Ammonium iron(III) citrate

Endpoint summary

Description of key information

Repeated dose toxicity: Oral

The No Observed Adverse Effect Level (NOAEL) of the test chemical in the rat via oral route of exposure is considered to be in the dose range of 601 -1200 mg/kg bw/day.4 mg/kg body weight in male and female animals.

Repeated dose toxicity: Inhalation

a short-term toxicity study does not need to be conducted because exposure of humans via inhalation in production and/or use is not likely as based on the provided thorough and rigorous exposure assessment.Test chemical has very low vapor pressure of 3.29e-09 Pa.( 2.4677e-11 mmHg). Also, the test chemical has a particle size distribution of 52-147 micron, so the potential for the generation of inhalable vapours is very low. Also the normal conditions of use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalatory route will be highly unlikely and therefore this end point for repeated dose toxicity by inhalation route is considered for waiver.

Repeated dose toxicity: Dermal

a short-term toxicity study does not need to be conducted because exposure of humans via the dermal route in production and/or use is not likely as based on the provided thorough and rigorous exposure assessment. The acute dermal toxicity value for Ferric ammonium citrate (as provided in section 7.2.3) is >2000 mg/kg body weight. The substance was also found to be not sensitizing to the skin. Based on these considerations, the end point for repeated dermal toxicity is considered as waiver.

Key value for chemical safety assessment

Toxic effect type:

dose-dependent

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

repeated dose toxicity: oral, other

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

Experimental data from various test chemicals

Justification for type of information:

Data for the target chemical is summarized based on data from various test chemicals.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

WoE for the target CAS is summarized based on data from various test chemicals.

GLP compliance:

not specified

Limit test:

no

Species:

rat

Strain:

other: 2.not specified 3.Fischer 344/DuCrj

Details on species / strain selection:

No data

Sex:

male/female

Details on test animals or test system and environmental conditions:

2.No data

3.
TEST ANIMALS
- Source: Charles River Japan (Yokohama, Japan)
- Age at study initiation: 5 weeks old when purchased
- Weight at study initiation: No data
- Fasting period before study:no data
- Housing: The animals were housed 3–4 rats per polycarbonate cage with
soft chip bedding. The cages and chip bedding were exchanged twice a week.
- Diet (e.g. ad libitum): Basal diet ad libitum
- Water (e.g. ad libitum): Tap water ad libitum
- Acclimation period: No data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23 ± 2°C
- Humidity (%): 55 ± 5%
- Air changes (per hr): 18 times/hr
- Photoperiod (hrs dark / hrs light): 12 hr light dark cycle

IN-LIFE DATES: From: To: No data

Route of administration:

other: 2. oral: feed 3.oral: feed

Details on route of administration:

No data

Vehicle:

other: 3.Feed

Details on oral exposure:

2. Details on oral exposure
PREPARATION OF DOSING SOLUTIONS: The test chemical was mixed with diet at dose level of 3 or 5% (1200 or 2000 mg/Kg bw/day)

DIET PREPARATION
- Rate of preparation of diet (frequency): No data
- Mixing appropriate amounts with (Type of food): No data
- Storage temperature of food: No data

VEHICLE
- Justification for use and choice of vehicle (if other than water): Feed
- Concentration in vehicle: 3 or 5% (1200 or 2000 mg/Kg bw/day)
- Amount of vehicle (if gavage): No data
- Lot/batch no. (if required): No data
- Purity: No data

3.
PREPARATION OF DOSING SOLUTIONS: The test chemical was mixed with diet at dose level of 0%, 0.25%, 1.0%, and 4.0% (w/w) (0, 125, 500 or 2000 mg/Kg bw/day)

DIET PREPARATION
- Rate of preparation of diet (frequency): No data
- Mixing appropriate amounts with (Type of food): No data
- Storage temperature of food: No data

VEHICLE
- Justification for use and choice of vehicle (if other than water): Basal diet
- Concentration in vehicle: 0%, 0.25%, 1.0%, and 4.0% (w/w) (0, 125, 500 or 2000 mg/Kg bw/day)
- Amount of vehicle (if gavage): No data
- Lot/batch no. (if required): No data
- Purity: No data

