Reaction mass of iron chelate of sodium salt N-[1,2 dicarboxyethyl] D,L aspartic acid and sodium chloride

Endpoint summary

Description of key information

- Acute oral toxicity, OECD 420, rats, LD50 > 2000 mg/kg bw
- Acute dermal toxicity, OECD 402, rats, LD50 > 2000 mg/kg bw

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

Study period:

2006-12-20 to 2007-01-23

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Statement of GLP Compliance No. G 024 (Slovak National Accreditation Service); Statement of GLP Compliance No. 4/2006/DPL

Test type:

fixed dose procedure

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Institute of Occupational Medicine in Łódź, Poland
- Age and weight at study initiation: Pilot study: 10 week (dose of 2,000 mg/kg bw). Main experiment: 11 weeks (average body weight of 188.5 g).
- Fasting period before study: yes. The day before the experiment was due to commence, some 18 hours before administration of the analysed material, the animals were deprived of feed, being left with only water. Feed was made available again 3 hours after administration of the analysed substance.
- Housing: The animals were kept in plastic cages with metal wire covers, with the following dimensions (length x width x height): 58 x 37 x 21 cm.
During the experiment, the animals were kept in cages individually (initial study) and in groups of four (main study).
- Diet (e.g. ad libitum): ad libitum (standard granulated "Murigran" laboratory feed, manufactured by Wytwórnia Koncentratów i Mieszanek Paszowych AGROPOL of Motycz
- Water (e.g. ad libitum): ad libitum (tap water)
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-21
- Humidity (%): 45-71
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES:
The analysed substance was administered to the rats on the following days: 20.12.2006. (1 female – initial experiment, dose of 2,000 mg/kg of body mass), 04.01.2007 (4 females – experiment proper, dose of 2000 mg/kg of body mass). The experiment was terminated on the following days: 03.01.2007. (1 female – initial experiment, dose of 2000 mg/kg of body mass), 23.01.2007. (4 females – experiment proper, dose of 2000 mg/kg of body mass), respectively.

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 1 ml of the water solution of the analysed substance contained: 400 mg of the substance (dose of 2,000 mg/kg of body mass).
- Amount of vehicle (if gavage): 0.5 mL per 100 g of the body weight.

- Rationale for the selection of the starting dose: During the initial experiment, one female received a dose of 2,000 mg/kg of body mass of the analysed substance. During the 14 day observation period were not observed the symptoms of toxicity. The female survived the 14-day observation period.
On the basis of the initial experiment, during the experiment proper the analysed substance was administered to four successive females in a dose of 2000 mg/kg of body mass.

Doses:

Initial experiment: 2000 mg/kg bw;
Main strudy: 2000 mg/kg bw

No. of animals per sex per dose:

Initial experiment: 2000 mg/kg bw - one animal;
Main strudy: 2000 mg/kg bw - four animals

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: An assessment of the general condition of the animals, i.e. observation of all animals in terms of incidence and mortality, was performed twice daily throughout the 14-day duration of the experiment. Detailed clinical observations were performed on the day of administration of the analysed substance (day 0), 10, 30 and 60 minutes after administration, and subsequently every hour over a period of 5 hours from the time of administration. On successive days of the 14-day period of the experiment – once daily.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight.

Preliminary study:

Following the single administration of the analysed substance at the dose of 2,000 mg/kg of body weight to one female (initial experiment), no symptoms of toxicity were observed during the 14-day period of observation. The animal survived the 14-day period of observation.

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: based on mortality, clinical signs and findings at necropsy at 2000 mg/kg bw

Mortality:

No mortality occured during the 14-day observation period.

Clinical signs:

other: After a single administration of test substance to one female at a dose of 2000 mg / kg b.w. (preliminary experience), during the 14-day period of observation there was no diagnosed with symptoms of toxicity. The female survived the 14-day observation per

Gross pathology:

Macroscopic studies did not find any pathological changes in the test animals.

Table 1. Clinical signs.

Dose (mg/kg bw)	Day following administra tion	Number of live animals			Rat No
			1 *	2	3	4	5
	0	5	BZ	BZ	BZ	BZ	BZ
	1	5	BZ	BZ	BZ	BZ	BZ
	2	5	BZ	BZ	BZ	BZ	BZ
	3	5	BZ	BZ	BZ	BZ	BZ
2000	4	5	BZ	BZ	BZ	BZ	BZ
	5	5	BZ	BZ	BZ	BZ	BZ
	6	5	BZ	BZ	BZ	BZ	BZ
	7	5	BZ	BZ	BZ	BZ	BZ
	8	5	BZ	BZ	BZ	BZ	BZ
	9	5	BZ	BZ	BZ	BZ	BZ
	10	5	BZ	BZ	BZ	BZ	BZ
	11	5	BZ	BZ	BZ	BZ	BZ
	12	5	BZ	BZ	BZ	BZ	BZ
	13	5	BZ	BZ	BZ	BZ	BZ
	14	5	BZ	BZ	BZ	BZ	BZ

* females from the initial experiment

BZ = without change

Table 2. Body weights

Dose mg/kg of body mass	Rat no.	Day of experiment			Difference 14 – 0
		0	7	14
2000	1* 2 3 4 5	198 179 174 189 174	233 206 225 211 222	241 225 232 216 232	52 35 42 33 41

* females from the initial experiment

Interpretation of results:

Toxicity Category V

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Based on the mortality, clinical signs and findings at necropsy in five animal dosed by 2000 mg/kg bw, LD50 is considered to be over 2000 mg/kg bw that corresponds to Cat. 5 (not clasified) in accordance with the ECHA guidance on the Application of the CLP Criteria (2013).

