kSIST FprEN 12353:2020
(Main)Chemical disinfectants and antiseptics - Preservation of test organisms used for the determination of bactericidal (including Legionella), mycobactericidal, sporicidal, fungicidal and virucidal (including bacteriophages) activity
Chemical disinfectants and antiseptics - Preservation of test organisms used for the determination of bactericidal (including Legionella), mycobactericidal, sporicidal, fungicidal and virucidal (including bacteriophages) activity
This document specifies methods for keeping test organisms used and defined in European Standards for the determination of bactericidal (incl. Legionella pneumophila), mycobactericidal, sporicidal, fungicidal and virucidal (incl. bacteriophages) activity of chemical disinfectants and antiseptics drawn up by CEN/TC 216. These methods for keeping test organisms can only be carried out in connection with at least one of those standards where a reference to this document is established.
NOTE 1 Annex A (informative) contains a non-exhaustive list of test organisms for which this document can be applied.
NOTE 2 European Standards (EN and prEN) where this document is referenced are listed in the Bibliography.
NOTE 3 A specific part on the preservation of bacterial spores could be added once the results of the ongoing ring trials are available.
Chemische Desinfektionsmittel und Antiseptika - Aufbewahrung von Prüforganismen für die Prüfung der bakteriziden (einschließlich Legionella), mykobakteriziden, sporiziden, fungiziden und viruziden (einschließlich Bakteriophagen) Wirkung
Dieses Dokument legt Verfahren zur Haltung von Prüforganismen fest, wie sie in den Europäischen Normen des CEN/TC 216 zur Bestimmung der bakteriziden (einschließlich Legionella pneumophila), myko-bakteriziden, sporiziden, fungiziden und viruziden (einschließlich Bakteriophagen) Wirkung chemischer Desinfektionsmittel und Antiseptika verwendet und festgelegt werden. Diese Verfahren zur Haltung von Prüforganismen können nur in Verbindung mit mindestens einer der Normen erfolgen, in denen auf dieses Dokument verwiesen wird.
ANMERKUNG 1 Anhang A (informativ) enthält eine nicht abschließende Liste von Prüforganismen, auf die dieses Dokument angewendet werden kann.
ANMERKUNG 2 Europäische Normen (EN), die auf dieses Dokument verweisen, sind in den Literaturhinweisen aufgelistet.
ANMERKUNG 3 Eine spezifische Beschreibung über die Aufbewahrung von bakteriellen Sporen könnten hinzugefügt werden, sobald die Ergebnisse der laufenden Ringversuche vorliegen.
Antiseptiques et désinfectants chimiques - Conservation des micro organismes d'essai utilisés pour la détermination de l'activité bactéricide (Legionella incluses), mycobactéricide, sporicide, fongicide et virucide (bactériophages inclus)
Le présent document spécifie les méthodes de conservation des micro-organismes d’essai utilisées et définies dans les Normes européennes relatives à la détermination de l’activité bactéricide (Legionella pneumophila incluse), mycobactéricide, sporicide, fongicide et virucide (bactériophages inclus) des désinfectants et antiseptiques chimiques établies par le CEN/TC 216. Ces méthodes de conservation des micro-organismes d’essai peuvent être mises en oeuvre uniquement en association avec au moins une des normes citant en référence le présent document.
NOTE 1 L’Annexe A (informative) dresse une liste non exhaustive des micro-organismes d’essai auxquels s’applique le présent document.
NOTE 2 Les Normes européennes (EN) faisant référence au présent document sont indiquées dans la bibliographie.
NOTE 3 Une description spécifique sur la conservation des spores bactériennes peut être ajoutée dès que les résultats des essais interlaboratoires en cours seront disponibles.
