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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 guidelien study with acceptable deviations

Data source

Referenceopen allclose all

Reference Type:
publication
Title:
Salmonella Mutagenicity Test results for 250 Chemicals
Author:
Haworth S, Lawlor T, Mortelmans K, Speck W, and Zeiger E
Year:
1983
Bibliographic source:
Environmental Mutagenesis Supplement 1:3-142
Reference Type:
secondary source
Title:
European Union Risk Assessment Report - 1,2,4-Trichlorobenzene
Author:
European Commission - European Chemicals Bureau
Year:
2003
Bibliographic source:
Office for Official Publications of the European Communities
Reference Type:
secondary source
Title:
1,2,4-Trichlorobenzene. BUA-Report 17
Author:
BUA. GDCH-Advisory Committee on Existing Chemicals of Environmental Relevance (Beratergremium für Umweltrelevante Altsoffe).
Year:
1987
Bibliographic source:
VCH

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
Only 4 strain tested instead of 5. Positive control: 2-aminoantracene was only indicator of efficiency of S9-mix. Individual plate reading not reported.
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): 1,2,4-Trichlorobenzene
- Analytical purity: 95%
- Lot/batch No.: B3J08
-Chemical source: MC/B

Method

Target gene:
his
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
microsomal enzyme reaction mix (S9-mix) prepared from Sprague-Dawley rats and male Syrian hamsters
Test concentrations with justification for top dose:
0.0, 3.3, 10.0, 33.3, 100.0, 333.3 ug/plate
Vehicle / solvent:
- Vehicle/solvent used: DMSO
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene (2-AA): in all strains in presence of rat and hamster S-9. 4-Nitro-o-phenylenediamine (NOPD): on TA98, without S-9. Sodium azide (SA) : on TA100 and TA 1535, without S-9. 9-aminoacridine (9AAD) was tested on 1537, without S-9.
Remarks:
The actual concentration for each positive control chemical used for each strain and activation condition was selected by the individual laboratory based on dose-response curves generated at the beginning of the testing program (see table 1)
Details on test system and experimental conditions:
METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 20 min
- Exposure duration: 48h
- Selection time (if incubation with a selection agent): 48h
- Concentration of S9 mix: 10% for both Hamster S9 mix and Rat S9 mix

SELECTION AGENT (mutation assays): L- histidine

NUMBER OF REPLICATIONS: 2 trial per strain and 3 dishes per dose

DETERMINATION OF CYTOTOXICITY
- Method: other: viability on complete medium and reduced numbers of revertant colonies per plate and/or thinning or absence of the bacterial lawn








Evaluation criteria:
A positive response was indicated by a reproducible, dose-related increase, whether it be twofold over background or not.
An equivocal response was defined as an increase in revertants which was not dose-related, not reproducible, or was of insufficient magnitude to support a determination of mutagenicity. A negative response was obtained when no increase in revertant colonies is observed following chemical treatment.

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:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
RANGE-FINDING/SCREENING STUDIES: To select the dose range for the mutagenetic assay, the test chemicals were checked for toxicity to TA 100 up to a concentration of 10 mg/plate or the limit of solubility, both in the presence and absence of S-9 mix. If toxicity was not apparent in the preliminary toxicity determination, the highest dose tested was 10 mg/plate; otherwise the upper limit of solubility was used. If toxicity was observed, the doses of test chemical were chosen so that the high dose exhibited some degree of toxicity. Occasionally, in the earlier tests, the high dose was greater than 10 mg/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

 

TA 100

TA 1535

TA 1537

TA 98

Dose

NA

RLI

HLI

NA

RLI

HLI

NA

RLI

HLI

NA

RLI

HLI

0.0

99±5.2

113±2.0

115±8.7

20±1.0

13±3.0

13±0.6

17±1.5

14±1.9

12±2.7

37±3.5

35±2.3

37±4.7

3.3

104±4.0

107±5.5

124±2.0

16±1.5

15±2.5

13±2.9

18±3.8

10±1.5

18±5.2

38±6.4

34±2.3

31±0.9

10.0

86±6.7

115±9.5

132±5.8

18±4.2

9±0.0

10±2.7

23±2.7

11±1.2

23±3.7

28±1.2

36±4.8

35±2.7

33.3

61±10.1 s

123±2.2

124±7.3

19±1.2

13±3.2

11±1.7

11±1.7

9±0.3

17±0.7

24±1.7

29±0.9

43±2.3

100.0

t

112±6.7

100±6.9

6±6.0s

9±1.8

12±2.7

9±1.8

8±1.7

17±4.0

24±1.3

34±4.1

38±6.4

333.3

t

114±9.4

21±21.5s

t

13±0.7s

6±1.5s

5±2.3

7±1.2

0±0.0s

17±2.5

34±3.7

9±8.5s

POS

625±28.8

460±15.3

963±104.3

444±23.0

305±3.2

268±34.8

308±8.0

158±10.6

244±5.8

850±18.0

288±3.0

1258±130.5

Table 1 Mutagenic responses of Salmonella strains TA100, TA 1535, TA 1537, and TA 98 (mean±SEM) to 1,2,4-trochlorobenzene.

 Abbreviations:NA, not activated; RLI, rat liver S-9, Aroclor1254 induced; HLI, hamster liver S-9, Aroclor1254 induced, t=complete clearing background

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative with metabolic activation
negative without metabolic activation
Executive summary:

Haworth (1983)

Ames test was performed with the Salmonella typhimurium strains TA1535, TA1537, TA98, and TA100 at 0, 3.3, 10.0, 33.3, 100.0, and 333.3µg 1,2,4-TCB/plate with and without S-9 from Aroclor 1254 induced rat and hamster livers using the preincubation procedure. Test doses were chosen following checks for toxicity: in the absence of toxicity a maximum of 10 mg/plate was used.

The test resulted was negative with and without metabolic activation.

The test was conducted according to OECD guideline 471 with deviations (Only 4 strains tested instead of 5. Positive control: 2-aminoantracene was only indicator of efficiency of S9-mix. Individual-plate-readings were not reported).