Executive summary:

A study was conducted to test oral toxicity potential of Fe(III)IDHA in rats. Following the single administration of the analysed substance in a dose of 2,000 mg/kg b.w. to a single female, no symptoms of toxicity were observed during the 14-day period of observation. The female survived the 14-day period of observation. Following the single administration of the analysed substance in a dose of 2,000 mg/kg bw to four successive females, no symptoms of toxicity were observed during the 14-day period of observation. The females survived the 14-day observation period. All the animals were put down following the 14-day period of observation and subsequently underwent autopsies and macroscopic studies. No pathological changes were found during the macroscopic study in the studied animals.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 ng/kg bw

Quality of whole database:

The study is conducted in accordance with OECD 420, is GLP compliant and has Klimish score 1.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

May, 29 - June, 12, 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: well documented GLP Guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

GLP compliance:

yes (incl. QA statement)

Remarks:

Statement of GLP Compliance No G-024 issued by Slovac National Accreditation Service; Statement of GLP Compliance No 4/2006/DPL issued by Bureau for Chemical Substances and Preparations

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Institute of Occupational Medicine in Łódz kept in conventional type.
- Age at study initiation: 10-week-old male rats and 11-weekold females
- Weight at study initiation: male rats with the average body weight of 287 g and females with the average body weight of 194 g
- Fasting period before study: no
- Housing:
- Diet (e.g. ad libitum): standard granulated "Murigran" fodder produced by Wytwórnia Koncentratów i Mieszanek Paszowych AGROPOL, Motycz
- Water (e.g. ad libitum): tap water
- Acclimation period: yes

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 - 23 °C;
- Humidity (%): 48 - 80
- Photoperiod (hrs dark / hrs light): artificial, fluorescent lighting: 12 hours light / 12 hours darkness

The animals were kept in cages with plastic bottom and wired lid, with dimensions: (length x width x height) 58 x 37 x 21 cm. Following administration of the test material on animals skin, each one was kept individually per cage. After removal of the test material from animals skin, during the following days of experiment the rats were kept five per cage, each sex separately.

Wooden UV-sterilized shavings were used as a litter. Each cage was equipped with label containing information on name of test material, study code, used dose, start date and planned ending date of the experiment, animal sex and animal numbers.

IN-LIFE DATES: From: To:
The experiment was started on May 29th, 2007 (males and females) and ended on June 12th, 2007.

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

The day before the test back and sides of animals were shaved with electric razor on skin surface area of about 4 x 6 cm. Only those animals that showed no macroscopically visible irritation or damages of skin were used for the experiments. The animals were not starved prior to the experiment.

The undiluted test material was applied to the dorsal skin of 10 rats (5 males and 5 females) in dose of 2000 mg/kg b.w. The area of skin treated with the test material was about 6 cm2. The test material was applied to gauze patches and then laid on the prepared skin. The gauze patches were covered with PCV foil and elastic bandage was used to make circular protecting band. After 24 hours the band and gauze patches were taken off and the residual test material was removed using water.

Duration of exposure:

24 hours

Doses:

2000 mg/kg b.w.

No. of animals per sex per dose:

5 males and 5 females

Control animals:

no

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 in regards to mortality and morbidity was conducted twice a day during 14-day observation period. Following administration of the test material, detailed clinical observations were performed in hour intervals during the day of administration (day 0). Since the first till the 14th day of observation period detailed clinical observations were performed once a day.
Body weight of animals was determined individually for each animal directly before administration of the test material (day 0) and then on seventh andfourteenth day - before the finishing of the experiment.
- Necropsy of survivors performed: yes
- Other examinations performed: After opening of thorax and abdomen macroscopic examination of internal organs was performed.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

All rats survived the 14-day observation period.

Clinical signs:

other: During 14-day experiment period no changes on skin or in behavior of test animals were stated.

Gross pathology:

No pathological changes were stated in animals during macroscopic examination.

