Registration Dossier
Registration Dossier
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EC number: - | CAS number: -
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
Zinc monoglycinate sulfate is not considered to be mutagenic in the bacterial reverse mutation assay with and without metabolic activation.
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- 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)
- Version / remarks:
- The study was conducted in 1981.
- GLP compliance:
- no
- Type of assay:
- bacterial reverse mutation assay
- Specific details on test material used for the study:
- Zinc sulfate was obtained from Merck Co., Darmstadt, Germany.
- Target gene:
- histidine-operon
- Species / strain / cell type:
- S. typhimurium TA 1537
- Species / strain / cell type:
- S. typhimurium TA 98
- Species / strain / cell type:
- S. typhimurium TA 1538
- Species / strain / cell type:
- S. typhimurium TA 100
- Species / strain / cell type:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 liver fraction from Aroclor-pretreated rats
- Test concentrations with justification for top dose:
- 5 doses up to 3600 µg/plate
- Vehicle / solvent:
- Hanks' balanced salt solution (HBSS)
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- benzo(a)pyrene
- Details on test system and experimental conditions:
- S9 containing about 25 mg protein per ml and S9-mix (10% S9) were prepared according to the standard procedure by Ames et al., 1975.
- Statistics:
- Statistical significance of results was debited by use of the tables of Kastenbaum and Bowman.
- Key result
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- True negative controls validity:
- not specified
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- True negative controls validity:
- not specified
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- True negative controls validity:
- not specified
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1538
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- True negative controls validity:
- not specified
- Positive controls validity:
- valid
- Conclusions:
- The test item is not mutagenic in the bacterial reverse mutation assay with and without metabolic activation.
- Executive summary:
In the study by Gocke et al. (1981), the mutagenic potential of zinc sulfate was examined according to the method described by Ames et al. (1975) in a bacterial reverse mutation assay.
5 tester strains of Salmonella typhimurium which were TA1535, TA100, TA1538, TA98 and TA1537 were treated with the test material at minimal five dose levels up to 3600 µg/plate, both with and without metabolic activation. S9 liver fraction was prepared from Aroclor-pretreated rats.
No significant increases in the frequency of revertant colonies were observed for any of the bacterial strains at any dose of zinc sulfate, either with or without metabolic activation. The positive control was benzo(a)pyrene. Validity of the results from vehicle and positive controls are not clearly stated.
In conclusion, zinc sulfate was not mutagenic in the bacterial reverse mutation assay.
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2019-12-03 to 2020-02-14
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Version / remarks:
- adopted July 21st, 1997
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Version / remarks:
- 2008
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- his locus
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
- Metabolic activation:
- with and without
- Metabolic activation system:
- Type and composition of metabolic activation system:
S9 Mix
- source of S9 :S9 was obtained by Trinova Biochem GmbH, Gießen. Batch nos. 4115
- method of preparation of S9 mix :produced from the livers of male Sprague-Dawley rats which were treated with 500 mg Aroclor 1254/kg body weight intraperitoneally.
- concentration or volume of S9 mix and S9 in the final culture medium:
S9-Mix
Phosphate buffer 22.5 mL
0.1M NADP-solution 1.0 mL
1M G6P-solution 0.125 mL
Salt solution 0.5 mL
Rat liver S9 1.0 mL
500 µL per 2000 µL top-agar (preincubation method) or 500 µL per 2700µL (plate incorporation method) - Test concentrations with justification for top dose:
- Nominal concentrations: 50, 150, 500, 1500, 5000 µg/plate for the first experiment (plate incorporation)
Nominal concentrations: 78, 156, 313, 625, 1250, 2500, 5000 µg/plate for the second experiment (preincubation method)
Concentrations were based on preliminary tests - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: aqueous solvents (water)
- Justification for choice of solvent/vehicle: Based on the non-GLP pre-test, a test item suspension in demin. water was used, be-
cause this solvent shows the most stable suspension with the test item and does not have any effects on the viability of the bacteria or the number of spontaneous revertants in the tested concentrations.