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

No data

Duration of treatment / exposure:

2. 2 years
3. 13 week

Frequency of treatment:

2. Daily
3. Daily

Remarks:

2.
3 or 5% (1200 or 2000 mg/Kg bw/day)

Remarks:

3.
0%, 0.25%, 1.0%, and 4.0% (w/w) (Males: 0, 143.9, 595.9 or 2834.7 mg/Kg bw/day; Females: 0, 147.7, 601.4 or 2845.6 mg/Kg bw/day)

No. of animals per sex per dose:

2.No data

3.
Total: 40 males and 40 females

0 mg/Kg bw/day: 10 males and 10 females

143.9 mg/Kg bw/day for males and 147.7 mg/Kg bw for females: 10 males and 10 females

595.9 mg/Kg bw/day for males and 601.4 mg/Kg bw for females: 10 males and 10 females

2834.7 mg/Kg bw/day for males and 2845.6 mg/Kg bw/day: 10 males and 10 females

Control animals:

yes, concurrent vehicle

not specified

Details on study design:

2. No data
3.
- Dose selection rationale: In a preliminary 4-week palatability study of test chemical with administration at the highest dose of 5.0% (w/w), significant decrease of body weight gain (>20% compared to the controls) was observed in males and females (data not shown). From these results, doses of test chemical were decided as 0%, 0.25%, 1.0%, and 4.0% (w/w) in both sexes for the present 13-week toxicity study, with diet replacement every week.
- Rationale for animal assignment (if not random): At the beginning of the experiment, the animals were randomly allocated to four groups of 10 male and 10 female rats each, based on their body weights measured just before starting the test chemical treatment.
- Rationale for selecting satellite groups: No data
- Post-exposure recovery period in satellite groups: No data
- Section schedule rationale (if not random): No data

Positive control:

2. No data
3. No data

Observations and examinations performed and frequency:

2.
CAGE SIDE OBSERVATIONS: No data
- Time schedule: No data
- Cage side observations checked in table [No.?] were included. No data

DETAILED CLINICAL OBSERVATIONS: No data
- Time schedule: No data

BODY WEIGHT: Yes
- Time schedule for examinations: No data

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): No data
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No data

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
- Time schedule for examinations: No data

OPHTHALMOSCOPIC EXAMINATION: No data
- Time schedule for examinations:
- Dose groups that were examined: No data

HAEMATOLOGY: No data
- Time schedule for collection of blood: No data
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: No data
- Parameters checked in table [No.?] were examined. No data

CLINICAL CHEMISTRY: No data
- Time schedule for collection of blood: No data
- Animals fasted: No data
- How many animals: No data
- Parameters checked in table [No.?] were examined. No data

URINALYSIS: No data
- Time schedule for collection of urine: No data
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters checked in table [No.?] were examined. No data

NEUROBEHAVIOURAL EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data
- Battery of functions tested: sensory activity / grip strength / motor activity / other: No data

OTHER: No data

3.
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily
- Cage side observations checked in table [No.?] were included. General condition and mortality

DETAILED CLINICAL OBSERVATIONS: No data
- Time schedule: No data

BODY WEIGHT: Yes
- Time schedule for examinations: body weights were measured once a week

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes, The amounts of supplied and residual diet were weighed weekly in order to calculate the average daily food intake and test chemical intake through the entire treatment period.
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

FOOD EFFICIENCY: No data
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No data

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
- Time schedule for examinations: No data

OPHTHALMOSCOPIC EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data

HAEMATOLOGY: Yes
- Time schedule for collection of blood: At the completion of treatment
- Anaesthetic used for blood collection: No data
- Animals fasted: Yes, the animals were fasted overnight
- How many animals: All animals
- Parameters checked in table [No.?] were examined. White blood cell count (WBC), red blood cell count (RBC), hemoglobin concentration (HGB), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and platelet count (PLT), counting of erythroblasts (Ebl) and differential
leukocytes

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: At the completion of treatment
- Animals fasted: Yes, the animals were fasted overnight
- How many animals: All animals
- Parameters checked in table [No.?] were examined. Total protein (TP), albumin/globulin ratio (A/G), albumin (Alb), total bilirubin (Bil), glucose, triglyceride (TG), total cholesterol (T-Chol), urea nitrogen (BUN), creatinine (Cre), sodium (Na), chlorine (Cl), potassium (K), calcium (Ca), inorganic phosphorus (IP), aspartate aminotransferase
(AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), γ-glutamyl transpeptidase (γ-GTP), transferrin, and iron (Fe).

