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EC number: 215-264-4 | CAS number: 1317-34-6
- 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
No data is available for the target substance. Thus, to assess the genotoxic potential of dimanganese trioxide, data from manganese dichloride (source substance) was used in read-across approach. The source substance was tested in an in vitro genotoxicity testing battery as required by Annex VIII of the REACH regulation 1907/2006 (OECD 471, OECD 473 and OECD 490, GLP).
In a bacterial reverse gene mutation test conducted according to OECD 471, the source substance did not induce mutagenicity. In a mammalian cell mutation assay conducted according to OECD 490, the source substance was also tested negative for inducing mutagenic effects. Moreover, the manganese dichloride was tested negative in an in vitro cytogenicity study in human lymphocytes conducted according to OECD 473. Based on the available data from the read-across partner, the target substance dimanganese trioxide can be considered to be non-genotoxic.
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Justification for type of information:
- For justification of read-across please refer to the read-across statement in IUCLID section 13.
- Reason / purpose for cross-reference:
- read-across source
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- Test material-induced toxicity was noted at the highest dose level employed in the test.
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: Precipitate of the test material was observed at and above 10 µg/mL in the 4-hour exposure groups in the absence of metabolic activation and at and above 20 µg/mL in the 4-hour exposure group in the presence of metabolic activation.
RANGE-FINDING/SCREENING STUDIES: All three exposure groups employed in the screening test exhibited a marked reduction in the Relative Suspension Growth (%RSG) of cells treated with the test material when compared to the concurrent vehicle controls. A precipitate of the test material was observed at and above 78.75 µg/mL in the 4-hour exposure group in the absence of metabolic activation, at and above 39.98 µg/mL in the 4-hour exposure group in the presence of metabolic activation, and at and above 19.69 µg/mL in the 24-hour exposure group in the absence of metabolic activation. In the mutagenicity experiments the maximum dose level was limited by test-material-induced toxicity. - Conclusions:
- The test material did not induce any toxicologically significant increases in the mutant frequency at the TK +/- locus in L5178Y cells and is therefore considered to be non-mutagenic under the conditions of the test.
- Executive summary:
In a mammalian cell gene mutation assay conducted in accordance to OECD guideline 476 (nowadays OECD 490), L5178Y mouse lymphoma cells cultured in vitro were exposed to manganese dichloride (> 95% purity), solved in R0 medium. Test concentrations chosen for the main experiments were based on the results from a preliminary toxicity test. In the first main experiment, the test item was tested in the presence of metabolic activation up to 160 µg/mL and in the absence of metabolic activation up to 120 µg/mL. The incubation time was 4 hours. In the second main experiment, the test item was tested in the absence of metabolic activation up to 15 µg/mL. The incubation time was 24 hours. Cytotoxicity were observed in all main experiments. The positive controls did induce the approriate response. The test material did not induce any toxicologically significant increases in the mutant frequency at the TK +/- locus in L5178Y cells and is therefore considered to be non-mutagenic under the conditions of the test.
This information is used in a read-across approach in the assessment of the target substance.
For justification of read-across please refer to the attached read-across statement (see IUCLID section 13).
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Justification for type of information:
- For justification of read-across please refer to the read-across statement in IUCLID section 13.
- Reason / purpose for cross-reference:
- read-across source
- Species / strain:
- lymphocytes: human, peripheral
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: Test material was noted to precipitate at 315 µg/mL in the 4(20)-hour cultures without S9, and above 630 µg/mL in the 4(20)-hour cultures in the presence of S9. No precipitate was observed at the end of the exposure period in the 24-hour cultures.
RANGE-FINDING/SCREENING STUDIES: The preliminary toxicity test dose range was 4.92 to 1260 µg/mL. The maximum dose was based on the maximum recommended 10 mM concentration. A precipitate of the test material was observed in the cultures at the end of the exposure, at and above 157.5 µg/mL in the 4(20)-hour exposure in the absence of S9 at and above 78.75 µg/mL in the 4(20)-hour exposure in the presence of s(, and at above 315 µg/mL in the 24 hour continuous exposure group. Haemolysis was observed at and above 315 µg/mL at harvesting in all three exposure groups. Microscopic assessment of the slides prepared from the exposed cultures showed that metaphase cells were present up to 1260 µg/mL in the 4(20)-hour exposure in the presence of metabolic activation and up to 157.5 µg/mL in the 4(20)-hour exposure in the absence of S9. The maximum dose with metaphases present in the 24-hour continuous exposure was 39.38 µg/mL. The test material induced clear evidence of toxicity in all of the exposure groups.
