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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:
Not available
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Read across study hence maximum reliability rating of 2 assigned according to ECHA guidance, although study was well documented, meets generally accepted scientific principles.
Justification for data waiving:
other:
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
secondary source
Title:
Unnamed
Year:
1999

Materials and methods

Principles of method if other than guideline:
Cells deficient in thymidine kinase (TK) due to the mutation of TK+/- to TK-/- are resistant to the cytotoxic effects of trifluorothymidine (TFT). Thymidine kinase proficient cells (TK+/-) are sensitive to TFT, which causes the inhibition of cellular metabolism and halts further cell division. Thus mutant cells are able to proliferate in the presence of TFT, whereas normal cells, which contain thymidine kinase, are not able to proliferate.
GLP compliance:
no
Type of assay:
mammalian cell gene mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Lauric Acid Diethanolamine Condensate
- Composition of test material, percentage of components: 90 % for lauric acid diethanolamine condensate, 5 % amine (probably diethanolamine) and approximately 5% other organic impurities, polar nitrosamine, nitrosodiethanolamine, was detected at a concentration of 3,600 ppb
- Lot/batch No.: CH1E952
- Stability under test conditions: Unstable when stored in glass vials for 2 wk at 60°C but very little at 25°C or less
- Storage condition of test material: Room temperature, protected from light, in amber glass bottles sealed with Teflon-lined caps

Method

Target gene:
Not applicable
Species / strain
Species / strain / cell type:
mouse lymphoma L5178Y cells
Details on mammalian cell type (if applicable):
- Type and identity of media: Supplemented Fischer’s medium
- Properly maintained: Yes
- Periodically "cleansed" against high spontaneous background: Yes, by exposing to medium containing thymidine, hypoxanthine, methotrexate, and glycine for 1 d; to medium containing thymidine, hypoxanthine, and glycine for 1 d; and to normal medium for 3 to 5 d
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
S9 from the livers of Aroclor 1254-induced male Fischer 344 rats
Test concentrations with justification for top dose:
Trial 1:
Without metabloic actvation: 2.5, 5, 10, 20, 30, 40 and 50 μg/mL
With metabloic actvation: 5, 10, 20, 30, 40, 50 and 60 μg/mL
Trial 2:
Without metabloic actvation: 5, 10, 20 and 30 μg/mL
With metabloic actvation: 5, 10, 20, 30 and 40 μg/mL
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Ethanol
Controlsopen allclose all
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
Ethanol
True negative controls:
no
Positive controls:
yes
Positive control substance:
methylmethanesulfonate
Remarks:
Migrated to IUCLID6: at 5 μg/mL in both trials 1 and 2
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
Ethanol
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: Methyl cholanthrene at 2.5 μg/mL in both trials 1 and 2
Details on test system and experimental conditions:
The experimental protocol is presented in detail by Myhr et al, 1985

METHOD OF APPLICATION: In medium


DURATION
- Exposure duration: 4 h
- Expression time (cells in growth medium): 48 h
- Selection time (if incubation with a selection agent): 10 to 12 d (at 37 °C in 5 % CO2)


SELECTION AGENT (mutation assays): Yes, Trifluorothymidine (TFT)



NUMBER OF REPLICATIONS: Duplicate (all treatment levels within an experiment, including concurrent positive and solvent controls, were replicated)


NUMBER OF CELLS EVALUATED: 6 × 10(6) cells in 10 mL medium


DETERMINATION OF CYTOTOXICITY
- Method: Cloning efficiency


Evaluation criteria:
Minimum criteria for accepting an experiment as valid and a detailed description of the statistical analysis and data evaluation are presented by Caspary et al. (1988). Both responses would have to be significant (P≤0.05) for test material to be considered positive, i.e., capable of inducing TFT resistance. A single significant response would lead to a call of “questionable,” and the absence of both a trend and peak response results in a “negative” call.
Statistics:
Not reported

Results and discussion

Test results
Species / strain:
mouse lymphoma L5178Y cells
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
No increase in the frequency of mutant colonies of L5178Y mouse lymphoma cells was noted after exposure to test material, with or without S9.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

None

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

Under the conditions of the test, no increase in the frequency of mutant colonies of L5178Y mouse lymphoma cells was noted after exposure to test material, with or without S9
Executive summary:

A study was conducted to assess the mutagenicity potential of lauric acid diethanolamine condensate, (LDEA, CAS No.120-40-1)

in mouse lymphoma cell line L5178Y.

The mouse lymphoma cells were treated with test material at 2.5, 5, 10, 20, 30, 40 and 50 μg/mL without metabloic actvation and at 5, 10, 20, 30, 40, 50 and 60 μg/mL with metabolic activation in trial 1 for 4 h. Cells were treated with test material at 5, 10, 20, 30, 40 μg/mL with metabloic actvation and 5, 10, 20, 30 without metabolic activation in trial 2 for 4 h. After the 48-hour expression period, cells were plated in medium and soft agar supplemented with TFT for selection of TFT-resistant cells and cells were plated in nonselective medium and soft agar to determine cloning efficiency. No increase in the frequency of mutant colonies of L5178Y mouse lymphoma cells was noted after exposure to test material, with or without S9 under the conditions of the test.