Kemična razkužila in antiseptiki - Shranjevanje preskusnih organizmov za določanje baktericidnega (vključno Legionella), mikobaktericidnega, sporocidnega, fungicidnega in virucidnega (vključno bakteriofagi) delovanja
General Information
RELATIONS
Standards Content (sample)
SLOVENSKI STANDARD
oSIST prEN 12353:2019
01-februar-2019
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GRORþDQMHEDNWHULFLGQHJD YNOMXþQR/HJLRQHOOD PLNREDNWHULFLGQHJD
VSRURFLGQHJDIXQJLFLGQHJDLQYLUXFLGQHJD YNOMXþQREDNWHULRIDJL GHORYDQMD
Chemical disinfectants and antiseptics - Preservation of test organisms used for the
determination of bactericidal (including Legionella), mycobactericidal, sporicidal,
fungicidal and virucidal (including bacteriophages) activityChemische Desinfektionsmittel und Antiseptika - Aufbewahrung von Testorganismen für
die Prüfung der bakteriziden (einschließlich Legionella), mykobakteriziden, sporiziden,
fungiziden und viruziden (einschließlich Bakteriophagen) WirkungTa slovenski standard je istoveten z: prEN 12353
ICS:
07.100.99 Drugi standardi v zvezi z Other standards related to
mikrobiologijo microbiology
71.100.35 Kemikalije za dezinfekcijo v Chemicals for industrial and
industriji in doma domestic disinfection
purposes
oSIST prEN 12353:2019 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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oSIST prEN 12353:2019
DRAFT
EUROPEAN STANDARD
prEN 12353
NORME EUROPÉENNE
EUROPÄISCHE NORM
January 2019
ICS 11.080.20; 71.100.35 Will supersede EN 12353:2013
English Version
Chemical disinfectants and antiseptics - Preservation of
test organisms used for the determination of bactericidal
(including Legionella), mycobactericidal, sporicidal,
fungicidal and virucidal (including bacteriophages) activity
Chemische Desinfektionsmittel und Antiseptika -
Aufbewahrung von Testorganismen für die Prüfung
der bakteriziden (einschließlich Legionella),
mykobakteriziden, sporiziden, fungiziden und
viruziden (einschließlich Bakteriophagen) Wirkung
This draft European Standard is submitted to CEN members for enquiry. It has been drawn up by the Technical Committee
CEN/TC 216.If this draft becomes a European Standard, CEN members are bound to comply with the CEN/CENELEC Internal Regulations
which stipulate the conditions for giving this European Standard the status of a national standard without any alteration.
This draft European Standard was established by CEN in three official versions (English, French, German). A version in any other
language made by translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC
Management Centre has the same status as the official versions.CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland,
Turkey and United Kingdom.Recipients of this draft are invited to submit, with their comments, notification of any relevant patent rights of which they are
aware and to provide supporting documentation.Warning : This document is not a European Standard. It is distributed for review and comments. It is subject to change without
notice and shall not be referred to as a European Standard.EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2019 CEN All rights of exploitation in any form and by any means reserved Ref. No. prEN 12353:2019 E
worldwide for CEN national Members.---------------------- Page: 3 ----------------------
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Contents Page
European foreword ....................................................................................................................................................... 5
Introduction .................................................................................................................................................................... 6
1 Scope .................................................................................................................................................................... 7
2 Normative references .................................................................................................................................... 7
3 Terms and definitions ................................................................................................................................... 7
4 Requirements ................................................................................................................................................... 7
5 Methods .............................................................................................................................................................. 8
5.1 Principle ............................................................................................................................................................. 8
5.2 Materials and reagents .................................................................................................................................. 8
5.2.1 Test organisms ................................................................................................................................................. 8
5.2.2 Culture media and reagents ........................................................................................................................ 8
5.2.3 Cell cultures .................................................................................................................................................... 16
5.2.4 Host strains for dairy bacteriophages (Lactococcus lactis) ........................................................... 18
5.3 Apparatus and glassware .......................................................................................................................... 18
5.3.1 General ............................................................................................................................................................. 18
5.3.2 Usual microbiological laboratory equipment .................................................................................... 18
5.4 Procedure for preservation of test organisms – General .............................................................. 20
5.4.1 Handling of freeze-dried / frozen test organisms from culture collections ........................... 20
5.4.2 Choice of incubation procedure, agar medium, cell culture/cell line ....................................... 20
5.5 Procedure for preservation of bacteria (incl. Legionella, spore-forming bacteria,
excl. mycobacteria and bacterial spores) and yeasts ...................................................................... 20
5.5.1 Reconstitution of the freeze-dried test organisms .......................................................................... 20
5.5.2 Preparation for storage ............................................................................................................................. 20
5.5.3 Preparation of stock culture / working cultures .............................................................................. 21
5.6 Procedure for preservation of mycobacteria .................................................................................... 21
