Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Administrative data

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
Type of genotoxicity: chromosome aberration
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Research paper. Study well documented meeting generally accepted scientific principles.

Data source

Reference Type:
Nuclear aberrations and micronuclei induction in the digestive tract of mice treated with different iron salts.
Bianchini F, Caderni G, Dolara P, Tanagnelli E
Bibliographic source:
J. App. Tox. 8(3): 179-183

Materials and methods

Test guideline
no guideline followed
Principles of method if other than guideline:
Method: other: Wargovich et al, J Natl Cancer Inst 71, 133-137 (1983)
GLP compliance:
not specified
Type of assay:
micronucleus assay

Test material

Constituent 1
Reference substance name:
Iron sulphate
EC Number:
EC Name:
Iron sulphate
Cas Number:
iron(2+) sulfate
Test material form:
solid: crystalline
Details on test material:
Test substance obtained from Merck and reportedly containing ca 20% Fe.
The material tested was the heptahydrate of Ferrous sulphate CAS No 7782-63-0 FeS04.7H20 but this does not affect the chemical species available to the test organisms.

Test animals

other: C57BL/6J
Details on test animals or test system and environmental conditions:
- Source: Charles river, Calco, Como Italy
- Age at study initiation: no information
- Weight at study initiation: 17-18 g
- Assigned to test groups randomly: no information
- Fasting period before study: 12 h, or not fasted
- Diet (e.g. ad libitum): ad libitum for non-fasting animals
- Water (e.g. ad libitum): ad libitum (after treatment)

Administration / exposure

Route of administration:
other: oral gavage or intrarectal
- Vehicle(s)/solvent(s) used: saline
- Amount of vehicle (if gavage or dermal): 0.2 ml
Details on exposure:

Duration of treatment / exposure:
24 hours
Frequency of treatment:
Single treatment
Post exposure period:
24 hours
Doses / concentrations
Doses / Concentrations:
2, 6.5 and 13 mg Fe/kg

No. of animals per sex per dose:
4-9 female animals per dose
Control animals:
Positive control(s):
- 2-amino-3-methylimidazo(4,5-f)quinoline) (IQ)
- Justification for choice of positive control(s): IQ is a genotoxic chemical with specific intestinal action.
- Route of administration: orally to non-fasting mice
- Doses / concentrations: 100 and 200 mg/kg bw


Tissues and cell types examined:
Samples of forestomach, duodenum and colon were taken following oral administration and colon samples after intrarectal administration
Details of tissue and slide preparation:
CRITERIA FOR DOSE SELECTION: The lowest dose tested was equivalent to a therapeutic dose in iron deficiency anaemia.

DETAILS OF SLIDE PREPARATION: colon and duodenum where opened longitudinally and rolled. The forestomach was excised. Specimens were fixed in 10% buffered formalin and processed for histology. Slides with 5 micron thick paraffin sections were stained with Feulgen-fast green.

Slides were examined and scored for a) micronuclei defined as nuclear fragments not larger than ¼ -1/3 of the diameter of the primary nucleus b) nuclear aberrations comprised of micronuclei, pyknotic nuclei, cytolysosomes and disintegrated nuclei. The forestomach was analysed by observing around 400 mucosal cells per animal. Mucosal cells lining 20 crypts for colon and 5 crypts for duodenum were analysed per animal beginning from the anal end of the colon and the pyloric end of the duodenum.
Evaluation criteria:
Evaluation criteria are not described.
No statistical evaluation was described.

Results and discussion

Test results
Vehicle controls validity:
Negative controls validity:
not applicable
Positive controls validity:
Additional information on results:
The reference compound IQ caused an increase of nuclear aberrations and micronuclei, results being presented for effects in the colon.  Ferrous sulphate did not significantly increase the incidence of micronuclei in the stomach, duodenum or colon following oral 
administration to fasting or non-fasting mice.  In the colon nuclear aberrations indicating toxicity were increased in fasting and non-fasting mice receiving oral ferrous sulphate.  Following intrarectal administration ferrous sulfate showed slight  toxicity to the colon producing a slight but statistically 
significant (p<0.01) increase in micronuclei and in chromosome aberrations.  The authors consider that the iron salts do not appear to have a significant genotoxic effect on the gastrointestinal tract and that the toxic effects are 
due to generalized toxicity rather than a genotoxic  effect.

Any other information on results incl. tables

The reference compound IQ caused an increase of nuclear aberrations and micronuclei, results being presented for effects in the colon. Ferrous sulphate did not significantly increase the incidence of micronuclei in the stomach, duodenum or colon following oral administration to fasting or non-fasting mice. In the colon nuclear aberrations indicating toxicity were increased in fasting and non-fasting mice receiving oral ferrous sulphate. 

Following intrarectal administration ferrous sulfate showed slight toxicity to the colon producing a slight but statistically significant (p<0.01) increase in micronuclei and in chromosome aberrations. 

The authors consider that the iron salts do not appear to have a significant genotoxic effect on the gastrointestinal tract and that the toxic effects are due to generalized toxicity rather than a genotoxic effect.

Applicant's summary and conclusion

Interpretation of results (migrated information): negative
Ferrous sulphate is not considered to have a significant genotoxic effect on the mouse gastrointestinal tract as measured by induction of micronuclei. An increased incidence of nuclear aberrations was considered evidence of general toxicity rather than mutagenic activity.
Executive summary:

 This study was undertaken to assess the effects of iron status on the gastrointestinal tract using the nuclear aberrations assay of Wargovich, Medline and Bruce, 1983. 

The current authors interpreted the results by differentiating between micronuclei and nuclear aberrations considering that micronuclei are a specific sign of genetic damage while nuclear aberrations are more unspecific effects related to cellular toxicity. This was because nuclear aberrations may also be associated with substances acting through non-genotoxic mechanisms.