TEST RESULTS:
All vehicle (solvent) controls had frequencies of cells with aberrations within the range expected for normal human lymphocytes. All the positive control materials induced statistically significant increases in the frequency of cells with aberrations indicating the satisfactory performance of test and of the activity of the metabolising system. The test material was found to be toxic to lymphocytes, and did not induce any toxicologically significant increases in the frequency of cells with aberrations, in any of the exposure conditions, using a dose range that included dose levels that induced approximately 50% mitotic inhibition. - Conclusions:
- The test material did not induce any toxicologically significant increases in the frequency of cells with aberrations in either of the 4(20)-hour exposure groups in the absence or presence of a liver enzyme metabolising system or following 24 hours continuous exposure. The test material was therefore considered to be non-clastogenic to human lymphocytes in vitro.
- Executive summary:
In a mammalian cell cytogenetic assay conducted according to OECD guideline 473, human peripheral lymphocyte cultures were exposed to manganese dichloride (> 95% purity), solved in MEM cell culture medium. Concentrations chosen for the main experiments were based on the results from a preliminary toxicity test. In the first main experiment, cells were treated for 4 hours in the presence and absence of metabolic activation. In the main second experiment cells were treated for 24 hours with metabolic activation. Afterwards, slides were prepared, stained with 5% Giemsa and where possible the first 100 consecutive well-spread metaphases from each culture were counted. All solvent controls had frequencies of cells with aberrations within the range expected for normal human lymphocytes. All the positive control materials induced statistically significant increases in the frequency of cells with aberrations indicating the satisfactory performance of test and of the activity of the metabolising system. The test material was found to be toxic to lymphocytes, and did not induce any toxicologically significant increases in the frequency of cells with aberrations, in any of the exposure conditions, using a dose range that included dose levels that induced approximately 50% mitotic inhibition. Based on the results, manganese dichloride is considered to be not clastogenic in human peripheral lymphocytes. This study is classified as acceptable and satisfies the requirement for the in vitro mammalian chromosomal aberration test according to OECD 473.
This information is used in a read-across approach in the assessment of the target substance.
For justification of read-across please refer to the attached read-across statement (see IUCLID section 13).
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Justification for type of information:
- For justification of read-across please refer to the read-across statement in IUCLID section 13.
- Reason / purpose for cross-reference:
- read-across source
- Species / strain:
- other: S. typhimurium TA98, TA100, TA1535 and TA1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS: Not reported
RANGE-FINDING/SCREENING STUDIES: The test substance was found to be non-toxic in the range-finding study.
COMPARISON WITH HISTORICAL CONTROL DATA: The historical control values were found to concur with the results from the study for both positive and vehicle controls.
RESULTS OF THE STUDY:
The test material was found to cause no visible reduction in growth of the bacterial background lawn at any dose and was therefore tested up to the maximum dose level of 5000 µg/plate A particulate precipitate was at 1500 µg/plate and above. This was considered not to prevent the scoring of revertant colonies. No toxicologically significant increases in the frequency of revertant colonies were recorded for and of the bacterial strains, with any dose of the test material, with or without metabolic activation. In the TA100 revertant colony, a small but statistically significant increase was observed on the 1500 µg/plate in Experiment 2 (increase of less than 1.5 times). However, these were within the range specified by the Standard Test Method, this increase proved non-reproducible over two separate experiments. This was concluded to have no biological or toxicological relevance. All of the positive control substances induced marked increases in the frequency of revertant colonies, confirming the activity of the S9-mix and the sensitivity of the bacterial strains. - Conclusions:
- In conclusion, manganese chloride is not genotoxic in the bacterial reverse gene mutation assay (OECD 471) in the presence and absence of mammalian metabolic activation.
- Executive summary:
In a reverse gene mutation assay in bacteria conducted according to OECD guideline 471, S. typhimurium strains TA1535, TA1537, TA98, TA100 and E. coli strain WP2uvrA were exposed to manganese dichloride (> 95% purity) at concentrations of 0, 50, 150, 500, 1500 and 5000 µg/plate in the presence and absence of mammalian metabolic activation.
The test item was tested up to the limit concentration of 5000 µg/plate. The positive controls induced the appropriate responses in the corresponding strains. There was no evidence of induced mutant colonies over background.
