temperatur und Menge, sind abhängig von dem jeweils zu trocknenden Material, der Dicke der Membran sowie der Größe und Anzahl ihrer Poren und müssen durch übliches und einfaches Ausprobieren für das jeweilige Material und die Verpackung bestimmt werden.The most favorable drying conditions, such as pressure, temperature and quantity, depend on the material to be dried, the thickness of the membrane as well as the size and number of its pores and must be determined for the respective material and the packaging by simple and customary testing.

Im folgenden wird die Erfindung anhand einiger Ausführungsbeispiele näher erläutert.The invention is explained in more detail below with the aid of a few exemplary embodiments.

Beispiel 1example 1

Die Prüfung der Keimdichte einer Membran wurde gemäß DIN 58 953 so durchgeführt, daß Mikroorganismen in Wassertropfen auf die Probestücke gebracht wurden und nach dem Antrocknen der Wassertropfen untersucht wurde, ob Mikroorganismen auf die Unterseite der Probestücke durchgetreten sind.The germ density of a membrane was tested in accordance with DIN 58 953 in such a way that microorganisms in water drops were placed on the test pieces and after the water drops had dried on, it was examined whether microorganisms had passed through on the underside of the test pieces.

Die zu prüfende Membranfolie wurde in Quadrate von etwa 50 mm Kantenlänge geschnitten. Die Probestücke wurden sterilisiert und getrocknet. Jedes Probestück der sterilisierten Membran wurde mit der Seite, die bei der Anwendung kontaminiert werden kann, nach oben auf eine sterilisierte Unterlage gelegt und mit 5 Tropfen zu je 0,1 ml (entsprechend 10⁶ bis 10⁷ Keimen) beimpft. Die Probestücke wurden bei Raumtemperatur von 20 bis 25 °C unter einer relativen Luftfeuchte von 40 bis 60 % gelagert. Die Tropfen müssen innerhalb von 6 h vollständig getrocknet sein. Jedes Probestück wurde mit der beimpften Fläche nach oben auf die Oberfläche einer Blutagarplatte (1,5 % Agar) gelegt, so daß die ganze Folienfläche mit dem Agar in Kontakt kam. Nach 5 bis 6 sec wurde das Papier entfernt. Die Platten wurden 16 bis 25 h bei 37 °C bebrütet. Weisen die mit solchen Folienproben behandelten Agarplatten kein Wachstum auf, gilt die Folie als ausreichend keimdicht. Weitere Angaben über die Prüfung der Keimdichtigkeit von Membranen, insbesondere die Herstellung von Testkeimsuspensionen, können dem Teil 6 der DIN-Norm 58 953 entnommen werden.The membrane film to be tested was cut into squares with an edge length of approximately 50 mm. The test pieces were sterilized and dried. Each test piece of the sterilized membrane was placed with the side that can be contaminated during use upwards on a sterilized surface and inoculated with 5 drops of 0.1 ml (corresponding to 10⁷ to 10⁷ germs). The test pieces were stored at room temperature from 20 to 25 ° C under a relative humidity of 40 to 60%. The drops must be completely dry within 6 hours. Each specimen was placed with the inoculated area up on the surface of a blood agar plate (1.5% agar) so that the entire film area came into contact with the agar. After 5 to 6 seconds the paper was removed. The plates were incubated at 37 ° C for 16 to 25 h. If the agar plates treated with such film samples show no growth, the film is considered to be sufficiently germ-tight. Further information on the testing of the germ tightness of membranes, in particular the production of test germ suspensions, can be found in part 6 of DIN standard 58 953.

Beispiel 2Example 2

Es wurde eine Nährlösung hergestellt, die aus 10 g Pepton, 5 g Glucose, 5 g NaCl, 0,084 g KH₂PO₄, 0,187 g Na₂-HPO₄x2H₂O und pyrogenfreiem Wasser ad 1,0 l bestand und die auf pH 7,0 eingestellt wurde. Anschließend wurde sie in einer verschlossenen Durchstichflasche endsterilisiert.A nutrient solution was prepared which consisted of 10 g peptone, 5 g glucose, 5 g NaCl, 0.084 g KH₂PO₄, 0.187 g Na₂-HPO₄x2H₂O and pyrogen-free water ad 1.0 l existed and which was adjusted to pH 7.0. It was then sterilized in a closed vial.

