Reaction mass of disodium [2,2'-(imino-κN)dibutanedioato-κ2O1,O4(4-)]cuprum(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 2010-11-10 to 2010-11-24

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:

2010

Report date:

2010

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:

Slovak National Accreditation Service (No. G-024)

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

disodium 2-[7-(carboxylatomethyl)-4,8-dioxo-1,3-dioxa-6-aza-2-cupracyclooctan-5-yl]acetate

EC Number:

938-868-6

Cas Number:

666828-79-1

Molecular formula:

not applicable

IUPAC Name:

disodium 2-[7-(carboxylatomethyl)-4,8-dioxo-1,3-dioxa-6-aza-2-cupracyclooctan-5-yl]acetate

Constituent 2

Reference substance name:

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

IUPAC Name:

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

Constituent 3

Reference substance name:

Cu (II) IDHA

IUPAC Name:

Cu (II) IDHA

Test material form:

other: powdered

Details on test material:

- Name of test material (as cited in study report): Cu (II) IDHA
- Substance type: chelate
- Physical state: solid (odourless, blue microgranules)
- Analytical purity: the test item contains 9.82% of copper
- Lot/batch No.: 1/10, date of production: April, 2010
- Expiration date of the lot/batch: April, 2013
- Storage condition of test material: in tightly closed packed at temperature from -10°C to +30°C

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: conventional husbandry of laboratory animals of Institute of Occupational Medicine, Łódź
- Age at study initiation: 10 weeks old
- Weight at study initiation: 295.8 g (males) and 218.2 g (females)
- Fasting period before study: none
- Housing: in cages with dimensions (length x width x height): 58 x 37 x 21 cm; with plastic bottom and wired lid. After application of the test item each animal was kept individually in cage. After removal of test item from animals’ skin during the following days of experiment, the rats were kept five per cage, each separately. UV-sterilized 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: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 - 23 °C;
- Humidity (%): 47 – 63 %
- 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 applied to gauze patches, moistened with few drops of water and then laid on prepared skin.)

VEHICLE
- Amount(s) applied (volume or weight with unit): a few drops of water were applied to the powdered amount of chemical on the gauze patch

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 were performed in all animals daily during the entire experiment. Body weight of animals was individually determined for each animal: on day 0 (directly before administration of test item) and on 7th and 14th day.
- Necropsy of survivors performed: yes (All animals were sacrificed after 14-day observation period, dissected and subjected to detailed gross necropsy. )
- 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:

no mortality occurred

Clinical signs:

other: no general signs of toxicity were stated in the animals. Changes in form of erythema, dryness of epidermis, desquamation of epidermis and scabs were stated on the skin in the site of test item application. Dryness of epidermis and scabs were stated in fiv

Gross pathology:

No pathological changes were stated in animals at gross necropsy.

Other findings:

no other findings reported

Any other information on results incl. tables

Following single application of test item no general signs of toxicity were stated in animals. Changes in form of erythema, dryness of epidermis, desquamation of epidermis and scabs were stated on skin in the site of test item application. Dryness of epidermis and scabs were stated in five males whereas desquamation of epidermis – in one male. Erythema and scabs were stated in four females, dryness of epidermis – in five females, desquamation of epidermis – in two females. These were transient changes except dryness of epidermis in three females which was observed till the end of experiment. All animals survived period of experiment. Slight body weight loss was stated in one female No 2 (6g) and three males No 1 (3g), No 4 (6g) and No 5 (8g) during 1st week of experiment. Body weight gains were stated in all animals after 14-day period of experiment. No pathological changes were stated in animals at gross necropsy.

Table 1: Summary of findings

Cu (II) IDHA: acute dermal toxicity study on rats
Administered dose of test item(mg/kg b.w.)	2000
Sex	males	females
Mortality of animals	0/5	0/5
Clinical signs	no general clinical signs;	no general clinical signs;
	on treated skin: dryness of epidermis and scabs (in 5 males)	on treated skin: erythema and scabs (in 4 females)
	on treated skin: desquamation of epidermis (in 1 male)	on treated skin: dryness of epidermis (in 5 females)
		on treated skin: desquamation of epidermis (in 2 females)

Clinical signs

Following single application of test item no general signs of toxicity were stated in animals. Changes in form of dryness of epidermis from 2nd – 10th to 5th – 13th day and scabs from 2nd to 5th – 9th day after administration were stated in five males on skin in the site of test item application. Furthermore, desquamation of epidermis on 2nd day after administration was stated in one male (No 1). Changes in form of erythema on 2nd day and scabs from 2nd to 9th – 14th day after administration were stated in four females (No 1, No 2, No 3 and No 5). Changes in form of dryness of epidermis from 2nd – 3rd to 4th – 5th day after administration were stated in five females. Furthermore, dryness of epidermis formed in place of scabs was observed in four females (No 1, No 2, No 3 and No 5) from 8th – 9th to 11th – 14th day after administration. Desquamation of epidermis on 2nd day after administration was stated in two females (No 1 and No 4). All animals survived period of experiment. The overall list of results of clinical observations is presented in Table 2.

