Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
11 May 2017 - 30 May 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
GLP compliance:
yes
Test type:
fixed dose procedure

Test material

Reference
Name:
Unnamed
Test material form:
liquid
Remarks:
clear colourless
Details on test material:
UVCB
At room temperature protected from light
31 March 2018 (expiry date)

Test animals

Species:
rat
Strain:
Wistar
Remarks:
Crl: WI(Han)
Sex:
male/female
Details on test animals and environmental conditions:
Source: Charles River UK, Kent, England
Outbred, SPF-quality
Young adult animals (approximately 10 weeks)
Weight at the Initiation of Dosing: 187 to 338 g.

On arrival, animals were group housed (up to 5 animals of the same sex together) in polycarbonate cages (Makrolon MIV type; height 18 cm.) and following assignment to the study, animals were individually housed in polycarbonate cages (Makrolon MIII type; height 18 cm.) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) equipped with water bottles.

The animals were allowed to acclimate to the Test Facility toxicology accommodation for at least 5 days before the commencement of dosing.
Target temperatures of 18 to 24°C with a relative target humidity of 40 to 70% were maintained. The actual daily mean temperature during the study period was 21 to 22°C with an actual daily mean relative humidity of 43 to 69%. A 12 hour light/12 hour dark cycle was maintained. Ten or greater air changes per hour with 100% fresh air (no air recirculation) were maintained in the animal rooms.

For psychological/environmental enrichment, animals were provided with paper (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom), except when interrupted by study procedures/activities.

Administration / exposure

Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
A single dose of test item was administered to the appropriate animals by dermal application on Day 1. One day before dosing, an area of approximately 5x7 cm on the back of the animals was clipped. The test item was applied in an area of approximately 10% of the total body surface, i.e. approximately 25 cm² for males and 18 cm² for females. The test item was held in contact with the skin with a dressing, consisting of a surgical gauze patch (Surgy 1D), successively covered with aluminum foil and Coban elastic bandage. A piece of Micropore tape was additionally used for fixation of the bandages in females only.
Duration of exposure:
The application period was 24 hours, after which the dressing was removed and the skin cleaned of residual test item using water or an appropriate vehicle.
Doses:
The dose level was 2000 mg/kg body weight.
No. of animals per sex per dose:
5 males and 5 females (females were nulliparous and non-pregnant)
Control animals:
no
Details on study design:
Throughout the study, animals were observed for general health/mortality and moribundity twice daily, in the morning and at the end of the working day.

Post-dose observations were performed at periodic intervals on the day of dosing (at least three times) and once daily thereafter. The observation period was 14 days. All the animals were examined for reaction to dosing. The onset, intensity and duration of these signs was recorded (if appropriate), particular attention being paid to the animals during and for the first hour after dosing.

Animals were weighed individually on Day 1 (pre-dose), 8 and 15.

All moribund animals and animals surviving to the end of the observation period were sacrificed by oxygen/carbon dioxide procedure. All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities were recorded.

Results and discussion

Effect levels
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Mortality:
One female was found dead on Day 2, no further mortality occurred.
Clinical signs:
Tremor, piloerection, red (snout), ptosis and/or hypothermia were noted for the majority of animals on Days 1 and/or 2.
Focal erythema, scales and/or scabs were seen in the treated skin-area of two males and one female. These local effects were considered not to have affected the conclusion of the study.
Body weight:
The mean body weight gain shown by the surviving animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain.
Gross pathology:
Beginning autolysis was noted for the animal that died during the study, at macroscopic post mortem examination. Macroscopic post mortem examination of the other animals at termination did not reveal any abnormalities.

Any other information on results incl. tables

TABLE 1 MORTALITY DATA


 
TEST DAY
HOURS AFTER TREATMENT
 

 
1
0
 

 
1
2
 

 
1
4
 

 
2

 

 
3

 

 
4

 

 
5

 

 
6

 

 
7

 

 
8

 

 
9

 

 
10

 

 
11

 

 
12

 

 
13

 

 
14

 

 
15

 

'

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

MALES 2000 MG/KG

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

.

FEMALES 2000 MG/KG

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

.

TABLE 2 CLINICAL SIGNS


 
TEST DAY
HOURS AFTER TREATMENT
 

 

MAX
GRADE
 

 
1
0

 

 
1
2

 

 
1
4

 

 
2

 

 
3

 

 
4

 

 
5

 

 
6

 

 
7

 

 
8

 

 
9

 

 
10

 

 
11

 

 
12

 

 
13

 

 
14

 

 
15

 

'


MALES 2000 MG/KG

ANIMAL 1

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

Skin / fur

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

 Piloerection

(1)

-

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

.