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- benzo(a)pyrene
- other: 4-Nitro-1,2-phenylene diamine; without metabolic activation; 20 µg in DMSO for TA 98 and 30 µg for TA102 and TA1537; 2-Amino-anthracene; with metabolic activation; 1 µg in DMSO for TA100, TA1535 and 2.4 µg in DMSO for TA 102, TA1537
- Details on test system and experimental conditions:
- NUMBER OF REPLICATIONS:
- Number of cultures per concentration (single, duplicate, triplicate) : Treatment with test item: triplicates; Spontaneous revertants: triplicates; Determination of titre: duplicates; Toxicity control: duplicates; Sterility control: four replicates; Positive controls: triplicates
- Number of independent experiments : two
METHOD OF TREATMENT/ EXPOSURE:
- Cell density at seeding (if applicable): at least E+09 cells/mL
- Test substance added in medium; in agar (plate incorporation, experiment one) and preincubation (experiment two)
TREATMENT AND HARVEST SCHEDULE:
- Preincubation period, if applicable: 20 min at 37 ± 1°C
METHODS FOR MEASUREMENT OF CYTOTOXICITY
- Method, e.g.: background growth inhibition - Rationale for test conditions:
- As recommended by OECD guideline 471
- Evaluation criteria:
- Five different analysable and non-toxic concentrations should be used for the evaluation of the mutagenic potential of the test item.
A substance is considered to be mutagenic, if a reproducible increase with or without metabolic activation of revertant colonies per plate exceeding an increase factor of 2 for the bacteria strains TA98, TA100, TA102, TA1535 and TA1537 compared to vehicle controls
in at least one strain can be observed.
A concentration-related increase over the range tested is also taken as a sign of mutagenic activity.
A substance is not mutagenic if it does not meet these criteria. If the criteria listed above are not clearly met, the results will be assessed as equivocal and will be discussed. - Key result
- Species / strain:
- S. typhimurium TA 1535
- 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
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1537
- 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
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 98
- 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
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium 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
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 102
- 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
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Water solubility: sufficient for using demin. water as vehicle
- Precipitation and time of the determination: Not observed
RANGE-FINDING/SCREENING STUDIES (if applicable): Preliminary non-GLP test to determine the solubility of the test item
STUDY RESULTS
- Concurrent vehicle negative and positive control data
Please refer to the 'Any other information on results incl. tables' section
For all test methods and criteria for data analysis and interpretation:
- Concentration-response relationship where possible : Due to lack of toxicity and mutagenicity not possible.
Ames test:
- Signs of toxicity : No signs of toxicity
HISTORICAL CONTROL DATA (with ranges, means and standard deviation, and 95% control limits for the distribution as well as the number of data)
- Positive historical control data: Please refer to the 'Any other information on results incl. tables' section
- Negative (solvent/vehicle) historical control data: Please refer to the 'Any other information on results incl. tables' section - Conclusions:
- Based on the results of this studyconducted according to OECD guideline 471 it is concluded that the test item Zinc bisglycinate is not mutagenic in the Salmonella typhimurium strains TA98, TA100, TA102, TA1535 and TA1537 in the presence and absence of metabolic activation under the experimental conditions in this study.
- Executive summary:
In a reverse gene mutation assay in bacteria according to OECD guideline 471 (1997), Salmonella typhimurium strains TA98, TA100, TA102, TA1535 and TA1537 in the presence and absence of metabolic activation were exposed to zinc bisglycinate in demin. water and DMSO, respectively, at concentrations of 50, 150, 500, 1500 and 5000 µg/plate (pre-incubation method) and to 78, 156, 313, 625, 1250, 2500 and 5000 µg/plate (plate incorporation method).
Zinc bisglycinate was tested up to limit concentration of 5000 µg/plate, there was no evidence of induced mutant colonies over background. The positive controls induced the appropriate responses in the corresponding strains.
This study is classified as acceptable. This study satisfies the requirement for Test Guideline OECD 471 for in vitro mutagenicity (bacterial reverse gene mutation) data. Thus, the test item is not classified as mutagen according to Regulation (EC) No. 1272/2008 (CLP) or the Globally Harmonized System of Classification and Labelling of Chemicals (GHS).
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- 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)
- Version / remarks:
- The study was conducted in 1984.
- GLP compliance:
- no
- Type of assay:
- bacterial reverse mutation assay
- Specific details on test material used for the study:
- Test material was supplied from the Japan Food Additives Association, Tokyo, at the request of the Ministry of Health and Welfare of Japan, where the purity and quality of each sample were checked.
