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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 213-139-9 | CAS number: 926-63-6
- 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
Ames: negative
HPRT: negative
MNT in vitro: negative
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 16 June 1999 - 14 July 1999
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Version / remarks:
- 1997
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Version / remarks:
- 1992
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- his operon for S. typhimurium strains
trp operon for the E. coli strain - Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
- Additional strain / cell type characteristics:
- other: TA 98: rfa-, uvrB-, R-factor; TA 100: rfa-, uvrB-, R-factor; TA 1535: rfa-, uvrB-; TA 1537: rfa-, uvrB; WP2: trp-; uvr A-
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 liver mix prepared from Sprague-Dawley rats treated with Aroclor 1254
- Test concentrations with justification for top dose:
- First experiment (standard plate test, with and without metabolic activation): 0, 20, 100, 500, 2500 and 5000 µg/plate
Second experiment (preincubation test with and without metabolic activation): 0, 20, 100, 500, 2500 and 5000 µg/plate
Third experiment (preincubation test without metabolic activation): 0, 125, 250, 500, 1000 and 1500 µg/plate - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: water
- Untreated negative controls:
- yes
- Remarks:
- (sterility control)
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Remarks:
- with S9-mix
- Positive control substance:
- other: 2-aminoanthracene (2-AA)
- Remarks:
- 2.5 µg/plate, in DMSO, for TA 1535, TA 1537, TA 100, TA 98; 60 µg/plate, in DMSO, for E. coli WP2 uvrA
- Positive controls:
- yes
- Remarks:
- without S9-mix
- Positive control substance:
- other: N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)
- Remarks:
- 5 µg/plate, in DMSO, for TA 1535 and TA 100
- Positive controls:
- yes
- Remarks:
- without S9-mix
- Positive control substance:
- 9-aminoacridine
- Remarks:
- 100 µg/plate, in DMSO, for TA 1537 Migrated to IUCLID6: (AAC)
- Positive controls:
- yes
- Remarks:
- without S9-mix
- Positive control substance:
- 4-nitroquinoline-N-oxide
- Remarks:
- 5 µg/plate, in DMSO, for E. coli WP2 uvrA Migrated to IUCLID6: (4-NQO)
- Positive controls:
- yes
- Remarks:
- (without S9-mix)
- Positive control substance:
- other: 4-nitro-o-phenylendiamine (NOPD)
- Remarks:
- 10 µg/plate, in DMSO, for TA 98
- Details on test system and experimental conditions:
- In the standard plate test, tubes were filled with 2mL portions of soft agar and kept in a water bath at 45°C. This soft agar consisted of 100 mL agar and 10 mL amino acid solution. As amino acid solution for the soft agar was used 0.5 mM histidine and 0.5 mM biotin for TA strains and 0.5 mM tryptophan for the E. coli strain.
Then following components are added:
0.1 ml test solution or vehicle
0.1 ml fresh bacterial culture
0.5 ml S9 -mix or phosphate buffer
After mixing samples were poured onto Vogel-Bonner (minimal glucose agar plates) plate and incubated for 48 - 72 hrs in the dark at 37°C.
For the preincubation test 0.1 mL test solution or vehicle, 0.1 mL bacterial suspension and 0.5 mL of the S9 mix were incubated at 37°C for 20 minutes. After addition of 2 mL soft agar samples were poured onto agar plates and incubated again at 37°C for 48 to 72 hrs.
For the E. coli strain, plate test differed again in mixture of amino acid solution of the soft agar, the histidine component used for the TA strains being replaced by tryptophan.
Triplicate testing is done. - Evaluation criteria:
- An assay is accepted when the following criteria are met:
1.) number of colonies in the negative control is in the historical control range
2.) no indication of bacterial contamination (checked by sterility control)
3.) number of colonies in the positive controls are in the range of historical control data
4.) titer of viable bacteria is ≥ 10 E+9/mL
Toxicity is detected by:
1.) decrease in the number of revertants
2.) titer reduction
3.) clearing or diminution of the background lawn
Precipitation:
As long as no interference between precipitation and colony counting occurs is 5 mg/plate set as maximum dose even for relatively insoluble compounds.
