Registration Dossier
Registration Dossier
Data platform availability banner - registered substances factsheets
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 443-010-4 | CAS number: 53641-10-4
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
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
- Study period:
- 1999-12-06 to 2000-01-28
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 000
- Report date:
- 2000
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Version / remarks:
- adopted: July 21st, 1997
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- -
- EC Number:
- 443-010-4
- EC Name:
- -
- Cas Number:
- 53641-10-4
- Molecular formula:
- C14H15ClN2O4
- IUPAC Name:
- N-[3-chloro-4-(3-oxobutanamido)phenyl]-3-oxobutanamide
- Test material form:
- solid
Constituent 1
Method
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:
- S9-mix from rat liver
- Test concentrations with justification for top dose:
- plate incorporation test:
with and without metabolic activation:
50, 160, 500, 1600 and 5000 µg/plate
preincubation test:
with and without metabolic activation:
16, 50,160,500, 1600 and 5000 µg/plate
The test compound did not precipitate on the plates up to the highest investigated dose
of 5000 µg/plate.
In the plate incorporation test toxicity was observed with and without metabolic
activation with the strain TA 100 at a concentration of 5000 µg/plate.
In the preincubation test toxicity was not observed with and without metabolic
activation. - Vehicle / solvent:
- Solvent: DMSO
Controls
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- 4-nitroquinoline-N-oxide
- 9-aminoacridine
- 2-nitrofluorene
- sodium azide
- other: 2-aminoanthracene
- Details on test system and experimental conditions:
- Two independent mutation tests were performed unless clearly positive or dose-related
activity was observed in the first test. Where results were negative or equivocal, a
second test was conducted. This included a pre-incubation step if the first test was
clearly negative. Pre-incubation involved incubating the test substance, S9-mix and
bacteria for a short period before pouring this mixture onto plates of minimal agar.
Each test was performed in both the presence and absence of S9-mix using all
bacterial tester strains and a range of concentrations of the test substance. Positive
and negative controls as well as solvent controls were included in each test. Triplicate
plates were used.
The highest concentration in the first mutation experiment was usually 50 mg/ml of the
test substance in the chosen solvent, which provided a final concentration of
5000 µg/plate. Further dilutions of 1600, 500, 160 and 50 µg/plate were used. Suitable
dose levels used in the second experiment may be different depending on any toxicity
seen in the first experiment. A reduction in the number of spontaneously occurring
colonies and visible thinning of the bacterial lawn were used as toxicity indicators.
Thinning of the bacterial lawn was evaluated microscopically.
In both tests top agar was prepared which, for the Salmonella strains, contained 100 ml
agar (0.6 % (w/v) agar, 0.5 % (w/v) NaCl) with 10 ml of a 0.5 mM histidine-biotin
solution. For E. coli histidine was replaced by tryptophan (2.5 ml, 0.5 mM). The
following ingredients were added (in the following order) to 2 ml of molten top agar at
approx. 48 °C:
0.5 ml S9-mix (if required) or buffer
0.1 ml of an overnight nutrient broth culture of the bacterial tester strain
0.1 ml test compound solution (dissolved in DMSO)
In the second mutagenicity test if appropriate these top-agar ingredients were
preincubated by shaking for approximately 20 minutes at approx. 30°C.
After mixing, and preincubation if appropriate, the liquid was poured into a petri dish
containing a 25 ml layer of minimal agar (1.5 % (w/v) agar, Vogel-Bonner E medium
with 2 % (w/v)glucose). After incubation for approximately 48 hours at approx. 37 °C in
the dark, colonies (his+ and trp+ revertants) were counted by hand or by a suitable
automatic colony counter.
The counter was calibrated for each test by reading a test pattern plate to verify the
manufacturer's requirements for the counter's sensitiveness. - Evaluation criteria:
- Criteria for a valid assay
The assay is considered valid if the following criteria are met:
- the solvent control data are within the laboratory's normal control range for the
spontaneous mutant frequency
- the positive controls induce increases in the mutation frequency which are
significant and within the laboratory's normal range
Criteria for a positive response
A test compound is classified as mutagenic if it has either of the following effects:
a) it produces at least a 2-fold increase in the mean number of revertants per plate of
at least one of the tester strains over the mean number of revertants per plate of
the appropriate vehicle control at complete bacterial background lawn
b) it induces a dose-related increase in the mean number of revertants per plate of at
least one of the tester strains over the mean number of revertants per plate of the
appropriate vehicle control in at least two to three concentrations of the test
compound at complete bacterial background lawn.
If the test substance does not achieve either of the above criteria, it is considered to
show no evidence of mutagenic activity in this system.
Results and discussion
Test resultsopen allclose all
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- (> 5000 µg/plate)
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- (>5000 µg/plate)
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- (> 5000 µg/plate)
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- ( 5000 µg/plate)
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- E. coli WP2 uvr A pKM 101
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- (>5000 µg/plate)
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
Applicant's summary and conclusion
- Conclusions:
- The results lead to the conclusion that the test item is not mutagenic in these bacterial
test systems either in the absence or in the presence of an exogenous metabolizing
system. - Executive summary:
The test item was tested for mutagenicity with the strains TA 100,
TA 1535, TA 1537 and TA 98 of Salmonella typhimurium and with Escherichia coli
WP2uvrA.
Two independent mutagenicity studies were conducted (one plate incorporation test
and one preincubation test), each in the absence and in the presence of a metabolizing
system derived from a rat liver homogenate.
For both studies, the compound was dissolved in DMSO, and each bacterial strain was
exposed to 5 dose levels, in the preincubation test to 6 dose levels.
The test compound did not precipitate on the plates up to the highest investigated dose
of 5000 μg/plate.
The concentrations for the plate incorporation test were 50, 160, 500, 1600 and
5000 μg/plate.
Because of toxicity in the plate incorporation test dose levels from 16 to 5000 μg/plate
were chosen for the pre incubation test.
Control plates without mutagen showed that the number of spontaneous revertant
colonies was within the laboratory's historical control range. All the positive control
compounds showed the expected increase in the number of revertant colonies.
Toxicity: In the plate incorporation test toxicity was observed with and without metabolic
activation with the strain TA 100 at a concentration of 5000 μg/plate.
In the preincubation test toxicity was not observed with and without metabolic
activation.
Mutagenicity: In the absence and in the presence of the metabolic activation system
the test item did not result in relevant increases in the number of revertants in any of the
bacterial strains.
Summarizing, it can be stated that the test item was not mutagenic in this bacterial
mutation test at any dose level either in the absence or presence of an exogenous
metabolic activation.
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.