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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 214-874-8 | CAS number: 1204-28-0
- 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:
- 20 May 2015-30 July 2015
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Remarks:
- The study has been performed according to OECD and/or EC guidelines and according to GLP principles.
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 015
- Report date:
- 2015
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Version / remarks:
- Adopted July 21, 1997
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- 4-chloroformylphthalic anhydride
- EC Number:
- 214-874-8
- EC Name:
- 4-chloroformylphthalic anhydride
- Cas Number:
- 1204-28-0
- Molecular formula:
- C9H3ClO4
- IUPAC Name:
- 1,3-dioxo-1,3-dihydro-2-benzofuran-5-carbonyl chloride
- Test material form:
- solid
- Details on test material:
- - Name of test material (as cited in study report): Trimellitic anhydride chloride
For more details see Confidential details on test material
Constituent 1
- Specific details on test material used for the study:
- No correction was made for the purity/composition of the test substance. Trimellitic anhydride chloride was dissolved in dimethyl sulfoxide (DMSO, SeccoSolv, Merck, Darmstadt, Germany). The stock solutions were treated with ultrasonic waves until the test substance had completely dissolved. Except in the dose range finding test where the stock solution was already dissolved after vortexing only. Test substance concentrations were used within 2 hours of preparation.
Method
- Target gene:
- - S. typhimurium: Histidine gene
- E. coli: Tryptophan gene
Species / strainopen allclose all
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Details on mammalian cell type (if applicable):
- not applicable
- Additional strain / cell type characteristics:
- not applicable
- Species / strain / cell type:
- E. coli WP2 uvr A
- Details on mammalian cell type (if applicable):
- not applicable
- Additional strain / cell type characteristics:
- not applicable
- Cytokinesis block (if used):
- not applicable
- Metabolic activation:
- with and without
- Metabolic activation system:
- Rat liver S9-mix induced by Aroclor 1254
- Test concentrations with justification for top dose:
- Dose range finding test (without and with 5% (v/v) S9-mix ; TA100 and WP2uvrA): 1.7, 5.4, 17, 52, 164, 512, 1600 and 5000 µg/plate.
The highest concentration of the test substance used in the subsequent mutation experiment was 5000 μg/plate or the level at which the test substance inhibited bacterial growth.
Mutation experiments:
Experiment 1 (with pre incubation):
Without and with 5% (v/v) S9-mix ; TA1535, TA1537, TA98, TA100 and WP2uvrA: 17, 52, 164, 512, 1600 and 5000 μg/plate
Experiment 2 (with pre incubation):
Without and with 10% (v/v) S9-mix ; TA1535, TA1537, TA98, TA100 and WP2uvrA: 154, 275, 492, 878, 1568 and 2800 μg/plate
Experiment 3 (with pre incubation):
Without S9-mix ; TA1535: 17, 52, 164, 512, 1600 and 5000 μg/plate - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: dimethyl sulfoxide (DMSO, SeccoSolv, Merck, Darmstadt, Germany)
- Justification for choice of solvent/vehicle:
Test compound was soluble in DMSO.
Controlsopen allclose all
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- Remarks:
- without S9-mix: 5 µg/plate in saline for TA1535
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: ICR-191
- Remarks:
- without S9-mix: 2.5 µg/plate in DMSO for TA1537
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 2-nitrofluorene
- Remarks:
- without S9-mix: 10 µg/plate in DMSO for TA98
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- methylmethanesulfonate
- Remarks:
- without S9-mix: 650 µg/plate in DMSO for TA100
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 4-nitroquinoline-N-oxide
- Remarks:
- without S9-mix: 10 µg/plate in DMSO for WP2uvrA
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 2-aminoanthracene in DMSO for all tester strains
- Remarks:
- with S9-mix
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: plate incorporation (in agar) and preincubation
DOSE FINDING RANGE TEST (no pre-incubation was performed during the dose range finding test)
S9-mix was prepared immediately before use and kept on ice. S9-mix contained per 10 ml: 30 mg NADP (Randox Laboratories Ltd., Crumlin, United Kingdom) and 15.2 mg glucose-6-phosphate (Roche Diagnostics, Mannheim, Germany) in 5.5 ml or 5.0 ml Milli-Q water (first or second experiment respectively) (Millipore Corp., Bedford, MA., USA); 2 ml 0.5 M sodium phosphate buffer pH 7.4; 1 ml 0.08 M MgCl2 solution (Merck); 1 ml 0.33 M KCl solution (Merck). The above solution was filter (0.22 μm)-sterilized. To 9.5 ml of S9-mix components 0.5 ml S9-fraction was added (5% (v/v) S9-fraction) to complete the S9-mix in the first experiment and to 9.0 ml of S9-mix components 1.0 ml S9-fraction was added (10% (v/v) S9-fraction) to complete the S9-mix in the second experiment.