This study is classified as acceptable and satisfies the requirement for Test Guideline OECD 471 for in vitro mutagenicity (bacterial reverse gene mutation assay).
This information is used in a read-across approach in the assessment of the target substance.
For justification of read-across please refer to the attached read-across statement (see IUCLID section 13).
Referenceopen allclose all
Table 2: Results from the preliminary toxicity test
Dose (µg/mL) |
%RSG (-S9) 4-Hour Exposure |
%RSG (=S9) 4-Hour Exposure |
%RSG (-S9) 24-Hour Exposure |
0 |
100 |
100 |
100 |
4.92 |
102 |
89 |
33 |
9.84 |
96 |
100 |
11 |
19.69 |
89 |
89 |
1 |
39.38 |
72 |
75 |
0 |
78.75 |
2 |
57 |
0 |
157.5 |
9 |
1 |
0 |
315 |
1 |
0 |
0 |
630 |
0 |
0 |
0 |
1260 |
0 |
0 |
0 |
Table 3: Summary of results for main experiment, 4 hour exposure
Treatment (µg/mL) |
4-Hours –S9 |
Treatment (µg/mL) |
4-Hours +S9 |
||||
%RSG |
RTG |
MF§ |
%RSG |
RTG |
MF§ |
||
0 |
100 |
1.00 |
81.37 |
0 |
100 |
1.00 |
74.20 |
2.5† |
97 |
|
|
20 |
91 |
1.04 |
64.08 |
5 |
95 |
1.06 |
73.26 |
40 |
68 |
0.86 |
78.58 |
10 |
91 |
0.94 |
96.72 |
60 |
43 |
0.41 |
116.75 |
20 |
101 |
1.24 |
90.28 |
80 |
37 |
0.32 |
96.12 |
40 |
44 |
0.46 |
104.53 |
100 |
32 |
0.22 |
104.65 |
60 |
19 |
0.08 |
128.81 |
120 |
26 |
0.15 |
104.12 |
80 |
17 |
0.13 |
91.23 |
140 |
25 |
0.18 |
100.39 |
120† |
14 |
|
|
160‡ |
13 |
0.04 |
42.87 |
Linear trend |
NS |
Linear trend |
|||||
EMS |
|
|
|
CP |
|
|
|
400 |
71 |
0.62 |
656.51 |
2 |
55 |
0.22 |
1662.80 |
† Not plated for viability or 4-TFT resistance MF§ 5-TFT resistant mutants/106viable cells 2 days after treatment NS Not significant ‡ Treatment excluded from test statistics due to toxicity |
Table 4: Summary of results for main experiment, 24 hour exposure
Treatment (µg/mL) |
24-Hours –S9 |
||
%RSG |
RTG |
MF§ |
|
0 |
100 |
1.00 |
103.16 |
0.31 |
97 |
0.99 |
91.33 |
0.63 |
105 |
0.97 |
79.60 |
1.25 |
95 |
1.07 |
50.22 |
2.5 |
76 |
0.81 |
100.92 |
5 |
41 |
0.38 |
173.75* |
7.5 |
14 |
0.11 |
372.63* |
10† |
7 |
|
|
15† |
4 |
|
|
Linear trend |
*** |
||
EMS |
|
|
|
150 |
54 |
0.32 |
1211.25 |
† Not plated for viability or 4-TFT resistance * p< 0.05 *** p< 0.001 |
Table 2: Mitotic index – 4(20)-Hours
Dose Level µg/mL |
4 Hours Treatment Without S9 |
4 Hours Treatment With S9 |
||||||
A |
B |
Mean |
% of Control |
A |
B |
Mean |
% of Control |
|
0 |
5.70 |
7.10 |
6.40 |
100 |
5.00 |
3.80 |
4.40 |
100 |
9.84 (78.75)a |
- |
- |
- |
- |
- |
- |
- |
- |
19.69 (157.5)a |
5.25 |
5.95 |
5.60 |
88 |
4.10 |
3.00 |
3.55 |
81 |
39.38 (315)a |
5.70 |
5.35 |
5.53 |
86 |
3.50 |
2.85 |
3.18 |
72 |
78.75 (630)a |
3.30 |
3.40 |
3.35 |
52 |
1.90 P |
3.10 P |
2.50 |
57 |
157.5 (945)a |
2.25 |
1.80 |
2.03 |
32 |
1.20 P |
2.40 P |
1.80 P |
41 |
315 (1260)a |
NM P |
NM P |
NM P |
0 |
1.15 P |
2.40 P |
1.78 P |
40 |
MMC 0.4 |
1.75 |
2.20 |
1.98 |
31 |
NA |
NA |
NA |
NA |