5.6.1 Reconstitution of the freeze-dried test organisms .......................................................................... 21
5.6.2 Preparation for storage ............................................................................................................................. 21
5.6.3 Preparation of working cultures ............................................................................................................ 22
5.7 Procedure for preservation of moulds (e.g. Aspergillus brasiliensis) ....................................... 22
5.7.1 Reconstitution of the freeze-dried test organism ............................................................................ 22
5.7.2 Preparation for storage ............................................................................................................................. 22
5.7.3 Preparation of stock culture / working cultures .............................................................................. 23
5.8 Procedure for preservation of viruses (except dairy bacteriophages) .................................... 23
5.8.1 Reconstitution of frozen virus ................................................................................................................. 23
5.8.2 Preparation for storage of stock virus suspension .......................................................................... 23
5.8.3 Preparation of test virus suspension .................................................................................................... 24
5.9 Procedure for preservation of bacteriophages ................................................................................. 24
5.9.1 Reconstitution of frozen bacteriophages ............................................................................................ 24
5.9.2 Preparation for storage ............................................................................................................................. 24
5.9.3 Preparation of bacteriophages working suspensions .................................................................... 25
5.10 Verification of the purity and identity of test organisms .............................................................. 25
5.10.1 General ............................................................................................................................................................. 25
5.10.2 Information on source of strains ............................................................................................................ 25
5.10.3 Purity ................................................................................................................................................................ 25
5.10.4 Identity ............................................................................................................................................................. 25
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5.11 Documentation .............................................................................................................................................. 25
5.11.1 General ............................................................................................................................................................. 25
5.11.2 Freeze-dried test organism / frozen viruses ...................................................................................... 26
5.11.3 Cryovials of frozen test organism ........................................................................................................... 26
5.11.4 Stock culture ................................................................................................................................................... 26
5.11.5 Verification of purity and identity .......................................................................................................... 26
5.11.6 Storage of documentation ......................................................................................................................... 26
Annex A (informative) Test organisms – Culture collection references and relation to
CEN/TC 216 standards and prENs .......................................................................................................... 27
A.1 Bacteria (except mycobacteria and spore-forming bacteria) ...................................................... 27
A.1.1 Enterobacter cloacae ................................................................................................................................... 27
A.1.2 Enterococcus hirae ....................................................................................................................................... 27
A.1.3 Enterococcus faecium ................................................................................................................................. 27
A.1.4 Escherichia coli (1) ....................................................................................................................................... 27
A.1.5 Escherichia coli (2) K12 .............................................................................................................................. 27
A.1.6 Lactobacillus brevis ..................................................................................................................................... 27
A.1.7 Legionella pneumophila, subsp. Pneumophila .................................................................................... 27
A.1.8 Proteus hauseri ............................................................................................................................................. 27
A.1.9 Pseudomonas aeruginosa .......................................................................................................................... 28
A.1.10 Salmonella enterica subsp. enterica, Serotype typhimurium ........................................................ 28
A.1.11 Staphylococcus aureus subsp. aureus .................................................................................................... 28
A.2 Mycobacteria .................................................................................................................................................. 28
A.2.1 Mycobacterium avium subsp. avium ...................................................................................................... 28
A.2.2 Mycobacterium terrae ................................................................................................................................ 28
A.3 Spore-forming bacteria .............................................................................................................................. 28
A.3.1 Bacillus cereus ............................................................................................................................................... 28
A.3.2 Bacillus subtilis subsp. spizizenii ............................................................................................................. 28
