Hexakis[μ-(acetato-O:O')]-μ3-oxo-triangulo-triruthenium acetate

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Ruthenium acetate was not mutagenic in bacterial reverse mutation assays involving five strains of Salmonella typhimurium (Thompson, 1996) and one strain of Escherichia Coli (Parnham, 1996), in the absence or presence of S9.

In a study equivalent to OECD guideline 473, and to GLP, a significant increase in the frequency of chromosome aberrations was reported in vitro in Chinese hamster lung cells, in the presence of S9 only (Durward, 1997a). Note that rodent derived cell lines with defective p53 are considered to give high rates of false positives.

Link to relevant study records

Reference

Reference 1

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

07-Nov-95 to 19-Dec-95

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Guideline study (OECD 471), to GLP, using five Salmonella strains. None of the strains utilised has the capacity to detect certain types of mutagens (e.g. oxidising or cross-linking agents). The results of this study are supplemented by those of the study reported by Parnham (1996), which utilised such a strain (E.coli).

Reason / purpose for cross-reference:

reference to other study

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Remarks:

(but no strain capable of detecting cross-linking mutagens was included)

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

no

Remarks:

(but no strain capable of detecting cross-linking mutagens was included)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Target gene:

histidine

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not applicable

Species / strain / cell type:

S. typhimurium TA 1538

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

S9 from Aroclor 1254-induced male rat liver

Test concentrations with justification for top dose:

0, 50, 150, 500, 1500 and 5000 ug/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: water

- Justification for choice of solvent/vehicle: not stated, but well-known solvent

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

water

True negative controls:

no

Positive controls:

yes

Remarks:

for TA100 and TA1535 -S9

Positive control substance:

other: N-ethyl-N'-nitro-N-nitrosoguanidine, 3 and 5 ug/plate

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

water

True negative controls:

no

Positive controls:

yes

Remarks:

for TA1537 -S9

Positive control substance:

9-aminoacridine

Remarks:

Migrated to IUCLID6: 80 ug/plate

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

water

True negative controls:

no

Positive controls:

yes

Remarks:

for TA98 -S9

Positive control substance:

4-nitroquinoline-N-oxide

Remarks:

Migrated to IUCLID6: 0.2 ug/plate

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

water

True negative controls:

no

Positive controls:

yes

Remarks:

for TA1538 -S9

Positive control substance:

other: 4-Nitro-o-phenylenediamine, 5 ug/plate

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

water

True negative controls:

no

Positive controls:

yes

Remarks:

5 for all strains +S9

Positive control substance:

other: 2-aminoantracene, 0.5-2 ug/plate

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 hours

NUMBER OF REPLICATIONS: In triplicate


DETERMINATION OF CYTOTOXICITY
- Method: other: thinning of the background lawn

Evaluation criteria:

For a substance to be considered positive, it would have induced a concentration-related and statistically significant increase in mutation rate in at least one strain in the presence and/or absence of S9 in both experiments at sub-toxic concentrations; to be considered negative, the number of induced revertants at each concentration would be less than 2x the number of spontaneous revertants.

Statistics:

Using the methods recommended by the United Kingdom Environmental Mutagen Society (UKEMS), normally Dunnett's method of linear regression.

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

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 examined

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 1538

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 examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS: none

RANGE-FINDING/SCREENING STUDIES: yes, preliminary toxicity study using TA100 exposed to the test material at 5000 ug/plate

COMPARISON WITH HISTORICAL CONTROL DATA: yes, within historical control range for the test laboratory

ADDITIONAL INFORMATION ON CYTOTOXICITY: no cytotoxicity

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Conclusions:

Ruthenium acetate was not mutagenic in a guideline Ames assay with five strains of Salmonella typhimurium, when tested at up to 5 mg/plate, with and without S9.

Executive summary:

The mutagenicity of ruthenium acetate (CAS 55466-76-7) was evaluated in an Ames test, conducted according to OECD Test Guideline 471 and to GLP. Five strains of bacteria (Salmonella typhimurium TA 1535, TA 1537, TA 1538, TA 98 and TA 100) were exposed to the test material at concentrations up to 5 mg/plate, in the presence and absence of an exogenous metabolising system (S9). No increase in the number of revertants was seen in any strain at any concentration either with or without S9. The test material was not mutagenic under the conditions of the test.