MUTATION EXPERIMENT
At least five different doses (increasing with approximately half-log steps) of the test substance were tested in triplicate in each strain. In the first experiment, the test substance was tested both in the absence and presence of 5% (v/v) S9-mix in the tester strains TA1535, TA1537, TA98, TA100 and WP2uvrA in a pre-incubation experiment. In a follow-up experiment with additional parameters, the test substance was tested both in the absence and presence of 10% (v/v) S9-mix in all tester strains in a pre-incubation experiment. An additional experiment was performed with tester strain TA1535 in the absence of S9-mix.
The negative control (vehicle) and relevant positive controls were concurrently tested in each strain in the presence and absence of S9-mix.
Direct plate experiment: Top agar in top agar tubes was melted by heating to 45 ± 2°C. The following solutions were successively added to 3 ml molten top agar: 0.1 ml of a fresh bacterial culture (10E9 cells/ml) of one of the tester strains, 0.1 ml of a dilution of the test substance in dimethyl sulfoxide and either 0.5 ml S9-mix (in case of activation assays) or 0.5 ml 0.1 M phosphate buffer (in case of non-activation assays). The ingredients were mixed on a Vortex and the content of the top agar tube was poured onto a selective agar plate.
Pre-incubation experiment: Top agar in top agar tubes was melted by heating to 45 ± 2°C. The following solutions were pre-incubated for 30 minutes by 70 rpm at 20°C, either 0.5 ml S9-mix (in case of activation assays) or 0.5 ml 0.1 M phosphate buffer (in case of non-activation assays), 0.1 ml of a fresh bacterial culture (10E9 cells/ml) of one of the tester strains, 0.1 ml of a dilution of the test substance in DMSO. After the pre-incubation period the solutions were added to 3 ml molten top agar. The ingredients were mixed on a Vortex and the content of the top agar tube was poured onto a selective agar plate.
After solidification of the top agar, the plates were inverted and incubated in the dark at 37°C for 48 ± 4 h. After this period revertant colonies (histidine independent (His+) for Salmonella typhimurium bacteria and tryptophan independent (Trp+) for Escherichia coli were counted.
COLONY COUNTING
The revertant colonies were counted automatically with the Sorcerer Colony Counter. Plates with sufficient test article precipitate to interfere with automated colony counting were counted manually.
Evidence of test article precipitate on the plates and the condition of the bacterial background lawn were evaluated when considered necessary, macroscopically and/or microscopically by using a dissecting microscope.
ACCEPTABILITY OF THE ASSAY
A Salmonella typhimurium reverse mutation assay and/or Escherichia coli reverse mutation assay is considered acceptable if it meets the following criteria:
a) The vehicle control and positive control plates from each tester strain (with or without S9-mix) must exhibit a characteristic number of revertant colonies when compared against relevant historical control data generated at Test Facility.
b) The selected dose range should include a clearly toxic concentration or should exhibit limited solubility as demonstrated by the preliminary toxicity range-finding test or should extend to 5 mg/plate.
c) No more than 5% of the plates are lost through contamination or some other unforeseen event. If the results are considered invalid due to contamination, the experiment will be repeated. - Evaluation criteria:
- A test substance is considered negative (not mutagenic) in the test if:
a) The total number of revertants in the tester strain TA100 or WP2uvrA is not greater than two (2) times the concurrent control, and the total number of revertants in tester strains TA1535, TA1537 or TA98 is not greater than three (3) times the concurrent vehicle control.
b) The negative response should be reproducible in at least one follow-up experiment.
A test substance is considered positive (mutagenic) in the test if:
a) The total number of revertants in the tester strain TA100 or WP2uvrA is greater than two (2) times the concurrent control, or the total number of revertants in tester strains TA1535, TA1537, TA98 is greater than three (3) times the concurrent vehicle control.
b) In case a follow up experiment is performed when a positive response is observed in one of the tester strains, the positive response should be reproducible in at least one follow up experiment.
Results and discussion
Test resultsopen allclose all
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- positive
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Species / strain:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: Precipitation of Trimellitic anhydride chloride on the plates was not observed at the start or at the end of the incubation period. Except in the first experiment where at the end of the incubation period Trimellitic anhydride chloride slightly precipitated at the top concentration of 5000 μg/plate in the presence of S9-mix in strains TA1535, TA1537 and TA98.
RANGE-FINDING/SCREENING STUDIES:
- No precipitation was observed at the start of the incubation period in both tester strain. No mutagenicity was observed up to and including the top dose of 5000 µg/plate. Toxicity: In tester strain TA100, a slight reduction of the bacterial background lawn was observed at the test substance concentration of 1600 μg/plate in the absence and presence of S9-mix. A complete lack of any micro-colony background lawn and no revertant colonies were present at 5000 μg/plate in the absence and presence of S9-mix. In tester strain WP2uvrA, an extreme reduction of the bacterial background lawn and an increase in the size of the microcolonies compared to the solvent control plate was observed at the test substance concentration of 5000 μg/plate in the absence and presence of S9-mix.
COMPARISON WITH HISTORICAL CONTROL DATA:
- The negative and strain-specific positive control values were within the laboratory historical control data ranges indicating that the test conditions were adequate and that the metabolic activation system functioned properly.
