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EC number: 221-374-3 | CAS number: 3081-01-4
- 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
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
The data from the bacterial mutation assays indicated no genotoxic potential of 7PPD (WTR 1990, Monsanto Co. 1976, 1977a, 1977b). This negative finding is confirmed by the results from a mammalian cell mutation assays (Monsanto Co. 1988). An in vitro chromosome aberration assay with CHO cells revealed weak clastogenic effects of 7PPD (Monsanto Co. 1989). Under in vivo conditions the potential clastogenic activity in vitro could not be confirmed. In an in vivo bone marrow chromosome aberration assay with Sprague-Dawley rats 7PPD did not significant increase the frequency of chromosome aberrations (Monsanto Co. 1989). In conclusion, based on the findings discussed above, the weight of evidence indicates a low or even a non-genotoxic potential of 7PPD in vitro and no genotoxicity in vivo and thus no classification is required.
Link to relevant study records
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Version / remarks:
- no mitotic index determined but MTD reached; 3 times 50 instead of 2 times 100 cells scored
- GLP compliance:
- yes
- Type of assay:
- in vitro mammalian chromosome aberration test
- Target gene:
- chromosome aberrations
- Species / strain / cell type:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9-mix, S9 from Aroclor 1254-induced rat liver homogenate (S9)
- Test concentrations with justification for top dose:
- pre-test I(+/-S9): 5, 20, 50, 100, 200, 500, 1000, 2000, 5000 µg/mL; pre-test II (+/-S9): 1, 2.5, 5, 7.5, 10, 12.5, 15, 17.5, 20 µg/mL,
main experiment I (+/-S9): 0, 7.5, 10, 15 µg/mL, main experiment II (-S9): 7.5, 10 µg/mL; - Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- methylmethanesulfonate
- Species / strain:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Genotoxicity:
- positive
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- other: solvent aceton induced a relative high number of aberrant cells in main experiment I
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other: strain/cell type: Chinese hamster Ovary (CHO)
- Conclusions:
- Interpretation of results: positive
Reference
1. Range-Finding Experiments:
CHO cells were treated with 5 to 5000 µg/mL of SANTOFLEX 14 both in the presence and absence of activation. The cells were scored for both mitotic index and average cell generation time and compared to the solvent (acetone) control. The average cell generation time for the solvent control was approximately 12 hours for both with and without activation, with a mitotic index of 5 to 8 %. Cytotoxic effects were observed at 20 µg/ml both in the absence and presence of activation as indicated by a sharp reduction in mitotic index and an extended average generation time.
The cytotoxicity of SANTOFLEX 14 was further defined in a second range-finding experiment where concentrations ranged from 1 to 20 µg/mL with and without activation. The cells were scored for both mitotic index and average cell generation time. Cytotoxic effects were observed at 12.5 µg/ml and higher concentrations in the absence and presence of activation as indicated by an extended average generation time (-S9 20.3 h at 12.5 µg/ml vs. 12.0 control; +S9: at 12.5 µg/ml 15.2 h vs. 12.7 control).
Based on the range-finding experiments, the highest concentrations of SANTOFLEX 14 tested were 15 µg/mL with and without activation. The harvest times were 12 and 24 hours for 1.5, 5, 7.5 and 10 µg/mL with and without activation, 18 and 36 hours for 15 µg/mL with activation and 24 and 48 hours for 15 µg/mL without activation. The harvest times were chosen to represent approximately 1X and 2X that of the cell generation times.
2. Cytogenetics Studies:
A. Without activation: The SANTOFLEX 14 levels of 7.5, 10 and 15 µg/mL were chosen as the highest three scorable doses.
Statistically significant increases in number of cells with structural aberrations (12 cells with aberrations vs. 3 cells with aberrations solvent control) and average structural aberrations per cell (0.060 vs. 0.015 solvent control) were observed at the 15 µg/mL dose level for the 48 hour harvest time (% aberrant cells: 6 % vs. 1.5 % solvent control) and for average structural aberrations per cell for the 24 hour harvest time (0.120 vs. 0.045 solvent control). A significant dose-response was not observed.