- Target gene:
- histidine operon
- Species / strain / cell type:
- S. typhimurium TA 1537
- Species / strain / cell type:
- S. typhimurium TA 1535
- Species / strain / cell type:
- S. typhimurium TA 98
- Species / strain / cell type:
- S. typhimurium TA 100
- Species / strain / cell type:
- S. typhimurium, other: TA92
- Species / strain / cell type:
- S. typhimurium, other: TA94
- Metabolic activation:
- with and without
- Metabolic activation system:
- Type and composition of metabolic activation system:
- method of preparation of S9 mix: The liver microsome fraction (S-9) was prepared from the liver of Fischer rats (Charles River Japan Co.) pre-treated 5 days before with polychlorinated biphenyls (500 mg/kg body weight of Kanechlor KC-400 in olive oil, ip). The reaction mixture (S-9 mix) contained 5 mM-glucose 6-phosphate, 4mM-NADPH, 4mM-NADH, 33mM-KCl, 8 mM-MgCI2, 100 mM-phosphate buffer (pH 7.4) and 3.75 ml S-9 (129 mg protein) in a total volume of 12.5 ml. - Test concentrations with justification for top dose:
- 35000 µg/plate (crystal) and 33000 µg/plate (powder), 6 doses. The maximum dose represents the highest non-cytotoxic dose used in the experiment.
- Vehicle / solvent:
- phosphate buffer
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- other: Fourteen out of 200 tested substances in the Ames assay were identified as mutagenic, which proves the validity of the system.
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: preincubation
DURATION:
-Preincubation period: 20 minutes
-Exposure duration: 48 hours
NUMBER OF PLATES : 2 - Evaluation criteria:
- The result was considered positive, if the number of colonies found was twice the number in the control.
- Key result
- Species / strain:
- S. typhimurium, other: TA94
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Key result
- Species / strain:
- S. typhimurium, other: TA92
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Key result
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Conclusions:
- The test item was not mutagenic in the bacterial reverse mutation assay.
- Executive summary:
In the study by Ishidate et al. (1984), the mutagenic potential of glycine was examined in a bacterial reverse mutation assay according to the method described by Ames et al. (1975).
6 tester strains of Salmonella typhimurium (TA92, TA1535, TA100, TA1537, TA94 and TA98) were treated with the test material, both with and without metabolic activation. 6 doses were applied, and the maximum doses were 35000 µg/plate (crystal) and 33000 µg/plate (powder), which represent the highest non-cytotoxic dose used in the experiment.
No significant increases in the frequency of revertant colonies were observed for any of the bacterial strains at any dose of glycine, either with or without metabolic activation. No information on the validity of vehicle and positive controls is reported.
In conclusion, glycine was not mutagenic in the bacterial reverse mutation assay.
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Justification for type of information:
- REPORTING FORMAT FOR THE ANALOGUE APPROACH
1. HYPOTHESIS FOR THE ANALOGUE APPROACH
This read-across hypothesis is based on transformation of the target and source substances to common compounds (scenario 1 of the RAAF). The target substance zinc monoglycinate sulfate and the source substances zinc sulfate and zinc bisglycinate consist of the Zn2+ cation and the respective anion. The amino acid glycine is constituent of both the target substance zinc monoglycinate sulfate and the source substance zinc bisglycinate.
It is generally accepted that the Zn2+ cation (as measure for dissolved zinc species) is the determining factor for toxicity and ecotoxicity, but not sulfate or glycine.
2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
The target substance zinc monoglycinate sulfate is a chelate-complex which consists of the divalent zinc ion as centre-ion and glycine as ligand. The remaining sulfate group stabilizes the center ion within the complex.
Zinc monoglycinate sulfate and the source substance zinc sulfate are ionic and consist of the Zn2+ cation and the respective anions. It is generally accepted that the zinc cation is the determining factor for toxicity and ecotoxicity. Therefore, this read-across approach is based on the assumption that the metal cation of both the target and the source substance, zinc, is the relevant component for assessment of toxicity and ecotoxicity.
The anion of the target substance is the essential amino acid glycine and the sulfate anion. In the source substance, it is the sulfate anion. These anions are not considered as (eco)toxicologically relevant at the given concentrations.
Please refer to the justification for read-across analogue approach in Chapter 13.2 for more detailed information.
