Reaction mass of disodium [2,2'-(imino-kappaN)dibutanedioato-kappa2O1,O4(4-)]manganese(2-) and sodium sulphate

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 2007-01-18

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: conventional farm of laboratory animals run by the Institute of Occupational Medicine in Łódź, Poland.
- Age at study initiation: Pilot study: 9 week; Main experiment: 11 weeks
- Weight at study initiation: Pilot study: 166 g; Main experiment: average body weight of 190.5 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: 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 (study proper). The litter comprised dedusted wood shavings, sterilised with ultraviolet radiation. Each cage was fitted with a signboard containing the study code, the dose applied, the date of commencement and planned termination of the experiment, as well as the sex and individual numbers of animals.
- 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) and 04.01.2007. (4 females – experiment proper, dose of 2,000 mg/kg of body mass). The experiment was terminated on the following days: 03.01.2007 (1 female – initial experiment, dose of 2,000 mg/kg of body mass) and 18.01.2007 (4 females – experiment proper, dose of 2,000 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 b.w.);
- Amount of vehicle (if gavage): 0.5 mL per 100 g of the body weight.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

Initial experiment: one animal
Main strudy: 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, this in a dose of 2,000 mg/kg of body mass, to one female (initial experiment), 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, this in a dose of 2,000 mg/kg of body mass, to four successive females (experiment proper), no symptoms of toxicity were observed during the 14-day period of observation for three of the animals. The females survived the 14-day observation period.

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:

All females survived the 14-day observation period.

Clinical signs:

other: Following the single administration of the analysed substance, this in a dose of 2,000 mg/kg of body mass, to one female (initial experiment), no symptoms of toxicity were observed during the 14-day period of observation. Following the single administrati

Gross pathology:

No pathological changes were found during the macroscopic study in the analysed animals.

Table 1. Mn (II) IDHA Acute oral toxicity study conducted on rats – clinical symptoms – summary breakdown 

Dose(mg/kg b.w.)	Day following administration	Number of live animals	Rat no.
			1*	2	3	4	5
2,000	0 1 2 3 4 5 6 7 8 9 10 11 12 13 14	5 5 5 5 5 5 5 5 5 5 5 5 5 5 5	BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ	BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ	BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ	BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ	BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ BZ

* female from the initial experiment

BZ = without change

Table 2. Mn (II) IDHA Acute oral toxicity study conducted on rats - animal body mass (g)

Dose mg/kg b.w.	Rat no.	Day of experiment			Difference 14 – 0
		0	7	14
2,000	1* 2 3 4 5	166 183 198 188 193	215 210 227 206 228	236 219 229 216 232	70 36 31 28 39

* females from the initial experiment

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

On the grounds of the study, it may be stated that the median lethal dose (LD50) of Mn (II) IDHA is greater than 2000 mg/kg b.w.

Executive summary:

A study was conducted to test oral toxicity potential of Mn (II) 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 mg/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 dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from 2013-10-01 to 2013-10-16

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)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: SOP/T/21: „Acute dermal toxicity study”

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Bureau of Chemical Substances

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: husbandry of laboratory animals of the Experimental Medicine Centre at the Medical University in Białystok kept behind the breeding barrier (number in the register of units entitled to the husbandry of laboratory animals: 0043).
- Age at study initiation: 8 weeks old
- Weight at study initiation: 282.8 g (males) and 211.4 g (females)
- Fasting period before study: none
- Housing: in plastic cages covered with wi re bar lids. The dimensions of the cages were 58 x 37 x 21 cm (length x width x height). After the application of the test item, each animal was housed individually. After the removal of the test item from the animals’ skin, there were five rats per cage. Each sex was kept separately. UV-sterilized wood shavings were used as bedding. Each cage was equipped with a label containing the study code, the dose, the dates of the commencement and the expected termination of the experiment, and the animals’ sex and numbers
- Diet (e.g. ad libitum): ad libitum to “Murigran” standard granulated fodder produced by Wytwórnia Koncentratów i Mieszanek Paszowych AGROPOL, Motycz
- Water (e.g. ad libitum): ad libitum, tap water
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 - 23 °C;
- Humidity (%): 35 – 58 %
- Air changes (per hr): about 16 times/hour
- Photoperiod (hrs dark / hrs light): 12 / 12

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: ca. 41 cm² (males) ca. 31 cm² (females)
- Type of wrap if used: gauze patches were covered with PCV foil and elastic bandage was used to make circular protecting band

REMOVAL OF TEST SUBSTANCE
- Washing (if done): water
- Time after start of exposure: 24 hours after application of chemical and immediately after removal of the gauze patch

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bw
- Constant volume or concentration used:no
- For solids, paste formed: no (The test item was ground, applied to gauze patches, and moistened with a few drops of water. Then, the patches were laid on the prepared skin.)