Chelate with name: Fe (III) IDHA Acute dermal toxicity study for rats – clinical signs – overall list

Dose (mg/kg b.w.)	Sex	Day after administration	No of alive animals	Rat No
				1	2	3	4	5
2000	Males	0	5	NC	NC	NC	NC	NC
		1	5	NC	NC	NC	NC	NC
		2	5	NC	NC	NC	NC	NC
		3	5	NC	NC	NC	NC	NC
		4	5	NC	NC	NC	NC	NC
		5	5	NC	NC	NC	NC	NC
		6	5	NC	NC	NC	NC	NC
		7	5	NC	NC	NC	NC	NC
		8	5	NC	NC	NC	NC	NC
		9	5	NC	NC	NC	NC	NC
		10	5	NC	NC	NC	NC	NC
		11	5	NC	NC	NC	NC	NC
		12	5	NC	NC	NC	NC	NC
		13	5	NC	NC	NC	NC	NC
		14	5	NC	NC	NC	NC	NC
	Females	0	5	NC	NC	NC	NC	NC
		1	5	NC	NC	NC	NC	NC
		2	5	NC	NC	NC	NC	NC
		3	5	NC	NC	NC	NC	NC
		4	5	NC	NC	NC	NC	NC
		5	5	NC	NC	NC	NC	NC
		6	5	NC	NC	NC	NC	NC
		7	5	NC	NC	NC	NC	NC
		8	5	NC	NC	NC	NC	NC
		9	5	NC	NC	NC	NC	NC
		10	5	NC	NC	NC	NC	NC
		11	5	NC	NC	NC	NC	NC
		12	5	NC	NC	NC	NC	NC
		13	5	NC	NC	NC	NC	NC
		14	5	NC	NC	NC	NC	NC

NC – no changes

Chelate with name: Fe (III) IDHA Acute dermal toxicity study for rats – body weight of animals (g)

Dose (mg/kg b.w.)	Sex	Rat No	Day			Difference
			0	7	14	14 - 0
2000	Males	1	260	281	302	42
		2	299	308	326	27
		3	285	293	315	30
		4	300	316	345	45
		5	293	309	332	39
		1	191	206	209	18
	Females
		2	191	203	215	24
		3	194	203	211	17
		4	198	207	216	18
		5	195	199	218	23

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Taking into account the obtained results one may state that the median dermal acute dose (LD50) for Chelate with name: Fe (III) IDHA is higher than 2000 mg/kg b.w.

Executive summary:

Acute dermal toxicity study of Chelate with name: Fe (III) IDHA for rats was performed according to the OECD Guideline for Testing of Chemicals No 402/ Method B.3 as well as Principles of Good Laboratory Practice (GLP - OECD, 1997). The test material was given to 10 rats (5 males and 5 females) to shaved dorsal skin in single dose of 2000 mg/kg b.w. for 24 hours. During 14-day observation period no changes on skin or in behavior were stated in test animals. All animals survived 14- day experiment period. All animals which were euthanized after 14-day observation period were dissected and studied macroscopically. Macroscopic examination showed no pathological changes in test animals. Taking into account the obtained results one may state that median dermal acute dose (LD50) for Chelate with name: Fe (III) IDHA is higher than 2000 mg/kg b.w.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

The study is conducted in accordance with OECD 420, is GLP compliant and has Klimish score 1.

Additional information

Acute toxicity: oral

A study was conducted to test oral toxicity potential of Fe(III)IDHA in rats.Following the single administration of the analysed substance in a dose of 2,000 mg/kg b.w. to a single female, no symptoms of toxicity were observed during the 14-day period of observation. The female survived the 14-day period of observation. Following the single administration of the analysed substance in a dose of 2,000 mg/kg bw to four successive females, no symptoms of toxicity were observed during the 14-day period of observation. The females survived the 14-day observation period. All the animals were put down following the 14-day period of observation and subsequently underwent autopsies and macroscopic studies. No pathological changes were found during the macroscopic study in the studied animals.

Acute toxicity dermal

Acute dermal toxicity study of Fe(III)IDHA for rats was performed according to the OECD Guideline for Testing of Chemicals No 402/ Method B.3 as well as Principles of Good Laboratory Practice (GLP - OECD, 1997). The test material was given to 10 rats (5 males and 5 females) to shaved dorsal skin in single dose of 2000 mg/kg b.w. for 24 hours. During 14-day observation period no changes on skin or in behavior were stated in test animals. All animals survived 14- day experiment period. All animals which were euthanized after 14-day observation period were dissected and studied macroscopically. Macroscopic examination showed no pathological changes in test animals. Taking into account the obtained results one may state that median dermal acute dose (LD50) for Chelate with name: Fe (III) IDHA is higher than 2000 mg/kg b.w.

Justification for selection of acute toxicity – oral endpoint
Only one study is available.

Justification for selection of acute toxicity – dermal endpoint
Only one study is available

Justification for classification or non-classification

Based on LD50 of greater than 2000 mg/kg bw established in the oral acute toxicity study, Fe(III)IDHA is not subject to classification and labelling for acute toxic effects by oral route of exposure according to European Regulation (EC) No. 1272/2008.

Based on LD50 of greater than 2000 mg/kg bw established in the dermal acute toxicity study, Fe(III)IDHA is not subject to classification and labelling for acute toxic effects by dermal route of exposure according to European Regulation (EC) No. 1272/2008.