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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in mammalian cells
Remarks:
Type of genotoxicity: gene mutation
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
From 2001-11-27 to 2002-03-05
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP guideline study but some reporting deficiencies
Cross-reference
Reason / purpose for cross-reference:
read-across source
Reference
Endpoint:
in vitro gene mutation study in mammalian cells
Remarks:
Type of genotoxicity: gene mutation
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
For details on endpoint specific justification please see read-across report in section 13 or find a link in cross-reference “assessment report”.
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
assessment report
Species / strain:
mouse lymphoma L5178Y cells
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
Without S9-mix: 5.56 µg/mL at 3 h treatment; 0.56 µg/mL at 24 h treatment; With S9-mix >5.56 µg/mL
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Additional information on results:
Preliminary toxicity test:
-The test item was freely soluble in the vehicle (DMSO) at 380 mg/mL (expressed as active item). In the culture medium, the dose-level of 3800 µg/mL showed a marked emulsion. At this dose-level, the pH was approximately 7.9 (7.4 for the vehicle control) and no increase in the osmolality due to the test item was noted. Consequently, with a treatment volume of 200 µL/20 mL (1% v/v) culture medium, the dose-levels for the preliminary toxicity test were: 7.6, 76, 380, 760, 1900 and 3800 µg/mL, both with and without S9.

-A slight to marked emulsion was observed at the end of the treatment period at dose-levels ≥ 760 µg/mL.

-Without S9 mix, the test item was markedly to strongly toxic at dose-levels ≥ 7.6 µg/mL (93-100% decrease in the cloning efficiency immediately after treatment (CE0) and in the relative survival (RS)).

-With S9 mix, the test item was markedly to strongly toxic at dose-levels ≥ 76 µg/mL (97-100% decrease in the CE0 and RS).
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Based on the preliminary toxicity test the selected dose-levels for treatment were expressed as active item (40.37%).

Conclusions:
Interpretation of results (migrated information):
negative

The test material was considered to be non mutagenic in mouse lymphoma assay.
Executive summary:

The study used as source investigated genetic toxicity in vitro (gene mutation in mammalian cells).The study results of the source compound were considered applicable to the target compound. Justification and applicability of the read-across approach (structural analogue) is outlined in the read-across report in section 13 or find a link in cross reference “assessment report”.

 

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2002
Report date:
2002

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.17 (Mutagenicity - In Vitro Mammalian Cell Gene Mutation Test)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of assay:
mammalian cell gene mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
Didecyldimethylammonium chloride
EC Number:
230-525-2
EC Name:
Didecyldimethylammonium chloride
Cas Number:
7173-51-5
Molecular formula:
C22H48N.Cl
IUPAC Name:
N-decyl-N,N-dimethyldecan-1-aminium chloride
Details on test material:
- Name of test material (as cited in study report): Didecyldimethylammonium chloride (DDAC)
- Composition of test material, percentage of components: ca. 40% Didecyldimethylammonium chloride (CAS no.: 7173-51-5) in water
- Physical state: Clear viscous liquid
- Analytical purity: 40.37% active ingredient in water
- Stability: Stable

Method

Species / strain
Species / strain / cell type:
mouse lymphoma L5178Y cells
Details on mammalian cell type (if applicable):
TK (Thymidine kinase ) -/+, not able to grow in Trifluorothimidine medium
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
S9 mix , from Aroclor 1254 induced (500 mg/kg i.p.) rat livers.
Test concentrations with justification for top dose:
Experiments without S9 mix:
0.07, 0.21, 0.62, 1.85, 2.78 and 5.56 µg/mL (1st experiment),
0.06, 0.19, 0.56, 1.67, 2.5 and 5 µg/mL (2nd experiment)
Experiments with S9 mix:
0.21, 0.62, 1.85, 5.56, 16.7 and 50 µg/mL (1st experiment),
0.19, 0.56, 1.67, 5, 7.5 and 10 µg/mL (2nd experiment).
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
Controlsopen allclose all
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
methylmethanesulfonate
Remarks:
Migrated to IUCLID6: without S9 mix
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
cyclophosphamide
Remarks:
Migrated to IUCLID6: with S9 mix
Details on test system and experimental conditions:
DURATION
- Exposure duration: Without S9-mix: 1st experiment: 3 h; 2nd experiment: 24 h
With S9-mix: 1st experiment: 3 h; 2nd experiment: 3 h

NUMBER OF REPLICATIONS: Two plates/dose-level for test and four plates for control

NUMBER OF CELLS EVALUATED: 2000 cells/well


Evaluation criteria:
No data
Statistics:
none

Results and discussion

Test results
Species / strain:
mouse lymphoma L5178Y cells
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
Without S9-mix: 5.56 µg/mL at 3 h treatment; 0.56 µg/mL at 24 h treatment; With S9-mix >5.56 µg/mL
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Additional information on results:
Preliminary toxicity test:
-The test item was freely soluble in the vehicle (DMSO) at 380 mg/mL (expressed as active item). In the culture medium, the dose-level of 3800 µg/mL showed a marked emulsion. At this dose-level, the pH was approximately 7.9 (7.4 for the vehicle control) and no increase in the osmolality due to the test item was noted. Consequently, with a treatment volume of 200 µL/20 mL (1% v/v) culture medium, the dose-levels for the preliminary toxicity test were: 7.6, 76, 380, 760, 1900 and 3800 µg/mL, both with and without S9.

-A slight to marked emulsion was observed at the end of the treatment period at dose-levels ≥ 760 µg/mL.

-Without S9 mix, the test item was markedly to strongly toxic at dose-levels ≥ 7.6 µg/mL (93-100% decrease in the cloning efficiency immediately after treatment (CE0) and in the relative survival (RS)).

-With S9 mix, the test item was markedly to strongly toxic at dose-levels ≥ 76 µg/mL (97-100% decrease in the CE0 and RS).
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Based on the preliminary toxicity test the selected dose-levels for treatment were expressed as active item (40.37%).

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

The test material (40% didecyldimethylammonium chloride in water) was considered to be non mutagenic in mouse lymphoma assay.
Executive summary:

A study was conducted to evaluate the potential of the test material (40% didecyldimethylammonium chloride (DDAC) in water) to induce mutations at the TK (thymidine kinase) locus in L5178Y mouse lymphoma cells. The study was conducted according to OECD guideline 476 and EU method B. 17.

After a preliminary toxicity test, test material was tested in two independent experiments, with and without a metabolic activation system, the S9 mix.

The test item was dissolved in DMSO. Since the test material was toxic in the preliminary test, the choice of the highest dose level for the main test was based on the level of toxicity according to the criteria specified in the international guidelines. Test material was tested at a concentration range of 0.07- 5.56 (without S9 mix) and 0.21- 50 (with S9 mix) µg DDAC /mL in the main test expressed as active item (40.37%). A slight to strong toxicity was induced, depending on the dose-levels and the treatment duration. No increase in the mutation frequency was induced. Under the test conditions, the test material didecyldimethylammonium chloride (40% DDAC in water) was considered to be non mutagenic in mouse lymphoma assay.