Table 2 - Clinical signs - overall list
Cu (II) IDHA: acute dermal toxicity study on rats
Dose (mg/kg b.w.)	Sex	Day after administration	Number 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	SIGNS	SIGNS	SIGNS	SIGNS	SIGNS
		3	5	SIGNS	SIGNS	SIGNS	SIGNS	SIGNS
		4	5	SIGNS	SIGNS	SIGNS	SIGNS	SIGNS
		5	5	SIGNS	SIGNS	SIGNS	SIGNS	SIGNS
		6	5	NC	SIGNS	SIGNS	SIGNS	SIGNS
		7	5	NC	SIGNS	SIGNS	SIGNS	SIGNS
		8	5	NC	SIGNS	SIGNS	SIGNS	SIGNS
		9	5	NC	SIGNS	SIGNS	NC	SIGNS
		10	5	NC	SIGNS	SIGNS	NC	SIGNS
		11	5	NC	SIGNS	SIGNS	NC	SIGNS
		12	5	NC	SIGNS	SIGNS	NC	SIGNS
		13	5	NC	NC	SIGNS	NC	SIGNS
		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	SIGNS	SIGNS	SIGNS	SIGNS
		3	5	SIGNS	SIGNS	SIGNS	SIGNS	SIGNS
		4	5	SIGNS	SIGNS	SIGNS	SIGNS	SIGNS
		5	5	SIGNS	SIGNS	SIGNS	SIGNS	SIGNS
		6	5	SIGNS	SIGNS	SIGNS	NC	SIGNS
		7	5	SIGNS	SIGNS	SIGNS	NC	SIGNS
		8	5	SIGNS	SIGNS	SIGNS	NC	SIGNS
		9	5	SIGNS	SIGNS	SIGNS	NC	SIGNS
		10	5	SIGNS	SIGNS	SIGNS	NC	SIGNS
		11	5	SIGNS	SIGNS	SIGNS	NC	SIGNS
		12	5	SIGNS	SIGNS	SIGNS	NC	NC
		13	5	SIGNS	SIGNS	SIGNS	NC	NC
		14	5	SIGNS	SIGNS	SIGNS	NC	NC
NC - no changes
SIGNS - clinical signs

Body weight of animals

Slight body weight loss was stated in one female No 2 (6g) and three males No 1 (3g), No 4 (6g) and No 5 (8g) during 1st week of experiment. Body weight gains were stated in all animals after 14-day period of experiment.

Table 2- Bodyweight of animals (g)-overall list
Cu(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	308	305	310	2
		2	292	295	312	20
		3	299	302	322	23
		4	287	281	296	9
		5	293	285	306	13
	females	1	218	220	237	19
		2	216	210	230	14
		3	219	220	233	14
		4	223	231	235	12
		5	215	228	232	17

Gross examination

At necropsy no pathological changes were stated in males and females sacrificed after 14-day experiment period.

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

On the ground of the study, one may state that the median dermal lethal dose (LD50) for Cu (II) IDHA is greater than 2000 mg/kg b.w.

Executive summary:

A study was conducted to test the dermal toxicity potential of Cu (II) IDHA in rats (Kropidlo, A., 2010). Following the single administration of the analysed substance at dose level of 2000 mg/kg bw (semi-occlusive application to the shaved skin for 24 hours) to 5 female and 5 male Wistar rats, no symptoms of general toxicity were observed within the 14 day observation period. No mortality was observed. Changes in form of erythema, dryness of epidermis, desquamation of epidermis and scabs were stated on skin in the site of test item application. Dryness of epidermis and scabs were stated in five males whereas desquamation of epidermis – in one male. Erythema and scabs were stated in four females, dryness of epidermis – in five females, desquamation of epidermis – in two females. These were transient changes except dryness of epidermis in three females which was observed till the end of experiment. Slight body weight loss was stated in one female No 2 (6g) and three males No 1 (3g), No 4 (6g) and No 5 (8g) during 1st week of experiment. Body weight gains were stated in all animals after 14-day period of experiment. No pathological changes were stated in animals at gross necropsy. On the ground of the study, one may state that the median dermal lethal dose (LD50) for Cu (II) IDHA is greater than 2000 mg/kg b.w.