CP 5 |
NA |
NA |
NA |
NA |
0.75 |
0.85 |
0.80 |
18 |
a= dose levels with S9
MMC= mitomycin C
CP= cyclophosphamide
P= precipitate
NA= not applicable
-= not assessed for mitotic index
NM= no scoreable metaphases
Table 3: Mitotic Index – 24 Hours
Dose Level µg/mL |
24 Hours Treatment Without S9 |
|||
A |
B |
Mean |
% of Control |
|
0 |
3.45 |
2.65 |
3.05 |
100 |
1.23 |
3.85 |
3.70 |
3.78 |
124 |
2.46 |
3.35 |
3.85 |
3.60 |
118 |
4.92 |
2.90 |
2.65 |
2.78 |
91 |
9.84 |
1.85 |
1.80 |
1.83 |
60 |
19.69 |
NM |
NM |
NM |
0 |
30.38 |
NM |
NM |
NM |
0 |
MMC 0.2 |
1.25 |
0.95 |
1.10 |
36 |
NM = No scorable metaphases
MMC = Mitomycin C
Table 4: Results of Chromosome Aberration Test – 4(20)-Hours Without Metabolic Activation (S9)
Treatment Group |
Replicable |
Mitotic Index (%) |
Number of Cells Scored |
Number of Aberrations |
Total Number of Aberrations |
Frequency of Aberrant Cells (%) |
|||||||
Gaps |
Chromatid |
Chromosome |
Others X |
(+ Gaps) |
(-Gaps) |
(+ Gaps) |
(-Gaps) |
||||||
Breaks |
Exchanges |
Breaks |
Exchanges |
||||||||||
Vehicle Group |
A |
5.70 |
100 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
B |
7.10 |
100 |
1 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
1 |
0 |
|
Total |
(100) |
200 |
1 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
1 (0.5) |
0 (0.0) |
|
19.59 µg/mL |
A |
5.25 |
100 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
B |
5.95 |
100 |
0 |
1 |
0 |
0 |
0 |
0 |
1 |
1 |
1 |
1 |
|
Total |
(88) |
200 |
0 |
1 |
0 |
0 |
0 |
0 |
1 |
1 |
1 (0.5) |
1 (0.5) |
|
39.38 µg/mL |
A |
5.70 |
100 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
B |
5.35 |
100 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
|
Total |
(86) |
200 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 (0.0) |
0 (0.0) |
|
78.75 µg/mL |
A |
3.30 |
100 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
B |
3.40 |
100 |
0 |
1 |
0 |
0 |
0 |
0 |
1 |
1 |
1 |
1 |
|
Total |
(52) |
200 |
0 |
1 |
0 |
0 |
0 |
0 |
1 |
1 |
1 (0.5 |
1 (0.5) |
|
Positive Control MMC 0.4 µg/mL |
A |
1.75 |
50a |
2 |
24 |
17 |
2 |
0 |
0 |
45 |
43 |
25 |
28 |
B |
2.20 |
50a |
7 |
18 |
15 |
2 |
1 |
0 |
13 |
36 |
26 |
25 |
|
Total |
(31) |
100 |
9 |
42 |
32 |
4 |
1 |
0 |
88 |
79 |
54 (54.0) |
53*** (53.0) |
MMC = Mitomycin C
a = Slide evaluation terminated at 50 cells because approximately 30-50 % of cells with aberrations had been observed
*** = P < 0.001
Table 5: Results of Chromosome Aberration Test – 4(20)-Hours With Metabolic Activation (S9)
Treatment Group |
Replicable |
Mitotic Index (%) |
Number of Cells Scored |
Number of Aberrations |
Total Number of Aberrations |
Frequency of Aberrant Cells (%) |
|||||||
Gaps |
Chromatid |
Chromosome |
Others X |
(+ Gaps) |
(-Gaps) |
(+ Gaps) |
(-Gaps) |
||||||
Breaks |
Exchanges |
Breaks |
Exchanges |
||||||||||
Vehicle Group |
A |
5.00 |
100 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