A test chemical is to be considered as mutagenic when:
1.) increase of number of revertant colonies is reproducible and dose-related.
2.) in at least 1 tester strain doubling of colony counts with or without S-9 mix or after adding a metabolizing system is seen.
A test chemical is to be considered as non-mutagenic when:
1.) the number of revertants is inside the range of historical negative control data in 2 experiments performed independently from each other. - Species / strain:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Remarks:
- no increase in number of revertants was observed
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- in preincubation test only (for detail see 'additional information on results')
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- Precipitation: no precipitation detected
Cytotoxicity: no cytotoxic effects were seen in the standard plate-incorporation test with and without metabolic activation. In the tests with preincubation but without metabolic activation cytotoxicty occurred at
1.) 1500 µg/ plate (ratio of number of revertants for test and control data < 0.5 ) for strain TA98
2.) 5000 µg/ plate (ratio of number of revertants for test and control data < 0.5 ) for strain E. coli WP2
3.) 2500 µg/ plate (reduced background growth) for strains TA 1535, TA 100, TA 1537 and TA 98
In the tests with preincubation and metabolic activation cytotoxicty occurred at 5000 µg/plate (reduced background growth) for strains TA 1535, TA 100, TA 1537 and TA 98 - Conclusions:
- A test of bacterial gene mutagenicity was conducted for DMPA according to the OECD TG 471 (1997), with following strains: TA 98, TA 100, TA 1535 and TA 1537 and E. coli WP2 uvrA . The test concentrations were 20, 100, 500, 2500 and 5000 µg/plate for the standard plate test with and without S9 mix, and for the first preincubation test with and without S9 mix. The concentrations were 125, 250, 500, 1000 and 1500 µg/plate for second preincubation test conducted without S9 mix.
In none of the tests any mutagenic effect could be detected. Therefore the substance is to be considered as non-mutagenic in bacteria. - Endpoint:
- in vitro cytogenicity / micronucleus study
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 487 (In vitro Mammalian Cell Micronucleus Test)
- Qualifier:
- according to guideline
- Guideline:
- other: EU method B.49 (In vitro Mammalian Cell Micronucleus Test)
- Principles of method if other than guideline:
- A series of in-house non-GLP validation experiments was performed to get distinct responses of statistical significance when using the specified positive controls. To achieve such response the test design, specifically for the treatment, the recovery phase and harvest time, was slightly modified comparing the current proposal given in the OECD Guideline 487. The optimum in responses was found with the time schedule stated in the Summary.
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- in vitro mammalian cell micronucleus test
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Source and lot/batch number of test material: BASF, D035-2017-DMPA
- Expiration date of the lot/batch: 2020-09 26
- Purity test date: Jun 28 - Aug 29, 2017
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature
- Stability under storage conditions: guaranteed over the study period
OTHER SPECIFICS
- measurement of pH, osmolality, and precipitate in the culture medium to which the test chemical is added: yes, pH was adjusted, if necessary, in the stock solutions to physiological values using a small amount of 2M HCL. - Species / strain / cell type:
- lymphocytes: human
- Details on mammalian cell type (if applicable):
- CELLS USED
- Type and source of cells: primary human lymphocytes (collected from healthy non-smoking donors)
- Normal cell cycle time: app. 16h
For lymphocytes:
- Sex, age and number of blood donors: 1male (20 years), 1 female (29 years)
- Whether whole blood or separated lymphocytes were used: whole blood in medium
- Whether blood from different donors were pooled or not: no
- Mitogen used for lymphocytes: PHA
MEDIA USED
- Type and composition of media, CO2 concentration, humidity level, temperature, if applicable:
Dulbecco's Modified Eagles Medium/Ham's F12 (DMEM/F12, mixture 1:1) already supplemented with 200 mM GlutaMAX™. Additionally, the medium was supplemented with penicillin/streptomycin (100 U/mL/100 μg/mL), the mitogen PHA (3 μg/mL), 10 % FBS (fetal bovine serum), 10 mM HEPES and the anticoagulant heparin (125 U.S.P.-U/mL).
