Reaction mass of N,N-dimethyldecan-1-amide and N,N-dimethyloctanamide

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:

other: Well conducted reliable study according to current guidelines.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1992

Report date:

1992

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

less strains

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Decanamide, N,N-dimethyl-, mixt. with N,N-dimethyloctanamide

Cas Number:

67359-57-3

IUPAC Name:

Decanamide, N,N-dimethyl-, mixt. with N,N-dimethyloctanamide

Details on test material:

- Name of test material (as cited in study report):Hallcomid M-8-10
- Substance type: Mixture containing of N,N-Dimethyl-octanoic acid amide, N,N-Dimethyl decanoic acid amide (major compounds) and N,N-Dimethyl-hexanoic acid amide, N,N-Dimethyl-dodecanoic acid amide (minor compounds)
- Physical state: clear, yellowish liquid
- Analytical purity: see below
- Impurities (identity and concentrations): see below
- Composition of test material, percentage of components: see below
- Lot/batch No.:002949
- Expiration date of the lot/batch:until January 21, 1993
- Stability under test conditions:A stability test in the solvent did not detect a relevant change in the percent
active ingredient.
- Storage condition of test material: at room temperature

Method

Target gene:

his operon

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254 induced SD rat S9mix

Test concentrations with justification for top dose:

5000, 1000, 200, 40, 8 µg/plate
Due to the substance's toxicity, doses ranging from 25 µg to 800 µg per plate were chosen for the repeat tests:
800, 400, 200, 100, 50, 25 µg/plate

Vehicle / solvent:

The solvent employed for Hallcomid M-8-10 was ethanol and for the positive controls DMSO.
No "untreated" negative control was set up for ethanol, since sufficient evidence was available in the literature

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

no

Remarks:

No "untreated" negative control was set up for ethanol, since sufficient evidence was available in the literature

Positive controls:

yes

Positive control substance:

other: Sodium azide for TA 1535; Nitrofurantoin for TA 100; 4-nitro-1,2-phenylene diamine for TA 1537 and TA 98; 2-aminoanthracene for all strains (with S9)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration:48h
- Selection time (if incubation with a selection agent):48h

NUMBER OF REPLICATIONS: performed in quadruplicates (two experiments)

NUMBER OF CELLS EVALUATED:

DETERMINATION OF CYTOTOXICITY
- Method:The toxicity of the substance was assessed in three ways: Background growth, by marked and dose-dependent reduction in the mutant count per
plate compared to the negative control and bacterial counts were taken on two plates for each concentration studied with S9 mix. However,
if an evaluation was performed only without S9 mix, the bacterial count was taken without S9 mix.

Evaluation criteria:

The following criteria determined the acceptance of an assay:
a) The negative controls had to be within the expected range, as defined by published data (e.g. Maron and Ames, 1983) and
the laboratories' own historical data (see Chapter 8).
b) The positive controls had to show sufficient effects, as defined by the laboratories' experience (see Chapter 8).
c) Titer determinations had to demonstrate sufficient bacterial density in the suspension.
An assay which did not comply with at least one of the above criteria was not used for assessment.

Assessment Criteria
A reproducible and dose-related increase in mutant counts of at least one strain is considered to be a positive result.
For TA 1535, TA 100 and TA 98 this increase should be about twice that of negative controls, whereas for TA 1537, at
least a threefold increase should be reached. Otherwise, the result is evaluated as negative. However, these guidelines
may be overruled by good scientific judgement.

Results and discussion

Test results

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

but tested up to the limit, reduced concentrations in second experiment

Vehicle controls validity:

not applicable

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

As may be seen, there was no indication of a bacteriotoxic effect of Hallcomid M-8-10 at doses of up to and including 100 µg per plate. The total
bacteria counts consistently produced results comparable to the negative controls, or differed only insignificantly. No inhibition of growth was noted as well. Higher doses had a strong, strain-specific bacteriotoxic effect. Therefore, they could only be used to a limited extent up to 1000 µg per plate for assessment purposes.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

None of the four strains concerned showed a dose-related and biologically relevant increase in mutant counts over those of the negative controls.The positive controls sodium azide, nitrofurantoin, 4-nitro-1,2-phenylene diamine and 2-aminoanthracene increased mutant

counts to well over those of the negative controls, and thus demonstrated the system's sensitivity and the activity of the

S9 mix.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

The registered substance was considered to be non-mutagenic without and with S9 mix in the Salmonella/microsome test.

Executive summary:

The registered substance was investigated using the Salmonella/microsome test for point mutagenic effects in doses of up to

5000 µg per plate on four Salmonella typhimurium LT2 mutants according to OECD 471. These comprised the histidine-auxotrophic strains TA 1535, TA 100, TA 1537 and TA 98.

Doses of up to and including 100 µg per plate did not cause any bacteriotoxic effects: Total bacteria counts remained unchanged

and no inhibition of growth was observed. At higher doses, the substance had a strong, strain-specific bacteriotoxic

effect, so that this range could only be used to a limited extent up to 1000 µg per plate for assessment purposes.

Evidence of mutagenic activity was not seen. No biologically relevant increase in the mutant count,

in comparison with the negative controls, was observed.

Therefore, the test substance was considered to be non-mutagenic without and with S9 mix in the Salmonella/microsome test.

The positive controls sodium azide, nitrofurantoin, 4-nitro-1,2-phenylene diamine and 2-aminoanthracene had a marked

mutagenic effect, as was seen by a biologically relevant increase in mutant colonies compared to the corresponding negative

controls.