Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Effect on fertility: via oral route
Endpoint conclusion:
no study available
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

Short description of key information:

Sodium ferrocyanide is a food additive known as E 535 in the EU and added to salt as an anticaking agent with an ADI for man set at 0-0.025 mg/kg bw/day. Exposure to Sodium ferrocyanide is negligible for workers, as the substance is used in a closed batch process. Calculations show that, for the general population, lifelong intake of products containing Sodium ferrocyanide has to be unrealistically high to exceed ADI. Consumer use of Sodium ferrocyanide in anti-freeze and de-icing products is expected to result in exposure well below ADI. As the exposure assessment using ADI indicates safe use, it is also considered safe for reproductive and developmental toxicity.

Based on the low potential for toxicity as derived from the toxicity data available and limited systemic exposure to Sodium ferrocyanide, no hazard and no risk is considered present for reproductive toxicity as well as developmental toxicity. Data from a developmental toxicity study (OECD 414) in rat demonstrate the absence of developmental toxicity, and there are no scientific data indicating rabbit developmental toxicity data (in the absence of systemic toxicity data) being more relevant for human risk assessment than rat data.

The available toxicity data have no indication for reproductive toxicity. As Sodium ferrocyanide is a chemical without specific working mechanism it is unlikely that this substance will specifically demonstrate reproduction toxicity in the absence of any systemic toxicity. Based on these considerations, additional toxicity data on reproduction toxicity is considered not scientifically justified and the Extended one-generation reproductive toxicity test (OECD 443) can be waived.

Effects on developmental toxicity

Description of key information
In an OECD 414 oral developmental toxicity study , rats were dosed up to 1000 mg/kg/ day of sodium ferrocyanide via gavage, resulting in no treatment related effects and thus an NOAEL of ≥ 1000 mg/kg/day could be derived (both maternal and developmental).
Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
16Jan 1992 -02Apr 1992
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Performed according to GLP principles and OECD test guide line, although not according to current guideline. Some minor deviations are not expected to have influence on the study.
Qualifier:
according to guideline
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Version / remarks:
Version May 1981
Deviations:
yes
Remarks:
Different exposure duration
Principles of method if other than guideline:
In this study, pregnant rats were only exposed on day 6-15 of pregnancy, whereas recent guideline recommends exposure day 5 - end of pregnancy. However, guideline from 2001: dosing only during organogenesis, not up to caesarian section.
In addition the gravid uteri and cervices are not weighed. These deviations were not considered to influence the study.
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
other: Crl: CD®(SD) BR VAF/Plus
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River UK Limited, Kent, UK
- Age at study initiation: 8-10 weeks
- Weight at study initiation: 173-248 g
- Housing: Rats were housed five to a cage in suspended galvanised metal cages
- Diet: free access to Biosure Laboratory animal diet No.1
- Water: tap water at libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21± 3
- Humidity (%): 47 ± 3
- Air changes (per hr): not described
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:
oral: gavage
Vehicle:
water
Remarks:
distilled and deionised
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
The highest concentration was prepared by dissolving an appropriate weight of sodium ferrocyanide in ditilled, deionised water. Lower concentrations were prepared by serial dilution of the highest concentration in distilled, deionized water. Formulations were prepared weekly and stored at 4°C in the dark when not in use.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
UV-spectroscopy was used as a method, which was also validated. Concentrations of substance in test solutions prepared for day 1 of dosing were determined. Chemical stability was determined.
Details on mating procedure:
Female rats were time mated to identified males of the same strain at Charles River and as such obtained. Day 0 = day of mating, as judged by the appearance of sperm in the vaginal smear or by the presence of a vaginal plug.
Duration of treatment / exposure:
Day 6-15 (including) of pregnancy.
Frequency of treatment:
Daily
Duration of test:
Day 1-20 of pregnancy.
No. of animals per sex per dose:
25, females only
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: In a preliminary study with less animals the highest dose (1000mg/kg/day) was shown to be well-tolerated.
- Rationale for animal assignment: Animals were weighed and assigned to four groups by computerised stratified randomisation to give approximately equal initial group mean body weights within each batch.
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily observation
- Cage side observations as described in test guideline were included.

BODY WEIGHT: Yes
- Time schedule for examinations: all animals weighed on day 1, 2, 3, 6, 8, 10, 12, 14, 16, 18 and 20

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Food consumption was measured daily for each cage of animals from day 3 to weigh day commencing on day 3 and represented as group mean values (g/rat/day).

WATER CONSUMPTION: Yes
- Water consumption was measured daily for each cage of animals from Day 3 through to termination and represented as group mean values (g/rat/day).

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Rats examined for congenital abnormalities and macroscopic pathological changes in maternal organs.
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes

Fetal examinations:
- External examinations: Yes, all foetuses
- Foetal weight
- Soft tissue examinations: Yes: half the foetuses in each litter
- Skeletal examinations: Yes: half the foetuses in each litter
Statistics:
Significance tests, employing analysis of variance followed by an intergroup comparison with the control, were performed on several parameters. Dependent on the heterogeneity of variance between the treatment groups, parametric tests (analysis of variance followed by Williams' test) or non-parametric tests (Kruskal-Wallis followed by Shirley's test) were used to analyse the data, as appropriate. For litter data and foetal changes, the basic sample unit was the litter and, due to the preponderance of non-normal distributions, non-parametric analyses were routinely used. Where 75% or more of the values for a given variable were the same, a Fisher's exact test was performed.
Indices:
Pre implantation loss (as percentage): ((# corpora lutea -# implantations)/ # corpora lutea)x100;
Post implantation loss (as percentage): ((# implantations - # live young)/ # implantations)x 100.
Historical control data:
Recent historical control groups showed lower incidence in renal lesions than seen in the 500 and 1000 mg/kg/day groups.
No other information given on historical control data.
Details on maternal toxic effects:
Maternal toxic effects:no effects

Details on maternal toxic effects:
Number of pregnant rats/ group did not change with treatment:
Control: 21/25
100 mg/kg/day: 22/25
500 mg/kg/day: 21/25
1000 mg/kg/day: 23/25
One total resorption in control group.