All incubations were done at 37 °C with 5.5 % CO2 in humidified air. - Cytokinesis block (if used):
- Cytochalasin B
- Metabolic activation:
- with and without
- Metabolic activation system:
- Phenobarbital/β-naphthoflavone induced rat liver S9
- Test concentrations with justification for top dose:
- 285, 489, 872 µg/ml
Dose selection was performed according to the current OECD Guideline for the in vitro micronucleus test. If no precipitate or limiting cytotoxicity is observed, the highest test concentration should correspond to 10 mM, 2 mg/mL or 2 μl/mL, whichever is the lowest. With regard to the molecular weight of the test item, 872 μg/mL (approx. 10 mM) were applied as top concentration, as no cytotoxicity or precipitation was observed. - Vehicle / solvent:
- culture medium
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- mitomycin C
- other: Demecolcine
- Details on test system and experimental conditions:
- NUMBER OF REPLICATIONS:
- Number of cultures per concentration: duplicate
- Number of independent experiments: 2 (4h, and 20h exposure)
TREATMENT AND HARVEST SCHEDULE:
- Preincubation period, if applicable: 48h, stimulation with PHA
- Exposure duration/duration of treatment: 4h (with and without S9) or 20h (only without S9)
- Harvest time after the beginning of treatment: 40h (2 - 2.5 cell cycles)
4h treatment, 16h recovery + 20h incubation with Cytochalasin B after removal of test substance
20h treatment + 20h incubation with Cytochalasin B after removal of test substance
FOR CHROMOSOME ABERRATION AND MICRONUCLEUS:
- If cytokinesis blocked method was used for micronucleus assay: indicate the identity of cytokinesis blocking substance (e.g. cytoB), its concentration, and duration and period of cell exposure: Cytochalasin, 4µg/ml, 20h
- Methods of slide preparation and staining technique used including the stain used (for cytogenetic assays): The cells were resuspended in 5 mL KCl solution (0.0375 M) and incubated at 37 °C for 20 minutes. 1 mL of ice-cold fixative mixture of methanol and glacial acetic acid (19 parts plus 1 part, respectively) was added to the hypotonic solution and the cells were resuspended carefully. After removal of the solution by centrifugation the cells were resuspended for 2 x 20 minutes in fixative and kept cold. The slides were prepared by dropping the cell suspension in fresh fixative onto a clean microscope slide. The cells were stained with Giemsa, mounted after drying and covered with a coverslip.
- Number of cells spread and analysed per concentration: at least 1000 binucleated cells per culture
- Criteria for scoring micronucleated cells: According to Countryman and Heddle (1976). The micronuclei have to be stained in the same way as the main nucleus. The area of the micronucleus should not extend the third part of the area of the main nucleus.
METHODS FOR MEASUREMENT OF CYTOTOXICITY
- Cytokinesis-block proliferation index
- Any supplementary information relevant to cytotoxicity: at least 5000 cells per cultrue used for evaluation - Evaluation criteria:
- A test item can be classified as non-clastogenic and non-aneugenic if:
− None of the test item concentrations exhibits a statistically significant increase compared with the concurrent solvent control
− There is no concentration-related increase
− The results in all evaluated test item concentrations should be within the range of the laboratory historical solvent control data (95% control limit realized as 95% confidence interval).
A test item can be classified as clastogenic and aneugenic if:
− At least one of the test item concentrations exhibits a statistically significant increase compared with the concurrent solvent control
− The increase is concentration-related in at least one experimental condition
− The results are outside the range of the laboratory historical solvent control data (95% control limit realized as 95% confidence interval). - Species / strain:
- lymphocytes: human
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- True negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- In this study, no precipitation of the test item in the culture medium was observed at the end of treatment.
