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BS EN 16437:2014 Chemical disinfectants and antiseptics. Quantitative surface test for the evaluation of bactericidal activity of chemical disinfectants and antiseptics used in veterinary area on porous surfaces without mechanical action. Test method and , 2014
- 1 Scope
- 2 Normative references
- 3 Terms and definitions
- 4 Requirements
- 5 Test method [Go to Page]
- 5.1 Principle
- 5.2 Materials and reagents [Go to Page]
- 5.2.1 Test organisms
- 5.2.2 Culture media and reagents [Go to Page]
- 5.2.2.1 General
- 5.2.2.2 Water
- 5.2.2.3 Tryptone Soya Agar (TSA)
- 5.2.2.4 Diluent
- 5.2.2.5 Neutraliser
- 5.2.2.6 Hard water for dilution of products
- 5.2.2.7 Interfering substances
- 5.2.2.8 Tryptone Soya Broth (TSB) with neutraliser
- 5.2.3 Test surface)
- 5.3 Apparatus and glassware [Go to Page]
- 5.3.1 General
- 5.3.2 Usual microbiological laboratory equipment) [Go to Page]
- 5.3.2.1 Apparatus for sterilisation (moist and dry heat)
- 5.3.2.2 Water bath, capable of being controlled at 4 °C ± 1 °C, 10 °C ± 1 °C, 20 °C ± 1 °C, 40 °C ± 1 °C (5.5.1) and 45 °C ± 1 °C (to maintain melted TSA, 5.2.2.3, 5.5.2.2 and 5.5.2.3).
- 5.3.2.3 Incubator, capable of being controlled at 36 °C ± 1 °C or 37 °C ± 1 °C (5.2.1). The same temperature shall be used for incubation performed during a test and its control and validation.
- 5.3.2.4 pH-meter, having an inaccuracy of calibration of no more than ± 0,1 pH units at 20 °C ± 1 °C.
- 5.3.2.5 Stopwatch
- 5.3.2.6 Shakers
- 5.3.2.7 Membrane filtration apparatus, constructed of a material compatible with the substances to be filtered with a filter holder of at least 50 ml volume and suitable for use with filters of diameter 47 mm to 50 mm and 0,45 µm pore size for sterilisation of hard water (5.2.2.6) and bovine albumin (5.2.2.7).
- 5.3.2.8 Refrigerator, capable of being controlled at 2 °C to 8 °C.
- 5.3.2.9 Graduated pipettes of nominal capacities 10 ml, 1 ml, 0,1 ml and 0,05 ml or calibrated automatic pipettes.
- 5.3.2.10 Petri dishes, (plates) of size 90 mm to 100 mm.
- 5.3.2.11 Glass beads (diameter 3 mm to 4 mm).
- 5.3.2.12 Volumetric flasks
- 5.3.2.13 Temperature controlled cabinet, capable of being controlled at 10 °C ± 1 °C.
- 5.3.2.14 Ultrasonic bath, capable of operating at a frequency 30 kHz to 55 kHz, maximal output 1 000 W.
- 5.4 Preparation of test organism suspension and product test solutions [Go to Page]
- 5.4.1 Test organism suspension (test and validation suspension) [Go to Page]
- 5.4.1.1 General
- 5.4.1.2 Preservation and stock cultures of test organisms
- 5.4.1.3 Working culture of test organisms
- 5.4.1.4 Test suspension (“N”)
- 5.4.1.5 Incubation and counting of the test suspension
- 5.4.2 Product test solution
- 5.5 Procedure for assessing the bactericidal activity of the product [Go to Page]
- 5.5.1 General [Go to Page]
- 5.5.1.1 Experimental conditions (obligatory and additional)
- 5.5.1.2 Neutralisation
- 5.5.1.3 General instructions for the validation and control procedures
- 5.5.1.4 Equilibration of temperature
- 5.5.2 Test procedure (Dilution-neutralisation method)) [Go to Page]
- 5.5.2.1 General
- 5.5.2.2 Test “Na” - determination of bactericidal concentrations
- 5.5.2.3 Water control “Nw”
- 5.5.2.4 Neutraliser control “B” - verification of the absence of toxicity of the neutraliser
- 5.5.2.5 Method validation “C” - dilution-neutralisation validation
- 5.5.2.6 Incubation and counting of the test mixture and the control and validation mixtures
- 5.6 Experimental data and calculation [Go to Page]
- 5.6.1 Explanation of terms and abbreviations [Go to Page]
- 5.6.1.1 Overview of the different suspensions and test mixtures
- 5.6.1.2 Vc values
- 5.6.2 Calculation [Go to Page]
- 5.6.2.1 General
- 5.6.2.2 Determination of Vc values
- 5.6.2.3 Calculation of N
- 5.6.2.4 Calculation of Na and Nw
- 5.6.2.5 Calculation of B and C
- 5.7 Verification of methodology [Go to Page]
- 5.7.1 General
- 5.7.2 Control of weighted mean counts
- 5.7.3 Basic limits
- 5.8 Expression of results and precision [Go to Page]
- 5.8.1 Reduction
- 5.8.2 Control of active and non-active product test solution (5.4.2)
- 5.8.3 Limiting test organism and bactericidal concentration
- 5.8.4 Precision, repetitions
- 5.9 Interpretation of results – conclusion [Go to Page]
- 5.9.1 General
- 5.9.2 Bactericidal activity for general purposes
- 5.9.3 Qualification for certain fields of application
- 5.10 Test report
- Annex A (informative)Referenced strains in national collections
- Annex B (informative)Neutralisers - Examples of neutralisers of the residual antimicrobial activity of chemical disinfectants and antiseptics
- Annex C (informative)Graphical representations of dilution-neutralisation method
- Annex D (informative)Example of a typical test report [Go to Page]