Signs
No treatment-related signs, except treatment at 1000 mg/kg/day was associated with occasional instances of post dosing salivation in 12/23 pregnant rats with greatest incidence on day 9 of pregnancy. Salivation occasionally brown stained, ceased within half an hour of dosing. No differences between pregnant rats in different groups in bodyweight change. No mortalities occurred.

Water and food consumption
An increase in water consumption was noted for all treated groups (cumulative water consumption during treatment period showed dosage-related increase in all treated groups (p<0.05). No effect on food consumption was found.

Macroscopic examination
No adverse effects of treatment were revealed
Dose descriptor:
NOAEL
Effect level:
>= 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: maternal toxicity
Dose descriptor:
NOAEL
Effect level:
>= 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: developmental toxicity
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
No adverse effect of treatment was apparent, as assessed by embryofoetal loss, litter and mean foetal weight or sex ratio.
Incidence, type and distribution of malformation and skeletal anomalies and the percentage of foetuses with variant sternebrae were considered not to reflect any adverse effect of treatment.
In groups dosed with 500 or 1000 mg/kg/day a marginal but not statistically significant increase in the number of foetuses with increased dilation of the renal pelvis/ureter was found (4 and 5 foetuses from 4 and 4 litters resp.; in control group: 2 foetuses from 2 litters).
Dose descriptor:
NOAEL
Effect level:
>= 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: developmental toxicity
Abnormalities:
not specified
Developmental effects observed:
not specified

The mean concentrations of sodium ferrocyanide in test solutions were all within 4% of nominal concentrations. The chemical stability of sodium ferrocyanide in aqueous solutions was confirmed during storage for 4 hours at ambient temperature and at +4 °C for 8 days. The method is confirmed to be precise and accurate.

Conclusions:
In an OECD 414 oral developmental toxicity study , rats were dosed up to 1000 mg/kg/ day of sodium ferrocyanide via gavage, resulting in no treatment related effects and thus an NOAEL of ≥ 1000 mg/kg/day could be derived (both maternal and developmental).

Executive summary:

In a prenatal developmental toxicity study, pregnant rats were orally exposed to different dosages of sodium ferrocyanide up to 1000 mg/kg/day. The animals were exposed daily, from day 6 to (and including) day 15 of the pregnancy. Rats were sacrificed at day 20. Treatment with sodium ferrocyanide at 1000 mg/kg/day elicited occasional instances of post-dosing salivation in about half of the animals with the greatest incidence occuring after 4 days of dosing. Over the dosing period as a whole, water intake of all three groups was significantly higher than of the control value, the greatest difference being recorded at 1000 mg/kg. This effect was not considered adverse. No other treatment or dosage-related responses were observed in the dams and litter parameters appeared to be unaffected at all of the dosages investigated, as judged by in utero survival, foetal growth and morphological development. Although a small number of foetuses from the two highest dosages showed increased dilation of the renal pelvis/ ureter, this increase was marginally and not statistically significant and thus not considered to be an effect of treatment. This results in a maternal NOAEL of ≥1000 mg/kg/day and a developmental NOAEL of ≥1000 mg/kg/day.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
Performed according to GLP principles and OECD test guide line, although not according to current guideline. Some minor deviations are not expected to have influence on the study.
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information

In a prenatal developmental toxicity study, pregnant rats were orally exposed to different dosages of sodium ferrocyanide up to 1000 mg/kg/day. The animals were exposed daily, from day 6 to (and including) day 15 of the pregnancy. Rats were sacrificed at day 20. Treatment with sodium ferrocyanide at 1000 mg/kg/day elicited occasional instances of post-dosing salivation in about half of the animals with the greatest incidence occuring after 4 days of dosing. Over the dosing period as a whole, water intake of all three groups was significantly higher than of the control value, the greatest difference being recorded at 1000 mg/kg. This effect was not considered adverse. No other treatment or dosage-related responses were observed in the dams and litter parameters appeared to be unaffected at all of the dosages investigated, as judged by in utero survival, foetal growth and morphological development. Although a small number of foetuses from the two highest dosages showed increased dilation of the renal pelvis/ ureter, this increase was marginally and not statistically significant and thus not considered to be an effect of treatment. This results in a maternal NOAEL of ≥1000 mg/kg/day and a developmental NOAEL of ≥1000 mg/kg/day.

 

Based on the low potential for toxicity as derived from the toxicity data available and limited systemic exposure to Sodium ferrocyanide, no hazard and no risk is considered present for developmental toxicity. Data from a developmental toxicity study (OECD 414) in rat demonstrate the absence of developmental toxicity, and there are no scientific data indicating rabbit developmental toxicity data (in the absence of systemic toxicity data) being more relevant for human risk assessment than rat data. As Sodium ferrocyanide is a chemical without specific working mechanism it is unlikely that this substance will specifically demonstrate reproduction toxicity in the absence of any systemic toxicity. Based on these considerations, additional toxicity data on reproduction toxicity is considered not scientifically justified and the developmental toxicity study, second species (OECD 414) can be waived.

Justification for selection of Effect on developmental toxicity: via oral route:

One study available with Klimisch reliability 1.

Justification for classification or non-classification

Based on the available data, sodium ferrocyanide is not classified for reproduction toxicity according to CLP Regulation (No) EC 1272/2008.

Additional information