No relevant influence on osmolarity was observed. The pH was adjusted to physiological values using a small amount of 2 M HCl
In this study in the absence and presence of S9 mix, no cytotoxicity was observed up to the highest applied concentration. - Conclusions:
- Dimethyl(propyl)amine did not induce micronuclei in primary human lymphocytes in vitro.
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test using the Hprt and xprt genes)
- Version / remarks:
- July 2016
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.17 (Mutagenicity - In Vitro Mammalian Cell Gene Mutation Test)
- Qualifier:
- according to guideline
- Guideline:
- EPA OTS 798.5300 (Detection of Gene Mutations in Somatic Cells in Culture)
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- in vitro mammalian cell gene mutation test using the Hprt and xprt genes
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Batch number of test material: D035-2017-DMPA
- Expiration date of the lot/batch: 2020-09-26
- Purity - study number: 17L00248
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature - Target gene:
- HPRT
- Species / strain / cell type:
- Chinese hamster Ovary (CHO)
- Details on mammalian cell type (if applicable):
- - Absence of Mycoplasma contamination:
yes
- Cell cycle length: 12-16h
- Modal number of chromosomes: 20
- Periodically ‘cleansed’ of spontaneous mutants: yes
MEDIA USED
Ham's F12 medium containing stable glutamine and hypoxanthine (PAN Biotech; Cat. No. P04-15500) supplemented with 10% (v/v) fetal calf serum (FCS). All media were supplemented with:
- 1% (v/v) penicillin/streptomycin (stock solution: 10000 IU / 10000 μg/mL)
- 1% (v/v) amphotericine B (stock solution: 250 μg/mL)
Cells were grown with 5% (v/v) CO2 at 37°C and ≥ 90% relative humidity up to approximate confluence and subcultured twice weekly. - Metabolic activation:
- with and without
- Metabolic activation system:
- phenobarbital and β-naphthoflavone induced rat liver S9
- Test concentrations with justification for top dose:
- 47.6, 85.7, 154.3, 277.8, 500, 900µg/mL
Repeat experiment: 300, 450, 600, 750, 900µg/mL
The top dose equals 10mM, the maximum concentration according to current guidelines, since no cytotoxicity, changes in pH value or osmolality, nor precipitation was observed in a pre-test up to 9.2mM (800µg/mL) - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: culture medium
- Justification for choice of solvent/vehicle: good solubility - Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 7,12-dimethylbenzanthracene
- ethylmethanesulphonate
- Details on test system and experimental conditions:
- NUMBER OF REPLICATIONS:
- Number of cultures per concentration:2
- Number of independent experiments : 3 (2 with S9, 2 w/out S9)
METHOD OF TREATMENT/ EXPOSURE:
- Cell density at seeding (if applicable): 20x10^6 / 40mL
- Test substance added in medium
TREATMENT AND HARVEST SCHEDULE:
- Preincubation period, if applicable: 20-24h
- Exposure duration/duration of treatment: 4h
FOR GENE MUTATION:
- Expression time (cells in growth medium between treatment and selection): 5-7 days
- Selection time (if incubation with a selective agent): app. 6-7 days
- Fixation time (start of exposure up to fixation or harvest of cells): 15 days
- Selective agent: 6-thioguanine (10µg/mL)
- Number of cells seeded and method to enumerate numbers of viable and mutants cells: 2x10^6 cells / 20mL
METHODS FOR MEASUREMENT OF CYTOTOXICITY
- Method: cloning efficiency; relative survival (RS) - Evaluation criteria:
- Acceptance criteria
The HPRT assay is considered valid if the following criteria are met:
• The absolute cloning efficiencies of the negative controls should not be less than 50% (with and without S9 mix).
• The background mutant frequency in the negative controls should be within our historical negative control data range (95% control limit). Weak outliers can be judged acceptable if there is no evidence that the test system is not “under control”.
• Concurrent positive controls both with and without S9 mix should induce responses that are compatible with those generated in the historical positive control data base and produce a statistically significant increase in mutant frequencies compared with the concurrent negative control.
