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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline with acceptable restrictions (missing E. coli/ TA102 strain)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1978
Report date:
1978

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
missing strain to detect crosslinking/ oxidizing agents
Principles of method if other than guideline:
According to Ames, B.N. et al.: Mutation Research 31, 347-364
GLP compliance:
no
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
Phthalonitrile
EC Number:
202-044-8
EC Name:
Phthalonitrile
Cas Number:
91-15-6
Molecular formula:
C8H4N2
IUPAC Name:
benzene-1,2-dicarbonitrile
Details on test material:
- Name of test material (as cited in study report): phthalonitrile
- Analytical purity: purity of the substance is not documented

Method

Target gene:
nis-
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254 induced rat liver S-9 mix
Test concentrations with justification for top dose:
3.15, 10, 31.5, 100, 315, 1000 and 3000 µg/plate (with S-9 mix); 10, 31.5, 100, 315, 1000 and 3000 µg/plate (without S-9 mix)
Vehicle / solvent:
DMSO
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
not specified
Positive controls:
yes
Positive control substance:
other: see freetext
Details on test system and experimental conditions:
Standard plate test
Test tubes containing 2 mL soft agar kept in a water bath at 45°C, and remaining
components added in the following order:
0 .1 mL test solution or vehicle
0 .1 mL bacterial suspension
0 .5 mL S-9 mix (in tests with metabolic activation) or 0 .5 ml KCl (in tests
without metabolic activation).
After mixing, the samples are poured onto Vogel-Bonner agar plates.
Where indicated the epoxide hydratase inhibitor and glutathione depletor 1, 1, 1 -trichloropropene 2,3-oxide (TCPO) was added together with the top agar (0.6/µL per plate) .
Two test plates per dose and controls were used; after incubation at 37°C for
48-72hours in the dark, the bacterial colonies ( his+ revertants) are counted.
Positive control:
with metabolic activation: 10 μg/plate Benzo(a)pryene, 10 μg/plate 2-aminoanthracene and 90 μg/plate 3-Methylcholanthrene for each strain;
without metabolic activation: 1 μg/plate Benzo(a)pryene-4,5-oxide and 3.15 μg/plate N-MethyI-N'-nitro-N-nitrosoguanidine, all substances were dissolved in DMSO for all strains.

Results and discussion

Test results
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Vehicle controls validity:
not specified
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
At the two highest concentrations the test compound partly precipitated on the plate .
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'. Remarks: Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537

Any other information on results incl. tables

Standard plate test (3.15 - 3000 µg/plate)

Strain

Metabolic activation system

Replicates

maximum revertant factor

dose dependency

Assessment

TA 98

no

2

1.2

no

negative

 

yes

2

1.1

no

negative

TA 98

not performed

 

 

 

with TCPO

yes

2

1.1

no

negative

TA 100

no

2

1.0

no

negative

Trial 1

yes

2

1.0

no

negative

TA 100

not performed

 

 

 

Trial 2

yes

2

1.0

no

negative

TA 1535

no

2

1.3

no

negative

 

yes

2

1.3

no

negative

TA 1537

no

2

1.1

no

negative

 

yes

2

1.2

no

negative

Applicant's summary and conclusion