3. ANALOGUE APPROACH JUSTIFICATION
Please refer to the justification for read-across analogue approach in Chapter 13.2 for more detailed information.
4. DATA MATRIX
Please refer to the justification for read-across analogue approach in Chapter 13.2 for more detailed information. - Reason / purpose for cross-reference:
- read-across source
- Reason / purpose for cross-reference:
- read-across source
- Reason / purpose for cross-reference:
- read-across source
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Remarks:
- Gocke et al., 1981
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Remarks:
- Gocke et al., 1981
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Remarks:
- Gocke et al., 1981
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Remarks:
- Gocke et al., 1981
- Species / strain:
- S. typhimurium TA 1538
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Remarks:
- Gocke et al., 1981
- Species / strain:
- S. typhimurium TA 102
- 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
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Remarks:
- Zinc-bisglycinate study
- Species / strain:
- S. typhimurium 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
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Remarks:
- Zinc-bisglycinate study
- Species / strain:
- S. typhimurium TA 98
- 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
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Remarks:
- Zinc-bisglycinate study
- Species / strain:
- S. typhimurium TA 1537
- 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
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Remarks:
- Zinc-bisglycinate study
- Species / strain:
- S. typhimurium TA 1535
- 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
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Remarks:
- Zinc-bisglycinate study
- Species / strain:
- S. typhimurium, other: TA94
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Remarks:
- Ishidate et al., 1984
- Species / strain:
- S. typhimurium, other: TA92
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Remarks:
- Ishidate et al., 1984
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Remarks:
- Ishidate et al., 1984
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Remarks:
- Ishidate et al., 1984
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Remarks:
- Ishidate et al., 1984
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Remarks:
- Ishidate et al., 1984
- Conclusions:
- The test item is not considered to be mutagenic in the bacterial reverse mutation assay with and without metabolic activation.
- Executive summary:
The mutagenicity of zinc monoglycinate sulfate is adressed with read-across to its components zinc sulfate and glycine as well as the source substance zinc bisglycinate.
In the study by Gocke et al. (1981), the mutagenic potential of zinc sulfate was examined according to the method described by Ames et al. (1975) in a bacterial reverse mutation assay. 5 tester strains of Salmonella typhimurium which were TA1535, TA100, TA1538, TA98 and TA1537 were treated with the test material at minimal five dose levels up to 3600 µg/plate, both with and without metabolic activation. S9 liver fraction was prepared from Aroclor-pretreated rats. No significant increases in the frequency of revertant colonies were observed for any of the bacterial strains at any dose of zinc sulfate, either with or without metabolic activation. The positive control was benzo(a)pyrene. Validity of the results from vehicle and positive controls are not clearly stated. In conclusion, zinc sulfate was not mutagenic in the bacterial reverse mutation assay.
In a reverse gene mutation assay in bacteria according to OECD guideline 471 (1997), Salmonella typhimurium strains TA98, TA100, TA102, TA1535 and TA1537 in the presence and absence of metabolic activation were exposed to zinc bisglycinate in demin. water and DMSO, respectively, at concentrations of 50, 150, 500, 1500 and 5000 µg/plate (pre-incubation method) and to 78, 156, 313, 625, 1250, 2500 and 5000 µg/plate (plate incorporation method). Zinc bisglycinate was tested up to limit concentration of 5000 µg/plate, there was no evidence of induced mutant colonies over background. The positive controls induced the appropriate responses in the corresponding strains. This study is classified as acceptable. This study satisfies the requirement for Test Guideline OECD 471 for in vitro mutagenicity (bacterial reverse gene mutation) data. Thus, zinc bisglycinate is not classified as mutagenic according to Regulation (EC) No. 1272/2008 (CLP) or the Globally Harmonized System of Classification and Labelling of Chemicals (GHS).
In the study by Ishidate et al. (1984), the mutagenic potential of glycine was examined in a bacterial reverse mutation assay according to the method described by Ames et al. (1975). 6 tester strains of Salmonella typhimurium (TA92, TA1535, TA100, TA1537, TA94 and TA98) were treated with the test material, both with and without metabolic activation. 6 doses were applied, and the maximum doses were 35000 µg/plate (crystal) and 33000 µg/plate (powder), which represent the highest non-cytotoxic dose used in the experiment. No significant increases in the frequency of revertant colonies were observed for any of the bacterial strains at any dose of glycine, either with or without metabolic activation. No information on the validity of vehicle and positive controls is reported. In conclusion, glycine was not mutagenic in the bacterial reverse mutation assay.
