Ammonium iron(III) citrate

Endpoint summary

Description of key information

Acute oral toxicity: 

The acute oral toxicity of test chemical was assessed in various experimental studies.Based on the available key and supporting studies,it can be concluded that LD50 of test chemical is >2000 mg/kg bw. Thus, comparing the above annotations with the criteria of CLP regulation, it cannot be classified for acute oral toxicity.

 

Acute Inhalation toxicity:

the study does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size (The acute inhalation toxicity study need not be conducted because exposure to humans via inhalation route is not likely taking into account due to the low vapour pressure of the test chemical, which is reported to be 3.29e-09 Pa.( 2.4677e-11 mmHg). Thus, exposure to inhalable dust, mist and vapour of the chemical is highly unlikely. Therefore this study is considered for waiver. )

 

Acute Dermal toxicity:

Data available for the structurally and functionally similar test chemicals have been reviewed to determine the acute dermal toxicity of the test chemical. Based on the summarized,it can be concluded that theLD50 value of test chemical is >2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, test chemical cannot be classified for acute dermal toxicity.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Data is from experimental study report.

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Qualifier:

according to guideline

Guideline:

other: EEC Guidelines (B.1)

Principles of method if other than guideline:

The purpose of the study was to evaluate the acute oral toxicity of the test article in rats.

GLP compliance:

no

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Remarks:

Cri: CD (SD) BR

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Italia S.p.A. Via Indipendenza, 11 22050 CALCO (Lecco) Shipping slips No.s 6603 (October 3. 1997) and 6984 (October 17, 1997)
- Age at study initiation: no more than three months
- Weight at study initiation: Males: 217-241 2 Females: 200-219 g
- Fasting period before study:
- Housing: 5 animals/sex/cage in air-conditioned room.
- Diet (e.g. ad libitum): GLP 4RF21 top certificate pelleted diet produced by Charles River Italia's feed licencee Mucedola S.rl.. Settimo Milanese,ad libitum
- Water (e.g. ad libitum): from the municipal water main system, ad libitum
- Acclimation period: at least 5 days before the start of the test.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C ±2
- Humidity (%): 55% ± 10
- Air changes (per hr): about 20 / hour filtered on HEPA 99.97%
- Photoperiod (hrs dark / hrs light): 12 hour cycle (7 a.m. - 7 p.m. )
-Animal identification: by appropriately coloring different areas of the limbs. Cage card gave experiment number, dosage group, sex and date of administration.

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 2000 mg/kg

DOSAGE PREPARATION (if unusual): one group of 5 rats/sex, randomly selected, was administered a dosage of 2000 mg/kg (limit dose) of the test article. The volume of administration was 1.44 ml/kg. The density of the test article is 1.39 g/ml.

Doses:

2000 mg/kg

No. of animals per sex per dose:

5 males + 5 females treated at 2000 mg/kg

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:twice pre-trial (at randomization and on day 1 just before administration) and on days 3, 8 and 14. On day 1 the animals were weighed after a 16-hour fasting period.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights

Statistics:

no data

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No mortality was observed.

Mortality:

No deaths occurred in the treated animals of either sex. The LD50 was not calculated and it was considered higher than 2000 mg/kg.

Clinical signs:

other: No clinical changes were seen in the animals.

Gross pathology:

At the autopsy carried out at the end of the observation period, no appreciable macroscopic findings were evident in any treated rat.

Interpretation of results:

other: Not Classified

Conclusions:

No deaths or clinical signs occurred in the treated animals. Body weight growth of the treated animals was normal during the observation period. At the final killing no appreciable macroscopic findings were evident in any animal.In conclusion, the LD50 of the test article, when administered to rats as a single dose by oral route, was higher than 2000 mg/kg. At this dose the compound did not induce mortality or apparent signs of toxicity in the treated rats.

Executive summary:

An acute oral toxicity study of test chemical was conducted in Sprague Dawley Crl:CD(SD) BR rats 5 males and 5 females) in accordance with the OECD Guideline 401 (Acute Oral Toxicity) and EEC Guidelines (B.1).

 

The test article was administered undiluted as supplied at the dosage of 2000 mg/kg. The volume of administration was 1.44 ml/kg (the density of the test article is 1.39 g/ml).