B. With activation: The SANTOFLEX 14 dose levels of 7.5, 10 and 15 µg/mL were selected as the three highest scorable doses. Statistically significant increases in the number of cells with structural aberrations (at 10 µg/mL 39 vs. 15 solvent control) and average structural aberrations per cell (10 µg/mL: 0.383 vs. 0.090 solvent control) were observed for the 10 µg/mL dose level (% aberrant cells: 19.9 % vs. 7.5 % solvent control) and for the number of cells with aberrations at the 7.5 µg/mL dose level for the 12 hour harvest time (at 7.5 µg/mL 29 cells with aberrations vs. 15 cells with aberration solvent control). A dose-related response was not observed.
C. Retest: Because of the relatively high aberration levels (-S9: 10 %, +S9: 7.5 %) for the 12 hour harvest time in the solvent control, the experiment was repeated using dose levels of 7.5 and 10 µg/mL with and without activation. The 24 and 48 hour harvests were not repeated as the solvent background was low and therefore the statistically significant response of the 15 µg/mL dose level without activation was concluded to be treatment induced. The 10 µg/mL dose level with activation was again statistically significant for the number of cells with aberrations (17 cells with aberrations vs. 6 cells solvent control) and for structural aberrations per cell (0.090 vs. 0.030 solvent control) A dose-response relationship was not observed.
Conclusion:
Without activation, a statistically significant increase in aberration frequency was observed for the 15 µg/mL dose level. In the presence of activation, a statistically significant response was observed at the 10 µg/mL dose level for the 12 hour harvest.
A significant dose-response relationship was not observed. The retest experiment confirmed the statistically significant response of the 10 µg/mL dose level in the presence of activation. A significant dose-response was not observed.
The authors suggested that the observed effects might be secondary because the effects were observed in the range of cytotoxicity indicated by an increased generation time (10 µg/mL +S9: pre-experiment: 13.7h vs. 12.7 h solvent control, 15 µg/mL -S9: 22.0 h vs. 12.1 h solvent control) and in addition because of a lack of a dose response relationship.
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
Additional information from genetic toxicity in vitro:
In vitro data : The mutagenic potential in bacteria of the test
substance 7PPD was evaluated in a GLP study (WTR 1990). Here, the tester
strains Salmonella typhimurium TA 98, TA 100, TA 1535 and TA 1537 were
used. Treatment by the pour-plate method was done. Cytotoxicity was
determinate in a preliminary toxicity test using Salmonella typhimurium
tester strain TA 98. The lowest level of 7PPD causing visible thinning
of the background lawn of non-revertant cells was 250 µg/ml; based on
this finding 250 µg/plate was selected as the top exposure level in the
main experiment. No increases in revertant colony numbers over control
counts were obtained with any tester strains following exposure to 7PPD
at concentration levels from 2.5 µg to 250µg/plate. Inhibition of
growth, observed as thinning of the background lawn of non-revertant
cells occurred in all strains following exposure to 7PPD at 250µg/plate,
and in strain TA 1535 only with 7PPD at 79 µg/plate. The authors
concluded, that 7PPD was negative in the Ames assay, under the
experimental conditions used. In addition, in several other earlier
bacterial mutation assays the test substance also indicated a
non-mutagenic potential (Monsanto Co. 1976, 1977a, 1977b). The negative
findings from the bacterial mutation assays are confirmed in a mammalian
cell mutation assay. The test substance 7PPD was negative in a HGPRT
assay, done with CHO cells (Monsanto Co. 1988). Initial cytotoxicity
experiments were conducted with 7PPD in CHO cells at different S9
concentrations. The concentrations that induced significant cytotoxicity