Assessment criteria
A test substance is considered to be clearly positive if all following criteria are met:
• A statistically significant increase in mutant frequencies is obtained.
• A dose-related increase in mutant frequencies is observed.
• The corrected mutation frequencies (MFcorr.) exceeds both the concurrent negative control value and the range of our laboratory’s historical negative control data (95% control limit).
Isolated increases of mutant frequencies above our historical negative control range or isolated statistically significant increases without a dose-response relationship may indicate a biological effect but are not regarded as sufficient evidence of mutagenicity.
A test substance is considered to be clearly negative if the following criteria are met:
• Neither a statistically significant nor dose-related increase in the corrected mutation frequencies is observed under any experimental condition.
• The corrected mutation frequencies in all treated test groups is close to the concurrent vehicle control value and within the range of our laboratory’s historical negative control data (95% control limit). - Species / strain:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- True negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Data on pH: small amounts of 37% HCl were added to the stock solution to adjust the pH value
- Data on osmolality: not affected by test substance
- Precipitation and time of the determination: none
- Cell morphology: not influenced by treatment
Repeat experiments:
Experiment 1 was aborted due to a severe shift in the pH value
In experiment 2, a significant increase in mutant frequency in a single concentration (not highest dose) compared to concurrent control was observed. This part was repeated though all obtained values were well within historical control values.
Experiment 3 (repeat experiment) was accidentally carried out without S9 mix
Experiment 4 was the correct repeat experiment. - Conclusions:
- In the absence and the presence of metabolic activation, Dimethyl(propyl)amine is not a mutagenic substance in the HPRT locus assay using CHO cells.
Referenceopen allclose all
Experiment 1: Standard plate-incorporation test
SPT without S9-Mix [mean no. of mutations/ plate] |
|||||
Dosage [µg/plate] | TA 1535 | TA 100 | TA 1537 | TA 98 | WP2 uvrA |
Solvent control | 18 | 120 | 11 | 32 | 27 |
20 | 17 | 119 | 12 | 28 | 25 |
100 | 19 | 133 | 13 | 28 | 25 |
500 | 17 | 118 | 13 | 25 | 26 |
2500 | 18 | 120 | 13 | 29 | 28 |
5000 | 20 | 116 | 17 | 27 | 33 |
Positive control | 689 | 1252 | 988 | 1218 | 1028 |
SPT with S9-Mix [mean no. of mutations/ plate] |
|||||
Dosage [µg/ plate] | TA 1535 | TA 100 | TA 1537 | TA 98 | WP2 uvrA |
Solvent control | 19 | 131 | 17 | 41 | 28 |
20 | 19 | 135 | 17 | 32 | 26 |
100 | 19 | 130 | 12 | 35 | 27 |
500 | 21 | 139 | 11 | 32 | 27 |
2500 | 15 | 118 | 11 | 36 | 25 |
5000 | 14 | 126 | 12 | 33 | 30 |
Positive control | 203 | 1088 | 141 | 1070 | 202 |
Experiment 2: Preincubation test PIT without S9-Mix [mean no. of mutations/ plate] |
|||||
Dosage [µg/ plate] | TA 1535 | TA 100 | TA 1537 | TA 98 | WP2 uvrA |
Solvent control | 20 | 125 | 11 | 28 | 27 |
20 | 17 | 132 | 10 | 27 | 24 |
100 | 18 | 144 | 13 | 25 | 36 |
500 | 17 | 136 | 9 | 24 | 32 |
2500 | B | B | B | B | 24 |
5000 | B | B | B | B | 13 |