A.3.3 Clostridium sporogenes ............................................................................................................................. 28
A.4 Fungi (moulds and yeasts) ........................................................................................................................ 29
A.4.1 Aspergillus brasiliensis (former “A. niger”) (mould) ...................................................................... 29
A.4.2 Candida albicans (yeast) ............................................................................................................................ 29
A.4.3 Saccharomyces cerevisiae (1) (yeast) ................................................................................................... 29
A.4.4 Saccharomyces cerevisiae (2) var. diastaticus (yeast) ................................................................... 29
A.5 Viruses .............................................................................................................................................................. 29
A.5.1 Adenovirus type 5, strain Adenoid 75 .................................................................................................... 29
A.5.2 Bovine Enterovirus Type 1(ECBO) ........................................................................................................... 29
A.5.3 Murine norovirus, strain S99 Berlin ....................................................................................................... 29
A.5.4 Murine Parvovirus, minute virus of mice, strain Crawford ............................................................ 29
A.5.5 Poliovirus type 1, Sabin strain LSc-2ab .................................................................................................. 29
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A.5.6 Vaccinia virus, strain Modified Vaccinia Virus Ankara (MVA) ...................................................... 30
A.5.7 Vaccinia virus, strain Elstree .................................................................................................................... 30
A.6 Bacteriophages ............................................................................................................................................. 30
A.6.1 Lactococcus lactis subsp. lactis bacteriophage P008 ....................................................................... 30
A.6.2 Lactococcus lactis subsp. lactis bacteriophage P001 ....................................................................... 30
Annex B (informative) Graphical representations........................................................................................ 31
Bibliography ................................................................................................................................................................. 36
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European foreword
This document (prEN 12353:2019) has been prepared by Technical Committee CEN/TC 216 “Chemical
disinfectants and antiseptics”, the secretariat of which is held by AFNOR.This document is currently submitted to the CEN Enquiry.
This document will supersede EN 12353:2013.
The document was revised to adapt it to the latest state of science, to correct errors and ambiguities.
The following are the significant technical changes since the last edition:— the methods of preservation of viruses are described more detailed (5.8.3);
— the description of BCYE Agar for Legionella was added (5.2.2.24);
— the information on source of strains was added (5.10.2);
— the information of the storage of documentation was added (5.11.6);
— the IP Number for fungi was deleted (see A.4);
— the used virus strains are re-drafted (see A.5).
The changes mentioned above have no impact on the test results obtained with reference to the
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Introduction
Standardized tests for the determination of bactericidal (incl. Legionella pneumophila),
mycobactericidal, sporicidal, fungicidal and virucidal (incl. bacteriophages) activity of chemical
disinfectants and antiseptics necessitate the use of test organisms whose purity and identity have been
verified and whose biological behaviour remains stable. Therefore it is essential to specify the storage
requirements.This document aims to describe methods for preservation of test organisms used for such purposes.
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1 Scope
This document specifies methods for keeping test organisms used and defined in European Standards
for the determination of bactericidal (incl. Legionella pneumophila), mycobactericidal, sporicidal,
fungicidal and virucidal (incl. bacteriophages) activity of chemical disinfectants and antiseptics drawn
up by CEN/TC 216. These methods for keeping test organisms can only be carried out in connection
with at least one of those standards where a reference to this document is established.
NOTE 1 Annex A (informative) contains a non-exhaustive list of test organisms for which this document can be
applied.NOTE 2 European Standards (EN and prEN) where this document is referenced are listed in the Bibliography.
NOTE 3 A specific part on the preservation of bacterial spores could be added once the results of the ongoing
ring trials are available.2 Normative references
The following documents are referred to in the text in such a way that some or all of their content
constitutes requirements of this document. For dated references, only the edition cited applies. For
undated references, the latest edition of the referenced document (including any amendments) applies.
EN 14885, Chemical disinfectants and antiseptics — Application of European Standards for chemical
disinfectants and antiseptics3 Terms and definitions
For the purposes of this document, the terms and definitions given in EN 14885 apply.
ISO and IEC maintain terminological databases for use in standardization at the following addresses:
• IEC Electropedia: available at http://www.electropedia.org/• ISO Online browsing platform: available at http://www.iso.org/obp
4 Requirements
Each test organism specified in a CEN/TC 216 European Standard and referred to in this document shall
be handled as described in this document.The purity and identity of the preserved test organism shall be verified during the preparation and
regularly during the storage, except for viruses where only the identity is checked before the stock virus
suspension is stored.The preserved test organism – except viruses - should be checked at regular intervals (at least in the
case of longer storage than 14 months) to ensure that its susceptibility to products has not changed. As
long as CEN/TC 216 has not developed specific tests for this purpose any suitable method can be used
e.g. EN 1040 for bacteria, EN 1275 for fungi, EN 14348 for mycobacteria, EN 13623 for Legionella
pneumophila, EN 14476 for viruses or EN 13610 for dairy bacteriophages.---------------------- Page: 9 ----------------------
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5 Methods
5.1 Principle
A sample of the test organism – in general in freeze-dried form - is obtained from a culture collection.
