Sodium [N-[2-[bis(carboxylatomethyl)amino]ethyl]-N-(2-hydroxyethyl)glycinato(3-)]zincate(1-)

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

The substance is a reaction mass of Zinc (Zn), EDTA, DTPA and HEEDTA. None of these substances have known mutagenic properties, based on literature, QSAR predictions and classification of individual components:

Zn: Zinc and zinc compounds such as ZnSO4 are not considered mutagenic (cfr. classification on ECHA dissemination website). Several in vitro and in vivo mutagenecity tests are included that found no mutagenetic properties for Zn compounds.

EDTA, DTPA and HEEDTA are classified as non-mutagenic on the ECHA dissemination website. To support this, a published negative Ames test for EDTA and QSAR results for EDTA, DTPA and HEEDTA are provided that predict negative outcomes of the Ames test with sufficient reliability.

Because of the lack of mutagenic potential in Zn compounds, EDTA, DTPA and HEEDTA, it is concluded that the reaction mass of ZnEDTA, ZnDTPA and ZnHEEDTA also should not be classified for genetic toxicity.

Link to relevant study records

Reference

Reference 1

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

(Q)SAR

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

results derived from a valid (Q)SAR model and falling into its applicability domain, with adequate and reliable documentation / justification

Justification for type of information:

1. SOFTWARE
VEGA v1.4.4

2. MODEL (incl. version number)
Mutagenicity (Ames test) model (CONSENSUS) 1.0.2.

3. SMILES OR OTHER IDENTIFIERS USED AS INPUT FOR THE MODEL
DTPA = C(CN(CC(=O)O)CC(=O)O)N(CCN(CC(=O)O)CC(=O)O)CC(=O)OC(CN(CC(=O)O)CC(=O)O)N(CCN(CC(=O)O)CC(=O)O)CC(=O)O

4. SCIENTIFIC VALIDITY OF THE (Q)SAR MODEL
[Explain how the model fulfils the OECD principles for (Q)SAR model validation. Consider attaching the QMRF or providing a link]
See attached QMRF

5. APPLICABILITY DOMAIN
[Explain how the substance falls within the applicability domain of the model]
See attached QMRF

6. ADEQUACY OF THE RESULT
[Explain how the prediction fits the purpose of classification and labelling and/or risk assessment]
An in-vitro gene mutation study in bacteria is required for REACH dossiers in the 1-10 tonnage band. This QSAR provides a prediciton for this endpoint.

Guideline:

other: REACH Guidance on QSARs R.6

Specific details on test material used for the study:

SMILES: C(CN(CC(=O)O)CC(=O)O)N(CCN(CC(=O)O)CC(=O)O)CC(=O)O

Remarks on result:

no mutagenic potential (based on QSAR/QSPR prediction)

Conclusions:

Based on the CAESAR model for Mutagenicity (Ames test) v2.1.13, DTPA is non-mutagenic.

Reference 2

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

(Q)SAR

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

results derived from a valid (Q)SAR model and falling into its applicability domain, with adequate and reliable documentation / justification

Justification for type of information:

1. SOFTWARE
VEGA v1.4.4

2. MODEL (incl. version number)
Mutagenicity (Ames test) model (CONSENSUS) 1.0.2.

3. SMILES OR OTHER IDENTIFIERS USED AS INPUT FOR THE MODEL
HEEDTA = C(CN(CC(=O)O)CC(=O)O)N(CCO)CC(=O)O

4. SCIENTIFIC VALIDITY OF THE (Q)SAR MODEL
[Explain how the model fulfils the OECD principles for (Q)SAR model validation. Consider attaching the QMRF or providing a link]
See attached QMRF

5. APPLICABILITY DOMAIN
[Explain how the substance falls within the applicability domain of the model]
See attached QMRF

6. ADEQUACY OF THE RESULT
[Explain how the prediction fits the purpose of classification and labelling and/or risk assessment]
An in-vitro gene mutation study in bacteria is required for REACH dossiers in the 1-10 tonnage band. This QSAR provides a prediciton for this endpoint.

Guideline:

other: REACH Guidance on QSARs R.6

Specific details on test material used for the study:

SMILES: C(CN(CC(=O)O)CC(=O)O)N(CCO)CC(=O)O

Remarks on result:

no mutagenic potential (based on QSAR/QSPR prediction)

Conclusions:

Based on the CAESAR model for Mutagenicity (Ames test) v2.1.13, HEEDTA is non-mutagenic.

