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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 283-042-4 | CAS number: 84539-54-8
- 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
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
Link to relevant study records
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- migrated information: read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Study period:
- 2000-09-04 to 2000-11-17
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: GLP guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- in vitro mammalian chromosome aberration test
- Target gene:
- Not applicable
The in vitro chromosome aberration test is able to identify substances that cause structural chromosome aberrations in cultured mammalian cells. - Species / strain / cell type:
- lymphocytes:
- Details on mammalian cell type (if applicable):
- - Type and identity of media: RPMI 1640 (containing 20% fetal calf serum)
- Additional strain / cell type characteristics:
- not applicable
- Remarks:
- stable karyotype with 46 chromosomes and an average cell cycle time of 12-14 hours
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix consisiting of enzymatic systems induced by Aroclor 1254 in rat liver microsomal fraction (S9 fraction) and the cofactors necessary for their function
- Test concentrations with justification for top dose:
- With a treatment volume of 100 µL/5.5 mL culture medium, the dose-levels were as follows:
First experiment: 78.125, 156.25, 312.5, 625, 1250, 2500, 3750 and 5000 µg/mL (with and without S9 mix)
Second experiment: 156.25, 312.5, 625, 1250, 2500 and 5000 mg/mL (with and without S9 mix) - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: culture medium
- Justification for choice of solvent/vehicle: The test substance was freely soluble in the culture medium. - Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- culture medium
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- mitomycin C
- Remarks:
- without S9 mix
Migrated to IUCLID6: 3 µg/mL (3h of treatment) or 0.2 µg/mL (continuous treatment) - Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- culture medium
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- Remarks:
- with S9 mix
Migrated to IUCLID6: 50 µg/mL or 25 µg/mL - Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 3 hours in the first experiment (with and without S9 mix) and in the second experiment (with S9 mix); 20 hours and 44 hours in the second experiment (without S9 mix)
- Expression time (cells in growth medium): 17 hours in the first experiment (with and without S9 mix) and in the second experiment (with S9 mix); 41 hours in the second experiment (with S9 mix)
- Fixation time (start of exposure up to fixation or harvest of cells): 20 hours in the first experiment (with and without S9 mix); 20 and 44 hours in the second experiment (with and without S9 mix).
SPINDLE INHIBITOR (cytogenetic assays): Colcemid (10 µg/mL)
STAIN (for cytogenetic assays): Giemsa
NUMBER OF CELLS EVALUATED:
200 metaphases/dose level (with 100 metaphases/culture whenever possible. Only 50 metaphases/culture were analysed when at least 10% cells with structural chromosome aberrations were observed)
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index
OTHER EXAMINATIONS:
- Determination of polyploidy: yes (were recorded when present)
- Determination of endoreplication: yes (were recorded when present) - Evaluation criteria:
- A reproducible and statistically significant increase in the frequency of cells with structural chromosome aberrations for at least one of the dose-levels and one of the two harvest times was considered as a positive result. Reference to historical data or other considerations of biological relevance, was also taken into account in the evaluation of the findings.
- Statistics:
- For each test and for each harvest time, the frequency of cells with structural chromosome aberrations (excluding gaps) in treated cultures was compared to that of the vehicle control cultures. If necessary, the comparison was performed using the X2 test, in which p = 0.05 was used as the lowest level of significance.
- Species / strain:
- lymphocytes:
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Remarks:
- in the first experiment
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- lymphocytes:
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- In the second experiment, after 20-hour exposure to the test substance, up to 44% decrease in the mitotic index was noted, mainly at dose-levels > 1250 µg/mL. After 44-hour exposure, up to 51% decrease in the mitotic index was observed.
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- lymphocytes:
- Metabolic activation:
- with
- Genotoxicity:
- negative
- Remarks:
- No significant increase in the frequency of cells with structural chromosomal aberrations was noted in both experiments.
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Remarks:
- No noteworthy toxicity was induced in both experiments
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: At the dose level of 5000 µg/mL, pH was about 7.6 (as for the vehicle control)
- Effects of osmolality: At the dose level of 5000 µg/mL, the osmolality was equal to 320 mOsm/kg H20 (292 for the vehicle control)
- Water solubility: The test substance was freely soluble in the culture medium.
- Precipitation: The final dose-level of 5000 µg/mL showed no precipitate in the culture medium.