were 7µg/ml, 10 µg/ml, 30 µg/ml, and 70 µg/ml in the absence of S9 and
in the presence of 1%, 2%, 5% and 10%, S9, respectively. An initial
experiment to determine the potential mutagenicity of the test material
was conducted using a range of S9 concentrations. In this experiment,
7PPD was significantly cytotoxic to the CHO cells at levels of 5µg/ml
and greater in the absence of exogenous metabolic activation. In the
presence of S9 activation, significant cytotoxicity was demonstrated for
treatment levels of 7 µg/ml, >10 µg/ml, 20 µg/ml and 70 µg/ml in the
presence of 1%, 2%, 5% and 10% S9, respectively. No statistically
significant increases in mutation frequency were observed in any of the
7PPD treated cultures. The non-mutagenicity of 7PPD was confirmed by a
subsequent experiment. In this experiment, 7PPD was tested at 1, 3, 5,
7, and 10 µg/ml in the absence of S9 and at 10, 15, 20, 25, and 30 µg/ml
in the presence of 5 % S9 activation. 7PPD was observed to be
significantly cytotoxic at levels of 5 µg/ml and greater in the absence
of S9. In the presence of 5% S9, 7PPD was not cytotoxic at 30µg/ml. No
statistically significant increases in mutant frequency were observed in
this experiment. A chromosomal aberration test with CHO cells was
performed to determine the clastogenic potential of 7PPD (Monsanto
1989). In a range-finding experiment CHO cells were treated with 5 to
5000 µg/ml of 7PPD both in the presence and absence of activation. The
cells were scored for both mitotic index and average cell generation
time and compared to the solvent (acetone) control. The average cell
generation time for the solvent control was approximately 12 hours for
both with and without activation, with a mitotic index of 5 to 8 %.
Cytotoxic effects were observed at 20µg/ml both in the absence and
presence of activation indicated by a sharp reduction in mitotic index
and an extended average generation time. The cytotoxicity of 7PPD was
further defined in a second range-finding experiment where
concentrations ranged from 1 to 20 µg/ml with and without activation.
The cells were scored for both mitotic index and average cell generation
time. Cytotoxic effects were observed at 12.5 µg/ml and higher
concentrations in the absence and presence of activation as indicated by
an extended average generation time (-S9 20.3 h at 12.5 µg/ml vs. 12.0
control; +S9: at 12.5 µg/ml 15.2 h vs. 12.7 control). Based on the
range-finding experiments, the highest concentrations of 7PPD tested
were 15µg/ml with and without activation. The harvest times were 12 and
24 hours for 1.5, 5, 7.5 and 10 µg/ml with and without activation, 18
and 36 hours for 15 µg/ml with activation and 24 and 48 hours for
15µg/ml without activation. In the main experiment without metabolic
activation 7PPD levels of 7.5, 10 and 15 µg/ml were chosen as the
highest three scorable doses. Statistically significant increases in
number of cells with structural aberrations (12 cells with aberrations
vs. 3 cells with aberrations solvent control) and average structural
aberrations per cell (0.060 vs. 0.015 solvent control) were observed at
the 15 µg/ml dose level for the 48 hour harvest time (% aberrant cells:
6 % vs. 1.5% solvent control) and for average structural aberrations per
cell for the 24 hour harvest time (0.120 vs. 0.045 solvent control). A
significant dose-response was not observed. In the main experiment with
metabolic activation 7PPD dose levels of 7.5, 10 and 15 µg/ml were
selected as the three highest scorable doses. Statistically significant
increases in the number of cells with structural aberrations (at 10
µg/ml 39 vs. 15 solvent control) and average structural aberrations per
cell (10 µg/ml: 0.383 vs. 0.090 solvent control) were observed for the
10 µg/ml dose level (% aberrant cells: 19.9 % vs. 7.5% solvent control)
and for the number of cells with aberrations at the 7.5 µg/ml dose level
for the 12 hour harvest time (at 7.5 µg/ml 29 cells with aberrations vs.