Reference 3

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

(Q)SAR

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

results derived from a valid (Q)SAR model and falling into its applicability domain, with adequate and reliable documentation / justification

Justification for type of information:

1. SOFTWARE
VEGA v1.4.4

2. MODEL (incl. version number)
Mutagenicity (Ames test) model (CONSENSUS) 1.0.2.

3. SMILES OR OTHER IDENTIFIERS USED AS INPUT FOR THE MODEL
EDTA = C(CN(CC(=O)O)CC(=O)O)N(CC(=O)O)CC(=O)O

4. SCIENTIFIC VALIDITY OF THE (Q)SAR MODEL
[Explain how the model fulfils the OECD principles for (Q)SAR model validation. Consider attaching the QMRF or providing a link]
See attached QMRF

5. APPLICABILITY DOMAIN
[Explain how the substance falls within the applicability domain of the model]
See attached QMRF

6. ADEQUACY OF THE RESULT
[Explain how the prediction fits the purpose of classification and labelling and/or risk assessment]
An in-vitro gene mutation study in bacteria is required for REACH dossiers in the 1-10 tonnage band. This QSAR provides a prediciton for this endpoint.

Guideline:

other: REACH Guidance on QSARs R.6

Specific details on test material used for the study:

SMILES: C(CN(CC(=O)O)CC(=O)O)N(CC(=O)O)CC(=O)O

Remarks on result:

no mutagenic potential (based on QSAR/QSPR prediction)

Conclusions:

Based on the CAESAR model for Mutagenicity (Ames test) v2.1.13, EDTA is non-mutagenic.

Reference 4

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Principles of method if other than guideline:

According to "Ames BN, McCann J & Yamasaki E (1975). Methods for detecting carcinogens and mutagens with the Salmonella/mammalian-microsome mutagenicity test. Mutat. Res., 31: 347-364".

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Specific details on test material used for the study:

Obtained from Merck Co., Darmstadt (Germany)

Species / strain / cell type:

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

Additional strain / cell type characteristics:

not applicable

Species / strain / cell type:

S. typhimurium TA 1538

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

S9 liver fraction from Aroclor-pretreated rats

Test concentrations with justification for top dose:

At least 5 doses, up to 3,600 µg/plate

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

Details on test system and experimental conditions:

All compounds were tested on 2 slightly different minimal media: one (in the following named ZLM medium) is a modified minimal medium for E. coli , and the other is the Vogel-Bonner (VB) medium. ZLM medium contained (in g/l): tri-Na- citrate.2H20 (0.82), K2HPO4-3H20 (4.60), KH2PO4 (1.50), (NH4)2SO4 (1.00), MgSO4.7H20 (0.10) and glucose (17.0).
The concentration of citrate was 3.5 times higher in VB medium than in ZLM medium. The concentrations of the other ions are up to 2-fold higher in VB medium.

Statistics:

No statistics

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Conclusions:

Mutagenicity of zinc sulfate was tested with a bacterial reverse mutation test. The outcome of the test was negative, both with and without metabolic activation.
Under the conditions of this test, zinc sulfate was considered to be non-mutagenic.

Executive summary:

Mutagenicity of zinc sulfate was tested with a bacterial reverse mutation test.

Salmonella typhimurium strains TA1535, TA1537, TA98, TA100 and TA1538 strain were treated with the test material in Vogel-Bonner (VB) & ZLM medium (modified minimal medium for E. coli) at minimal five dose levels, up to 3,600 µg/plate, both with and without activation by the S9 liver fraction from Aroclor-pretreated rats.

 

The positive control chemicals used in the test induced marked increases in the frequency of revertant colonies, both with or without metabolic activation. No significant increases in the frequency of revertant colonies were observed for any of the bacterial strains, with any dose of the test material, either with or without metabolic activation.

 

The test material was considered to be non-mutagenic under the conditions of this test.

Reference 5

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

(No data of negetive control)

Principles of method if other than guideline:

Not applicable

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Target gene:

No data

Species / strain / cell type:

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

Details on mammalian cell type (if applicable):

No data

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9 fraction from Aroclor induced male Sprague-Dawley rats

Test concentrations with justification for top dose:

1000-5000 µg/plate

Vehicle / solvent:

DMSO

Untreated negative controls:

yes

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

Migrated to IUCLID6: at 5 µg/plate

Untreated negative controls:

yes

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: 2-Aminoanthracene at 1 µg/plate

Untreated negative controls:

yes

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: 4-nitro-o-phenylendiamine at 5 µg/plate

Details on test system and experimental conditions:

METHOD OF APPLICATION: In agar (plate incorporation)
NUMBER OF REPLICATIONS: Triplicate

Evaluation criteria:

Positive response: Induction of reproducible dose related increases in the number of his+ revertants in the treated group compared to the control.

Statistics:

No data

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:

not determined

Vehicle controls validity:

not specified

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

None

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

None

Conclusions:

Interpretation of results (migrated information):
negative with metabolic activation
negative without metabolic activation

The test material was considered to be non-mutagenic under the condition of this test.

Executive summary:

A study was conducted to determine the potential mutagenicity of Zinc oxide using bacterial reverse mutation assay.

Salmonella typhimurium strains TA 1535, TA 1537, TA 98 and TA 100 were treated with the test material using the plate incorporation method at dose levels ranging from 1,000 to 5,000 µg/plate in triplicate in presence and absence of a metabolic activation system.

No significant increases in the frequency of his+ revertant colonies were recorded at the dose range tested.