 

All rats were treated after a 16-hour fasting period. The day of treatment was considered day 1 of the study. The animals were weighed twice before treatment (at randomization and on day 1 just before treatment) and on days 3, 8 and 14. They were clinically observed for 14 days following the treatment. On day 15 the rats were killed (fasted overnight) by excision of the femoral arteries after i.p. overdosage anesthesia with 5% sodium pentobarbital and were subjected to a thorough autopsy.

 

No deaths or clinical signs occurred in the treated animals. Body weight growth of the treated animals was normal during the observation period. At the final killing no appreciable macroscopic findings were evident in any animal.

 

In conclusion, the LD50 of the test article, when administered to rats as a single dose by oral route, was higher than 2000 mg/kg. At this dose the compound did not induce mortality or apparent signs of toxicity in the treated rats.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Data is Klimisch 1 and from experimental study report.

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

exposure considerations

Justification for data waiving:

the study does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size

Endpoint conclusion

Quality of whole database:

Waiver

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

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 the available information from various test chemicals.

Reason / purpose for cross-reference:

read-across: supporting information

Reason / purpose for cross-reference:

read-across: supporting information

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

Test type:

other: not specified

Limit test:

no

Species:

other: 2.rabbit 3. rat

Strain:

other: 2. New Zealand Albino 3. not specified

Sex:

male/female

Details on test animals or test system and environmental conditions:

2.
TEST ANIMALS
- Weight at study initiation: 2.30 to 2.86 kg
- Fasting period before study: Not fasted
- Housing: The animals were individually housed in metal cages elevated above the droppings
- Diet (e.g. ad libitum): Purina Rabbit Chow, ad libitium
- Water (e.g. ad libitum): Tap water, ad libitum

3.
not specified

Type of coverage:

other: 2. occlusive 3.dermal

Vehicle:

other: 2. 50%(w/w) isotonic saline 3.not specified

Details on dermal exposure:

2.
TEST SITE
- Area of exposure: intact and abraded skin of the trunk, free of hair
- % coverage: two or three centimeters longitudinally over the area of exposure.
- Type of wrap if used: A plastic binder was slipped onto each animal, which was then placed in a comfortable but immobilized position in a multiple animal holder. The binder was then fastened tightly to keep the preparation in close contact with the skin for 24 hours.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): At the end of the period of exposure, the binders were removed, the amount of unabsorbed material estimated, the skin reactions recorded, and the remaining test material wiped from the animals' bodies.
- Time after start of exposure:24 hours

3.
not specified

Duration of exposure:

2.
Single exposure for a period of 24 hours

3.
not specified

Doses:

2.
2000, 4000, 8000 mg/kg bw

3.
2000 mg/kg bw

No. of animals per sex per dose:

2.
Twelve rabbits, 6 animals per sex

3.
not specified

Control animals:

not specified

Details on study design:

2.
- Duration of observation period following administration: 14 days
- Necropsy of survivors performed: Yes, animals which succumbed were necropsied.
- Other examinations performed: The animals were observed for gross effects at regular intervals on the day of dosing and daily thereafter for 14 days.Following the observation period, all surviving animals were weighed, sacrificed and necropsied.

3.
not specified

Statistics:

not specified

Preliminary study:

not specified

Sex:

male/female

Dose descriptor:

LD50

Remarks:

2.

Effect level:

> 8 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: no mortality was observed.

Sex:

not specified

Dose descriptor:

LD50

Remarks:

3.

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No morality was observed

Mortality:

2.
No mortality at highest dose level.

3.
No morality was observed at dose 2000 mg/kg bw.

Clinical signs:

other: 2. Few systemic signs of toxicity were observed at any dose level. At all dose levels there was moderate to severe erythema with chemical burns and/or blanching especially along abrasions The intensity increased with the dose level.This was followed at 7

Gross pathology:

2.
Gross necropsy of animal which succumbed showed the skin to have severe erythema of sides and ventrum with severe congestion appearance of subcutaneous tissue. It has the appearance of chemical burns. The stomach was blanched with severe erosion of the mucosal surface. The small intestines showed severe scattered congestion.
Gross necropsy of the animals sacrificed at 14 days showed one animal at 8000 mg./kg with an approximate 90% loss of fat tissue. Other organs and tissues in all animals were not remarkable.