None of the strains utilised has the capacity to detect certain types of mutagens (e.g. oxidising or cross-linking agents). The results of this study are supplemented by those of the study reported by Parnham (1996), which utilised such a strain.

Reference 2

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

Study period:

22-Sep-1996 to 13-Jan-1997

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study (OECD 473) and to GLP, although no suitable positive control was included in the group tested in the absence of S9.

Remarks:

Note that the use of CHL cells is now not recommended due to a high incidence of false (misleading) positives (Fowler et al 2012 Reduction of misleading (“false”) positive results in mammalian cell genotoxicity assays. I. Choice of cell type, Mutation Research 742 (11-25)).

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

Deviations:

yes

Remarks:

(no suitable positive control -S9)

GLP compliance:

yes (incl. QA statement)

Type of assay:

in vitro mammalian chromosome aberration test

Target gene:

Not applicable

Species / strain / cell type:

mammalian cell line, other: Chinese hamster lung (CHL) cell line

Details on mammalian cell type (if applicable):

- Type and identity of media: Eagle's Minimal Essential Medium with Earle's Salts, supplemented with 10% foetal bovine serum and antibiotics

- Properly maintained: yes

- Periodically checked for Mycoplasma contamination: no data

- Periodically checked for karyotype stability: no data

- Periodically "cleansed" against high spontaneous background: no data

Additional strain / cell type characteristics:

other: doubling time: 11 hours

Metabolic activation:

with and without

Metabolic activation system:

S9 from Aroclor 1254-induced male rat liver

Test concentrations with justification for top dose:

6-hour treatment, +/-S9: 0, 313, 625, 1250, 1875, 2500 and 5000 ug/ml
24-hour treatment, -S9: 0, 39, 78, 156, 234, 323 and 625 ug/ml
48-hour treatment, -S9: 0, 10, 20, 39, 78, 156 and 313 ug/ml

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: culture medium

- Justification for choice of solvent/vehicle: test material was soluble at the required concentration.

Untreated negative controls:

yes

Negative solvent / vehicle controls:

no

Remarks:

culture medium

True negative controls:

no

Positive controls:

yes

Remarks:

+/-S9, 6-hour exposure

Positive control substance:

cyclophosphamide

Remarks:

inappropriate as a positive control -S9 Migrated to IUCLID6: 10 ug/ml

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

culture medium

True negative controls:

no

Positive controls:

yes

Remarks:

-S9, 24- and 48-hour exposure

Positive control substance:

mitomycin C

Remarks:

Migrated to IUCLID6: 0.05 and 0.025 ug/ml

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Exposure duration: 6 hours +/- S9 (also 24 and 48 hours -S9, but no chromsome aberration results presented for these exposures)

- Fixation time (start of exposure up to fixation or harvest of cells): 24 hours (after the 6-hr exposure cells were washed and left for an 18-hr recovery period in treatment free media before cell harvest)

SPINDLE INHIBITOR (cytogenetic assays): demecolcine (Colcemid, 0.1 ug/ml, 2 hours before harvest time)

STAIN (for cytogenetic assays): Gurr's Giemsa R66, 2%, 5 min

NUMBER OF REPLICATIONS: duplicate

NUMBER OF CELLS EVALUATED: 100/culture, 2 cultures per concentration (total of 200 cells/concentration)

DETERMINATION OF CYTOTOXICITY
- Method: growth inhibition estimated by counting number of cells at end of culture period on electronic cell counter (Coulter) and expressing cell count as percentage of concurrent vehicle control

OTHER EXAMINATIONS:
- Determination of polyploidy: yes

- Determination of endoreplication: no data, but presumably yes

Evaluation criteria:

Generally, a positive result was considered to be when the proportion of cells with aberrations (including gaps) was >=10%, equivocal if 5-10% and negative if <5%; for polyploidy a 10% incidence considered positive; guidance of the United Kingdon Environmental Mutagen Society also taken into consideration, requiring expert judgement on the incidence of rare chromosomal events (exchanges)

Statistics:

Fisher's exact test

Species / strain:

mammalian cell line, other: Chinese hamster lung (CHL) cell line

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

not valid

Species / strain:

mammalian cell line, other: Chinese hamster lung (CHL) cell line

Metabolic activation:

with

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS: none

RANGE-FINDING/SCREENING STUDIES: yes, cultures exposed to test material at up to 5000 ug/ml for 6 hrs (+/-S9), or 24 or 48-hrs (-S9)

