[No public or meaningful name is available]

Genetic toxicity: in vitro

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:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Remarks:

Only four bacterial strains used

Cross-reference

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1999

Materials and methods

Principles of method if other than guideline:

Suspensions of bacterial cells are exposed to the test substance by the plate incorporation method in the presence and in the absence of an exogenous metabolic activation system. In this method, the suspensions are mixed with an overlay agar and plated immediately onto the minimal medium and incubated for two days at 37˚C, The results are interpreted by counting the revertant colonies and comparing to the number of spontaneous revertant colonies on solvent-control plates.

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Reaction products of C14-C18 (even numbered) and C14-C18-unsatd. (even numbered) fatty acids or triglycerides with diethanolamine

Molecular formula:

not applicable gor UVCB

IUPAC Name:

Reaction products of C14-C18 (even numbered) and C14-C18-unsatd. (even numbered) fatty acids or triglycerides with diethanolamine

Test material form:

liquid: viscous

Details on test material:

see Section 1.2 for detailed composition

Reference

Constituent 1

Reference substance name:

Amides, C18-unsatd., N,N-bis(hydroxyethyl)

EC Number:

700-972-2

Molecular formula:

C22H43NO3

IUPAC Name:

Amides, C18-unsatd., N,N-bis(hydroxyethyl)

Test material form:

liquid

Details on test material:

- Name of test material (as cited in study report): Oleic acid diethanolamine condensate
- Source of the test material: Obtained from Henkel Corporation, Emery Group (Cincinnati, OH)
- Molecular weight: 387.68
- Physical state: Clear liquid
- Analytical purity: Purity of 47.5% was obtained when analyzed by HPLC
- Impurities (identity and concentrations): Analyzed by HPLC/mass spectrometry. Impurities identified were fatty acid alkanolamides (approximately 30%), other fatty acids or unidentified organic impurities, one polar nitrosamines (i.e., nitrosodiethanolamine was detected at a concentration of 68 ppb), free diethanolamine at 0.19%; no non-polar nitrosamine were found.
- Lot/batch No.: 1H01722285 (for purity determination by HPLC method) and DA-021 (for stability studies by gas chromatography)
- Stability under test conditions: Stable when stored in glass vials for 2 wk at 25 °C but unstable at 60 °C.
- Storage condition of test material: At room temperature, protected from ultraviolet light, in amber glass bottles with Teflon®-lined lids
- Other: Solubility: soluble in alcohols, glycols, ketones, chlorinated solvents, and other aliphatic hydrocarbon solvents.

Specific details on test material used for the study:

The discrepancy between originally usded identifier of the substance and new test material information is due to change in the main identifier through the Substance identity adaptation service offered by ECHA. The new identifier is covered by old identifier mentioned in Additional test material information section

Method

Target gene:

No data

Species / strain

Species / strain / cell type:

other: S. typhimurium TA97, TA98, TA100 and TA1535

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9 mix (metabolic activation enzymes and cofactors from Aroclor 1254-induced male Sprague-Dawley rat or Syrian hamster liver)

Test concentrations with justification for top dose:

Without metabolic activation: 0, 0.1, 0.3, 1, 3.3 and 10 µg/plate
With metabolic activation: 0, 3.3, 10, 33, 100 and 200 µg/plate

Vehicle / solvent:

Ethanol

Controls

Controls 1

Untreated negative controls:

yes

Remarks:

(solvent control)

Negative solvent / vehicle controls:

yes

Remarks:

(Ethanol)

True negative controls:

not specified

Positive controls:

yes

Remarks:

(for strains TA100 and TA1535)

Positive control substance:

sodium azide

Remarks:

absence of metabolic activation

Controls 2

Untreated negative controls:

yes

Remarks:

(solvent control)

Negative solvent / vehicle controls:

yes

Remarks:

(Ethanol)

True negative controls:

not specified

Positive controls:

yes

Remarks:

(for strain TA 98)

Positive control substance:

other: 4-nitro-o-phenylenediamine

Remarks:

absence of metabolic activation

Controls 3

Untreated negative controls:

yes

Remarks:

(solvent control)

Negative solvent / vehicle controls:

yes

Remarks:

(Ethanol)

True negative controls:

not specified

Positive controls:

yes

Remarks:

(for strain TA 97)

Positive control substance:

9-aminoacridine

Remarks:

absence of metabolic activation

Controls 4

Untreated negative controls:

yes

Remarks:

(solvent control)

Negative solvent / vehicle controls:

yes

Remarks:

(Ethanol)

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene (all strains)

Remarks:

metabolic activation

Details on test system and experimental conditions:

- Test medium: Top agar supplemented with L-histidine and d-biotin
- Method of application: In agar (plate incorporation)
- Duration of incubation: 2 d at 37 °C
- Number of replicates: Three

Evaluation criteria:

- Positive response: Reproducible, dose-related increase in histidine-independent (revertant) colonies in any one strain/activation combination.
- Equivocal response: An increase in revertants that are not dose related, is not reproducible, or is not of sufficient magnitude to support a determination of m utagenicity.
- Negative response: When no increase in revertant colonies is observed following chemical treatment.
- There is no minimum percentage or fold increase required for a chemical to be judged positive or weakly positive.

Statistics:

Not reported

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium, other: S. typhimurium TA97, TA98, TA100 and TA1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

(at ≥3.3 µg/plate without metabolic activation; at 200 µg/plate with metabolic activation) .

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

(at ≥3.3 µg/plate without metabolic activation; at 200 µg/plate with metabolic activation).

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

(at ≥3.3 µg/plate without metabolic activation; at 200 µg/plate with metabolic activation).

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

(at ≥3.3 µg/plate without metabolic activation; at 200 µg/plate with metabolic activation).

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

None

Applicant's summary and conclusion

Conclusions:

Under the conditions of the study, the test substance was not mutagenic in Salmonella typhimurium strain TA97, TA98, TA100 or TA1535, with or without S9 metabolic activation.

Executive summary:

A study was conducted to determine the mutagenic potential of the test substance in a bacterial mutation assay, in compliance with GLP. Salmonella typhimurium strains TA97, TA98, TA100 and TA1535 were treated with the test substance using the Ames plate incorporation method at up to five concentration levels for each bacterial strain, in triplicate, both with and without metabolic activation (S9 mix; Aroclor induced rat and hamster liver homogenate metabolising system). The concentration range was 0.1 to 10 µg/plate (-S9 -mix) and 3.3 to 200 µg/plate (+S9 -mix). Cytotoxicity was observed at ≥3.3 µg/plate without metabolic activation and at 200 µg/plate with metabolic activation. No significant increase in the frequency of revertant colonies was recorded for any of the bacterial strains with any concentration tested, either with or without metabolic activation. The vehicle (ethanol) control or the negative control plates produced counts of revertant colonies within the normal range. All the positive control chemicals used in the test produced marked increases in the frequency of revertant colonies, both with and without the S9 -mix. Under the conditions of the study, the test substance was not mutagenic in Salmonella typhimurium strain TA97, TA98, TA100 or TA1535, with or without S9 metabolic activation (Irwin, 1999).