3.
not specified

Other findings:

not specified

Interpretation of results:

other: not classified

Conclusions:

According to CLP regulation,the test chemical cannot be classified for acute dermal toxicity, as the LD50 value is >2000 mg/kg bw.

Executive summary:

Data available for the test chemical has been reviewed to determine the acute dermal toxicity of the test chemical.The studies are as mentioned below:

The acute dermal toxicity study was conducted by using the given test chemical in 12 New Zealand Albino rabbits, six males and six females, were used to evaluate the potential toxicity following dermal application by a single exposure for a period of 24 hours at the concentrations of 2000, 4000, 8000 mg/kg bw.Prior to exposure, the animals were prepared by clipping the skin of the trunk free of hair, and only those animals without observable skin defects or irritation were used. One-half of the animals in each group were further prepared by making epidermal abrasions every two or three centimeters longitudinally over the area of exposure. Abrasions were sufficiently deep to penetrate the stratum corneum but not to disturb the derma. A plastic binder was slipped onto each animal, which was then placed in a comfortable but immobilized position in a multiple animal holder. Test chemical in the form of a 50% w/w suspension in isotonic saline, was introduced under the plastic binder; the binder was then fastened tightly to keep the preparation in close contact with the skin for 24 hours. At the end of the period of exposure, the binders were removed, the amount of unabsorbed material estimated, the skin reactions recorded, and the remaining test material wiped from the animals' bodies. The animals were housed in their respective cages. The animals were observed for gross effects at regular intervals on the day of dosing and daily thereafter for 14 days. Animals which succumbed were necropsied. Following the observation period, all surviving animals were weighed, sacrificed and necropsied.No mortality at highest dose level. Few systemic signs of toxicity were observed at any dose level. At all dose levels there was moderate to severe erythema with chemical burns and/or blanching especially along abrasions.The intensity increased with the dose level.This was followed at 7 and 14 days with desquamation and drying. At 2000 mg/kg bw, two animals exhibited a generalized weakness for 24 to 48 hours after dosing. At 8000 mg/kg bw, one animal exhibited generalized weakness for 72 hours and three animals were observed to be thin after 72 hours and until day 10 of the study. Final body weight of surviving animals at termination showed two with intact skin and one with abraded skin at 2000 mg/kg bw to have significant (100% or greater) weight gains. At 4000 mg/kg bw, one (abraded skin) has a significant weight gain. Other surviving animals had weight gains or losses which were less than 10%. Gross necropsy of animal which succumbed showed the skin to have severe erythema of sides and ventrum with severe congestion appearance of subcutaneous tissue. It has the appearance of chemical burns. The stomach was blanched with severe erosion of the mucosal surface. The small intestines showed severe scattered congestion. Gross necropsy of the animals sacrificed at 14 days showed one animal at 8000 mg./kg with an approximate 90% loss of fat tissue. Other organs and tissues in all animals were not remarkable.Under the condition of the study, the acute dermal toxicity dose (LD50) was considered to be >8000 mg/kg, when 12 New Zealand Albino rabbits, six males and six females, were treated with the given test chemical following dermal application by a single exposure for a period of 24 hours.

In another acute dermal toxicity study, rats were treated with the given test chemical at the concentration of 2000 mg/kg bw by dermal application.Animals were observed for mortality. No morality was observed at dose 2000 mg/kg bw.Hence, the LD50 value was considered to be >2000 mg/kg bw, when rats were treated with the given test chemical by dermal application according to OECD Guideline 402 (Acute Dermal Toxicity).

Thus, based on the above summarised studies of test chemical, it can be concluded that LD50 value is greater than 2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation,test chemical cannot be classified for acute dermal toxicity.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Data is Klimisch 2 and from prediction report.