URINALYSIS: No data
- Time schedule for collection of urine: No data
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters checked in table [No.?] were examined. No data

NEUROBEHAVIOURAL EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data
- Battery of functions tested: sensory activity / grip strength / motor activity / other: No data

OTHER: No data

Sacrifice and pathology:

2.
GROSS PATHOLOGY: No data

HISTOPATHOLOGY: Yes, tissues were observed microscopically

3.
GROSS PATHOLOGY: Yes, Complete necropsy was performed for all animals, and the brain, thymus, lungs, heart, spleen, liver, adrenal glands, kidneys, and gonads were weighed.

HISTOPATHOLOGY: Yes, the brain, thymus, lungs, heart, spleen, liver, adrenal glands, kidneys, and gonads, skin, mammary gland, sternum with marrow, femur with marrow, mandibular and mesenteric lymph nodes, salivary glands, aorta, trachea, tongue, esophagus, stomach, small and large intestines, pancreas, urinary bladder, epididymides, seminal vesicles, prostate gland, bulbourethral glands, uterus, vagina, pituitary gland, thyroid glands, parathyroid glands, spinal cord with vertebrae, trigeminal nerve, sciatic nerve, harderian glands, femoral skeletal muscle, and nasal cavity were stained with hematoxylin and eosin for histopathological examination. The testes and eyes were fixed in Bouin’s fixative and Davidson’s solution, respectively. Bony tissues including the nasal cavity, vertebrae, sternum, and femur were decalcified with a mixture of 10% formic acid and 10% buffered formalin for up to 2 weeks. Histopathological assessment was first performed on all tissues of the control and highest dose groups. If a chemical treatment-related change appeared at the highest dose, the relevant tissues from the lower dose groups then also underwent examination.

Other examinations:

No data

Statistics:

2. No data
3. Variance in data for body and organ weights, as well as the results of hematology and serum biochemistry, was checked by Bartlett’s test for homogeneity. When the data were homogeneous, one-way analysis of variance (ANOVA) was conducted. In heterogeneous cases, the Kruskal–Wallis test was applied. When statistically significant differences were indicated, the Dunnett’s or non-parametric Dunnett’s multiple test were employed for comparisons between control and other groups. For incidences of histopathological findings, the Fisher’s exact probability test was applied. P values of < 0.05 were considered to be statistically significant.

Clinical signs:

not specified

Description (incidence and severity):

3.All animals were without any clinical signs

Mortality:

no mortality observed

Description (incidence):

3.No deaths were noted

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

2.
A slight decrease in growth was noted at 2000 mg/Kg bw/day

3.
Body weight gain of both sexes was significantly reduced with 4.0% (2834.7 mg/Kg bw/day for males and 2845.6 mg/Kg bw/day for females)test chemical treatment compared to the controls from the 1st week until the end of the experiment except in the 9th and 12th weeks in females. Comparing with the control groups, the average bodyweight gain during the experiment in 2834.7 mg/Kg bw/day males and 2845.6 mg/Kg bw/day females were reduced by 13.4% and 5.9%, respectively.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

2.
Food consumption was lower in 2000 mg/Kg bw/day dosed animals

3.
Food intake: Daily food intake of 2834.7 mg/Kg bw/day males and 2845.6 mg/Kg bw/day females was slightly higher than in the other groups. Since there was no avoidance of the experimental diet, average intakes of ferric citrate per body weight was dose-dependent

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

3.Significant decreases of RBC and lymphocytes and increases of PLT and eosinophils were detected in 2834.7 mg/Kg bw/day males and 2845.6 mg/Kg bw/day females. In males, significant increases of MCV and MCH were observed in the 1.0% and 4.0% groups and with 4.0%, respectively.