Zur Aufnahme der sterilen, zu lyophilisierenden Nährlösung wurde ein Klarsichtsterilbeutel, bestehend aus einer Klarsichtfolie und einem geeigneten Papier, angefertigt. Hierzu wurde von der handelsüblichen Klarsichtsterilisierungsbeutelfolie der Firma Wipak Medical, Typ Steri-King R 47, die sich schlauchförmig, d.h. beiderseits verschweißt, sonst offen, auf einer Rolle (Breite der Rolle 400 mm) befindet, ein Stück von einer Länge von 800 mm abgeschnitten. Dieser Schlauch wurde an den beiden offenen Seiten zu einem Beutel mit einem handelsüblichen Foliengerät verschweißt. Anschließend wurde dieser Beutel in einem Autoklaven mit Filterprogramm bei 123 °C und 2 bar Dampfdruck sterilisiert, der sterile Beutel mit der Klarsichtfolie nach unten zur besseren Handhabung in eine unsterile Blechwanne (VA-Blech, Abmessungen: Länge 800 mm, Breite 400 mm, Höhe 30 mm) gelegt und in einer Laminar-Flowbox unter sterilen Bedingungen mit einer desinfizierten Schere durch Abschneiden einer Ecke geöffnet. Durch diese Öffnung von etwa 30 mm zwischen Folie und Papier wurden 1,5 l der sterilen Nährlösung über einen in die Öffnung geschobenen sterilen Schlauch eingefüllt. Der so gefüllte Beutel wurde noch in der Laminar-Flowbox unter sterilen Bedingungen mittels eines handelsüblichen Folienschweißgeräts durch Verschweißen über Eck verschlossen.A transparent sterile bag consisting of a transparent film and a suitable paper was prepared to hold the sterile nutrient solution to be lyophilized. For this purpose, the commercially available transparent sterilization pouch film from Wipak Medical, type Steri-King R 47, which is tubular, i.e. welded on both sides, otherwise open, located on a roll (width of the roll 400 mm), cut off a piece with a length of 800 mm. This tube was welded to a bag on the two open sides with a commercially available film device. This bag was then sterilized in an autoclave with a filter program at 123 ° C and 2 bar steam pressure, the sterile bag with the transparent film facing downwards for better handling in an unsterile sheet metal tray (VA sheet, dimensions: length 800 mm, width 400 mm, height 30 mm) and opened in a laminar flow box under sterile conditions with disinfected scissors by cutting off a corner. Through this opening of approximately 30 mm between the film and the paper, 1.5 l of the sterile nutrient solution were introduced via a sterile tube pushed into the opening. The bag filled in this way was sealed in the laminar flow box under sterile conditions by means of a commercially available film sealing device by corner sealing.

Die gesamte Anordnung (Blechwanne, Beutel und sterile Näherlösung) wurde auf eine auf -45 °C vorgekühlte Platte einer handelsüblichen, nicht sterilisierbaren Gefriertrocknungsanlage der Firma Edwards + Kniese mit einer Gesamtstellfläche von 1,5 m² verbracht und die Lösung eingefroren. Nach vollständigem Einfrieren der Lösung unter nichtsterilen Bedingungen wurde bei einem Druck von 10⁻¹ torr und einer Plattentemperatur von 22 °C gefriergetrocknet und das Produkt bei 10⁻³ torr, ebenfalls unter nichtsterilen Bedingungen, nachgetrocknet. Die Gesamttrocknungsdauer betrug ca. 72 h.The entire arrangement (sheet tray, bag and sterile proximity solution) was placed on a plate of a commercially available, non-sterilizable freeze drying system from Edwards + Kniese pre-cooled to -45 ° C a total floor space of 1.5 m² and the solution frozen. After completely freezing the solution under non-sterile conditions, freeze-drying was carried out at a pressure of 10⁻¹ torr and a plate temperature of 22 ° C and the product was dried at 10⁻³ torr, also under non-sterile conditions. The total drying time was approx. 72 h.