This sample is cultured, prepared for storage, filled into storage vessels and placed in the deep freeze.
From this sample a stock culture is prepared and subsequently used to prepare working cultures for the
test procedure. In some cases the working cultures are directly prepared from the deep freeze samples.
5.2 Materials and reagents5.2.1 Test organisms
See Annex A for examples of test organisms.
The documentation on the test organisms should follow 5.10.2 and 5.11.3.
5.2.2 Culture media and reagents
5.2.2.1 General
The formulas of all media and reagents are given in case commercial ready-to-use material is not used.
It is to be checked that each commercial supplier has established an appropriate quality control system.
All weights of chemical substances given in this document refer to the anhydrous salts unless otherwise
stated. Hydrated forms may be used as an alternative, but the weights required shall be adjusted to
allow for consequent molecular weight differences.The reagents shall be of analytical grade and/or appropriate for microbiological purposes. They shall be
free from substances that are toxic or inhibitory to the test organisms.To improve reproducibility, it is recommended that whenever possible, commercially available
dehydrated material is used for the preparation of culture media. The manufacturer's instructions
relating to the preparation of these products should be rigorously followed.All specified pH values are measured at (20 ± 1) °C.
For each culture medium, cell culture and reagent a limitation for use should be fixed.
5.2.2.2 WaterThe water shall be freshly glass distilled water and not demineralized water. If distilled water of
adequate quality is not available, water for injections (see bibliographic reference [1]) can be used.
Sterilize in the autoclave (5.3.2.1a). Sterilization is not necessary if the water is used for e.g. preparation
of culture media and subsequently sterilized.5.2.2.3 Tryptone Soya Broth (TSB) for bacteria, except Legionella
Tryptone soya broth, consisting of:
Tryptone, pancreatic digest of casein 17,0 g
Soya peptone, papaic digest of soybean meal 3,0 g
Sodium chloride (NaCl) 5,0 g
Water (5.2.2.2) 800,0 ml
Dipotassium phosphate (K HPO ) 2,5 g
2 4
Glucose 2,5 g
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Water (5.2.2.2) to 1 000,0 ml
Sterilize in the autoclave (5.3.2.1a). After sterilization the pH of the medium shall be equivalent to
7,2 ± 0,2.5.2.2.4 Malt Extract Broth (MEB) for fungi
Malt extract broth, consisting of:
Malt extract (food grade, e.g. Christomalt powder from Difal or equivalent that is not highly purified
and not only based on maltose, e.g.20,0 g
malt extract from OXOID)
Water (5.2.2.2) to 1 000,0 ml
Sterilize in the autoclave (5.3.2.1a). After sterilization the pH of the medium shall be equivalent to
5,6 ± 0,2.5.2.2.5 Cryoprotectant solution for bacteria, spore-forming bacteria, fungi
Cryoprotectant solution, consisting of:
Beef extract 3,0 g
Tryptone, pancreatic digest of casein 5,0 g
Glycerol (C H O ) [2] 150,0 g
3 8 3
Water (5.2.2.2) to 1 000,0 ml
Dissolve the constituents in boiling water. Sterilize in the autoclave (5.3.2.1a). After sterilization the pH
of the solution shall be equivalent to 6,9 ± 0,2.Any commercially available cryoprotectant containing glycerol for preservation of test organisms
equivalent to the solution described above may be used.If justified, any other equivalent cryoprotectant solution may be used, e.g. for Legionella (5.5.2).