VEHICLE
- Amount(s) applied (volume or weight with unit): a few drops of water were applied to the ground test item, which was applied to gauze patches

Duration of exposure:

24 h

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: General and detailed clinical observations of all animals were performed daily during the entire experiment. Body weights of the animals were determined on days 0 (di rectly before the application of the test item), 7, and 14. After the 14-day observation period, the animals were euthanized, dissected, and subjected to detailed gross examinations.
- Other examinations performed: clinical signs, body weight, gross necropsy

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

All animals survived the experiment.

Clinical signs:

other: no general signs of toxicity were stated in the animals. No pathological skin changes in the site of the test item application were noticed in all males and three females (no. 2, 4, and 5). Skin dryness was stated in two females (no.1 and 3).Moreover, sca

Gross pathology:

Gross examinations did not reveal any pathological changes in the examined animals.

Other findings:

no other findings reported

Following single application of the test item, the animals did not exhibit any general clinical signs. No pathological skin changes in the site of the test item application were noticed in all males and three females (no. 2, 4, and 5). Skin dryness was stated in two females (no. 1 and 3). Moreover, scabs were stated in one female (no. 3). All animals survived the experiment. During the 14-day experiment, body weight gain was stated in all animals. Gross examinations did not reveal any pathological changes in the examined animals.

Table 1: Summary of results

Mn (II) IDHA: acute dermal toxicity study on rats
Dose of test item(mg/kg b.w.)	2000
Sex	males	females
Mortality	0/5	0/5
Clinical signs	no changes	no changes were stated (no 2, 4, and 5)
		skin dryness was stated (no 1 and 3)
		scabs were stated (no. 3)

Clinical signs

Following single application of the test item, the animals did not exhibit any general clinical signs. No pathological skin changes in the site of the test item application were noticed in all males and three females (no. 2, 4, and 5). Skin dryness was stated in two females (no. 1 and 3) between the 2nd and the 3rd day after the application. Moreover, scabs were stated in one female (no. 3) between the 2nd and the 3rd day after the application. All animals survived the experiment. An overall list of the results of the clinical observations is presented in Table 2.

Table 2 - Clinical signs - overall list
Mn (II) IDHA: acute dermal toxicity study on rats
Dose (mg/kg b.w.)	Sex	Day after application	Number of living animals	Rat number
				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	SIGNS	NC	SIGNS	NC	NC
		3	5	SIGNS	NC	SIGNS	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
SIGNS - clinical signs

Body weights of the animals

During the 14-day experiment, body weight gain was stated in all animals. The individual results of body weight measurements are presented in Table 3

Table 3- Bodyweight of animals (g)-overall list
Mn(II)IDHA:acute dermal toxicity study on rats
Dose(mg/kgb.w.)	Sex	Rat No	Day of experiment/Body weight(g)			Body weight gain(g)(0-14)
			0	7	14
2000	males	1	298	330	362	64
		2	285	305	326	41
		3	300	325	361	61
		4	243	262	280	37
		5	288	301	322	34
	females	1	203	220	230	27
		2	215	229	245	30
		3	208	223	231	23
		4	211	223	233	22
		5	220	237	252	32

Gross examinations

The gross examinations did not reveal any pathological changes in the examined animals.

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

On the grounds of the study, it may be stated that the median lethal dose (LD50) of Mn (II) IDHA is greater than 2000 mg/kg b.w.

Executive summary:

A study was conducted to test the dermal toxicity potential of Mn (II) IDHA in rats (Kropidlo, A., 2013). Following single application of the test item, the animals did not exhibit any general clinical signs. No pathological skin changes on the site of the test item application were noticed in all males and three females (no. 2, 4, and 5). Skin dryness was stated in two females (no. 1 and 3). Moreover, scabs were stated in one female (no. 3). All animals survived the experiment. During the 14-day experiment, body weight gain was stated in all animals. Gross examinations did not reveal any pathological changes in the examined animals. On the grounds of the study, it may be stated that the median lethal dose (LD50) of Mn (II) IDHA is greater 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 402, is GLP compliant and has Klimish score 1.

Additional information

Acute toxicity: oral

A study was conducted to test oral toxicity potential of Mn(2Na)IDHA in rats (Gruszka, 2007, Report No. OS-45/06, according to OECD 420). Following the single administration of the analysed substance in a dose of 2000 mg/kg bw 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 2000 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. LD50 of greater than 2000 mg/kg bw was established.

Acute toxicity: dermal

A study was conducted to test the dermal toxicity potential of Mn(2Na)IDHA in rats (Kropidlo, 2013, Report No. DER-11/13, according to OECD 402). Following single application of the test item, the animals did not exhibit any general clinical signs. No pathological skin changes in the site of the test item application were noticed in all males and three females (no. 2, 4, and 5). Skin dryness was stated in two females (no. 1 and 3). Moreover, scabs were stated in one female (no. 3). All animals survived the experiment. During the 14-day experiment, body weight gain was stated in all animals. Gross examinations did not reveal any pathological changes in the examined animals. On the grounds of the study, it may be stated that the median lethal dose (LD50) of Mn(2Na)IDHA is greater than 2000 mg/kg bw.

Justification for selection of acute toxicity – oral endpoint
Only one study 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, Mn(2Na)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, Mn(I2Na)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.