B |
3.80 |
100 |
1 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
1 |
0 |
|
Total |
(100) |
200 |
1 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
1 (0.5) |
0 (0.0) |
|
157.5 µg/mL |
A |
4.10 |
100 |
0 |
1 |
0 |
0 |
0 |
0 |
1 |
1 |
1 |
1 |
B |
3.00 |
100 |
1 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
1 |
0 |
|
Total |
(81) |
200 |
1 |
1 |
0 |
0 |
0 |
0 |
2 |
1 |
2 (1.0) |
1 (0.5) |
|
315 µg/mL |
A |
3.50 |
100 |
1 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
1 |
0 |
B |
2.85 |
100 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
|
Total |
(72) |
200 |
1 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
1 (0.5) |
0 (0.0) |
|
630 µg/mL |
A |
1.90 |
100 |
0 |
1 |
0 |
0 |
0 |
0 |
2 |
2 |
2 |
2 |
B |
3.10 |
100 |
1 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
1 |
0 |
|
Total |
(57) |
200 |
1 |
1 |
0 |
0 |
0 |
0 |
3 |
2 |
3 (1.5) |
2 (1.0) |
|
945 µg/mL |
A |
1.20 |
100 |
3 |
5 |
0 |
0 |
0 |
0 |
8 |
5 |
7 |
4 |
B |
2.40 |
100 |
1 |
2 |
0 |
0 |
0 |
0 |
3 |
2 |
3 |
2 |
|
Total |
(41) |
200 |
4 |
7 |
0 |
0 |
0 |
0 |
11 |
7 |
10 (5.0) |
6* (3.0) |
|
Positive Control CP 5 µg/mL |
A |
0.75 |
100 |
2 |
18 |
7 |
2 |
0 |
0 |
29 |
27 |
22 |
22 |
B |
0.85 |
100 |
6 |
30 |
2 |
11 |
0 |
0 |
49 |
43 |
31 |
30 |
|
Total |
(18) |
200 |
8 |
48 |
9 |
13 |
0 |
0 |
78 |
70 |
53 (26.5) |
52*** (26.0) |
CP = Cyclophosphamide
* = P < 0.05
*** = P < 0.001
Table 6: Results of Chromosome Aberration Test – 24-Hours Without Metabolic Activation (S9)
Treatment Group |
Replicable |
Mitotic Index (%) |
Number of Cells Scored |
Number of Aberrations |
Total Number of Aberrations |
Frequency of Aberrant Cells (%) |
|||||||
Gaps |
Chromatid |
Chromosome |
Others X |
(+ Gaps) |
(-Gaps) |
(+ Gaps) |
(-Gaps) |
||||||
Breaks |
Exchanges |
Breaks |
Exchanges |
||||||||||
Vehicle Group |
A |
3.45 |
100 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
B |
2.65 |
100 |
2 |
1 |
0 |
0 |
0 |
0 |
3 |
1 |
3 |
1 |
|
Total |
(100) |
200 |
2 |
1 |
0 |
0 |
0 |
0 |
3 |
1 |
3 (1.5) |
1 (0.5) |
|
19.59 µg/mL |
A |
3.35 |
100 |
1 |
0 |
0 |
1 |
0 |
0 |
2 |
1 |
2 |
0 |
B |
3.85 |
100 |
1 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
1 |
1 |
|
Total |
(118) |
200 |
2 |
0 |
0 |
1 |
0 |
0 |
3 |
1 |
3 (1.5) |
1 (0.5) |
|
39.38 µg/mL |
A |
2.90 |
100 |
0 |
2 |
0 |
3 |
0 |
0 |
5 |
5 |
3 |
3 |
B |
2.65 |
100 |
1 |
2 |
0 |
1 |
0 |
0 |
4 |
3 |
3 |
3 |
|
Total |
(91) |
200 |
1 |
4 |
0 |
4 |
0 |
0 |
9 |
8 |
6 (3.0) |
6 (3.0) |
|
78.75 µg/mL |
A |
1.85 |
100 |
1 |
8 |
0 |
0 |
0 |
0 |
9 |
8 |
7 |
7 |
B |
1.80 |
100 |
5 |
5 |
0 |
0 |
0 |
0 |
10 |
5 |
7 |
5 |
|
Total |
(60) |
200 |
6 |
13 |
0 |
0 |
0 |
0 |
19 |
13 |
14 (7.0) |
12** (6.0) |
|
Positive Control MMC 0.4 µg/mL |
A |
1.25 |
50a |
0 |
21 |
32 |
2 |
0 |
1 |
56 |
56 |
34 |
34 |
B |
0.96 |
50a |
4 |
32 |
14 |
6 |
0 |
0 |
56 |
51 |
33 |
32 |
|
Total |
