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Administrative data

acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2008-07-15 to 2008-10-24
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

Data source

Reference Type:
study report
Report date:

Materials and methods

Test guidelineopen allclose all
according to guideline
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
changes in air humidity
according to guideline
EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)
changes in air humidity
Principles of method if other than guideline:
Few times during experiment the relative air humidity was above 70%. It did not influence study course and results. No other deviations from guideline / method were stated during the entire experiment.
GLP compliance:
yes (incl. QA statement)
Test type:
fixed dose procedure
Limit test:

Test material

Constituent 1
Chemical structure
Reference substance name:
iron(3+) sodium 2-({2-[(carboxylatomethyl)[(2-oxidophenyl)methyl]amino]ethyl}[(2-oxidophenyl)methyl]amino)acetate
EC Number:
Cas Number:
Molecular formula:
iron(3+) sodium 2-({2-[(carboxylatomethyl)[(2-oxidophenyl)methyl]amino]ethyl}[(2-oxidophenyl)methyl]amino)acetate
Details on test material:
- Name of test material (as cited in study report): Fe (III) HBED
- Substance type: chelate
- Physical state: microgranulate
- Analytical purity: 8.6% of Fe3+
- Impurities (identity and concentrations): no data
- Composition of test material, percentage of components: no data
Data concerning identity, purity and stability of test item are responsibility of Sponsor.
- Purity test date: 2008-05-26
- Lot/batch No.: 215/P/Fe-HBED
- Expiration date of the lot/batch: not reported
- Stability under test conditions: up to 15.05.2011
- Storage condition of test material: at temp. from -10°C to +30°C
- Other:

Test animals

Details on test animals or test system and environmental conditions:
- Source: Institute of Occupational Medicine, Łódź.
- Age at study initiation: 11 week
- Weight at study initiation: first rat: 203 g; four rats females with the average body weight of 195 g
- Fasting period before study: The day before the start of experiment, about 18 hours before administration of test item, the food was withheld but water was still available. The food was restored 3 hours after administration of test item.
- Housing: individually (the sighting study) or four rats per cage (the main study) in cages with plastic bottom and wired lid, with dimensions: (length x width x height) 58 x 37 x 21 cm. UV-sterilised wooden shavings were used as a bedding.
- Diet (e.g. ad libitum): ad libitum; standard granulated “Murigran” fodder produced by Wytwórnia Koncentratów i Mieszanek Paszowych AGROPOL, Motycz.
- Water (e.g. ad libitum): ad libitum; tap water
- Acclimation period: 5 days

- Temperature (°C): 21-24
- Humidity (%): 44-90
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 2008-08-20 To: 2008-09-03 (one animal);
From: 2008-08-22 To: 2008-09-05 (four animals)

Administration / exposure

Route of administration:
oral: gavage
Details on oral exposure:
- Concentration in vehicle: 200 mg/mL

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: the dose level of 2000 mg/kg bw was chosen based on the information available for structurally related chemical Fe NaEDDHA (CAS No. 84539-55-9). No acute toxicity was observed. LD50 > 2000 mg/kg bw.
2000 mg/kg bw
No. of animals per sex per dose:
Control animals:
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Evaluation of general condition of animals, i.e. observation of all animals forcmorbidity and mortality during 14-day observation period was conducted twice a day or once a day (on days off). Detailed clinical observations were performed on day of administration (day 0), after 10, 30 and 60 minutes since administration and then in hour intervals – till the 5th hour since administration. From 1st to 14th day of observation period detailed clinical observations were performed once a day.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight
Body weight of animals was individually determined for each animal directly before administration of test item (day 0) and then on 7th and 14th day – before termination of experiment.
Not reported

Results and discussion

Preliminary study:
Following administration of test item in dose 2000 mg/kg b.w. to one female rat in the sighting study, no clinical changes were stated. The female survived 14-day period of experiment.
Effect levels
Dose descriptor:
Effect level:
> 2 000 mg/kg bw
One female rat (No. 4) died on the day 11.
Remaining animals survived all until day 14 of post-treatment period.
Clinical signs:
other: The following changes were stated in the female No 4: rounded back from 8th to 11th day of experiment, dejection on 8th and 9th day of experiment, bristled coat on 10th day of experiment. No clinical changes were observed in the remaining animals. The re
Gross pathology:
At necropsy slight congestion of lungs was stated in female No 2; slight congestion of lungs and abscess in left lung in female No 3; uneven congestion of lungs in female No 5. Pleural adhesions, marble-like and enlarged lungs as well as empty alimentary tract were stated in female which died spontaneously (No 4). These changes were caused by microbial infection. The gross changes stated in females were independent on test item. Diseases of respiratory system occur quite often in laboratory rats.
The results of gross examination are presented in Individual Data Sheets attached to this file (pages 20 - 29).
Other findings:
No other findings were reported

Applicant's summary and conclusion

Interpretation of results:
practically nontoxic
Migrated information Criteria used for interpretation of results: OECD GHS
On the ground of the study one may state that the median oral lethal dose (LD50) for Fe (III) HBED is greater than 2000 mg/kg b.w.
Executive summary:

The acute oral toxicity of the test substance Fe(III) HBED (CAS 1061328 -86 -6) was evaluated in Wistar rats according to the Fixed Dose Procedure (Gruszka, 2008). The study was conducted in compliance with the OECD TG 420. The test substance was prepared in purified water and was administered by oral route (gavage), under a volume of 1 mL/100g, to one fasted female rat. As the test substance was anticipated to be non-toxic at 2000 mg/kg, a limit test was performed by administering 2000 mg/kg to the first animal (sighting study). No clinical changes were observed in the treated animal. The animal survived the 14 -day post-treatment period. As no death occurred, the study was completed by the administration of the dose-level of 2000 mg/kg to four female animals (main study). Clinical signs, mortality and body weight gain were checked for a period of up to 14 days following the single administration of the test substance. All animals were subjected to necropsy.

Following the administration of the test item, rounded back, dejection and bristled coat were observed in one animal from day 8 of post-treatment period. The female died on 11th day of experiment. No clinical signs were stated in the remaining females. The other females survived 14-day period of experiment. At necropsy, slight congestion of lungs was stated in the female No 2; slight congestion of lungs and abscess in left lung in female No 3; uneven congestion of lungs in female No 5. Pleural adhesions, marble-like and enlarged lungs as well as empty alimentary tract were stated in the female No 4 which died spontaneously. These changes were caused by microbial infection. The clinical signs and the gross changes stated in the treated animals were considered to be unrelated to the test item. Excluding these effects and the death of one animal (as the consequence of the effects), the oral LD50 of the test substance is considered to be higher than 2000 mg/kg bw.