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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
Pyridine-2,6-dicarboxylic acid
EC Number:
207-894-3
EC Name:
Pyridine-2,6-dicarboxylic acid
Cas Number:
499-83-2
Molecular formula:
C7H5NO4
IUPAC Name:
2,6-Pyridinedicarboxylic Acid Pyridin-2,6-dicarbonsäure Pyridine-2,6-dicarboxylic Acid
Test material form:
solid: crystalline

Method

Target gene:
TA98:
his D 3052; rfa-; uvrB-; R-factor: frame shift mutations
TA 100:
his G 46; rfa-; uvrB-; R-factor: base-pair substitutions
TA 1535:
his G 46; rfa-; uvrB-: base-pair substitutions
TA 1537:
his C 3076; rfa-; uvrB-: frame shift mutations
TA 102:
his G 428 (pAQ1); rfa-; R-factor: base-pair substitutions
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with and without
Metabolic activation system:
S9 Homogenate
Test concentrations with justification for top dose:
The test item concentrations to be applied in the main experiments were chosen according to the results of the pre-experiment (see chapter 12.1 Pre-Experiment). 5000 µg/plate was selected as the maximum concentration. The concentration range
covered two logarithmic decades. Two independent experiments were performed with the following concentrations: Experiment I: 3.16, 10.0, 31.6, 100,316, 1000,2500 and 5000 µg/plate (all tester strains except TA 100 without metabolic activation)
0.100,0.316, 1.00,3.16, 10.0,31.6, 100,316, 1000and 2500 µg/plate (only TA 100 without metabolic activation) Experiment II: 8, 25, 70, 200, 600, 1000, 1800 and 5000 llg/plate (all tester strains except TA 98 and TA 100 without metabolic activation) 0.25, 0.8, 2.5, 8, 25, 70, 200, 600, 1000 and 1800 µg/plate (only TA 98 and TA 100 without metabolic activation). As the results of the pre-experiment TA 98 with and without metabolic activation and TA 100 with metabolic activation were in accordance with the criteria described above, these were reported as a part of the main experiment I.
Vehicle / solvent:
The test item was dissolved in EtOH and diluted prior to treatment. The solvent was compatible with the survival ofthe bacteria and the S9 activity.
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
sodium azide
methylmethanesulfonate
other: 4-nitro-o-pheny1ene-diamine; 2-aminoanthracene

Results and discussion

Test resultsopen allclose all
Key result
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
other: In tester strain TA 1535 toxic effects ofthe test item were noted at concentrations of 70 µg/plate and higher (without metabolic activation) and at concentrations of 200 µg/plate and higher (with metabolic activation).
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
other: TA 153 7 toxic effects of the test item were observed at concentrations of 70 µg/plate and higher (without metabolic activation) and at concentrations of 600 µg/plate and higher (with metabolic activation).
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
other: at concentrations of 31.6 µg/plate and higher (without metabolic activation).
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
other: In tester strain TA 100 toxic effects of the test item were noted at concentrations of 10 µg/plate and higher (without metabolic activation) and at concentrations of 100 µg/plate (with metabolic activation) and higher.
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid

