Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2011-10-10 to 2012-02-07
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report Date:
2012

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:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
(Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany)
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
- Full barrier in an air-conditioned room
- Temperature: 22 +/- 3 °C
- Relative humidity: 55 +/- 10%
- Artificial light, sequence being 12 hours light, 12 hours dark
- Air change: 10 x / hour
- Free access to Altromin 1324 maintenance diet for rats and mice (lot no. 1956)
- Free access to tap water, sulphur acidified to a pH value of approximately 2.8 (drinking water, municipal residue control, microbiological controls at regular intervals)
- The animals were kept individually in IVC cages, type III H, polysulphone cages on Altromin saw fibre bedding (lot no. 190711)
- Certificates of food, water and bedding are filed at BSL BIOSERVICE
- Adequate acclimatisation period (at least five days) under laboratory conditions

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
water
Remarks:
Aqua ad injectionem (AlleMan Pharma, lot no. 110147, expiry date: 02/2014)
Details on dermal exposure:
Preparation of the Animals:
The animals were marked for individual identification by tail painting.
Approximately 24 hours before the test, the fur was removed from the dorsal area of the trunk using an electric clipper. Care was taken to avoid abrading the skin, and only animals with healthy intact skin were used.
No less than 10% of the body surface was cleared for the application.
Prior to the application a detailed clinical observation was made of all animals.
Application:
The test item was applied at a single dose, uniformly over an area which was approximately 10% of the total body surface.
The test item was held in contact with the skin by a dressing throughout a 24-hour period. The dressing consisted of a gauze-dressing and non-irritating tape and was fixed with an additional dressing in a suitable manner.
Duration of exposure:
The test item was held in contact with the skin throughout a 24-hour period. At the end of the exposure period the residual test item was removed using. At the end of the exposure period the residual test item was removed using tap water.
Doses:
The test item was applied at a single dose of 2000 mg/kg body weight to each animal.
No. of animals per sex per dose:
5 male and 5 female
Control animals:
not required
Details on study design:
Observation period:
All animals were observed for 14 days after dosing

Weight Assessment:
The animals were weighed on day 1 (prior to the application) and on days 8 and 15.

Clinical Examination:
careful clinical examination was made several times on the day of dosing (at least once during the first 30 minutes and with special attention
given during the first 4 hours post-dose). As soon as symptoms were noticed they were recorded. Thereafter, the animals were observed for
clinical signs once daily until the end of the observation period. All abnormalities were recorded.
Cageside observations included changes in the skin and fur, eyes and mucous membranes. Also respiratory, circulatory, autonomic and central
nervous systems and somatomotor activity and behaviour pattern were examined. Attention was directed to observations of tremors,
convulsions, salivation, diarrhoea, lethargy, sleep and coma.

Pathology:
At the end of the observation period the surviving animals were sacrificed with an overdosage of pentobarbital injected intraperitoneally
(Narcoren®, Merial) at the dosage of approximately 8 mL/kg bw. All animals were subjected to gross necropsy. All gross pathological changes
were recorded and in case of findings the tissues were preserved for a possible histopathological evaluation. The preserved tissues of which
no histopathological evaluation was made will be discarded 3 months after the release of the final report unless otherwise agreed upon with
the sponsor.

Evaluation of Results:
Individual reactions of each animal were recorded at each time of observation.
Toxic response data were recorded by sex and dose level.
Nature, severity and duration of clinical observations were described.
The body weight changes were summarised in a tabular form.
Necropsy findings were described.
On the basis of the test results, the test item may be classified in one of the following classes in conformity with the criteria given in
Annex VI to Commission Directive 2001/59/EC:
• Very toxic
Substances and preparations shall be classified as very toxic, and assigned the symbol “T+” and indication of danger “very toxic” in
accordance with the criteria specified below:
R27 Very toxic in contact with skin
- LD50 dermal, rat or rabbit: <= 50 mg/kg
• Toxic
Substances and preparations shall be classified as toxic, and assigned the symbol “T” and indication of danger “toxic” in accordance
with the criteria specified below. Risk phrases shall be assigned in accordance with the following criteria:
R24 Toxic in contact with skin
- LD50 dermal, rat or rabbit: 50 < LD50 <= 400 mg/kg
• Harmful
Substances and preparations shall be classified as harmful, and assigned the symbol “Xn” and indication of danger “harmful” in accordance
with the criteria specified below. Risk phrases shall be assigned in accordance with the following criteria:
R21 Harmful in contact with skin
- LD50 dermal, rat or rabbit: 400 < LD50 <= 2000 mg/kg
On the basis of the test results, the following risk phrases may be assigned in conformity with the criteria given in Annex I of Regulation (EC) 1272/2008:
Category 1: LD50 <= 50 mg/kg. DANGER. Skull and crossbones in diamond. Fatal in contact with skin.
Category 2: LD50 > 50 mg/kg <= 200 mg/kg. DANGER. Skull and crossbones in diamond. Fatal in contact with skin.
Category 3: LD50 > 200 mg/kg <= 1000 mg/kg. DANGER. Skull and crossbones in diamond. Toxic in contact with skin.
Category 4: LD50 > 1000 mg/kg <= 2000 mg/kg. WARNING. Exclamation point in diamond. Harmful in contact with skin.
On the basis of the test results, the following risk phrases may be assigned in conformity with the criteria given in GHS - Globally Harmonized System of Classification and Labelling of Chemicals, third revised edition, July 2009:
Category 1: LD50 <= 50 mg/kg. DANGER. Skull and crossbones in diamond. Fatal in contact with skin.
Category 2: LD50 > 50 mg/kg <= 200 mg/kg. DANGER. Skull and crossbones in diamond. Fatal in contact with skin.
Category 3: LD50 > 200 mg/kg <= 1000 mg/kg. DANGER. Skull and crossbones in diamond. Toxic in contact with skin.
Category 4: LD50 > 1000 mg/kg <= 2000 mg/kg. WARNING. Exclamation point in diamond. Harmful in contact with skin.
Category 5: LD50 > 2000 mg/kg <= 5000 mg/kg. WARNING. No symbol. May be harmful in contact with skin.
Statistics:
According to OECD guidelines, the biological relevance of the results is the criterion for the interpretation of results,
a statistical evaluation of the results is not regarded as necessary.