(36) |
100 |
4 |
53 |
46 |
8 |
0 |
1 |
112 |
108 |
67 (67.0) |
66*** (66.0) |
MMC = Mitomycin C
a = Slide evaluation terminated at 50 cells because approximately 30-50 % of cells with aberrations had been observed
** = P < 0.01
*** = P < 0.001
Table 7: Mean Frequency of Polyploid Cells (%)
Dose Level µg/mL |
Harvest Time 24 Hours |
||
4 Hours Without S9 |
4 Hours With S9 |
24 Hours Without S9 |
|
0 |
1.0 |
0.5 |
0.5 |
2.46 (157.5)a |
NA |
0.0 |
0.0 |
4.92 (315)a |
NA |
0.0 |
0.0 |
9.84 (630)a |
- |
0.0 |
0.0 |
19.69 (945)a |
0.0 |
0.0 |
- |
39.38 |
0.0 |
NA |
- |
78.75 |
0.5 |
NA |
NA |
MMC 0.2 |
NA |
NA |
0.0 |
MMC 0.4 |
0.0 |
NA |
NA |
CP5 |
NA |
0.0 |
NA |
a = Dose levels with S9
MMC = Mitomycin C
CP = Cyclophosphamide
NA – Not applicable
- = Not determined
Table 1: Range finding study – toxicity assay
With (+) or Without (-) S9-mix |
Strain |
Dose (µg/plate) |
||||||||||
0 |
0.15 |
0.5 |
1.5 |
5 |
15 |
50 |
150 |
500 |
1500 |
5000 |
||
- |
TA100 |
75 |
81 |
78 |
96 |
89 |
77 |
85 |
74 |
70 |
86P |
84P |
+ |
TA100 |
77 |
96 |
84 |
85 |
74 |
75 |
91 |
88 |
74 |
70P |
73P |
- |
WP2uvrA- |
25 |
23 |
25 |
23 |
27 |
30 |
23 |
35 |
31 |
22P |
27P |
+ |
WP2uvrA- |
37 |
24 |
27 |
34 |
29 |
25 |
36 |
32 |
38 |
30P |
24P |
P - Precipitate |
Table 2: Spontaneous Mutation Rates (Concurrent Negative Controls), Experiment 1
Number of Revertants (mean number of colonies per plate) |
||||
Base-pair substitution type |
Frameshift type |
|||
TA100 |
TA1535 |
WP2uvrA- |
TA98 |
TA1537 |
93 |
15 |
22 |
14 |
10 |
96 (95) |
19 (18) |
20 (22) |
16 (17) |
13 (12) |
97 |
21 |
24 |
20 |
13 |
Table 3: Spontaneous Mutation Rates (Concurrent Negative Controls), Experiment 2
Number of Revertants (mean number of colonies per plate) |
||||
Base-pair substitution type |
Frameshift type |
|||
TA100 |
TA1535 |
WP2uvrA- |
TA98 |
TA1537 |
123 |
22 |
33 |
22 |
12 |
123 (118) |
25 (23) |
25 (29) |
25 (24) |
12 (12) |
109 |
21 |
29 |
24 |
13 |
Table 4: Test Results, Experiment 1 – Without Metabolic Activation
Test Period |
From 19 July 2009 |
To 22 July 2009 |
|||
Test Substance (µg/plate) |
Number of revertants (mean number of colonies per plate) |
||||
Base-pair substitution type |
Frameshift type |
||||
TA100 |
TA1535 |
WP2uvrA- |
TA98 |
TA1537 |
|
0 |
95 111 (102) 99 8.3# |
16 20 (18) 19 2.1 |
23 25 (23) 20 2.5 |
26 21 (23) 23 2.5 |
15 13 (14) 13 1.2 |
50 |
112 96 (104) 104 8.0 |
16 20 (19) 20 2.3 |
18 25 (22) 23 3.6 |
21 22 (22) 24 1.5 |
14 13 (12) 10 2.1 |
150 |
115 110 (109) 102 6.6 |
21 13 (18) 21 4.6 |
26 15 (20) 19 5.6 |
16 20 (21) 27 5.6 |
12 11 (12) 13 1.0 |
500 |
103 92 (95) 90 7.0 |
18 19 (17) 14 2.6 |
21 23 (25) 30 4.7 |
21 21 (21) 20 0.6 |
15 15 (12) 15 0.0 |
1500 |
110 P 95 P (103) 104P 7.5 |
19P 18P (18) 16P 1.5 |
22 24P (23) 22P 1.2 |
22P 23P (25) 26P 3.2 |
13P 15P (12) 9P 3.1 |
5000 |
88P 117P (105) 111P 15.3 |
18P 21P (19) 18P 1.7 |
