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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Study period:
2002
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducted according to the appropriate OECD test guideline, and in compliance with GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2002
Report Date:
2002

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to
Guideline:
JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
Test material form:
solid: crystalline
Details on test material:
- Name of test material (as cited in study report): Sulfuric acid, iron(2+) salt (1:1), heptahydrate
- Molecular formula (if other than submission substance): FeSO4. 7H2O
- Molecular weight (if other than submission substance): 278.01
- Physical state: powder, blue-green crystals
- Analytical purity: 91.6 wt%
- Impurities (identity and concentrations): Mg 0.22%, Mn 0.2%, Zn 87 ppm
- Stability under test conditions:
- Storage condition of test material: room temperature under Argon
- Other: MP 64°C, Water solubility: 32.8g/l at 0°C

Method

Target gene:
His Operon (S. typhimurium strains) and Trp Operon (E. coli strain)
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Metabolic activation system:
S9-mix obtained from SD rat liver, induced with phenobarbital and 5,6-benzoflavone
Test concentrations with justification for top dose:
0, 156, 313, 625, 1250, 2500, 5000 µg/plate
Vehicle / solvent:
distilled water
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: -S9: aminofluorene for TA100 and TA98; sodium azide for TA1535; TA1535: NaN3, N-ethyl-N'-nitrosogunanidine for WP2uvrA/pKM101 and 9-aminoacridine hydrochloride for TA1537 +S9: 2-amonoanthracene
Details on test system and experimental conditions:
METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 20 minutes
- Exposure duration: 48 hours

PLATES/TEST: 3
NUMBER OF REPLICATIONS: 2 tests

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth


Evaluation criteria:
No information but standard test guideline criteria apply
Statistics:
Colonies were counted for 3 plates. Mean values which are at least twice as high as the negative vehicle control are considered as positive. Otherwise it is considered as negative.

Results and discussion

Test results
Species / strain:
other: S. typhimurium TA 1535, TA 1537, TA 98 and TA 100; E. coli WP2 uvrA pKM 101
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
Toxicity was observed at a concentration of > 2500 µg/plate with or without metabolic activation
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
There was no precipitation at any test concentration.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1. Mutagenicity of ferrous sulfate heptahydrate in reverse mutation test in bacteria

 

 

 

number of revertants: mean value of negative control

 

 

number of revertants: mean value of positive control

 

max. number of revertants: mean value of test material [µg/plate]

 without S9-mix

Base-pair change type

TA100

101 ± 6

630 ± 29

113 ± 7 [5000]

 TA 1535

9.5 ± 3

436 ± 21

13 ± 3 [ 313]

WP2uvrA/pKM101

74.5 ± 4

3227 ± 107

92 ± 7 [2500]

Frame-shift type

 TA 98

15.5 ± 3

604 ± 30

20 ± 4 [156]

 TA1537

7.5 ± 2

211 ± 28

11 ± 1 [313]

 without S9-mix

Base-pair change type

TA100

103 ± 6

1897 ± 202

136 ± 18 [5000]

 TA 1535

12 ± 3

241 ± 17

14 ± 5 [ 2500]

WP2uvrA/pKM101

96 ± 7

831 ± 72

106 ± 1 [5000]

Frame-shift type

 TA 98

26.5 ± 7

479 ± 30

29 ± 3 [313]

 TA1537

13 ± 3

227 ± 17

21 ± 2 [1250]

Applicant's summary and conclusion

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

Ferrous sulfate heptahydrate was tested for bacterial mutagenicity according OECD 471 under GLP using the preincubation method. It did not induce an increase in reversions in any of the strains tested in either the initial or the repeat test. Solvent and positive controls gave the expected results. It is concluded that ferrous sulfate heptahydrate is not mutagenic to bacteria under the conditions of the test.
Executive summary:

Ferrous sulfate heptahydrate was tested for bacterial mutagenicity according OECD 471 under GLP using the preincubation method. It did not induce an increase in reversions in any of the strains tested in either the initial or the repeat test. Solvent and positive controls gave the expected results. It is concluded that ferrous sulfate heptahydrate is not mutagenic to bacteria under the conditions of the test.