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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: Acceptable publication which meets basic scientific principles

Data source

Referenceopen allclose all

Reference Type:
publication
Title:
The bacterial mutation test. Six tests for carcinogenicity
Author:
Anderson, D. and Styles, J.A.
Year:
1978
Bibliographic source:
Br J Cancer 37: pp. 924-30
Reference Type:
secondary source
Title:
4-Aminoazobenzene. BUA-Stoffbericht 217
Author:
Beratergremium für Altstoffe (BUA)
Year:
1998
Bibliographic source:
BUA der Gesellschaft dt. Chemiker, S. Hirzel Wissenschaftliche Verlagsgesellschaft, Stuttgart

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
only 4 strains used
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
no data

Method

Target gene:
his
Species / strain
Species / strain:
other: Salmonella typhimurium TA98, TA100, TA1535, TA1538
Details on mammalian cell lines (if applicable):
- Type and identity of media: Minimal Glucose Agar Medium (1.5% Difco Bacto agar in Vogel-Bonner medium with 2% glucose)
- Properly maintained: yes
- Periodically checked for: histidine requirements, deep rough, DNA excision-repair deficiency, R-factor plasmid, rate of reversion and purity
Additional strain characteristics:
not specified
Metabolic activation:
with
Metabolic activation system:
S9-mix
Test concentrations with justification for top dose:
4, 20, 100, 500, 2500 µg/plate
Vehicle:
- Vehicle(s)/solvent(s) used: DMSO; water
Controlsopen allclose all
Negative controls:
yes
Solvent controls:
yes
True negative controls:
yes
Positive controls:
yes
Positive control substance:
2-nitrofluorene
Remarks:
with metabolic activation

Migrated to IUCLID6: for strain TA1538 and TA98
Negative controls:
yes
Solvent controls:
yes
True negative controls:
yes
Positive controls:
yes
Positive control substance:
other: 2-(1-chloro-2-isopropylaminoethyl)naphthalene for strains TA1535 and TA100
Remarks:
with metabolic activation
Details on test system and conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 2 to 3 days
- Expression time (cells in growth medium): 2 to 3 days

Evaluation criteria:
The results were accepted as positve if:
- there was a 2-fold increase over the negative control count for any strain;
- the negative-control cultures had counts within about 50% of the mean value;
- the positive-control cultures had counts greater than twice the negative-control values;
- the correct strains responded to the appropriate positive-control compounds;
- there was a background lawn indicating at least 10% survival

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with
Genotoxicity:
positive
Remarks:
3fold at 500µg AAB/plate
Cytotoxicity:
not specified
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with
Genotoxicity:
positive
Remarks:
5fold at 100µg AAB/plate
Cytotoxicity:
not specified
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with
Genotoxicity:
positive
Remarks:
6fold at 500µg AAB/plate
Cytotoxicity:
not specified
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with
Genotoxicity:
positive
Remarks:
5fold at 20µg AAB/plate
Cytotoxicity:
not specified
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Mean Number of S. Typhimurium Revertant Colonies in Each Strain with No Treatment (C) or with 0.1ml/Plate Water or DMSO

TA1535 TA1538 TA98 TA100
  C water DMSO C water DMSO C water DMSO C water DMSO
mean 30.4 30.33 25.87 28.83 31.47 26.4 47.11 48.74 42.06 63.57 60.90 57.92
s.d. 17.15 21.82 14.82 15.00 15.58 11.12 31.27 33.27 30.48 43.44 31.4 25.46
s.e. 3.13 4.0 2.70 2.74 2.84 2.03 5.28 5.62 5.08 9.48 6.85 4.89
n 30 30 30 30 30 30 35 35 36 21 21 27

Induction of Revertants in 4 Test Strains of S. Typhimurium

TA1535 TA1538 TA98 TA100 Test result
maximum fold increase in Salmonella revertants above the concurrent negativ-control values 3 5 6 5 +
concentration of test substance (µg/plate) 500 100 500 20

Applicant's summary and conclusion