Therefore, zinc monoglycinate sulfate is not considered to be mutagenic in the bacterial reverse mutation assay with and without metabolic activation.
Referenceopen allclose all
Spontaneous Revertants demin. water (colonies per plate) Demin. water; experiment 1
Strain |
TA98 |
TA100 |
TA102 |
TA1535 |
TA1537 |
|||||
Induction |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
Repl. 1 |
15 |
18 |
62 |
70 |
280 |
280 |
9 |
17 |
8 |
9 |
Repl. 2 |
13 |
10 |
78 |
72 |
248 |
272 |
9 |
17 |
8 |
10 |
Repl. 3 |
18 |
17 |
72 |
76 |
264 |
272 |
12 |
19 |
6 |
8 |
Mean |
15 |
15 |
71 |
73 |
264 |
275 |
10 |
18 |
7 |
9 |
sd |
2.5 |
4.4 |
8.1 |
3.1 |
16.0 |
4.6 |
1.7 |
1.2 |
1.2 |
1.0 |
Spontaneous Revertants demin. water (colonies per plate) Demin. water; experiment 2
Strain |
TA98 |
TA100 |
TA102 |
TA1535 |
TA1537 |
|||||
Induction |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
Repl. 1 |
19 |
20 |
96 |
96 |
352 |
368 |
9 |
10 |
6 |
9 |
Repl. 2 |
20 |
19 |
92 |
92 |
384 |
384 |
12 |
10 |
5 |
7 |
Repl. 3 |
20 |
19 |
84 |
80 |
376 |
376 |
10 |
11 |
4 |
7 |
Mean |
20 |
19 |
91 |
89 |
371 |
376 |
10 |
10 |
5 |
8 |
sd |
0.6 |
0.6 |
6.1 |
8.3 |
16.7 |
8.0 |
1.5 |
0.6 |
1.0 |
1.2 |
Spontaneous Revertants demin. water (colonies per plate) DMSO, experiment 1
Strain |
TA98 |
TA100 |
TA102 |
TA1535 |
TA1537 |
|||||
Induction |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
Repl. 1 |
11 |
15 |
60 |
62 |
256 |
288 |
15 |
15 |
11 |
8 |
Repl. 2 |
14 |
13 |
62 |
80 |
272 |
264 |
17 |
10 |
8 |
9 |
Repl. 3 |
14 |
19 |
58 |
66 |
248 |
280 |
18 |
14 |
8 |
8 |
Mean |
13 |
16 |
60 |
69 |
259 |
277 |
17 |
13 |
9 |
8 |
sd |
1.7 |
3.1 |
2.0 |
9.5 |
12.2 |
12.2 |
1.5 |
2.6 |
1.7 |
0.6 |
Spontaneous Revertants demin. water (colonies per plate) DMSO, experiment 2
Strain |
TA98 |
TA100 |
TA102 |
TA1535 |
TA1537 |
|||||
Induction |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
Repl. 1 |
18 |
19 |
76 |
80 |
352 |
360 |
9 |
11 |
6 |
9 |
Repl. 2 |
18 |
20 |
80 |
84 |
360 |
368 |
8 |
15 |
6 |
6 |
Repl. 3 |
18 |
21 |
88 |
84 |
352 |
368 |
10 |
10 |
6 |
8 |
Mean |
18 |
20 |
81 |
83 |
355 |
365 |
9 |
12 |
6 |
8 |
sd |
0.0 |
1.0 |
6.1 |
2.3 |
4.6 |
4.6 |
1.0 |
2.6 |
0.0 |
1.5 |
Historical Data of Spontaneous Revertants
Strain |
TA98 |
TA100 |
TA102 |
TA1535 |
TA1537 |
||||||
Induction |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
|
Demin. Water |
Mean |
28 |
30 |
91 |
96 |
303 |
318 |
15 |
15 |
6 |
8 |
Min |
6 |
8 |
46 |
53 |
85 |
67 |
6 |
6 |
3 |
5 |
|
Max |
93 |
109 |
147 |
141 |
509 |
587 |
36 |
40 |
15 |
22 |
|
SD |
16 |
15 |
16 |
16 |
70 |
77 |
6 |
6 |
2 |
2 |
|
Exp1 |
15 |
15 |
71 |
73 |
264 |
275 |
10 |
18 |
7 |
9 |
|
Exp2 |
20 |
19 |
91 |