Additional information

Acute oral toxicity: 

In different studies, the given test chemical has been investigated for acute oral toxicity. The studies are summarized as below –

 

An acute oral toxicity study of test chemical was conducted in Sprague Dawley Crl:CD(SD) BR rats 5 males and 5 females) in accordance with the OECD Guideline 401 (Acute Oral Toxicity) and EEC Guidelines (B.1). The test article was administered undiluted as supplied at the dosage of 2000 mg/kg. The volume of administration was 1.44 ml/kg (the density of the test article is 1.39 g/ml). All rats were treated after a 16-hour fasting period. The day of treatment was considered day 1 of the study. The animals were weighed twice before treatment (at randomization and on day 1 just before treatment) and on days 3, 8 and 14. They were clinically observed for 14 days following the treatment. On day 15 the rats were killed (fasted overnight) by excision of the femoral arteries after i.p. overdosage anesthesia with 5% sodium pentobarbital and were subjected to a thorough autopsy. No deaths or clinical signs occurred in the treated animals. Body weight growth of the treated animals was normal during the observation period. At the final killing no appreciable macroscopic findings were evident in any animal. In conclusion, the LD50 of the test article, when administered to rats as a single dose by oral route, was higher than 2000 mg/kg. At this dose the compound did not induce mortality or apparent signs of toxicity in the treated rats.

 

The above result was supported by another acute oral toxicity study by using the given test chemical in groups of 5 male and female rabbits. The animals (overnight fasted) weighed between 2-3 kg were administered the given test chemical via oral gavage route. Necropsy of survivors performed (Necropsies with examination of all major organs followed as quickly as possible after death. Sections were cut of parts of the stomach, intestine, liver, and kidneys, and were stained for iron as well as with haematoxylin and eosin). Haematological examination was performed included differential and total counts of red and white cells and estimates of haemoglobin levels. Samples of urine were collected daily for 3 days. Slopes of the regression curves, and the errors of estimating the LD50 were obtained by the graphical method of de Beer (1945). 50% mortality was observed at dose 2800 mg/kg bw. Within a few minutes of receiving a toxic dose of test substance the rabbits became prostrated. They lay on their stomachs with limbs extended, their respiratory rate increased, micturition often occurred, and reflex movement of the hind legs was considerably retarded. Coma followed, with shallow breathing and gradual disappearance of reflex movements. The animal either died two to six hours after receiving the dose (according to its size), sometimes following convulsions, or recovered. At necropsy the stomachs usually showed congested areas with shedding of mucosa, especially at the greater curvature.The amount of damage was to a large extent determined by the size of dose: the smaller toxic doses caused only very slight damage to the stomach wall. The small intestines generally showed much hyperaemia in their upper regions, but here also haemorrhages were seen only after very large doses. Surviving animals showed no evidence of kidney damage, the urine being invariably devoid of abnormal constituents. There were no departures from normal blood counts. Recovery of surviving animals was usually rapid. Changes in the histology of stomach, liver, and kidney in rabbits killed by the iron compounds were very small, and appeared insufficient to account for the death of the animals. In the stomach there was only slight necrosis of the superficial layer of the villi and deposits of iron on the mucous membrane (occasionally also in the endothelium of the smaller blood vessels). There were deposits of iron in the bile ducts, but only slight hydropic changes in the liver. In rabbits surviving and killed three days after dosing we observed small foci of fatty degeneration, and necrosis was present in the peripheral parts of the lobuli, with deposits of iron, mainly in the Kupffer cells. The surviving animals appeared to be completely restored to normal health and activity after a few days. Under the condition of the study, the lethal concentration (LD50) value for acute oral toxicity test was considered to be 2800 mg/kg bw, when groups of 5 male and female rabbits were treated with the given test chemical orally via gavage.

 

Th overall result was further supported by another acute oral toxicity study of test chemical conducted in mice at the dose concentration of 5000 mg/kg bw. Animals were observed for mortality. 50% mortality was observed at dose 5000 mg/kg bw. Hence, the lethal concentration (LD50) value for acute oral toxicity test was considered to be 5000 mg/kg bw, when mice were treated with the given test chemical orally.

 

Thus, based on the above summarized studies of test chemical, it can be concluded that LD50 value is greater than 2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, test chemical cannot be classified for acute oral toxicity.