RANGE-FINDING/SCREENING STUDIES: No preliminary cytotoxicity test was performed. Dose-levels were selected on the basis of pH, osmolality and solubility.
COMPARISON WITH HISTORICAL CONTROL DATA: in the range - Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results (migrated information):
negative
Under the experimental conditions, the test substance EDDHAS Fe 3K did not induce any clear evidence of chromosome aberrations in cultured human lymphocytes. - Executive summary:
The objective of this study was to evaluate the potential of the test substance EDDHAS Fe 3K to induce chromosome aberrations in cultured human lymphocytes.
Methods
The test substance was tested in two independent experiments, both with and without a metabolic activation system, the S9 mix, prepared from a liver microsomal fraction (S9 fraction) of rats induced with Aroclor 1254. No preliminary cytotoxicity test was performed. Dose-levels were selected on the basis of pH, osmolality and solubility. A wide-range of treatment-levels was used for the first experiment and dose-levels for scoring of chromosomal aberrations were selected on the basis of cytotoxicity indicated by reduction of mitotic index (MI). For each culture, heparinised whole blood was added to culture medium containing a mitogen (phytohaemagglutinin) and incubated at 37°C in a humidified atmosphere of 5% CO2 /95% air, for 48 hours.
First experiment
Lymphocyte cultures were exposed to the test or control substances, with or without S9 mix, for 3 hours then rinsed. Cells were harvested 20 hours after the beginning of treatment, corresponding to approximately 1.5 normal cell cycles. One and a half hour before harvest, each culture was treated with a colcemid solution (10 µg/mL) to block cells at the metaphase-stage of mitosis.
Second experiment
- without S9 mix, cells were exposed continuously to the test or control substances,
- with S9 mix, cells were exposed to the test or control substances for 3 hours and then rinsed.
Cells were harvested 20 hours and 44 hours after the beginning of treatment, corresponding to approximately 1.5 normal cell cycles and 24 hours later, respectively. One and a half hour before harvest, each culture was treated with a colcemid solution (10 pg/ml) to block cells at the metaphase-stage of mitosis.
For both experiments, after hypotonic treatment (KCl 0.075 M), the cells were fixed in a methanol/acetic acid mixture (3/1; v/v), spread on glass slides and stained with Giemsa. All the slides were coded for scoring. The test substance EDDHAS Fe 3K was dissolved in culture medium.
The dose-levels of the positive controls were as follows:
- without S9 mix, mitomycin C: 3 µg/mL (3 hours of treatment) or 0.2 µg/mL (continuous treatment),
- with S9 mix, cyclophosphamide: 50 µg/mL or 25 µg/mL.
Results
The test substance was freely soluble in the culture medium. The final dose-level of 5000 mg/mL showed no precipitate in the culture medium and pH and osmolality values were equivalent to those of the vehicle control culture. With a treatment volume of 100 µL/5.5 mL culture medium, the dose-levels both with and without S9 mix were as follows:
- 78.125, 156.25, 312.5, 625, 1250, 2500, 3750 and 5000 µg/mL: for the first experiment,
- 156.25, 312.5, 625, 1250, 2500 and 5000 µg/mL: for the second experiment.
Cytotoxicity:
Except for a moderate toxicity induced at the highest dose-levels without S9 mix in the second experiment, no noteworthy toxicity was induced.
Chromosomal aberration analysis:
The metaphase analysis was performed at the following dose-levels, both with and without S9 mix:
- 2500, 3750 and 5000 µg/mL: for the first experiment,
-1250, 2500 and 5000 µg/mL: for the 20-hour harvest time in the second experiment,
- 5000 µg/mL: for the 44-hour harvest time in the second experiment.
No significant increase in the frequency of cells with structural chromosomal aberrations was noted in both experiments. The frequencies of cells with structural chromosome aberrations of the vehicle and positive controls were as specified in acceptance criteria. The study was therefore considered valid.
Conclusion
Under the experimental conditions, the test substance EDDHAS Fe 3K did not induce any clear evidence of chromosome aberrations in cultured human lymphocytes.
Reference
Chromosomal aberration analysis (experiments without S9 mix):
In the first experiment, after 3-hour exposure to the test substance, no significant increase in the frequency of cells with structural chromosomal aberrations was noted. After 20-hour exposure, a slight but a non significant increase (frequency of 3%) in the frequency of aberrant cells was noted at 5000 µg/mL. However, this increase was not considered as biologically relevant since it was neither dose-related nor reproducible between the two cultures. After 44-hour exposure, even though a slight increase in the frequency of aberrant cells was noted at 5000 µg/mL (3.5%), no statistically significant difference was obtained.