89 |
371 |
376 |
10 |
10 |
5 |
8 |
|
DMSO |
Mean |
28 |
29 |
88 |
91 |
302 |
311 |
15 |
15 |
6 |
7 |
Min |
7 |
8 |
37 |
42 |
79 |
80 |
6 |
6 |
4 |
4 |
|
Max |
104 |
108 |
143 |
199 |
531 |
499 |
35 |
37 |
15 |
20 |
|
SD |
16 |
15 |
16 |
17 |
70 |
69 |
6 |
6 |
4 |
4 |
|
Exp1 |
13 |
16 |
60 |
69 |
259 |
277 |
17 |
13 |
9 |
8 |
|
Exp2 |
18 |
20 |
81 |
83 |
355 |
365 |
9 |
12 |
6 |
8 |
|
Positive controls |
Mean |
433 |
188 |
529 |
829 |
1138 |
1209 |
289 |
151 |
124 |
111 |
Min |
77 |
39 |
218 |
273 |
491 |
408 |
55 |
45 |
79 |
100 |
|
Max |
s.g. |
s.g. |
1256 |
1912 |
2331 |
6083 |
s.g. |
s.g. |
165 |
123 |
|
SD |
219 |
159 |
214 |
275 |
364 |
473 |
119 |
106 |
17 |
7 |
|
Exp1 |
s.g. |
98 |
s.g. |
s.g. |
605 |
s.g. |
221 |
175 |
232 |
168 |
|
Exp2 |
s.g. |
111 |
s.g. |
s.g. |
765 |
872 |
248 |
167 |
152 |
160 |
|
Mean Revertants First Experiment (1)
Strain |
TA98 |
TA100 |
TA102 |
TA1535 |
TA1537 |
||||||
Induction |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
|
Demin. Water |
Mean |
15 |
15 |
71 |
73 |
264 |
275 |
10 |
18 |
7 |
9 |
SD |
2.5 |
4.4 |
8.1 |
3.1 |
16.0 |
4.6 |
1.7 |
1.2 |
1.2 |
1.0 |
|
DMSO |
Mean |
13 |
16 |
60 |
69 |
259 |
277 |
17 |
13 |
9 |
8 |
SD |
1.7 |
3.1 |
2.0 |
9.5 |
12.2 |
12.2 |
1.5 |
2.6 |
1.7 |
0.6 |
|
Positive controls |
Mean |
s.g. |
98 |
s.g. |
s.g. |
605 |
s.g. |
221 |
175 |
232 |
168 |
SD |
-- |
17.1 |
-- |
-- |
28.1 |
-- |
6.1 |
4.6 |
8.0 |
21.2 |
|
f(l) |
> 2 |
6.13 |
> 2 |
> 2 |
2.34 |
> 2 |
22.10 |
13.46 |
25.78 |
21.00 |
|
5000µg/plate |
Mean |
15 |
19 |
125 |
120 |
256 |
256 |
13 |
13 |
9 |
12 |
SD |
0.6 |
2.9 |
5.0 |
8.0 |
16.0 |
13.9 |
1.2 |
2.5 |
1.2 |
1.5 |
|
f(l) |
1.00 |
1.27 |
1.76 |
1.64 |
0.97 |
0.93 |
1.30 |
0.72 |
1.29 |
1.33 |
|
1500 µg/plate |
Mean |
17 |
17 |
94 |
107 |
253 |
269 |
11 |
14 |
12 |
9 |
SD |
3.8 |
2.5 |
7.2 |
4.6 |
18.5 |
20.1 |
1.5 |
2.3 |
2.5 |
0.6 |
|
f(l) |
1.13 |
1.13 |
1.32 |
1.47 |
0.96 |
0.98 |
1.10 |
0.78 |
1.71 |
1.00 |
|
500 µg/plate |
Mean |
12 |
15 |
67 |
89 |
272 |
264 |
11 |
12 |
9 |
10 |
SD |
2.5 |
3.1 |
1.2 |
2.3 |
16.0 |
8.0 |
2.1 |
0.6 |
0.6 |
1.5 |
|
f(l) |
0.80 |
1.00 |
0.94 |
1.22 |
1.03 |
0.96 |
1.10 |
0.67 |
1.29 |
1.11 |
|
150 µg/plate |
Mean |
13 |
13 |
62 |
65 |
267 |
261 |
12 |
13 |
9 |
8 |
SD |
3.1 |
4.0 |
2.0 |
5.0 |
12.2 |
18.5 |
1.7 |
0.6 |
1.2 |
1.0 |
|
f(l) |
0.87 |
0.87 |
0.87 |
0.89 |
1.01 |
0.95 |
1.20 |
0.72 |
1.29 |
0.89 |
|
50 g/plate |
Mean |
10 |
14 |
62 |
63 |
264 |
264 |
12 |
12 |
10 |
10 |
|
SD |
1.7 |
3.0 |
2.0 |
3.1 |
8.0 |
0.0 |
0.6 |
2.1 |
2.1 |
2.1 |
|
f(l) |
0.67 |
0.93 |
0.87 |
0.86 |
1.00 |
096 |
1.20 |
0.67 |
1.43 |
1.11 |
Mean Revertants Second Experiment (2)
Strain |
TA98 |
TA100 |
TA102 |
TA1535 |
TA1537 |