Any other information on results incl. tables

The test item 1, 1 0-Dichlorodecane was investigated for its potential to induce gene mutations according to the plate incorporation test ( experiment I and II) using Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537 and TA 102. In two independent experiments several concentrations of the test item were used. Bach assay was conducted with and without metabolic activation. The concentrations, including the controls, were tested in triplicate. The following concentrations of the test item were prepared and used in the experiments: Experiment 1: 3.16, 10.0, 31.6, 100,316, 1000,2500 and 5000 µg/plate (all tester strains except TA 100 without metabolic activation) 0.100, 0.316, 1.00, 3.16, 10.0, 31.6, 100,316, 1000 and 2500 µg/plate (only TA 100 without metabolic activation) Experiment II: 8, 25, 70, 200, 600, 1000, 1800 and 5000 µg/plate (all tested strains except TA 98 and TA 100 without metabolic activation) 0.25, 0.8, 2.5, 8, 25, 70, 200, 600, 1000 and 1800 µg/plate (only TA 98 and TA 100 without metabolic activation). No precipitation of the test item was observed in any tester strain used in experiment I and II (with and without metabolic activation). Toxic effects of the test item were noted in four of five tester strains evaluated in experiment I and II. In experiment I toxic effects of the test item were observed in tester strain TA 98 at concentrations of 31.6 µg/plate and higher (without metabolic activation). In tested strain TA 100 toxic effects of the test item were noted at concentrations of 10 µg/plate and higher (without metabolic activation) and at concentrations of 100 µg/plate (with metabolic activation) and higher. In tester strain TA 1535 toxic effects of the test item were noted at concentrations of 2500 µg/plate and higher (without metabolic activation) and at concentrations of 316 µg/plate (with metabolic activation). In tester strain TA 153 7 toxic effects of the test item were observed at concentrations of 316 µg/plate and higher (with and without metabolic activation). In experiment II toxic effects of the test item were noted in tester strain TA 98 at concentrations of 70 µg/plate and higher (without metabolic activation). In tester strain TA 100 toxic effects of the test item were noted at concentrations of 25 µg/plate and higher (without metabolic activation) and at concentrations of 200 µg/plate and higher (with metabolic activation). In tester strain TA 1535 toxic effects ofthe test item were noted at concentrations of 70 µg/plate and higher (without metabolic activation) and at concentrations of 200 µg/plate and higher (with metabolic activation). In tester strain TA 1537 toxic effects of the test item were observed at concentrations of 70 µg/plate and higher (without metabolic activation) and at concentrations of 600 µg/plate and higher (with metabolic activation). No biologically relevant increases in revertant colony numbers of any of the five tester strains were observed following treatment with 1,10-Dichlorodecane at any concentration level, neither in the presence nor absence of metabolic activation in experiment I and II. The reference mutagens induced a distinct increase of revetiant colonies indicating the validity ofthe experiments.

Applicant's summary and conclusion

Conclusions:
In conclusion, it can be stated that during the described mutagenicity test and under the
experimental conditions reported, 1,1 0-Dichlorodecane did not cause gene mutations by
base pair changes or frameshifts in the genome of the tester strains used.
Therefore, 1,1 0-Dichlorodecane is considered to be non-mutagenic in this bacterial
reverse mutation assay
Executive summary:

Summary Results

In order to investigate the potential of 1,10-Dichlorodecane for its ability to induce gene

mutations the plate incorporation test ( experiment I and II) was performed with the

Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537 and TA 102.

In two independent experiments several concentrations of the test item were used. Bach

assay was conducted with and without metabolic activation. The concentrations,

including the controls, were tested in triplicate. The following concentrations of the test

item were prepared and used in the experiments:

Experiment 1:

3.16, 10.0, 31.6, 100, 316, 1000,2500 and 5000 f.tg/plate

(all tester strains except TA 100 without metabolic activation)

0.100, 0.316, 1.00, 3.16, 10.0, 31.6, 100, 316, 1000 and 2500 f.tg/plate

( only TA 100 without metabolic activation)

Experiment II:

8, 25, 70, 200, 600, 1000, 1800 and 5000 f.tg/plate

(all tester strains except TA 98 and TA 100 without metabolic activation)

0.25, 0.8, 2.5, 8, 25, 70,200,600, 1000 and 1800 f.tg/plate

( only TA 98 and TA 100 without metabolic activation)

No precipitation of the test item was observed in all tester strains used in experiment I

and II (with and without metabolic activation).

Toxic effects of the test item were noted in four tester strains used in experiment I

and II:

• In experiment I toxic effects of the test item were observed at concentrations of

10.0 f.tg/plate and higher (without metabolic activation) and at concentrations of

100 f.tg/plate and higher (with metabolic activation), depending on the particular

tester strain.

• In experiment II toxic effects of the test item were noted at concentrations of

25 f.tg/plate and higher (without metabolic activation) and at concentrations of

200 f.tg/plate and higher (with metabolic activation), depending on the particular

tester strain.

No biologically relevant increases in revertant colony numbers of any of the five tester

strains were observed following treatment with 1,10-Dichlorodecane at any

concentration level, neither in the presence nor absence of metabolic activation in

experiment I and II.

The reference mutagens induced a distinct increase of revertant colonies indicating the

validity of the experiments.