26P 21P (24) 24P 2.5 |
25P 23P (25) 26P 1.5 |
9P 12P (12) 16P 3.5 |
Name Concentration No. colonies per plate |
ENNG |
ENG |
ENNG |
4NQO |
9AA |
3 |
5 |
2 |
0.2 |
80 |
|
475 526 (492) 476 29.2 |
99 150 (115) 95 30.7 |
145 145 (145) 145 0.0 |
123 118 (119) 115 4.0 |
345 462 (400) 394 58.8 |
|
ENNG – N-ethyl-N’-nitro-N-nitrosoguanidine 4NQO – 4-Nitroquinoline-1-oxide 9AA – 9-Aminoacridine P – Precipitate # - Standard deviation |
Table 5: Test Results, Experiment 1 – With Metabolic Activation
Test Period |
From 19 July 2009 |
To 22 July 2009 |
|||
Test Substance (µg/plate) |
Number of revertants (mean number of colonies per plate) |
||||
Base-pair substitution type |
Frameshift type |
||||
TA100 |
TA1535 |
WP2uvrA- |
TA98 |
TA1537 |
|
0 |
97 96 (98) 100 8.3# |
12 12 (11) 9 1.7 |
23 29 (28) 32 4.6 |
25 24 (23) 20 2.6 |
14 9 (12) 14 2.9 |
50 |
117 101 (106) 100 9.5 |
11 9 (10) 10 1.0 |
31 25 (25) 19 6.0 |
20 21 (20) 18 1.5 |
12 12 (12) 12 0.0 |
150 |
97 102 (101) 103 3.2 |
10 9 (9) 9 0.6 |
25 23 (25) 27 2.0 |
26 22 (24) 24 2.0 |
15 10 (12) 11 12.6 |
500 |
115 74 (94) 92 20.6 |
13 8 (10) 9 2.6 |
29 19 (23) 21 5.3 |
31 20 (24) 20 6.4 |
16 15 (15) 13 1.5 |
1500 |
90 P 82 P (88) 91P 4.9 |
12P 9P (11) 13P 2.1 |
26P 24P (23) 20P 3.1 |
19P 18P (24) 20P 6.4 |
11P 15P (12) 9P 3.1 |
5000 |
92P 104P (95) 90P 7.6 |
12P 9P (11) 11P 1.5 |
24P 24P (23) 20P 2.3 |
20P 24P (22) 22P 2.0 |
12P 9P (11) 13P 2.1 |
Name Concentration No. colonies per plate |
2AA |
2AA |
2AA |
BP |
2AA |
1 |
2 |
10 |
5 |
2 |
|
2248 2526 (2506) 2743 248.1 |
209 151 (179) 177 29.1 |
172 225 (184) 154 36.9 |
206 184 (197) 202 11.7 |
278 217 (247) 247 30.5 |
|
2AA – 2-Aminoanthracene BP – Benzo(a)pyrene P – Precipitate # - Standard deviation |
Table 6: Test Results, Experiment 2 – Without Metabolic Activation
Test Period |
From 19 July 2009 |
To 22 July 2009 |
|||
Test Substance (µg/plate) |
Number of revertants (mean number of colonies per plate) |
||||
Base-pair substitution type |
Frameshift type |
||||
TA100 |
TA1535 |
WP2uvrA- |
TA98 |
TA1537 |
|
0 |
101 95 (104) 115 10.3# |
20 16 (19) 20 2.3 |
21 24 (23) 25 2.1 |
26 26 (25) 23 1.7 |
11 16 (14) 15 2.6 |
50 |
106 97 (104) 106 5.2 |
21 21 (21) 21 0.0 |
18 26 (24) 29 5.7 |
20 26 (25) 2 9 4.6 |
8 15 (12) 13 3.6 |
150 |
113 107 (111) 114 3.8 |
24 20 (21) 20 2.3 |
22 29 (25) 23 3.8 |
20 19 (22) 27 4.4 |
10 8 (9) 10 1.2 |
500 |
96 104 (102) 106 5.3 |
16 22 (17) 14 4.2 |
23 24 (24) 25 1.0 |
23 25 (26) 30 3.6 |
15 8 (9) 10 1.2 |
1500 |
118P (118) 119P 1.0 117 * |
24P 20P (23) 26P 3.1 |
21P 24P (22) 22P 1.5 |
29P 21P (25) 26P 4.0 |
10P 11P (10) 9P 1.0 |
5000 |
101P 104P (105) 110P 4.6 |
22P 23P (22) 21P 1.0 |
27P 24P (24) 21P 3.0 |
26P 25P (25) 24P 1.0 |
12P 13P (11) 9P 2.1 |
Name Concentration No. colonies per plate |
ENNG |
ENG |
ENNG |
4NQO |
9AA |
3 |
5 |
2 |
0.2 |
80 |
|
295 312 (309) 320 12.8 |
209 222 (212) 206 8.5 |