||||||
Induction |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
|
Demin. Water |
Mean |
20 |
19 |
91 |
89 |
371 |
376 |
10 |
10 |
5 |
8 |
SD |
0.6 |
0.6 |
6.1 |
8.3 |
16.7 |
8.0 |
1.5 |
0.6 |
1.0 |
1.2 |
|
DMSO |
Mean |
18 |
20 |
81 |
83 |
355 |
365 |
9 |
12 |
6 |
8 |
SD |
0.0 |
1.0 |
6.1 |
2.3 |
4.6 |
4.6 |
1.0 |
2.6 |
0.0 |
1.5 |
|
Positive controls |
Mean |
s.g. |
111 |
s.g. |
s.g. |
765 |
872 |
248 |
167 |
152 |
160 |
SD |
-- |
2.3 |
-- |
-- |
12.2 |
21.2 |
13.9 |
2.3 |
18.3 |
13.9 |
|
f(l) |
> 2 |
5.55 |
> 2 |
> 2 |
2.15 |
2.39 |
24.80 |
13.92 |
25.33 |
20.00 |
|
5000µg/plate |
Mean |
19 |
24 |
108 |
108 |
368 |
349 |
10 |
12 |
7 |
8 |
SD |
1.5 |
1.0 |
4.0 |
4.0 |
8.0 |
4.6 |
0.6 |
2.1 |
1.5 |
1.0 |
|
f(l) |
0.95 |
1.26 |
1.19 |
1.21 |
0.99 |
0.93 |
1.00 |
1.20 |
1.40 |
1.00 |
|
2500 g/plate |
Mean |
19 |
21 |
92 |
88 |
363 |
349 |
13 |
12 |
7 |
8 |
SD |
1.0 |
1.0 |
4.0 |
6.9 |
23.1 |
12.2 |
1.5 |
1.7 |
2.1 |
0.6 |
|
f(l) |
0.95 |
1.11 |
1.01 |
0.99 |
0.98 |
0.93 |
1.30 |
1.20 |
1.40 |
1.00 |
|
1250µg/plate |
Mean |
18 |
20 |
89 |
97 |
373 |
355 |
10 |
11 |
6 |
6 |
SD |
1.0 |
1.2 |
8.3 |
6.1 |
12.2 |
18.5 |
0.6 |
1.0 |
0.6 |
1.0 |
|
f(l) |
0.90 |
1.05 |
0.98 |
1.09 |
1.01 |
0.94 |
1.00 |
1.10 |
1.20 |
0.75 |
|
625 µg/plate |
Mean |
19 |
22 |
87 |
95 |
360 |
368 |
10 |
8 |
7 |
6 |
SD |
0.6 |
1.0 |
2.3 |
2.3 |
8.0 |
16.0 |
1.0 |
1.0 |
1.5 |
1.7 |
|
f(l) |
0.95 |
1.16 |
0.96 |
1.07 |
0.97 |
0.98 |
1.00 |
0.80 |
1.40 |
0.75 |
|
313 µg/plate |
Mean |
19 |
18 |
87 |
92 |
355 |
347 |
9 |
10 |
5 |
6 |
SD |
1.2 |
0.6 |
2.3 |
4.0 |
16.7 |
16.7 |
1.0 |
2.0 |
0.6 |
0.6 |
|
f(l) |
0.95 |
0.95 |
0.96 |
1.03 |
0.96 |
0.92 |
0.90 |
1.00 |
1.00 |
0.75 |
|
156 g/plate |
Mean |
19 |
22 |
87 |
95 |
360 |
368 |
10 |
8 |
7 |
6 |
SD |
0.6 |
1.0 |
2.3 |
2.3 |
8.0 |
16.0 |
1.0 |
1.0 |
1.5 |
1.7 |
|
f(l) |
0.95 |
1.16 |
0.96 |
1.07 |
0.97 |
0.98 |
1.00 |
0.80 |
1.40 |
0.75 |
|
78 g/plate |
Mean |
19 |
18 |
87 |
92 |
355 |
347 |
8 |
10 |
5 |
6 |
SD |
1.2 |
0.6 |
2.3 |
4.0 |
16.7 |
16.7 |
0.6 |
2.0 |
0.6 |
0.6 |
|
f(l) |
0.95 |
0.95 |
0.96 |
1.03 |
0.96 |
0.92 |
0.80 |
1.00 |
1.00 |
0.75 |
|
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
Justification for classification or non-classification
The mutagenicity of zinc monoglycinate sulfate is adressed with read-across to its components zinc sulfate and glycine as well as the source substance zinc bisglycinate.