The test material was considered to be non-mutagenic under the conditions of this test.

Reference 6

Endpoint:

in vitro gene mutation study in mammalian cells

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 490 (In Vitro Mammalian Cell Gene Mutation Tests Using the Thymidine Kinase Gene)

Principles of method if other than guideline:

Cells deficient in thymidine kinase (TK) due to the mutation TK+/- to TK-/- are resistant to the cytotoxic effects of trifluorothymidine (TFT). Thymidine kinase proficient cells (TK+/-) are sensitive to TFT, which causes the inhibition of cellular metabolism and halts further cell division. Thus mutant cells are able to proliferate in the presence of TFT, whereas normal cells, which contain thymidine kinase, are not able to proliferate.

GLP compliance:

not specified

Type of assay:

mammalian cell gene mutation assay

Target gene:

Thymidine kinase locus/TK +/-

Species / strain / cell type:

mouse lymphoma L5178Y cells

Details on mammalian cell type (if applicable):

- Type and identity of media: RPMI 1640 medium
- Properly maintained: Yes
- Periodically checked for Mycoplasma contamination: Yes
- Periodically checked for karyotype stability: No data
- Periodically "cleansed" against high spontaneous background: Yes

Additional strain / cell type characteristics:

not specified

Metabolic activation:

without

Metabolic activation system:

No data

Test concentrations with justification for top dose:

1.21-12.13 µg/mL

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: Normal saline (1 %)
- Justification for choice of solvent/vehicle: Not reported

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

not specified

Positive control substance:

not specified

Remarks:

None

Details on test system and experimental conditions:

METHOD OF APPLICATION: In medium


DURATION
- Preincubation period: Not reported
- Exposure duration: 3 h
- Expression time (cells in growth medium): 48 h
- Selection time (if incubation with a selection agent): 7 d (at 37 °C in 5 % C02-95 % humidified air)



SELECTION AGENT (mutation assays): 5-trifluorothymidine (TFT) - 4 μg/mL


NUMBER OF REPLICATIONS: Duplicate cell culture for test material treatment while triplicate plates were prepared for both survival and mutation frequency determinations for each of the 2 replicate cultures


NUMBER OF CELLS EVALUATED: Cells densities were 30000/mL (1 x 100,000/plate) for mutant selection and 15/mL (500/plate) for viability detrmination


DETERMINATION OF CYTOTOXICITY
- Method: Cell survival for each culture was the product of growth in suspension culture and cloning efficiency in soft-agar medium, each relative to solvent controls

OTHER: Cell culture contained 6 x 100,000 cells each in 10 mL test medium
Light exposure was minimal during treatment of cells.
Test conducted at 37 °C
For the recovery and mutant expression, all cells were maintained at 37 °C for 48 h in log phase growth after treatment with test material

Evaluation criteria:

Colonies growing in the presence of triflurothymidine (TFT resistant) or its absence (viable count colonies) were counted. TFT Resistant colonies which were equivalent in size to colonies growing in the solvent control viable count plates ie., large, were scored as mutants.

Statistics:

Not reported

Species / strain:

mouse lymphoma L5178Y cells

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Remarks on result:

other: strain/cell type:

Remarks:

Migrated from field 'Test system'.

Conclusions:

Mutagenicity of ZnCl2 was tested using an in vitro gene mutation test in mouse lymphoma cells with the thymidine kinase gene. ZnCl2 was found to be not mutagenic under the test conditions.

Executive summary:

A study was conducted to assess the potential mutagenicity of test material in the mouse lymphoma thymidine kinase locus using the cell line L5178Y.

The mouse lymphoma cells (TK+/-) were treated with test material at 1.21-12.13 µg/mL for 3 h. 48 h after treatment, cells were treated with 4 µg/mL trifluorothymidine (TFT) for 7 d. Colonies growing in the presence of triflurothymidine (TFT resistant) or its absence (viable count colonies) were counted. TFT resistant colonies which were equivalent in size to colonies growing in the solvent control viable count plates i.e., large, were scored as mutants.

The test material was found to be non-mutagenic under the test conditions.

Reference 7

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Species / strain / cell type:

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

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

uninduced and arochlor induced liver S9 mix of male Fischer 344 rats, B6C3F1 mice, and Syrian hamsters

Test concentrations with justification for top dose:

10, 33, 100, 333, 1000, 3333, 10000 µg/plate

Vehicle / solvent:

distilled water

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

distilled water

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

N-ethyl-N-nitro-N-nitrosoguanidine

other:

Details on test system and experimental conditions:

All plates were prepared in triplicate, and concurrent positive and negative controls were run at all times.

Statistics:

not needed

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Conclusions:

Based on the Ames test, EDTA has no mutagenic properties.

Reference 8

Endpoint:

in vitro cytogenicity / micronucleus study

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

an in vitro cytogenicity study in mammalian cells or in vitro micronucleus study does not need to be conducted because adequate data from an in vivo cytogenicity test are available

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)