416 492 (462) 477 40.3 |
246 217 (228) 220 15.9 |
1078 2042 (1426) 1157 535.2 |
|
ENNG – N-ethyl-N’-nitro-N-nitrosoguanidine 4NQO – 4-Nitroquinoline-1-oxide 9AA – 9-Aminoacridine P – Precipitate # - Standard deviation * p≤0.05 |
Table 7: Test Results, Experiment 2 – With Metabolic Activation
Test Period |
From 19 July 2009 |
To 22 July 2009 |
|||
Test Substance (µg/plate) |
Number of revertants (mean number of colonies per plate) |
||||
Base-pair substitution type |
Frameshift type |
||||
TA100 |
TA1535 |
WP2uvrA- |
TA98 |
TA1537 |
|
0 |
108 106 (108) 109 1.5# |
10 9 (11) 13 2.1 |
25 27 (26) 26 1.0 |
22 21 (22) 22 0.6 |
16 16 (15) 12 2.3 |
50 |
93 91 (106) 107 8.7 |
15 9 (12) 13 3.1 |
22 22 (25) 31 5.2 |
21 26 (22) 22 0.6 |
13 15 (15) 16 1.5 |
150 |
98 89 (99) 110 10.5 |
13 8 (10) 13 3.1 |
22 23 (22) 31 0.6 |
19 24 (22) 22 2.5 |
14 14 (12) 7 4.0 |
500 |
91 106 (102) 110 10.0 |
12 14 (12) 9 2.5 |
30 29 (30) 30 0.6 |
26 21 (23) 22 2.6 |
16 14 (14) 12 2.0 |
1500 |
81P 100P (96) 110P 10.0 |
9P 13P (10) 9P 2.3 |
25P 31P (27) 25P 3.5 |
25P 25P (24) 22P 1.7 |
9P 16P (14) 16P 4.0 |
5000 |
90P 84P (92) 102P 9.2 |
11P 10P (10) 8P 1.5 |
26P 23P (25) 27P 2.1 |
24P 21P (22) 22P 1.5 |
12P 15P (12) 10P 2.5 |
Name Concentration No. colonies per plate |
2AA |
2AA |
2AA |
BP |
2AA |
1 |
2 |
10 |
5 |
2 |
|
2474 2427 (2510) 2629 105.7 |
374 332 (332) 291 41.5 |
342 264 (313) 333 42.7 |
261 620 (457) 490 181.8 |
497 494 (498) 504 5.1 |
|
2AA – 2-Aminoanthracene BP – Benzo(a)pyrene P – Precipitate # - Standard deviation |
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
No data is available for the target substance. Thus, to assess the genotoxic potential of dimanganese trioxide, data from manganese dichloride (source substance) was used in read-across approach. The source substance was tested in an in vitro genotoxicity testing battery as required by Annex VIII of the REACH regulation 1907/2006 (OECD 471, OECD 473 and OECD 490, GLP).
In a bacterial reverse gene mutation test conducted according to OECD 471, the source substance did not induce mutagenicity. In a mammalian cell mutation assay conducted according to OECD 490, the source substance was also tested negative for inducing mutagenic effects. Moreover, the manganese dichloride was tested negative in an in vitro cytogenicity study in human lymphocytes conducted according to OECD 473. Based on the available data from the read-across partner, the target substance dimanganese trioxide can be considered to be non-genotoxic.
Justification for classification or non-classification
Based on the lack of mutagenicity/cytogenicity in all conducted in vitro assays with the source substance, dimanganese trioxide is considered to be non-genotoxic and thus no classification is warranted in accordance with CLP Regulation 1272/2008.
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