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 was observed.
Clinical signs:
No treatment-related effects were observed.
Body weight:
The body weight development of all male and female animals was within the expected range.
A slight weight loss was recorded for 1 out of 5 female animals during the observation period. The effects on weight development might be
secondary to the dressing, and toxicological relevance of this finding cannot clearly be concluded.
The male animals showed weight gain during the first and the second week of the observation.
Gross pathology:
No treatment-related effects were observed.
Other findings:
Erythema grade 1 was observed in 0 of 5 male and 5 of 5 female animals. Eschar was observed in 0 of 5 male and 2 of 5 female animals.
Scratches were observed in 0 of 5 male and 2 of 5 female animals.
All signs of irritation were reversible within the observation period, except for eschar in one female animal.

Any other information on results incl. tables

Table Clinical Signs of Systemic Toxicity – Individual Data - Males:

Animal
No. / Sex / Dose

Time of
Observation (Post-Dose)

Observations

(for Signs of Dermal Irritation, see Table 5)

21/ male /
2000 mg/kg bw

during the whole observation period

no signs of toxicity

22/ male /
2000 mg/kg bw

during the whole observation period

no signs of toxicity

23/ male /
2000 mg/kg bw

during the whole observation period

no signs of toxicity

24/ male /
2000 mg/kg bw

during the whole observation period

no signs of toxicity

25/ male /
2000 mg/kg bw

during the whole observation period

no signs of toxicity

min = minute(s), h = hour(s), bw = body weight

Table Clinical Signs of Systemic Toxicity – Individual Data – Females:

Animal
No. / Sex / Dose

Time of
Observation (Post-Dose)

Observations

(for Signs of Dermal Irritation, see Table 6)

26/ female / 2000 mg/kg bw

during the whole observation period

no signs of toxicity

27/ female / 2000 mg/kg bw

during the whole observation period

no signs of toxicity

28/ female / 2000 mg/kg bw

during the whole observation period

no signs of toxicity

29/ female / 2000 mg/kg bw

day 1

no signs of toxicity

day 2

slight nasal discharge

day 3 until the end of the observation period

no signs of toxicity

30/ female / 2000 mg/kg bw

during the whole observation period

no signs of toxicity

min = minute(s), h = hour(s), bw = body weight

Table Skin Irritation – Individual Data – Males:

Day after Start of Application

Animal No. 21

Animal No. 22

Animal No. 23

Animal No. 24

Animal No. 25

E/O

Comments

E/O

Comments

E/O

Comments

E/O

Comments

E/O

Comments

day 2

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

day 3

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

day 4

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

day 5

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

day 6

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

day 7

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

day 8

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

day 9

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

0/0

nsf

0/0

nsf

day 10

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

0/0

nsf

0/0

nsf

day 11

0/0

*,nsf

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

day 12

0/0

*,nsf

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

day 13

0/0

*,nsf

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

day 14

0/0

*,nsf

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

day 15

0/0

*,nsf

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

 

Comments:

E = erythema; O = oedema; 1, 2, 3, 4 = scoring system laid down in OECD Guideline 404 (Table 2)

nsf = no specific findings, * = remainder of test substance

 

Table Skin Irritation – Individual Data – Females:

Day after Start of Application

Animal No. 26

Animal No. 27

Animal No. 28

Animal No. 29

Animal No. 30

E/O

Comments

E/O

Comments

E/O

Comments

E/O

Comments

E/O

Comments

day 2

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

day 3

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

day 4

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

0/0

*, es

0/0

*,nsf

day 5

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

0/0

*,nsf

day 6

1/0

*,nsf

1/0

*,nsf

1/0

*, s

1/0

*,nsf

1/0

*, s

day 7

1/0

*,nsf

1/0

*,nsf

1/0

*, s

1/0

*,nsf

1/0

*, s

day 8

0/0

*,nsf

0/0

*,nsf

1/0

*, s

0/0

*,nsf

0/0

*,nsf

day 9

0/0

*,nsf

0/0

*,nsf

0/0

*, s

0/0

*,nsf

0/0

*,nsf

day 10

0/0

*,nsf

0/0

*,nsf

0/0

*, s

0/0

*,nsf

0/0

*,nsf

day 11

0/0

*,nsf

0/0

*,nsf

0/0

*, s

0/0

*,nsf

0/0

*,nsf

day 12

0/0

*,nsf

0/0

*,nsf

0/0

*, s

0/0

*,nsf

0/0

*,nsf

day 13

0/0

*,nsf

0/0

*,nsf

0/0

*, s

0/0

*,nsf

0/0

*,nsf

day 14

0/0

*,nsf

0/0

*,nsf

0/0

*, s

0/0

*,nsf

0/0

*,nsf

day 15

0/0

*,nsf

0/0

*,nsf

0/0

*, es

0/0

*,nsf

0/0

*,nsf

 

Comments:

E = erythema; O = oedema; 1, 2, 3, 4 = scoring system laid down in OECD Guideline 404 (Table 2)

es = eschar; s = scratches; nsf = no specific findings, * = remainder of test substance

 

Table Absolute Body Weights in g and Body Weight Gain in %:

Dose: 2000 mg/kg body weight

Animal No. / Sex

g
Day 1

g
Day 8

g
Day 15

%
Day 1-15

21 / male

260

270

296

14

22 / male

263

270

296

13

23 / male

269

283

314

17

24 / male

266

271

302

14

25 / male

256

267

296

16

26 / female

221

219

228

3

27 / female

228

227

226

-1

28 / female

224

222

224

0

29 / female

215

220

228

6

30 / female

226

228

229

1

Table Macroscopic Findings - Individual Data – Males and Females:


Dose: 2000 mg/kg bw

Animal No. /
Sex

Organ

Macroscopic Findings

21 / male

-

nsf

22 / male

-

nsf

23 / male

-

nsf

24 / male

-

nsf

25 / male

-

nsf

26 / female

-

nsf

27 / female

-

nsf

28 / female

-

nsf

29 / female

lung

dark spots

30 / female

-

nsf

nsf = no specific findings

Table LD50:

Dose (Unit)

 

Number of Animals Investigated

Number of Intercurrent Deaths

LD50

2000 mg/kg bw

5 males

0

> 2000 mg/kg bw

2000mg/kg bw

5 females

0

> 2000 mg/kg bw

bw = body weight

Applicant's summary and conclusion

Interpretation of results:
study cannot be used for classification
Remarks:
Migrated information
Conclusions:
Under the conditions of the present study, single dermal application of the test item FAT 40853/A TE to rats at a dose of 2000 mg/kg body weight
was associated with no mortality and neither signs of toxicity, but signs of slight irritation.
The dermal LD50 was determined to be > 2000 mg FAT 40853/A TE / kg body weight.
In conformity with the criteria given in Annex VI to Commission Directive 2001/59/EC the test item FAT 40853/A TE has no obligatory
labelling requirement for percutaneous toxicity.
According to Annex I of Regulation (EC) 1272/2008 the test item FAT 40853/A TE has no obligatory labelling requirement for
percutaneous toxicity and is unclassified.
According to GHS (Globally Harmonized Classification System) the test item FAT 40853/A TE has no obligatory labelling requirement for
percutaneous toxicity and is not classified.

Executive summary:

LD50:  > 2000 mg /kg bw

Species/strain:  WISTAR Crl: WI(Han) rats

Vehicle (moistening):  Aqua ad injectionem

Number of animals: 5 male and 5 female

Duration of exposure: 24 hours

Method: OECD 402, EC 440/2008, Method B.3, OPPTS 870.1200 Signs of toxicity related to dose level used, time of onset and duration:

No treatment-related effects were observed.

Effect on organs (related to dose level):

No treatment-related effects were observed.

Signs of irritation: Erythema grade 1 was observed in 0 of 5 male and 5 of 5 female animals. Eschar was observed in 0 of 5 male and 2 of 5 female animals.Scratches were observed in 0 of 5 male and 2 of 5 female animals.

All signs of irritation were reversible within the observation period, except for eschar in one female animal.

Conclusion:

Under the conditions of the present study, single dermal application of the test itemFAT 40853/A TEto rats at a dose of 2000 mg/kg body weight was associated

with no mortality and neither signs of toxicity, but signs of slight irritation.

The dermal LD50 was determined to be > 2000 mgFAT 40853/A TE/ kg body weight.

In conformity with the criteria given inAnnex VI to Commission Directive 2001/59/EC the test itemFAT 40853/A TEhas no obligatory labelling requirement

for percutaneous toxicity.

According to Annex I of Regulation (EC) 1272/2008 the test itemFAT 40853/A TEhas no obligatory labelling requirement for percutaneous toxicity and is unclassified.

According to GHS (Globally Harmonized Classification System) the test itemFAT 40853/A TEhas no obligatory labelling requirement for percutaneous toxicity

and is not classified.