Registration Dossier

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 guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
Qualifier:
according to
Guideline:
other: SOP/T/21: „Acute dermal toxicity study”
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Slovak National Accreditation Service (No. G-024)
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
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 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:
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 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.
Body weight:
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.
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.