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
Salmonella and E.Coli tests in vitro (AmesTests):
Fe(3K)EDDHSA
(EC 462-490-6) was tested in the Bacterial Reverse Mutation Test in five
strains of Salmonella typhimurium (TA 1535, TA 1537, TA98, TA 100 and TA
102) (OECD Guideline 471) (CIT, 2000e; Report No. 20510 MMO). The test
substance Fe(3K)EDDHSA was dissolved in distilled water. The evaluation
of the toxicity was performed on the basis of the observation of the
decrease in the number of revertant colonies and/or a thinning of the
bacterial lawn. In a preliminary toxicity test the test substance did
not induce cytotoxicity up to the highest dose level of 5000 µg/plate
with and without a metabolic activation system. The test substance was
then tested in two independent experiments according to the direct plate
incorporation method except for the second test with S9 mix, which was
performed according to the preincubation method (60 minutes, 37°C). Each
strain was exposed to five dose-levels of the test substance: 312.5,
625, 1250, 2500 and 5000 µg/plate (three plates/dose-level), with and
without metabolic activation. After 48 to 72 hours of incubation at
37°C, the revertant colonies were scored.
Except for a slight thinning of the bacterial lawn noted in the TA 1535
strain in the second experiment at dose-levels > 2500 µg/plate, no
noteworthy toxicity was observed towards all the strains used, both with
and without S9 mix. The test substance did not induce any noteworthy
increase in the number of revertants, both with and without S9 mix, in
any of the five strains. The number of revertants for the vehicle and
positive controls fulfilled the acceptance criteria. The study was
therefore considered as valid. Under the experimental conditions, the
test substance Fe(3K)EDDHSA does not show mutagenic activity in the
bacterial reverse mutation test with Salmonella typhimurium.
Fe(Na)EDDHA (CAS 84539 -55 -9) was tested for the ability to induce mutagenic effect in histidine-requiring strains of Salmonella mutagenic (TA 98, TA 100, TA 102, TA 1535, TA 1537) and in a tryptophan-requiring strain of Escherichia coli (WP2 uvr A) (OECD Guideline 471) (CIBA-GEIGY, 1994b; Report No. 931146a). The test compound was dissolved in bidistilled water and tested at five concentrations ranging from 312.5 to 5000 µg/plate with and without metabolic activation.
Suitable positive controls were used for each strain. All experiments were repeated in order to confirm the results.
The results revealed no increased incidence of mutants by the test item with and without metabolic activation. Therefore, it was concluded that the test compound did not show mutagenic activity in S. typhimurium and E. coli. The positive controls induced mutagenic activity. In conclusion, Fe(Na)EDDHA provoked no mutagenic activity in this test system.
In-vitro Mammalian Chromosome Aberation Test
Fe(3K)EDDHSA
(EC 462-490-6) was tested for its potential to induce chromosome
aberrations in cultured human lymphocytes (OECD Guideline 473) (CIT,
2001b; Report No. 20677 MHL ). The test substance was tested in two
independent experiments, both with and without a metabolic activation.
No preliminary cytotoxicity test was performed. Dose-levels were
selected on the basis of pH, osmolality and solubility. A wide-range of
treatment-levels was used (for the first experiment: 78.125, 156.25,
312.5, 625, 1250, 2500, 3750 and 5000 µg/mL; and for the second
experiment: 156.25, 312.5, 625, 1250, 2500 and 5000 µg/mL) and
dose-levels for scoring of chromosomal aberrations were selected on the
basis of cytotoxicity, indicated by reduction of mitotic index (MI). In
the first experiment, lymphocyte cultures were exposed to the test or
control substances, with or without S9 mix, for 3 hours then rinsed.
Cells were harvested 20 hours after the beginning of treatment. In the
second experiment,cells
were exposed continuously to the test or control substances (without S9
mix) and for 3 hours and then rinsed (with S9 mix), respectively. Cells
were harvested 20 hours and 44 hours after the beginning of treatment,
corresponding to approximately 1.5 normal cell cycles and 24 hours
later, respectively. In both experiments, one and a half hour before
harvest, each culture was treated with a colcemid solution (10 µg/mL) to
block cells at the metaphase-stage of mitosis.