Positive control | 1174 | 1082 | 759 | 1230 | 558 |
PIT with S9-Mix [mean no. of mutations/ plate] |
|||||
Dosage [µg/ plate] | TA 1535 | TA 100 | TA 1537 | TA 98 | WP2 uvrA |
Solvent control | 19 | 136 | 10 | 40 | 37 |
20 | 17 | 140 | 10 | 37 | 35 |
100 | 18 | 136 | 9 | 31 | 39 |
500 | 16 | 132 | 11 | 32 | 45 |
2500 | 16 | 132 | 9 | 30 | 44 |
5000 | B | 75 | 3 | 25 | 35 |
Positive control | 100 | 640 | 96 | 722 | 244 |
Experiment 3: PIT without S9-mix | |||||
Dosage [µg/ plate] | TA 1535 | TA 100 | TA 1537 | TA 98 | |
Solvent control | 20 | 136 | 8 | 27 | |
125 | 19 | 124 | 10 | 30 | |
250 | 19 | 117 | 7 | 20 | |
500 | 15 | 118 | 8 | 19 | |
1000 | 16 | 110 | 7 | 23 | |
1500 | 15 | 92 | 7 | 14 | |
Positive control | 1027 | 1177 | 717 | 1158 | |
B: reduced background growth, cytotoxicity |
Table 1 Summary of results | |||||||
Exp. | Preparation | Test item | Proliferation | Cytostasis | Micronucleated | ||
interval | concentration | index | in %* | cells | 95% Ctrl limit | ||
in µg/mL | CBPI | in %** | |||||
Exposure period 4 h without S9 mix | |||||||
I | 40 h | Solvent control1 | 1.73 | 0.25 | 0.01 – 1.20 | ||
Positive control2 | 1.59 | 19.5 | 9.55S | 2.66 – 22.74 | |||
285 | 1.71 | 3.2 | 0.25 | ||||
498 | 1.80 | n.c. | 0.40 | ||||
872 | 1.73 | n.c. | 0.60 | ||||
Trend test: p-value 0.053 | |||||||
Exposure period 20 h without S9 mix | |||||||
II | 40 h | Solvent control1 | 1.86 | 0.30 | 0.00 – 1.14 | ||
Positive control3 | 1.45 | 48.0 | 4.00S | 1.15 – 6.44 | |||
93.0 | 1.72 | 15.6 | 0.20 | ||||
285 | 1.68 | 21.2 | 0.60 | ||||
872 | 1.66 | 23.1 | 0.50 | ||||
Trend test: p-value 0.414 | |||||||
Exposure period 4 h with S9 mix | |||||||
I | 40 h | Solvent control1 | 1.83 | 0.75 | 0.00 – 1.24 | ||
Positive control4 | 1.37 | 55.1 | 5.00S | 1.01 – 7.34 | |||
285 | 1.76 | 8.1 | 0.75 | ||||
498 | 1.67 | 19.6 | 1.10 | ||||
872 | 1.76 | 8.7 | 0.50 |
* For the positive control groups and the test item treatment groups the values are related to the solvent controls
** The number of micronucleated cells was determined in a sample of 2000 binucleated cells
S The number of micronucleated cells is statistically significantly higher than corresponding control values
n.c. Not calculated as the CBPI is equal or higher than the solvent control value
1 Culture medium 2 MMC 0.8 μg/mL 3 Demecolcine 125 ng/mL 4 CPA 17.5 μg/mL
Experiment 1: aborted due to pH shift
Experiment 2
Test groups [µg/mL] | S9 | # of colonies | Mutant frequency (per 10^6) | ||
flask 1 | flask 2 | uncorrected | corrected | ||
Negative control | - | 2 | 6 | 2.00 | 2.89 |
154.3 | - | 0 | 4 | 1.00 | 1.48 |
277.8 | - | 0 | 2 | 0.50 | 0.91 |
500.0 | - | 6 | 4 | 2.50 | 2.67 |
900.0 | - | 2 | 3 | 1.25 | 2.18 |
EMS 400.0 | - | 93 | 80 | 43.25 | 94.54 |
Negative control | + | 1 | 1 | 0.50 | 0.75 |
154.3 | + | 2 | 0 | 0.50 | 0.91 |
277.8 | + | 1 | 4 | 1.25 | 1.91 |
500.0 | + | 6 | 5 | 2.75 | 4.20 |
900.0 | + | 5 | 4 | 2.25 | 3.60 |
DMBA 1.25 | + | 55 | 58 | 28.25 | 55.67 |
Experiment 3
Test groups [µg/mL] | S9 | # of colonies | Mutant frequency (per 10^6) | ||
flask 1 | flask 2 | uncorrected | corrected | ||
Negative control | - | 2 | 6 | 2.00 | 3.42 |
450.0 | - | 1 | 1 | 0.50 | 0.65 |
600.0 | - | 3 | 1 | 1.00 | 1.77 |
750.0 | - | 3 | 1 | 1.00 | 1.62 |
900.0 | - | 3 | 1 | 1.00 | 1.70 |
EMS 400.0 | - | 92 | 102 | 48.50 | 127.63 |