Das so erhaltene, als hellbraunes Pulver im Klarsichtsterilisierbeutel vorliegende gefriergetrocknete Gut wurde einschließlich Beutel in eine Laminar-Flowbox verbracht und in 1,5 l sterilem Wasser gelöst. Hierzu wurde auf der Papierseite die vorgesehene Einstichstelle mit Alkohol desinfiziert, mittels einer sterilen Kanüle und geeigneten sterilen Spritze insgesamt 1,5 l steriles Wasser in den Beutel gegeben, das getrocknete Gut gelöst und die Lösung in eine sterile Flasche überführt. Diese Lösung wurde 4 Tage bei 37 °C bebrütet und anschließend die Keimzahl der bebrüteten Lösung nach der Membranfiltermethode bestimmt.The freeze-dried material thus obtained, which was present as a light brown powder in the transparent sterilization bag, was placed in a laminar flow box including the bag and dissolved in 1.5 l of sterile water. For this purpose, the intended puncture site on the paper side was disinfected with alcohol, a total of 1.5 l of sterile water was added to the bag using a sterile cannula and a suitable sterile syringe, the dried material was dissolved and the solution was transferred to a sterile bottle. This solution was incubated for 4 days at 37 ° C. and the germ count of the incubated solution was then determined using the membrane filter method.

Es zeigte sich, daß durch die Gefriertrockung keine Keime eingeschleppt werden.It was shown that no germs were introduced by freeze-drying.

Claims (10)

1. Container for freeze drying of especially biological or pharmaceutical material under sterile conditions, characterised in that its sides consist at least partly of two walls connected hermetically and pressure-proof with one another on their edges, one wall of which is made from liquid-tight material and the other wall of a hydrophobic, porous, germ-tight water vapour-permeable material.
2. Container according to claim 1, characterised in that the pores of the membrane have a size of ≦ 0.5 µm.
3. Container according to claim 2, characterised in that the pores of the membrane have a size of ≦ 0.2 µm.
4. Container according to one of claims 1 to 3, characterised in that the membrane is a film with the properties according to DIN 58 953.
5. Container according to one of claims 1 to 4, characterised in that the membrane consists of semi-permeable paper, preferably of cellulose and cellulose derivatives.
6. Container according to claim 5, characterised in that the membrane consists of cellulose acetate.
7. Container according to claim 1 to 4, characterised in that the membrane is a film of a polymer compound, preferably of polytetrafluoroethylene or polypropylene.
8. Container according to claim 1 to 7, characterised in that it is formed as tube or bag.
9. Container according to claim 1 to 7, characterised in that it is formed as flask, ampoule or vial which is closed with the membrane.
10. Container according to claim 1 to 7, characterised in that it is formed as trough which is connected pressure-proof with the membrane as covering.
EP89109246A 1988-05-26 1989-05-23 Container for freeze drying under sterile conditions Expired - Lifetime EP0343596B1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AT89109246T ATE73226T1 (en) 1988-05-26 1989-05-23 CONTAINERS FOR FREEZE DRYING IN STERILE CONDITIONS.

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE3817906A DE3817906A1 (en) 1988-05-26 1988-05-26 METHOD AND CONTAINER FOR FREEZING DRYING UNDER STERILE CONDITIONS
DE3817906 1988-05-26

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EP0343596A2 EP0343596A2 (en) 1989-11-29
EP0343596A3 EP0343596A3 (en) 1990-02-28
EP0343596B1 true EP0343596B1 (en) 1992-03-04

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EP (1) EP0343596B1 (en)
JP (1) JPH0229256A (en)
AT (1) ATE73226T1 (en)
CA (1) CA1337974C (en)
DD (1) DD283864A5 (en)
DE (2) DE3817906A1 (en)
DK (1) DK173643B1 (en)
ES (1) ES2030556T3 (en)
FI (1) FI91442C (en)
GR (1) GR3004584T3 (en)
HU (1) HU204126B (en)
IE (1) IE61012B1 (en)
PL (1) PL159938B1 (en)

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US5958778A (en) * 1995-09-22 1999-09-28 The United States Of America As Represented By The Department Of Health And Human Services Container for drying biological samples, method of making such container, and method of using same
US6312648B1 (en) 1998-01-12 2001-11-06 The United States Of America As Represented By The Department Of Health And Human Services Applicator system
US10793327B2 (en) 2017-10-09 2020-10-06 Terumo Bct Biotechnologies, Llc Lyophilization container and method of using same
US11604026B2 (en) 2019-03-14 2023-03-14 Terumo Bct Biotechnologies, Llc Lyophilization loading tray assembly and system