In the study by Gocke et al. (1981), the mutagenic potential of zinc sulfate was examined according to the method described by Ames et al. (1975) in a bacterial reverse mutation assay. 5 tester strains of Salmonella typhimurium which were TA1535, TA100, TA1538, TA98 and TA1537 were treated with the test material at minimal five dose levels up to 3600 µg/plate, both with and without metabolic activation. S9 liver fraction was prepared from Aroclor-pretreated rats. No significant increases in the frequency of revertant colonies were observed for any of the bacterial strains at any dose of zinc sulfate, either with or without metabolic activation. The positive control was benzo(a)pyrene. Validity of the results from vehicle and positive controls are not clearly stated. In conclusion, zinc sulfate was not mutagenic in the bacterial reverse mutation assay.
In a reverse gene mutation assay in bacteria according to OECD guideline 471 (1997), Salmonella typhimurium strains TA98, TA100, TA102, TA1535 and TA1537 in the presence and absence of metabolic activation were exposed to zinc bisglycinate in demin. water and DMSO, respectively, at concentrations of 50, 150, 500, 1500 and 5000 µg/plate (pre-incubation method) and to 78, 156, 313, 625, 1250, 2500 and 5000 µg/plate (plate incorporation method). Zinc bisglycinate was tested up to limit concentration of 5000 µg/plate, there was no evidence of induced mutant colonies over background. The positive controls induced the appropriate responses in the corresponding strains. This study is classified as acceptable. This study satisfies the requirement for Test Guideline OECD 471 for in vitro mutagenicity (bacterial reverse gene mutation) data. Thus, zinc bisglycinate is not classified as mutagen according to Regulation (EC) No. 1272/2008 (CLP) or the Globally Harmonized System of Classification and Labelling of Chemicals (GHS).
In the study by Ishidate et al. (1984), the mutagenic potential of glycine was examined in a bacterial reverse mutation assay according to the method described by Ames et al. (1975). 6 tester strains of Salmonella typhimurium (TA92, TA1535, TA100, TA1537, TA94 and TA98) were treated with the test material, both with and without metabolic activation. 6 doses were applied, and the maximum doses were 35000 µg/plate (crystal) and 33000 µg/plate (powder), which represent the highest non-cytotoxic dose used in the experiment. No significant increases in the frequency of revertant colonies were observed for any of the bacterial strains at any dose of glycine, either with or without metabolic activation. No information on the validity of vehicle and positive controls is reported. In conclusion, glycine was not mutagenic in the bacterial reverse mutation assay.
Therefore, zinc monoglycinate sulfate is not considered to be mutagenic in the bacterial reverse mutation assay with and without metabolic activation.
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