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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

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:
2002

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
July 1997
Qualifier:
according to guideline
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Version / remarks:
May 2000
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
Dibutyl carbonate
EC Number:
208-816-0
EC Name:
Dibutyl carbonate
Cas Number:
542-52-9
Molecular formula:
C9H18O3
IUPAC Name:
dibutyl carbonate
Constituent 2
Chemical structure
Reference substance name:
Butyl carbamate
EC Number:
209-751-0
EC Name:
Butyl carbamate
Cas Number:
592-35-8
Molecular formula:
C5H11NO2
IUPAC Name:
butyl carbamate
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: B323
- Purity test date: July 4, 2001

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature
- Solubility and stability of the test substance in the solvent/vehicle: confirmed

OTHER SPECIFICS: Colorless liquid

Method

Target gene:
his, trp
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254 induced rat liver S9
Test concentrations with justification for top dose:
Standard Test: 24, 120, 600, 3000, 6000µg/plate
Preinbuation assay: 4, 20, 100, 500, 2500µg/plate due to bacteriotoxicity.
Vehicle / solvent:
DMSO
Due to the Iimited solubility of the test substance in water, DMSO was selected as the vehicle, which had been demonstrated to be suitable in bacterial reverse mutation tests and for which historical control data are available.
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
4-nitroquinoline-N-oxide
9-aminoacridine
other: 2-aminoanthracene (with S9), N-methyl-N-nitro-N-nitrosoguanidine (TA 1535, TA 100, without S9), 4-nitro-o-phenylendiamine (TA 98, without S9)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation); preincubation; in suspension; as impregnation on paper disk

DURATION
- Preincubation period: 20min (pre-incubation assay only)
- Exposure duration: 48 - 72h

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
decrease in the number of revertants
clearing or diminution of the background lawn
reduction in the titer
Evaluation criteria:
Acceptance criteria
Generally, the experiment is considered valid if the following criteria are met:
- The number of revertant colonies in the negative controls was within the normal range of the historical control data for each tester strain.
- The sterility controls revealed no indication of bacterial contamination.
- The positive control articles both with and without S-9 mix induced a significant increase in the number of revertant colonies within the range of the historical control data.
- The titer of viable bacteria was at least 10E8/mL.

Evaluation criteria
The test chemical is considered positive in this assay if the following criteria are met:
-A dose-related and reproducible increase in the number of revertant colonies, i.e. about doubling of the spontaneous mutation rate in at least one tester strain either without S-9 mix or after adding a metabolizing system.

A test substance is generally considered nonmutagenic in this test if:
-The number of revertants for all tester strains were within the historical negative control range under all experimental conditions in two experiments carried out independently of each other.

Results and discussion

Test results
Species / strain:
bacteria, other: all strains tested
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
Toxicity:
A bacteriotoxic effect (reduced background growth, slight decrease in the number of revertants, reduction in the titer) was observed in the standard plate test depending on the strain and test conditions from about 600 µg - 3,000 µg/plate.
In the preincubatian assay bacteriotoxicity was observed depending on the strain and test conditions from about 100µg - 500µg/plate onward.

Precipitation:
Test substance precipitation was found at 6000µg/plate.

Applicant's summary and conclusion