The
test substance was freely soluble in the culture medium. The final
dose-level of 5000 mg/mL showed no precipitate in the culture medium and
pH and osmolality values were equivalent to those of the vehicle control
culture.Except
for a moderate toxicity induced at the highest dose-levels without S9
mix in the second experiment, no noteworthy toxicity was induced.
The
metaphase analysis was performed at the dose levels of 2500, 3750 and
5000 µg/mL in the first experiment and at the1250, 2500 and 5000 µg/mL
for the 20-hour harvest time in the second experiment, and at the 5000
µg/mL for the 44-hour harvest time in the second experiment both with
and without S9 mix. No significant increase in the frequency of cells
with structural chromosomal aberrations was noted in both experiments.
The frequencies of cells with structural chromosome aberrations of the
vehicle and positive controls fulfilled the acceptance criteria. The
study was therefore considered valid. In
conclusion, the test substance Fe(3K)EDDHSA did not induce any clear
evidence of chromosome aberrations in cultured human lymphocytes.
The test compound Fe(Na)EDDHA (CAS 84539 -55 -9) was tested for the ability to provoke clastogenic effects in Chinese hamster ovary cells (CCL61) in vitro (OECD TG 473) (CIBA-GEIGY, 1994c; Report No. 931147). The compound was dissolved in DMSO and tested without metabolic activation at concentrations of 0, 7.81,15.63 and 31.25 µg/mL for 18 and 42 hours. With metabolic activation (liver S9 fraction from Aroclor 1254 induced rat liver) concentrations of 0, 31.25, 62.5 and 125 µg/mL were applied for 3 hours followed by 15 hours recovery or 3 hours followed by 39 hours recovery. Higher concentrations could not be reached due to solubility limitations.
Three independent experiments of each with and without metabolic activation were performed. Two replicate culture per concentration and 200 cells per concentration were evaluated.
The results showed in both experiments with and without metabolic activation no increased number of metaphases with chromosomal aberrations. In contrast, the positive controls (Mitomycin 0.2 µg/mL and Cyclophosphamide 20 µg/mL) induced clastogenic effects. In conclusion, the test substance provoked no clastogenic activity in this test in vitro.
In-vitro Mammalian Cell Gene Mutation Test (Mouse Lymphoma Assay):
Fe(Na)EDDHA (CAS 84539 -55 -9) was tested for the ability to provoke mutations at the tk locus in L5178Y mouse lymphoma cells in vitro (OECD Guideline 476) (CIBA-GEIGY, 1994a; Report No. 931146b). The test compound was dissolved in DMSO. The range finding experiments showed that 1000 µg/mL was the highest concentration which could be used. Higher concentrations (greater than 100 mg/mL) produced precipitates in the vehicle.
In the presence of metabolic activation (liver S9 -fraction from Aroclor 1254 treated rats) the two highest concentrations revealed cytotoxicity. In absence of metabolic activation no toxicity was noted.
For the mutagenicity experiment, concentrations ranging from 0 to 125 µg/mL with metabolic activation and from 0 to 1000 µg/mL without metabolic activation were used. In the confirmatory experiment with metabolic activation concentrations ranging from 0 to 250 µg/mL were applied. The same concentrations (0 to 1000 µg/mL) were used in the confirmatory experiment without metabolic activation.
Corresponding positive controls (N-Nitrosodimethylamine, with metabolic activation and Ethylmethansulfonate without metabolic activation) were included. The mouse lymphoma cells were treated for 4 hours. After two days expression time, mutations at the tk locus were selected by resistance to 5-trifluorothymidine. Two types of colonies were selected, large colonies (base-pair substitutions and deletions) and small colonies (chromosome aberrations). The results showed no increase incidence of mutations at the tk locus of mouse lymphoma L5178Y cells in presence or absence of metabolic activation. Positive controls showed mutagenic activity. In conclusion, Fe(Na)EDDHA was not mutagenic in this test system in vitro.
Justification for selection of genetic toxicity endpoint
Key study conducted with the nearest analogue Fe(3K)EDDHSA (EC 462-490-6) in mammalian cells
Justification for classification or non-classification
Based on results of the three different in vitro genetic toxicity studies, performed with the nearest analogues Fe(3K)EDDHSA and Fe(Na)EDDHA, the target substance Fe(3Na)EDDHSA does not need to be classified and labelled as genotoxic according to Regulation 1272/2008/EC (CLP).
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.