15 cells with aberration solvent control). A dose-related response was
not observed. Because of the relatively high aberration levels (-S9:
10%, +S9: 7.5%) for the 12 hour harvest time in the solvent control, the
main experiment was repeated using dose levels of 7.5 and 10 µg/ml with
and without activation. The 24 and 48 hour harvests were not repeated as
the solvent background was low and therefore the statistically
significant response of the 15 µg/ml dose level without activation was
concluded to be treatment induced. The 10 µg/ml dose level with
activation was again statistically significant for the number of cells
with aberrations (17 cells with aberrations vs. 6 cells solvent control)
and for structural aberrations per cell (0.090 vs. 0.030 solvent
control) A dose-response relationship was not observed. The authors
concluded that 7PPD had a weak clastogenicity in CHO cells with and
without metabolic activation. Because the significant increases in
aberrant cells were observed in the range of cytotoxicity secondary
effects could not be excluded. In vivo data The genotoxic potential of
7PPD was evaluated in an in vivo bone marrow chromosome aberration assay
with Sprague-Dawley rats (Monsanto 1989). A dose range finding
experiment was performed to select the maximum tolerated dose. Rats were
dosed per gavage with 1050, 1100, 1200, 1500 and 2000 mg/kg bw.
Mortality was observed in animals treated with 1500 and 2000 mg/kg test
substance. Animals dosed with 1100 and 1200 mg/kg bw exhibited moderate
to severe clinical signs. No clinical signs were noted in animals which
were dosed with 1050 mg/kg bw. Based on these findings, a dose level of
1100 mg/kg was chosen for the main experiment. In the main experiment
male and female rats (five per sex and treatment) were administered
single oral with the 1100 mg/kg bw. The treated animals were sacrificed
6, 18 and 30 hours after dosing. Concurrent solvent controls were also
included for all preparation time points. An extra group of rats was
dosed with the positive control cyclophasphamid and sacrificed 18 hours
later. Approximately two hours prior to each sacrifice, animals were
administered colchicine to arrest cells in metaphase. At the appropriate
time, animals were sacrificed and both femurs were removed from each
animal and metaphase slides were prepared. A total of 50 metaphase cells
were analyzed for each animal. The occurrences of clinical signs were
observed during the study. No clinical signs were observed in animals
administered 7PPD immediately after dosing; whereas clinical signs were
noted prior colchicine administration in all treatment groups. In the 6
hour group all males and females had decreased body tone. In addition
all females had vocalization on touch and two females had arched back
and abnormal stance. In the 18 hour group two males had decreased body
tone. One male had vocalization when touched, abnormal stance and arched
back. Another male had piloerection and diarrhea. All females had
decreased body tone with four females exhibiting piloerection and
abnormal stance. Two females made vocalization when toughed. One female
had also arched back. In the 30 hour group all treated rats had
decreased body tone and piloerection. Three males and two females made
vocalizations when touched. One male and one female had an arched back.
In addition, one female had abnormal gait. Based on these findings the
authors suggested that rats dosed with 1100 mg/kg 7PPD exhibited
moderate to severe clinical signs indicating that these animals were
dosed at or near the maximum tolerated dose. One male of the positive
control group exhibited decreased body tone. No clinical signs were
observed in animals from the solvent control. No statistically
significant increase in number of aberrations or in the number of
aberrant metaphases were noted in 7PPD treated animals in any of the
three sacrifice times compared to the corresponding solvent control;
whereas significant increased were noted in the positive control. Based
on these findings the authors concluded that 7PPD (1100 mg/kg) was
negative in its ability to induce structural chromosomal aberrations to
the hemopoietic cells of the rat bone marrow under the experimental
conditions of this assay (Monsanto 1989)
Justification for classification or non-classification
No classification is required according to the classification criteria 67/548/EWG and regulation no. 1272/2008 (GHS).
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.
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