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IL90188A0 (en) * 1988-05-18 1989-12-15 Cryopharm Corp Process and medium for the lyophilization of erythrocytes
US5045446A (en) * 1988-08-26 1991-09-03 Cryopharm Corporation Lyophilization of cells
US5257983A (en) * 1991-04-12 1993-11-02 Cryopharm Corporation Blood bag for lyophilization
WO1995027180A1 (en) * 1994-04-04 1995-10-12 W.L. Gore & Associates, Inc. Improved method for minimizing contamination of freeze-dried products
WO1996006018A1 (en) * 1994-08-19 1996-02-29 W. L. Gore & Associates, Inc. Vented vial for freeze-drying and method of minimizing contamination of freeze-dried products
FR2738057B1 (en) * 1995-08-22 1997-11-07 Lab Francais Du Fractionnement WATERPROOF PACKAGING FOR DRYING, ESPECIALLY LYOPHILIZATION, AND DRYING METHOD, ESPECIALLY LYOPHILIZATION, USING SUCH PACKAGING
FR2740108B1 (en) * 1995-08-22 1998-03-27 Lab Francais Du Fractionnement WATERPROOF PACKAGING FOR DRYING, ESPECIALLY LYOPHILIZATION, AND DRYING METHOD, ESPECIALLY LYOPHILIZATION, USING SUCH PACKAGING
US5596814A (en) * 1995-11-06 1997-01-28 W. L. Gore & Associates, Inc. Vented vial stopper for processing freeze-dried products
AT1399U1 (en) 1995-11-29 1997-04-25 Immuno Ag METHOD AND DEVICE FOR LYOPHILIZING
DE19751031A1 (en) 1997-11-19 1999-06-24 Ingo Dipl Ing Heschel Process for the production of porous structures
DE19815993C2 (en) * 1998-04-09 2003-03-06 Schott Glas Freeze-drying containers and storage for medical products
EP1958618A1 (en) 2007-02-15 2008-08-20 Octapharma AG Method for freeze-drying with optimum reconstitution of biopolymers
JP2010124931A (en) * 2008-11-26 2010-06-10 Kanae Co Ltd Method for manufacturing package of freeze-dried medicine
EP2386399B8 (en) * 2010-04-23 2015-08-05 MC Beteiligungs-GmbH Method for making openings in a waterproof coating and base body with such coating
WO2015162273A1 (en) 2014-04-25 2015-10-29 Merck Sharp & Dohme Bv A method to dry multiple individual frozen bodies and a system for applying this method
JP2019090596A (en) * 2017-11-10 2019-06-13 エイブル株式会社 Method for producing freeze-dried product, freeze-drying bag, and freeze-drying device

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JPS6136942A (en) * 1984-07-28 1986-02-21 Sony Corp Apparatus for electronic part

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5958778A (en) * 1995-09-22 1999-09-28 The United States Of America As Represented By The Department Of Health And Human Services Container for drying biological samples, method of making such container, and method of using same
US6312648B1 (en) 1998-01-12 2001-11-06 The United States Of America As Represented By The Department Of Health And Human Services Applicator system
US10793327B2 (en) 2017-10-09 2020-10-06 Terumo Bct Biotechnologies, Llc Lyophilization container and method of using same
US11604026B2 (en) 2019-03-14 2023-03-14 Terumo Bct Biotechnologies, Llc Lyophilization loading tray assembly and system
US11609042B2 (en) 2019-03-14 2023-03-21 Terumo Bct Biotechnologies, Llc Multi-part lyophilization container and method of use

Also Published As

Publication number Publication date
DK252589A (en) 1989-11-27
GR3004584T3 (en) 1993-04-28
CA1337974C (en) 1996-01-23
EP0343596A2 (en) 1989-11-29
FI892563A (en) 1989-11-27
HU204126B (en) 1991-11-28
DK252589D0 (en) 1989-05-24
ATE73226T1 (en) 1992-03-15
DD283864A5 (en) 1990-10-24
ES2030556T3 (en) 1992-11-01
PL159938B1 (en) 1993-01-29
PL279609A1 (en) 1990-01-22
HUT52617A (en) 1990-07-28
FI91442B (en) 1994-03-15
FI91442C (en) 1994-06-27
DK173643B1 (en) 2001-05-14
IE891541L (en) 1989-11-26
DE3817906A1 (en) 1989-11-30
IE61012B1 (en) 1994-09-07
EP0343596A3 (en) 1990-02-28
JPH0229256A (en) 1990-01-31
JPH0450830B2 (en) 1992-08-17
FI892563A0 (en) 1989-05-25
DE58900902D1 (en) 1992-04-09

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