COMPARISON WITH HISTORICAL CONTROL DATA: yes, solvent controls reported to be within range for laboratory

ADDITIONAL INFORMATION ON CYTOTOXICITY: cell counts reduced and/or no metaphases at 625 ug/ml and above for 24-hour exposure (-S9), at 313 ug/ml and above for 48-hour exposure (-S9), and at 2500 ug/ml and above for 6-hour exposure (+/-S9)

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

In the section of the study from which metaphases were scored for chromosome aberrations (6 hour exposure), the positive control substance (cyclophosphamide) was suitable only for exposure +S9; no suitable positive control for exposure -S9 (e.g. mitomycin C) was included. Since a positive result was obtained for clastogeniciy +S9, this omission has not altered the outcome of the study.

Conclusions:

Ruthenium acetate induced a statistically significant increase in the frequency of chromosome aberrations in Chinese hamster lung cells (in vitro), in the presence of S9 only. It was therefore considered clastogenic under the conditions of the test.

Executive summary:

The genotoxic potential of ruthenium acetate (CAS 55466-76-7) was evaluated in an in vitro chromosome aberration test (assessing structural and numerical aberrations) in Chinese hamster lung (CHL) cells using a protocol similar to that of OECD Test Guideline 473. Cultures were exposed to the test material for 6 hours (with an 18-hr recovery period) at up to 5000 ug/ml (in the presence and absence of a rat liver metabolic activation system; S9), for 24-hrs at up to 625 ug/ml (without S9), or for 48-hrs at up to 313 ug/ml (without S9). Evidence of cytotoxicity was seen at the higher doses.

In the 6 hour exposures (with S9), a high incidence of cells with chromosome aberrations was seen, which reached statistical significance at 1250, 1875 and 2500 ug/ml when compared to the solvent controls. No such increases were seen in the studies performed in the absence of S9. The test material was therefore considered clastogenic, with S9, under the conditions of the test.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (positive)

Genetic toxicity in vivo

Description of key information

No clastogenic activity was observed in the bone marrow of mice following intraperitoneal injection of ruthenium triacetate at up to 200 mg/kg bw in a guideline in vivo micronucleus test, to GLP (Durward, 1997b).

Link to relevant study records

Reference

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Study period:

16-Jan-1997 to 10-Feb-1997

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: OECD Guideline study, to GLP, with acceptable deviation (only 1000 PCEs/animal scored for micronuclei)

Qualifier:

according to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Deviations:

yes

Remarks:

(only 1000 PCEs/animal scored for micronuclei)

Qualifier:

according to guideline

Guideline:

EU Method B.12 (Mutagenicity - In Vivo Mammalian Erythrocyte Micronucleus Test)

GLP compliance:

yes (incl. QA statement)

Type of assay:

micronucleus assay

Species:

mouse

Strain:

CD-1

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (UK) Ltd., Margate, Kent
- Age at study initiation: 5-7 weeks
- Weight at study initiation: males 24-30 g, females 21-26 g
- Assigned to test groups randomly: yes
- Fasting period before study: no data
- Housing: <=7/cage, solid floor polypropylene cages with woodflakes bedding
- Diet (e.g. ad libitum): Rat and Mouse Expanded Diet No. 1, ad libitum
- Water (e.g. ad libitum): mains water, ad libitum
- Acclimation period: >=5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-21
- Humidity (%): 47-52
- Air changes (per hr): ~15
- Photoperiod (hrs dark / hrs light): 12 / 12

IN-LIFE DATES: From: 16-Jan-1997 To: 10-Feb-1997

Route of administration:

intraperitoneal

Vehicle:

- Vehicle(s)/solvent(s) used: physiol. saline
- Justification for choice of solvent/vehicle: no data, but well-known
- Concentration of test material in vehicle: 5, 10 and 20 mg/ml (for 50, 100 and 200 mg/kg bw doses respectively)
- Amount of vehicle (if gavage or dermal): 10 ml/kg bw
- Type and concentration of dispersant aid (if powder): not applicable
- Lot/batch no. (if required): Gibco 18Q7563
- Purity: no data

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
- freshly prepared as required as a suspension in the vehicle

Duration of treatment / exposure:

single intraperitoneal administration

Frequency of treatment:

single administration

Post exposure period:

24, 48 and 72 hours

Dose / conc.:

0 mg/kg bw/day (nominal)

