Registration Dossier

Diss Factsheets

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

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: see 'Remark'
Remarks:
The authors tested the mutagenicity and recombinogenicity of benzylchloride following a methodology similar to the OECD guideline 471 (Bacterial Reverse Mutation Test). While GLP standards are not specified, materials and methodology as well as results are very well described and few minor deviations were observed. Thus the study can be considered a Klimisch 2.c, comparable to a guideline study with acceptable restrictions.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1976
Report date:
1976

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
maximum concentration tested is 10 µl/plate, no E. coli strain tested and four tested concentrations instead of five
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Reference substance name:
chloromethylbenzene
IUPAC Name:
chloromethylbenzene
Constituent 2
Chemical structure
Reference substance name:
α-chlorotoluene
EC Number:
202-853-6
EC Name:
α-chlorotoluene
Cas Number:
100-44-7
Molecular formula:
C7H7Cl
IUPAC Name:
(chloromethyl)benzene
Details on test material:
- Name of test material (as cited in study report): BIO-76-16, Benzyl Chlorine, Analytical reagent grade
- Physical state: colourless liquid

No more data available

Method

Target gene:
Histidine for Salmonella typhimurium
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
S. typhimurium TA 1538
Species / strain / cell type:
Saccharomyces cerevisiae
Metabolic activation:
with and without
Metabolic activation system:
rat liver (pretreatment with Aroclor 1254)
Test concentrations with justification for top dose:
0.1 µL/plate, 1 µL/plate, 5 µL/plate, 10 µL/plate
Vehicle / solvent:
No data
Controlsopen allclose all
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: methylnitrosoguanidine (MNNG)
Remarks:
solvent water or saline, without metabolic activation
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
2-nitrofluorene
Remarks:
solvent dimethylsulfoxide, without metabolic activation

Migrated to IUCLID6: (NF)
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: quinacrine mustard (QM)
Remarks:
solvent water or saline, without metabolic activation
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-anthramine (ANTH)
Remarks:
solvent dimethylsulfoxide, with metabolic activation
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
2-acetylaminofluorene
Remarks:
solvent dimethylsulfoxide, with metabolic activation

Migrated to IUCLID6: (AAF)
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 8-aminoquinoline (AMQ)
Remarks:
solvent dimethylsulfoxide, with metabolic activation
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: dimethylnitrosamine (DMNA)
Remarks:
solvent saline, with metabolic activation
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 to 72h

NUMBER OF CELLS EVALUATED: 10^9 cells from a log phase culture

No further data available
Evaluation criteria:
Amino-acid requiring strains of bacteria are used to detect reverse gene mutations. Based on the number of revertants, the mutagenicity of the test compound is assessed. Indeed, if a clear relation is established between the number of revertants and the tested non toxic concentrations (in presence or not of metabolic activativation), the test is interpretated to be positive.
Statistics:
No data

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
positive
Remarks:
for strain TA 100 with and without metabolic activation, strain TA 1535 weak response at highest dose with metabolic activation
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with and without
Genotoxicity:
not specified
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Species / strain:
Saccharomyces cerevisiae
Metabolic activation:
with and without
Genotoxicity:
positive
Remarks:
with and without metabolic activation
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Additional information on results:
No data
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Result for the mutagenicity (Salmonella strains) and recombinogenicity (Saccharomyces) experiment with benzylchloride without metabolic activation for all tested strains, including controls.

S. typhimurium

S. typhimurium

S. typhimurium

S. typhimurium

S. typhimurium

S. cerevisiae

TA-1535

TA-1537

TA-1538

TA-98

TA-100

D4

Solvent control

37

38

19

17

170

49

*

TA-1535

MNNG

10 µL/plate

Positive control*

>104

>104

821

>103

>104

460

TA-1537

QM

10 µL/plate

Test compound

0.1 µL/plate

50

44

24

192

37

TA-1538

NF

100 µg/plate

1 µL/plate

31

38

18

28

214

34

TA-98

NF

100 µg/plate

5 µL/plate

38

31

15

265

248

56

TA-100

MNNG

10 µL/plate

10 µL/plate

42

25

18

38

674

333

D4

MNNG

10 µL/plate

Table 2: Result for the mutagenicity (Salmonella strains) and recombinogenicity (Saccharomyces) experiment with benzylchloride with metabolic activation for all tested strains, including controls.

S. typhimurium S. typhimurium S. typhimurium S. typhimurium S. typhimurium S. cerevisiae
TA-1535 TA-1537 TA-1538 TA-98 TA-100 D4
Solvent control 29 68 48 35 97 37 **

TA-1535

ANTH

100 µg/plate
Positive control** >103 >103 >103 >103 >103 44 TA-1537

AMQ

100 µg/plate
Test compound 0.1 µL/plate 31 35 45 23 155 40 TA-1538 AAF 100 µg/plate
1 µL/plate 30 33 53 20 121 38 TA-98 AAF 100 µg/plate
5 µL/plate 11 19 47 23 174 44 TA-100 ANTH 100 µg/plate
10 µL/plate 69 27 30 22 391 134 D4 DMNA 100 µmoles/plate

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
positive with and without metabolizing system

The authors tested mutagenicity and recombinogenicity of benzylchloride in the absence and presence of a metabolizing system with a methodology similar to the OECD guideline 471. Under these test conditions, the test substance was considered mutagenic for Salmonella typhimurium and recombinogenic for Saccharomyces cerevisiae with and without metabolizing system.
Executive summary:

The authors tested the mutagenicity and recombinogenicity of benzylchloride (CAS n° 100-44-7) in the absence and presence of a metabolizing system with a methodology similar to the OECD guideline 471. The Salmonella typhimurium strains TA 1535, TA 1537, TA 1538, TA 98 and TA 100 and the Saccharomyces cerevisiae strain D4 were used and the metabolizing system was obtained from rat liver homogenates. These strains were exposed to 0.1, 1, 5 and 10 µL/plate with and without metabolic activation for 48 to 72 h. Solvent controls and positive controls were performed.

The test showed that benzylchloride was mutagenic for Salmonella typhimurium strain TA 100 and recombinogenic for Saccharomyces cerevisiae strain D4. This respons was observed in absence and presence of the metabolic activation system, demonstrating that metabolic activation is not essential for mutagenicity or recombinogenicity. Furthermore, there was also an indication of a weak response at the high dose of the

Salmonella typhimurium strain TA 1535. Effects were only seen at relatively high doses and dose-related responses at the lower end of the dose range were not observed. Hence, based on this information, further in vitro testing would be required with other systems to clarify if in vivo testing would be necessary.

No data was available on the GLP status of this study. However this study was similar to the OECD guideline 471 with minor deviations as following, the maximum concentration tested was 10 µl/plate, as no E. coli strain was tested and as four concentrations were tested instead of five.Therefore, this study should be considered reliable with restictions, a Klimisch 2.c study comparable to a guideline study with acceptable restrictions.

Categories Display