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

3.
An increase of serum Fe levels and decreases of TP and transferrin were noted in 2834.7 mg/Kg bw/day males and 2845.6 mg/Kg bw/day females. Significant decrease of transferrin was also detected in 601.4 mg/Kg bw/day females. In males, increase of Na and decreases of AST and ALT were observed in the 2834.7 mg/Kg bw/day group. In females, significant increase of IP and decreases of Alb and Cl were found in the 2845.6 mg/Kg bw/day group. Increase of ALT in 595.9 mg/Kg bw/day males and decrease of K levels in 147.7 mg/Kg bw/day females were also detected, without any dose-dependence.

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

3.
In males, absolute heart weights were decreased in the 4.0% group (2834.7 mg/Kg bw/day) compared to the controls. In addition, relativeweights of brain, spleen, adrenals, kidneys, and testes were significantly increased in 2834.7 mg/Kg bw/day males. In females, decrease of absolute and relative heart weights, increase of absolute and relative spleen weights and increase of relative liver weights were observed in the 2845.6 mg/Kg bw/day group. Absolute weights of liver were also increased in 147.7 mg/Kg bw/day females without dose-dependence.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

3.
At necropsy, one female rat in the control group had severe enlargement of the right kidney due to a histologically characteristic nephroblastoma. Although nephroblastoma is rare in F344 rats, it was considered to be an incidental case, because the rat was one of the untreated controls and no other animals had similar lesions. The animal was therefore excluded from the effective numbers for the hematology, serum biochemistry, organ weight and histopathology.

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

2.
No tissue abnormalities were found on examination of the major organs.

Histopathological findings: neoplastic:

effects observed, treatment-related

Description (incidence and severity):

3.
Treatment related pathological findings were noted in the colon, mesenteric lymph node, spleen and bone marrow. There was moderate to severe colitis in all 2834.7 mg/Kg bw/day males and 2845.6 mg/Kg bw/day females. In the affected colons, numbers of eosinophils as well as neutrophils and macrophages had infiltrated into the lamina propria and submucosa. Absorptive epithelial cells of colonic mucosa were replaced by hyperplastic mucous cells with frequent mitotic figures. Eosinophilic infiltration was also observed occasionally in the submucosa of stomach, duodenum and cecum in 4.0% males and females with a lower incidence than in the colon. Mesenteric lymph nodes of all animals of 2834.7 mg/Kg bw/day males and 2845.6 mg/Kg bw/day females were macroscopically enlarged and histologically showed dilated sinuses featuring aggregation of plasmacytes and hemosiderin deposits. Infiltration of eosinophils was noted not only in the sinuses of mesenteric lymph nodes but also in the surrounding adipose tissue. In spleens of 2834.7 mg/Kg bw/day males and 2845.6 mg/Kg bw/day females, the degree of hemosiderin deposition was higher than in control groups. Hematopoietic cells in the bone marrow were increased in 2834.7 mg/Kg bw/day males and 2845.6 mg/Kg bw/day females. Although several lesions in other organs were sporadically detected, no significant treatment dependent alteration in their incidences was apparent.

Other effects:

not specified

Dose descriptor:

NOAEL

Remarks:

2.

Effect level:

1 200 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male

Basis for effect level:

body weight and weight gain

food consumption and compound intake

histopathology: non-neoplastic

Remarks on result:

other: No toxic effects were observed

Dose descriptor:

NOAEL

Remarks:

3.

Effect level:

601.4 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

female

Basis for effect level:

body weight and weight gain

clinical biochemistry

clinical signs

food consumption and compound intake

gross pathology

haematology

histopathology: non-neoplastic

mortality

organ weights and organ / body weight ratios

Remarks on result:

other: No toxic effects were observed

Dose descriptor:

NOAEL

Remarks:

3.

Effect level:

595.9 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male

Basis for effect level:

body weight and weight gain

clinical biochemistry

clinical signs

food consumption and compound intake

gross pathology

haematology

histopathology: non-neoplastic

mortality

organ weights and organ / body weight ratios

Remarks on result:

other: No toxic effects were observed

Critical effects observed:

not specified

Conclusions:

The No Observed Adverse Effect Level (NOAEL) for test substance was considered to be in a dose range of 601-1200 mg/kg bw/day for males and female in the 2 years toxicity study performed.