Dose / conc.:

5 mg/kg bw/day (nominal)

Dose / conc.:

10 mg/kg bw/day (nominal)

Dose / conc.:

20 mg/kg bw/day (nominal)

No. of animals per sex per dose:

5/sex per dose at 24-hour timepoint;
also 5/sex at top dose at 48- and 72-hour timpoints

Control animals:

yes, concurrent vehicle

Positive control(s):

cyclophosphamide
- Justification for choice of positive control(s): "known to produce micronuclei under the conditions of the test"
- Route of administration: oral
- Doses / concentrations: 50 mg/kg bw

Tissues and cell types examined:

erythrocytes from bone marrow of femur

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION: based on range-finding study in which deaths were observed with doses of 250-2000 mg/kg bw, but not at 200 mg/kg bw or below

TREATMENT AND SAMPLING TIMES (in addition to information in specific fields): sampled at 24, 48 and 72 hours after dosing in top dose group (200 mg/kg bw) and at 24 hours in mid and low dose groups (100 and 50 mg/kg bw)

DETAILS OF SLIDE PREPARATION:
- immediately after sacrifice, 1 femur dissected/animal, aspirated with foetal calf serum
- centrifuged, resuspended and bone marrow smears prepared
- air-dried, fixed in absolute methanol, stained with May-Grunwald/Giemsa

METHOD OF ANALYSIS:
- slides coded prior to examination
- light microscopy at 1000x magnification
- incidence of micronucleated cells/1000 polychromatic erythrocytes (PCEs; blue-stained, immature cells) scored for each animal
- incidence of normochromatic erythrocytes (NCEs; pink-stained, mature cells)/1000 erythrocytes scored for each animal
- incidence of micronuclei in these NCEs scored for each animal
- ratio of PCE:NCE calculated

Evaluation criteria:

Positive mutagenic response: statistically significant increase in number of micronucleated PCEs at any timepoint
Positive bone marrow toxicity: statistically significant decrease in group mean PCE:NCE ratio

Statistics:

Data transformed, then analysed by 2-tailed Student's t-test; significant results confirmed using one-way analysis of variance (United Kingdom Environmental Mutagen Society (UKEMS) Sub-committee on Guidelines for Mutagenicity Testing Report, part III, 1989)

Sex:

male/female

Genotoxicity:

negative

Remarks:

at all doses at all timepoints

Toxicity:

yes

Remarks:

clinical signs of toxicity at top dose at all timepoints; bone marrow toxicity at top dose at 48-hour timepoint

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

RESULTS OF RANGE-FINDING STUDY
- Dose range:
- intraperitoneal: 125-2000 mg/kg bw
- oral: 1000-2000 mg/kg bw
- Solubility: administered as a suspension
- Clinical signs of toxicity in test animals:
- intraperitoneal: deaths at 250 mg/kg bw (1/2), 500 mg/kg bw (2/2), 1000 mg/kg bw (2/2) and 2000 mg/kg bw (2/2); clinical signs (including hunched posture, lethargy, decreased respiratory rate, laboured respiration, ataxia, ptosis and grey extremities) at 1000 and 2000 mg/kg bw
- oral: no deaths, no clinical signs
- Evidence of cytotoxicity in tissue analyzed: not examined in range-finding study
- Rationale for exposure: for the main study, the doses selected were the maximum tolerated dose of 200 mg/kg bw and 2 lower doses
- Harvest times: not examined in range-finding study
- High dose with and without activation: not applicable to in vivo study

RESULTS OF DEFINITIVE (MAIN) STUDY
- Clinical signs of toxicity in test animals: in top dose group (200 mg/kg bw), clinical signs of toxicity at all timepoints, including hunched posture, ptosis, dehydration, decreased respiratory rate, laboured respiration, ataxia, emaciation, lethargy, piloerection and grey extremities; deaths in 72-hour group (males 1/5, females 3/5)
- Induction of micronuclei (for Micronucleus assay): see Table 1
- Ratio of PCE/NCE (for Micronucleus assay): see Table 1
- Appropriateness of dose levels and route: appropriate; highest dose produced signs of toxicity such that higher dose levels would be expected to produce lethality; highest dose also produced some indication of toxicity in the bone marrow (i.e. a reduction in the PCE:NCE ratio at 48 hours)
- Statistical evaluation: no statistically significant increase in the number of PCE with micronuclei per 1000 PCE at any dose at any timepoint in either sex; positive control produced a highly significant increase

