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

Acute Toxicity: dermal

Administrative data

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

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 3

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

Test material

Test material information

Constituent 1

Reference substance name:

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

EC Number:

939-867-3

Cas Number:

666748-58-9

Molecular formula:

MnC8H7NO8Na2

IUPAC Name:

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

Constituent 2

Reference substance name:

Manganese chelate of sodium salt N-[1,2 dicarboxyethyl] D,L aspartic acid

IUPAC Name:

Manganese chelate of sodium salt N-[1,2 dicarboxyethyl] D,L aspartic acid

Constituent 3

Reference substance name:

Mn (II) IDHA

IUPAC Name:

Mn (II) IDHA

Test material form:

other: powdered

Details on test material:

- Name of test material (as cited in study report): Mn (II) IDHA
- Substance type: chelate
- Physical state: solid (odourless, white microgranules)
- Analytical purity: content 56.7% of MnIDHA, 40% of Na2SO4, and 3% of H2O (humidity)
- Lot/batch No.: 7/13, date of production: June, 2013
- Expiration date of the lot/batch: June, 2016
- Storage condition of test material: at temperature between 0 –30°C

Test animals

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

Administration / exposure

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

Results and discussion

Effect levels

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

Any other information on results incl. tables

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.

Applicant's summary and conclusion

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.