Executive summary:

The data available for the test chemical was reviewed to determine the toxic nature of test chemical when repeatedly exposed by oral route. The study is as mentioned below:

Chronic toxicity study was performed to determine the toxic nature of test substance. The study was performed using male rats at dose levels of 3 or 5% for 2 years. The animals were dosed by the feed route of exposure. During the study period, the animals were observed for changes in body weight and food consumption and the major organs were subjected to histopathology. A slight decrease in growth and food consumption was observed in the top-dose group of 5%), but no tissue abnormalities were found on examination of the major organs. Based on the observations made, the No Observed Adverse Effect Level (NOAEL) for test substance is considered to be 3.0% (1200 mg/kg bw/day for males) in the 2 years toxicity study performed.

 

In another 13 weeks subchronic toxicity study which was performed to determine the toxic nature of test substance . The study was performed using 40 male and 40 female F344/DuCrj rats. The test chemical was mixed with basal diet and used at dose level of 0%, 0.25%, 1.0%, and 4.0% (w/w) (Males: 0, 143.9, 595.9 or 2834.7 mg/Kg bw/day; Females: 0, 147.7, 601.4 or 2845.6 mg/Kg bw/day) for 13 weeks. During the study period, the animals were observed for general condition, mortality, changes in body weight and food intake, hemotology, clinical chemistry and were subjected to gross and histopathology.All animals were without any clinical signs and no deaths were reported. Reduction of body weight gain was noted in 2834.7 mg/Kg bw/day males and 2845.6 mg/Kg bw/day females.On hematology assessment, decreases of red blood cells and lymphocytes and increases of platelets and eosinophils were noted in 2834.7 mg/Kg bw/day males and 2845.6 mg/Kg bw/day females. Serum biochemistry demonstrated increased iron and decreased total protein and transferrin in both sexes treated with 2834.7 mg/Kg bw/day and 2845.6 mg/Kg bw/day ferric citrate respectively. In addition, an increase of serum inorganic phosphorus levels was noted in 2845.6 mg/kg bw females. Regarding organ weights, an increase of relative spleen weights was detected in 2834.7 mg/Kg bw/day males and 2845.6 mg/Kg bw/day females and a decrease of absolute and relative heart weights in 2845.6 mg/kg bw/day females. On histopathological assessment, colitis with infiltration of eosinophils and hyperplasia of mucosal epithelium, eosinophilic infiltration in mesenteric lymph nodes, and increased hemosiderosis in spleen were observed as treatment-related toxicological changes in 2834.7 mg/Kg bw/day males and 2845.6 mg/Kg bw/day females. Based on the observations made, the No Observed Adverse Effect Level (NOAEL) for test substance is considered to be 1.0% (595.9 mg/kg bw/day for males and 601.4 mg/kg bw/day for females) in the 13 weeks study performed.

Based on the above detailed studies available, the No Observed Adverse Effect Level (NOAEL) of the test chemical in the rat via oral route of exposure is considered to be in the dose range of 601.4 -1200 mg/kg body weight in male and female animals.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 200 mg/kg bw/day

Study duration:

chronic

Species:

rat

Quality of whole database:

Data is Klimisch 2 and from prediction report.

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records

Reference

Endpoint:

repeated dose toxicity: inhalation, other

Data waiving:

exposure considerations

Justification for data waiving:

a short-term toxicity study does not need to be conducted because exposure of humans via inhalation in production and/or use is not likely as based on the provided thorough and rigorous exposure assessment

Endpoint conclusion

Quality of whole database:

Waiver

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records

Reference

Endpoint:

repeated dose toxicity: dermal, other

Data waiving:

exposure considerations

Justification for data waiving:

a short-term toxicity study does not need to be conducted because exposure of humans via the dermal route in production and/or use is not likely as based on the provided thorough and rigorous exposure assessment

Endpoint conclusion

Quality of whole database:

Waiver

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Repeated dose toxicity: Oral

The data available for the test chemical was reviewed to determine the toxic nature of test chemical when repeatedly exposed by oral route. The study is as mentioned below:

Chronic toxicity study was performed to determine the toxic nature of test substance. The study was performed using male rats at dose levels of 3 or 5% for 2 years. The animals were dosed by the feed route of exposure. During the study period, the animals were observed for changes in body weight and food consumption and the major organs were subjected to histopathology. A slight decrease in growth and food consumption was observed in the top-dose group of 5%), but no tissue abnormalities were found on examination of the major organs. Based on the observations made, the No Observed Adverse Effect Level (NOAEL) for test substance is considered to be 3.0% (1200 mg/kg bw/day for males) in the 2 years toxicity study performed.

In another 13 weeks subchronic toxicity study which was performed to determine the toxic nature of test substance . The study was performed using 40 male and 40 female F344/DuCrj rats. The test chemical was mixed with basal diet and used at dose level of 0%, 0.25%, 1.0%, and 4.0% (w/w) (Males: 0, 143.9, 595.9 or 2834.7 mg/Kg bw/day; Females: 0, 147.7, 601.4 or 2845.6 mg/Kg bw/day) for 13 weeks. During the study period, the animals were observed for general condition, mortality, changes in body weight and food intake, hemotology, clinical chemistry and were subjected to gross and histopathology.All animals were without any clinical signs and no deaths were reported. Reduction of body weight gain was noted in 2834.7 mg/Kg bw/day males and 2845.6 mg/Kg bw/day females.On hematology assessment, decreases of red blood cells and lymphocytes and increases of platelets and eosinophils were noted in 2834.7 mg/Kg bw/day males and 2845.6 mg/Kg bw/day females. Serum biochemistry demonstrated increased iron and decreased total protein and transferrin in both sexes treated with 2834.7 mg/Kg bw/day and 2845.6 mg/Kg bw/day ferric citrate respectively. In addition, an increase of serum inorganic phosphorus levels was noted in 2845.6 mg/kg bw females. Regarding organ weights, an increase of relative spleen weights was detected in 2834.7 mg/Kg bw/day males and 2845.6 mg/Kg bw/day females and a decrease of absolute and relative heart weights in 2845.6 mg/kg bw/day females. On histopathological assessment, colitis with infiltration of eosinophils and hyperplasia of mucosal epithelium, eosinophilic infiltration in mesenteric lymph nodes, and increased hemosiderosis in spleen were observed as treatment-related toxicological changes in 2834.7 mg/Kg bw/day males and 2845.6 mg/Kg bw/day females. Based on the observations made, the No Observed Adverse Effect Level (NOAEL) for test substance is considered to be 1.0% (595.9 mg/kg bw/day for males and 601.4 mg/kg bw/day for females) in the 13 weeks study performed.

Based on the above detailed studies available, the No Observed Adverse Effect Level (NOAEL) of the test chemical in the rat via oral route of exposure is considered to be in the dose range of 601 -1200 mg/kg bw/day.4 mg/kg body weight in male and female animals.

Repeated dose toxicity: Inhalation

a short-term toxicity study does not need to be conducted because exposure of humans via inhalation in production and/or use is not likely as based on the provided thorough and rigorous exposure assessment.Test chemical has very low vapor pressure of 3.29e-09 Pa.( 2.4677e-11 mmHg). Also, the test chemical has a particle size distribution of 52-147 micron, so the potential for the generation of inhalable vapours is very low. Also the normal conditions of use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalatory route will be highly unlikely and therefore this end point for repeated dose toxicity by inhalation route is considered for waiver.

Repeated dose toxicity: Dermal

a short-term toxicity study does not need to be conducted because exposure of humans via the dermal route in production and/or use is not likely as based on the provided thorough and rigorous exposure assessment. The acute dermal toxicity value for Ferric ammonium citrate (as provided in section 7.2.3) is >2000 mg/kg body weight. The substance was also found to be not sensitizing to the skin. Based on these considerations, the end point for repeated dermal toxicity is considered as waiver.

Justification for classification or non-classification

Considering the above discussion and applying the weight of evidence approach with no toxic effects noted, the test chemical is not likely to be toxic as per the criteria mentioned in CLP regulation.