5.2.2.6 Middlebrook 7 H 9 broth with 10 % ADC enrichment and glycerol as reconstituent and
cryoprotectant solution for mycobacteria (MADC)Middlebrook 7 H 9 broth, consisting of:
Middlebrook 7 H 9 broth powder 4,7 g
Glycerol (C H O ) [2] 100,0 ml
3 8 3
Water (5.2.2.2) 800,0 ml
Treat in the autoclave (5.3.2.1a) for a holding time of only 10 min and cool to 45 °C. Add under aseptic
conditions 100 ml Middlebrook ADC enrichment to obtain approximately 1 000,0 ml. The pH of the
medium shall be equivalent to 6,6 ± 0,2.This information is given for the information of users of this standard and does not constitute an endorsement of the
products named. Corresponding products supplied by other manufacturers may be used if they can be shown to lead to the
same results.---------------------- Page: 11 ----------------------
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5.2.2.7 Polysorbate 80 solution
Polysorbate 80 solution, consisting of:
Polysorbate 80 0,5 g
Water (5.2.2.2) to 1 000,0 ml
Sterilize in the autoclave (5.3.2.1a).
5.2.2.8 DMSO as cryoprotectant for cell culture freezing
Dimethyl sulphoxide (DMSO) is used to help protect the cells from rupture by the formation of ice
crystals.Since DMSO is toxic it should be handled with care. It can be absorbed through the skin and may cause
irritation and/or burns. It is teratogenic and an allergen. Latex gloves should be worn when handling it.
5.2.2.9 Glutamine solution, 3 %Dissolve 12 g Glutamine in 400 ml of water (5.2.2.2) and sterilize by membrane filtration. The solution
is stored at (−20 ± 1) °C.5.2.2.10 TV (Trypsin-Versene)
Dissolve 0,05 g Trypsin in 100 ml of 0,53 mM EDTA (Ethylene diamine tetra acetic acid) and sterilize by
membrane filtration. Store at (4 ± 1) °C.5.2.2.11 Antibiotic suspension
Chemicals
50 million units Penicillin-G
(eg Sigma PEN-K )
50 g Streptomycin sulphate (approx. equal to 750 IU/mg) 2
(eg Sigma Cat: 56501 )
25 × 500,000 units Mycostatin 2
(eg Nystatin: E R Squibb 59150 )
Water (5.2.2.2) to 2,5 l.
Preparation
Dissolve vial contents of antibiotics in water (5.2.2.2) and fill up to 2,5 l.
Dispense aseptically into 50 ml and 5 ml aliquots.
Store at − 20 °C. Shake the bottle after thawing.
Use 5 ml per litre of medium to give a final concentration of:
Penicillin 100 units/ml
Streptomycin 100 µg/ml
Mycostatin 25 units/ml
This information is given for the information of users of this standard and does not constitute an endorsement of the
products named. Corresponding products supplied by other manufacturers may be used if they can be shown to lead to the
same results.---------------------- Page: 12 ----------------------
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5.2.2.12 Phosphate-buffered saline solution (PBS)
Sodium chloride (NaCl) 8,00 g
Potassium chloride (KCl) 0,20 g
Disodium hydrogen phosphate, 12-hydrate (Na HPO x 12H O) 2,89 g
2 4 2
Potassium phosphate, monobasic (KH PO ) 0,20 g
2 4
Water (5.2.2.2) to 1 000,0 ml
5.2.2.13 Foetal calf serum (FCS)
FCS has to be certified free of viruses and mycoplasma.
Extraneous viruses and mycoplasma may interfere with cell and virus growth resulting in false results.
5.2.2.14 Earle’s BSSSodium chloride (NaCl) 68,0 g
Potassium chloride (KCl) 4,0 g
Calcium chloride (CaCl ) 2,0 g
Magnesium sulphate, 7-hydrate (MgSO x 7H O) 2,0 g
4 2
Sodium hydrogenphosphate, 2-hydrate (NaH PO x 2H O) 1,4 g
2 2 2
Glucose 10,0 g
Phenol red, 1 % (5.2.2.15) 20,0 ml
Water (5.2.2.2) to 1 000,0 ml
CaCl should be dissolved separately in 100 ml of water (5.2.2.2) and added to the other dissolved
reagents just before the solution is brought to its final volume. The solution is 10-fold concentrated. It is
sterilized by membrane filtration through a 0,22 µm Millipore or Seitz-type filter and can be stored at
(4 ± 1) °C for 4 weeks....
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