Water quality. Evaluation of genotoxicity by measurement of the induction of micronuclei. Part 2: Mixed population method using the cell line V79 (ISO 21427-2:2006)
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ISO 21427-2:2006 specifies a method for the determination of genotoxicity of water and waste water using a mammalian in vitro test which detects damage, induced by water-soluble substances, to the chromosomes or the mitotic apparatus of V79 cells from the Chinese hamster. The micronucleus test allows the identification of substances that cause cytogenetic damage which results in the formation of micronuclei containing lagging chromosome fragments and/or whole chromosomes. The assay is based on the increase in the frequency of micronucleated cells after incubation with and without metabolic activation.
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Sisällysluettelo
Foreword
1 Scope
2 Normative references
3 Terms and definitions
4 Principle
5 Interferences
6 Reagents and media
6.1 Water.
6.2 Reagents.
6.3 Preparation of culture media
6.4 Cell system.
6.5 Metabolic activation
7 Apparatus
8 Test facility criteria
9 Procedure
9.1 Sampling and samples
9.2 Experimental size
9.3 Negative controls
9.4 Positive controls
9.5 Time schedule
9.6 Preparatory steps
9.7 Preparation of test cultures
9.8 Treatment of test cultures
10 Evaluation and assessment
10.1 Cell morphology
10.2 Selection of concentrations to be analysed
10.3 Reading of slides
10.4 Mitotic index
10.5 Survival index
10.6 Proliferation index
10.7 Validity criteria
10.8 Assessment criteria
10.9 Determination of the decisive D value
11 Precision
12 Test report
Annex A Bromodeoxyuridine (BrdU) method (informative)