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

Genetic toxicity: in vitro

Administrative data

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).

Cross-reference

Reason / purpose for cross-reference:

reference to other study

Reference

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:

18-Sep-1996 to 11-Oct-1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Study does not stand alone, since only one bacterial strain was utilised, but it provides supplementary data of a comparable reliability to that in another study (Thompson, 1996), which utilised five Salmonella strains.

Reason / purpose for cross-reference:

reference to other study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

(only one bacterial strain used)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Target gene:

Tryptophan

Species / strain / cell type:

E. coli WP2 uvr A

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

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

water

True negative controls:

no

Positive controls:

yes

Remarks:

-S9

Positive control substance:

other: N-ethyl-N'-nitro-N-nitrosoguanidine, 2 ug/plate

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

water

True negative controls:

no

Positive controls:

yes

Remarks:

+S9

Positive control substance:

other: 2-aminoanthracene, 10 ug/plate

Species / strain:

E. coli WP2 uvr A

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

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Conclusions:

Ruthenium acetate was not mutagenic in a GLP bacterial reverse mutation assay in Escherichia coli WP2 uvrA, 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 bacterial reverse mutation assay using a single strain of Escherichia coli WP2 uvrA, following a protocol similar to OECD guideline 471 and to GLP. The bacteria were exposed to the test material at concentrations of 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 at any concentration either with or without S9. The test material was not mutagenic under the conditions of the test. This bacterial strain is utilised for its capacity to detect certain types of mutagen, such as cross-linking agents, not detected by Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 or TA1538. The results of this study supplement those of the study reported by Thompson (1996), which omitted such a strain.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Report date:

1996

Materials and methods

Test guideline

Test guideline 1

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)

Test guideline 2

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

Test material

Test material information

Constituent 1

Reference substance name:

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

EC Number:

259-653-7

EC Name:

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

Cas Number:

55466-76-7

Molecular formula:

C12H18O13Ru3.C2H3O2

IUPAC Name:

hexamethyl-2λ³-oxa-4λ³-oxa-6λ³-oxa-8λ³-oxa-10λ³-oxa-12λ³-oxa-13λ³-oxa-15λ³-oxa-16λ³-oxa-18λ³-oxa-19λ³-oxa-21λ³,22λ¹-dioxa-1,5,9-triruthenahexacyclo[7.3.3.3¹,⁵.3⁵,⁹.1¹,⁵.0⁹,²²]docosa-3,7,11,14,17,20-hexaene-1,1,1,5,5,5,9,9,9-nonakis(ylium)-2,6,10,13,16,19-hexaide-22,22-diuide acetate

Details on test material:

- Name of test material (as cited in study report): ruthenium acetate

- Substance type: black crystalline solid

- Physical state: solid

- Analytical purity: not stated

- Impurities (identity and concentrations): not stated

- Composition of test material, percentage of components: not stated

- Isomers composition: not stated

- Purity test date: not stated

- Lot/batch No.: 60350

- Expiration date of the lot/batch: not stated

- Stability under test conditions: not stated

- Storage condition of test material: room temperature

- Other:
- Date received: 23-Oct-1995

Method

Target gene:

histidine

Species / strain

Species / strain 1

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 2

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

Controls

Controls 1

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

Controls 2

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

Controls 3

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

Controls 4

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

Controls 5

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.

Results and discussion

Test results

Test results 1

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

Test results 2

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'.

Applicant's summary and conclusion

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.