ADDITIONAL INFORMATION ON CYTOTOXICITY:
- Toxicity to all strains exihbited at 5000 µg/plate in the first mutation experiment without S9-mix only. Toxicity to majority of strains exhibited at 2800 µg/plate with or without S9-mix. - Remarks on result:
- other: Mutagenic
Applicant's summary and conclusion
- Conclusions:
- Based on the results of this study it is concluded that Trimellitic anhydride chloride is mutagenic in the Salmonella typhimurium reverse mutation assay (strain TA100 only) and is not mutagenic in the Escherichia coli reverse mutation assay.
- Executive summary:
The mutagenic activity of Trimellitic anhydride chloride was investigated using the Salmonella typhimurium reverse mutation assay and the Escherichia coli reverse mutation assay in accordance with OECD Guideline No. 471 and the EU Method B.13/14 and under GLP compliance.
The test substance was tested in the Salmonella typhimurium reverse mutation assay with four histidine-requiring strains of Salmonella typhimurium (TA1535, TA1537, TA100 and TA98) and in the Escherichia coli reverse mutation assay with a tryptophan-requiring strain of Escherichia coli (WP2uvrA). At first a dose range finding test was performed in the strains TA100 and WP2uvrA in a direct plate experiment both in the absence and presence of S9-mix (rat liver S9-mix induced Aroclor 1254). After that the test was performed with the inclusion of the pre-incubation step in two independent experiments both in the absence and presence of S9-mix (rat liver S9-mix induced Aroclor 1254). An additional experiment was performed with tester strain TA1535 in the absence of S9-mix.
In the dose range finding test, the test substance was initially tested up to concentrations of 5000 μg/plate in the absence and presence of 5% (v/v) S9-mix in the strains TA100 and WP2uvrA in the direct plate experiment. The test substance did not precipitate on the plates at this dose level. In tester strain TA100, toxicity was observed at dose levels of 1600 and 5000 μg/plate in the absence and presence of S9-mix. In tester strain WP2uvrA, toxicity was observed at the dose level of 5000 μg/plate in the absence and presence of S9-mix. No biologically relevant increase in the number of revertants was observed upon treatment with the test substance.
Based on the results of the dose range finding test, the test substance was tested in the first mutation experiment at a concentration range of 17 to 5000 μg/plate in the absence and presence of 5% (v/v) S9-mix in the tester strains TA1535, TA1537, TA98, TA100 and WP2uvrA with the inclusion of a pre-incubation step. Test substance precipitated on the plates in the presence of S9-mix in the tester strains TA1535, TA1537 and TA98. Toxicity was observed in all tester strains. In tester strain TA1535, test substance induced up to 5.1- and 3.4-fold increases in the number of revertant colonies compared to the solvent control in the absence and presence of S9-mix, respectively. In tester strain TA100, test substance induced up to 1.9- and 2.7-fold dose related increases in the number of revertant colonies compared to the solvent control in the absence and presence of S9-mix, respectively.
In a follow-up experiment with additional parameters, the test substance was tested at a concentration range of 154 to 2800 μg/plate in the absence and presence of 10% (v/v) S9-mix in the tester strains TA1535, TA1537, TA98, TA100 and WP2uvrA in the pre-incubation experiment. Toxicity was observed in all tester strains. In tester strain TA100, test substance induced up to 2.0- and 4.0-fold dose related increases in the number of revertant colonies compared to the solvent control in the absence and presence of S9-mix, respectively.
To verify the mutagenic response of tester strain TA1535 in the first experiment, an additional pre-incubation experiment was performed with this tester strain at a concentration range of 17 to 5000 μg/plate in the absence of S9-mix. Toxicity was observed and no biologically relevant increase in the number of revertants was observed.
In tester strain TA100, test substance induced up to 2.0- and 4.0-fold dose related, increases in the number of revertant colonies compared to the solvent control in the absence and presence of S9-mix, respectively in both pre-incubation experiments. Since the increases observed in tester strain TA100 were above the historical control data range and the results were reproducible in the repeat experiment, these increases are biologically relevant and test substance is considered to be mutagenic in the absence and presence of S9-mix.
The test substance showed increases in the number of revertant colonies with tester strain TA1535. Although the up to 5.1 fold increase in the absence of S9-mix was above the historical control data range, this increase was not seen in two repeat experiments and only observed at one dose level. Therefore, this increase is considered to be not biologically relevant. The up to 3.4-fold increase in the presence of S9-mix was not above the historical control data range and was observed in only one experiment. Therefore, this increase is considered to be not biologically relevant.
All other bacterial strains showed negative responses over the entire dose range, i.e. no dose-related, increase in the number of revertants in follow-up experiments.
In this study, the negative and strain-specific positive control values were within the laboratory historical control data ranges indicating that the test conditions were adequate and that the metabolic activation system functioned properly.
Based on the results of this study it is concluded that Trimellitic anhydride chloride is mutagenic in tester strain TA100 of the Salmonella typhimurium reverse mutation assay. Trimellitic anhydride chloride is not mutagenic in the other Salmonella typhimurium tester strains (TA1535, TA1537 or TA98) or Escherichia coli strain reverse mutation assay using strain WP2uvrA.
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.