Table 1:Results of in vivo micronucleus test with ruthenium acetate

(5 males and 5 females/group; mean ± standard deviation)

Treatment group		Number of PCE with micronuclei per 1000 PCE		PCE:NCE ratio
		Group mean	SD	Group mean	SD
Dose	Sampling time (hours)
Vehicle control	72	0.7	1.3	1.62	0.35
	48	0.8	0.9	1.53	0.67
	24	1.3	0.8	1.49	0.49
Positive control	24	28.9***	13.1	1.44	0.5
High dose (200 mg/kg bw)	72^a	0.3	0.5	1.25	0.93
	48	0.7	0.9	0.81*	0.52
	24	0.6	0.7	1.17	0.31
Mid dose (100 mg/kg bw)	24	0.5	0.5	1.55	0.39
Low dose (50 mg/kg bw)	24	0.6	0.7	1.33	0.53

 

PCE, polychromatic erythrocyte, NCE, normochromatic erythrocyte, SD, standard deviation, *** p<0.001, * p<0.05, a data from 6 animals

 

Conclusions:

In a reliable in vivo study, conducted according to OECD Test Guideline 474 and to GLP, no genotoxicity was seen in the bone marrow of mice administered ruthenium acetate by intraperitoneal injection at up to 200 mg/kg bw; clinical signs of toxicity and toxicity in the bone marrow were seen at the highest dose.

Executive summary:

Groups of 5 male and 5 female mice were administered a single intraperitoneal dose of ruthenium acetate at 50, 100 or 200 mg/kg bw and evaluated for the incidence of micronuclei in the bone marrow 24 hours after dosing. Two further groups were administered 200 mg/kg bw and evaluated 48 and 72 hours after dosing. Vehicle control groups were evaluated at each timepoint and a positive control group was orally administered cyclophosphamide at 50 mg/kg bw and evaluated at 24 hours.

Clinical signs of toxicity were seen in all groups dosed with 200 mg/kg bw and 4/10 mice died in the 72 -hour group at this dose level. Bone marrow toxicity, indicated by a decrease in the ratio of normochromatic to polychromatic erythrocytes (NCE:PCE), was seen in the 48 -hour group. There was no increase in the incidence of micronuclei at any dose at any timepoint in either sex. A clear positive response was seen in the positive control group.

In a reliable GLP study, conducted according to OECD Test Guideline 474, no genotoxicity was seen in the bone marrow of mice administered ruthenium acetate by intraperitoneal injection at up to 200 mg/kg bw; clinical signs of toxicity and toxicity in the bone marrow were seen at the highest dose.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Mode of Action Analysis / Human Relevance Framework

No data identified.

Additional information

Good quality unpublished in vitro and in vivo genotoxicity studies with ruthenium (III) acetate (CAS RN 55466‑76‑7) are available.

 

The test substance, at up to 5 mg/plate, was not mutagenic in GLP bacterial reverse mutation assays involving five strains of Salmonella typhimurium (Thompson, 1996) and one strain of Escherichia Coli (Parnham, 1996), in the absence or presence of S9.

 

A significant increase in the frequency of chromosome aberrations was reported in vitro in Chinese hamster lung cells, in the presence of S9 only, in a GLP study conducted using a protocol similar to OECD Test Guideline 473 (Durward, 1997a).

 

However, no clastogenic activity was observed in the bone marrow of mice (10/group) following intraperitoneal injection of ruthenium triacetate at up to 200 mg/kg bw in an in vivo micronucleus test conducted according to OECD Test Guideline 474 and to GLP. Clinical signs of toxicity and toxicity in the bone marrow were seen at the highest dose (Durward, 1997b).

Justification for classification or non-classification

No evidence of genotoxic activity was seen in reliable mutagenicity assays in bacterial cells. The observation of clastogenic activity in vitro was not apparent in a reliable in vivo study assessing chromosome damage in somatic cells. No studies specifically assessing the mutagenic activity in germ cells were identified. However, due to the generally reassuring genotoxicity data on this compound, including a negative in vivo micronucleus test, classification of hexakis[mu-(acetato-O:O')]-μ3-oxo-triangulo-triruthenium acetate / ruthenium acetate for germ cell mutagenicity is not warranted, according to EU CLP criteria (EC 1272/2008).