Registration Dossier

Administrative data

acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
18 of September - 6 of December 2013
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Well conducted and well documented study undner GLP conditions.

Data source

Reference Type:
study report
Report date:

Materials and methods

Test guidelineopen allclose all
according to guideline
OECD Guideline 402 (Acute Dermal Toxicity)
according to guideline
EU Method B.3 (Acute Toxicity (Dermal))
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method

Test material

Constituent 1
Reference substance name:
Constituent 2
Chemical structure
Reference substance name:
disodium [2,2'-(ethane-1,2-diylbis{[2-(hydroxy-kO)benzyl]imino-kN})diacetato-kO(4-)]zinc(2-)
Cas Number:
Molecular formula:
C20 H20 N2 Na2 O6 Zn
disodium [2,2'-(ethane-1,2-diylbis{[2-(hydroxy-kO)benzyl]imino-kN})diacetato-kO(4-)]zinc(2-)
Test material form:
solid: particulate/powder
migrated information: powder

Test animals

Details on test animals or test system and environmental conditions:
The experiment was conducted on the Wistar male and female rats (Crl: WI(Han); outbred) obtained from the 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). The animals were quarantined and observed daily for 5 days. A general medical-veterinary examination was performed on the day of the introduction of the animals to the quarantine, whereas a detailed medical-veterinary examination was performed before the beginning of the experiment. Only animals without any clinical signs were introduced to the experiment. The animals were randomized to the experiment. Thei r sex and body weights were taken into account. All animals were marked individually. Five 8-week-old males whose average body weight was 278.0 g and five 8-week-old females whose average body weight was 210.0 g were used in theexperiment.

Housing conditions
During the quarantine and the experiment, the animals were kept in air-conditioned rooms under the following conditions:
- air temperature: 21 – 23 °C
- relative ai r humidity: 35 – 58%
- artificial fluorescent lighting; lighting cycle: 12 hours light/12 hours dark
- facility air exchange: about 16 times/hour.
The animals were housed 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 thestudy code, the dose, the dates of the commencement and the expectedtermination of the experiment, and the animals’ sex and numbers.

Food and water
The animals were given ad libitum access to the “Murigran” standard granulated fodder and drinking tap water.

Administration / exposure

Type of coverage:
Details on dermal exposure:
The area of skin treated with the test item was about 41 cm2 (males) and 31 cm2 (females), i.e. about 10% of the body surface area. The test item in the form of a powder was applied to gauze patches and moistened with a few drops of water. Then, the patches were laid on the prepared skin and covered with PVC foil. An elastic bandage and a sticking plaster were used to make a circular protecting band. After 24 hours, the band and the gauze patches were taken off and the residual test item was removed using water
Duration of exposure:
24 h
2 000 mg/kg b.w.
No. of animals per sex per dose:
5 males and 5 females. All animals received the same dose 2 000 mg/kg b.w.
Control animals:
Details on study design:
The test item at a single dose of 2000 mg/kg b.w. was applied to the shaved dorsal area of the trunks
of 5 males and 5 females for 24 hours.
After the application of the test item, the animals were observed for 14 days. 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.

Results and discussion

Preliminary study:
Not concern
Effect levels
Dose descriptor:
Effect level:
ca. 2 000 mg/kg bw
Based on:
test mat.
No mortaliy, all animals survived the test.
Clinical signs:
No pathological skin changes in the site of the test item application were noticed in all males. As for females, desquamation of the epidermis was stated in all females,i.e. on day 5 in female no. 1, from day 5 to day 7 in female no. 2, from day 5 to day 8 in female no. 3, from day 5 to day 6 in female no. 4, and on day 6 in female no. 5. Skin dryness was also noticed in all females, i.e. from day 5 to day 7 in female no. 1, from day 3 to day 8 in female no. 2, from day 5 to day 8 in female no. 3, from day 1 to day 8 in female no. 4, and from day 3 to day 7 in female no. 5. Moreover, scabs were stated in three females, i.e. from day 3 to day 6 in female no. 2, from day 1 to day 4 in female no. 3, and from day 1 to day 5 in female no. 4.
Body weight:
Body weight loss was stated in two females (no. 2 and 5) during the second week of the experiment.
After the 14-day experiment period, body weight gain was stated in all animals.
Gross pathology:
The gross examination did not reveal any pathological changes in the examined animals.
Other findings:
No other findings.

Applicant's summary and conclusion

Interpretation of results:
not classified
Migrated information Criteria used for interpretation of results: EU
On the grounds of the study, it may be stated that the median lethal dose (LD50) of Zn (II) HBED is greater than 2000 mg/kg b.w.
According to the Regulation (EC) No. 1272/2008 of the European Parliament and of the Council of December 16, 2008 on classification, labelling, and packaging of substances and mixtures, amending and repealing Directives 67/548/EEC and 1999/45/EC and amending Regulation (EC) No. 1907/2006, it may be concluded that the test item, i.e. Zn (II) HBED is beyond categorization.