Experiment 4
Test groups [µg/mL] | S9 | # of colonies | Mutant frequency (per 10^6) | ||
flask 1 | flask 2 | uncorrected | corrected | ||
Negative control | + | 1 | 2 | 0.75 | 0.89 |
450.0 | + | 0 | 0 | 0.00 | 0.00 |
600.0 | + | 3 | 8 | 2.75 | 3.29 |
750.0 | + | 2 | 2 | 1.00 | 1.33 |
900.0 | + | 3 | 1 | 1.00 | 1.65 |
DMBA 1.25 | + | 67 | 72 | 34.75 | 59.91 |
correction on the basis of the absolute cloning efficiency 2 at the end of the expression period
significant values indicated in bold
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
In vitro genotoxicity:
A test of bacterial gene mutagenicity was conducted for DMPA according to the OECD TG 471 (1997), with following strains: TA 98, TA 100, TA 1535 and TA 1537 and E. coli WP2 uvrA (BASF AG 40M0164/994067). The test concentrations were 20, 100, 500, 2500 and 5000 µg/plate for the standard plate test with and without S9 mix, and for the first preincubation test with and without S9 mix. The concentrations were 125, 250, 500, 1000 and 1500 µg/plate for the second preincubation test conducted without S9 mix. In none of the tests any mutagenic effect could be detected, indicating that DMPA is not mutagenic in bacteria.
In a study according to OECD 476 (BASF SE 2020), DMPA was assessed for its potential to induce gene mutations at the hypoxanthine-guanine phophoribosyl transferase (HPRT) locus in Chinese hamster ovary (CHO) cells. The highest tested concentration was based on the molecular weight of the test substance corresponding to 10mM. THe cells were treated with DMPA for 4 hours in the absence or presence of rat liver S9. No relevant cytotoxicity was observed. DMPA did not increase the mutant frequencies with or without S9. Positive and negative controls were valid. Thus, DMPA is not mutagenic in the HPRT locus assay.
The potential of DMPA to induce micronuclei in human lymphocytes was assessed in two independent experiments in vitro according to OECD 487 (BASF 2020). Cells were either exposed without metabolic activation (rat liver S9) for 4 or 24 hours, or in the presence of S9 mix for 4 hours. For each experimental group, 1000 binucleated cells each from two parallel cultures were analyzed for cytogenetic damage. The highest applied concentration was based on the molecular weight of the test substance and corresponds to app. 10mM. No cytotoxicty was observed up to the highest concentration. No relevant increase in the numbers of micronucleated cells was observed in the presence or absence of metabolic activation. Appropriate positive control substances led to the expected significant increase in micronuclei.
Data for the structurally related substance DMEA (598 -56 -1) are comparable:
Ames: neg.
HPRT: neg.
CA: neg.
Justification for classification or non-classification
Classification, Labeling, and Packaging Regulation (EC) No. 1272/2008
The available experimental test data are reliable and suitable for the purpose of classification under Regulation (EC) No 1272/2008. All mutagenicity / genotoxicity tests conducted with DMPA (Ames, HPRT, MNT) gave negative results. As a result, DMPA does not need to be classified as mutagenic under Regulation (EC) No 1272/2008, as amended for the thirteenth time in Regulation (EU) No 2018/1480.
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.