 

Acute inhalation toxicity:

the study does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size (The acute inhalation toxicity study need not be conducted because exposure to humans via inhalation route is not likely taking into account due to the low vapour pressure of the test chemical, which is reported to be 3.29e-09 Pa.( 2.4677e-11 mmHg).Thus, exposure to inhalable dust, mist and vapour of the chemical is highly unlikely. Therefore this study is considered for waiver. )

 

 

Acute Dermal toxicity:

Data available for the test chemical has been reviewed to determine the acute dermal toxicity of the test chemical. The studies are as mentioned below:

 

The acute dermal toxicity study was conducted by using the given test chemical in 12 New Zealand Albino rabbits, six males and six females, were used to evaluate the potential toxicity following dermal application by a single exposure for a period of 24 hours at the concentrations of 2000, 4000, 8000 mg/kg bw.Prior to exposure, the animals were prepared by clipping the skin of the trunk free of hair, and only those animals without observable skin defects or irritation were used. One-half of the animals in each group were further prepared by making epidermal abrasions every two or three centimeters longitudinally over the area of exposure. Abrasions were sufficiently deep to penetrate the stratum corneum but not to disturb the derma. A plastic binder was slipped onto each animal, which was then placed in a comfortable but immobilized position in a multiple animal holder. Test chemical in the form of a 50% w/w suspension in isotonic saline, was introduced under the plastic binder; the binder was then fastened tightly to keep the preparation in close contact with the skin for 24 hours. At the end of the period of exposure, the binders were removed, the amount of unabsorbed material estimated, the skin reactions recorded, and the remaining test material wiped from the animals' bodies. The animals were housed in their respective cages. The animals were observed for gross effects at regular intervals on the day of dosing and daily thereafter for 14 days. Animals which succumbed were necropsied. Following the observation period, all surviving animals were weighed, sacrificed and necropsied.No mortality at highest dose level. Few systemic signs of toxicity were observed at any dose level. At all dose levels there was moderate to severe erythema with chemical burns and/or blanching especially along abrasions.The intensity increased with the dose level.This was followed at 7 and 14 days with desquamation and drying. At 2000 mg/kg bw, two animals exhibited a generalized weakness for 24 to 48 hours after dosing. At 8000 mg/kg bw, one animal exhibited generalized weakness for 72 hours and three animals were observed to be thin after 72 hours and until day 10 of the study. Final body weight of surviving animals at termination showed two with intact skin and one with abraded skin at 2000 mg/kg bw to have significant (100% or greater) weight gains. At 4000 mg/kg bw, one (abraded skin) has a significant weight gain. Other surviving animals had weight gains or losses which were less than 10%. Gross necropsy of animal which succumbed showed the skin to have severe erythema of sides and ventrum with severe congestion appearance of subcutaneous tissue. It has the appearance of chemical burns. The stomach was blanched with severe erosion of the mucosal surface. The small intestines showed severe scattered congestion. Gross necropsy of the animals sacrificed at 14 days showed one animal at 8000 mg./kg with an approximate 90% loss of fat tissue. Other organs and tissues in all animals were not remarkable.Under the condition of the study, the acute dermal toxicity dose (LD50) was considered to be >8000 mg/kg, when 12 New Zealand Albino rabbits, six males and six females, were treated with the given test chemical following dermal application by a single exposure for a period of 24 hours.

 

In another acute dermal toxicity study, rats were treated with the given test chemical at the concentration of 2000 mg/kg bw by dermal application.Animals were observed for mortality. No mortality was observed at dose 2000 mg/kg bw.Hence, the LD50 value was considered to be >2000 mg/kg bw, when rats were treated with the given test chemical by dermal application according to OECD Guideline 402 (Acute Dermal Toxicity).

 

Thus, based on the above summarized studies of test chemical, it can be concluded that LD50 value is greater than 2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, test chemical cannot be classified for acute dermal toxicity.

 

Justification for classification or non-classification

Based on the above studies on test chemical, it can be concluded that LD50 value is >2000 mg/kg bw for acute oral and acute dermal toxicity .Thus, comparing these values with the criteria of CLP regulation, the given test chemical cannot be classified for acute oral and dermal toxicity. For Acute inhalation toxicity waiver was added so, not possible to classify.