Various

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

 Ptosis

(3)

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

.

 Red (Snout)

(1)

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

.

ANIMAL 2

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

Skin / fur

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

 Erythema focal (Treated skin)

(4)

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

.

Various

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

 Ptosis

(3)

1

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

.

ANIMAL 3

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

Skin / fur

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

 Piloerection

(1)

-

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

.

 Scales (Flank right)

(3)

-

-

-

-

-

-

1

1

1

1

1

-

-

-

-

-

-

.

Various

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

 Ptosis

(3)

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

.

 Red (Snout)

(1)

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

.

ANIMAL 4

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

Skin / fur

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

 Erythema focal (Treated skin)

(4)

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

.

 Piloerection

(1)

-

-

1

1

1

-

-

-

-

-

-

-

-

-

-

-

-

.

Various

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

 Ptosis

(3)

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

.

ANIMAL 5

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

No clinical signs noted

 

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

.


FEMALES 2000 MG/KG

ANIMAL 6

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

Skin / fur

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

 Piloerection

(1)

-

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

.

Various

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

 Ptosis

(3)

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

.

 Red (Snout)

(1)

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

.

ANIMAL 7

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

Skin / fur

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

 Piloerection

(1)

-

-

1

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

Various

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

 Ptosis

(3)

-

-

1

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

ANIMAL 8

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

Spasms

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

 Tremor

(3)

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

.

Skin / fur

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

 Piloerection

(1)

-

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

.

Various

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

 Ptosis

(3)

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

.

 Hypothermia

(1)

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

.

 

- = sing not observed

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

TABLE 2 CLINICAL SIGNS (continued)


 
TEST DAY
HOURS AFTER TREATMENT
 

 

MAX
GRADE
 

 
1
0

 

 
1
2

 

 
1
4

 

 
2

 

 
3

 

 
4

 

 
5

 

 
6

 

 
7

 

 
8

 

 
9

 

 
10

 

 
11

 

 
12

 

 
13

 

 
14

 

 
15

 

'


FEMALES 2000 MG/KG

ANIMAL 9

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

Skin / fur

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

 Piloerection

(1)

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

.

Various

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

 Ptosis

(3)

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

.

ANIMAL 10

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

Skin / fur

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

.

 Erythema focal (Treated skin)

(4)

-

-

-

1

1

1

-

-

-

-

-

-

-

-

-

-

-

.

 Piloerection

(1)

-

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

.

 Scabs (Treated skin)

(3)

-

-

-

-

1

1

1

1

1

1

1

-

-

-

-

-

-

.

 

- = sign not observed

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

TABLE 3 BODY WEIGHTS (GRAM)


 

 
SEX/DOSE LEVEL
 

 
ANIMAL
 

 
DAY 1
 

 
DAY 8
 

 
DAY 15
 

'


MALES 2000 MG/KG

 

 

1

301

299

324

.

 

2

321

336

360

.

 

3

313

321

351

.

 

4

323

321

343

.

 

5

338

347

378

.

 

 

 

 

 

.

 

MEAN

319

325

351

.

 

ST.DEV.

14

18

20

.

 

N

5

5

5

.

 

 

 

 

 

.


FEMALES 2000 MG/KG

 

 

6

217

218

240

.

 

7

185*

---

---

.

 

8

196

202

218

.

 

9

214

223

240

.

 

10

197

201

213

.

 

 

 

 

 

.

 

MEAN

202

211

228

.

 

ST.DEV.

13

11

14

.

 

N

5

4

4

.

 

 

 

 

 

.

* Animal was found dead on Day 2, terminal body weight was 189 grams.

 

 

 

TABLE 4 MACROSCOPIC FINDINGS


 
ANIMAL
 

 
ORGAN
 

 
FINDING
 

 
DAY OF DEATH
 

'


MALES 2000 MG/KG

1

 

No  findings noted

Scheduled necropsy

.

 

 

 

Day 15 after treatment

.

2

 

No  findings noted

Scheduled necropsy

.

 

 

 

Day 15 after treatment

.

3

 

No  findings noted

Scheduled necropsy

.

 

 

 

Day 15 after treatment

.

4

 

No  findings noted

Scheduled necropsy

.

 

 

 

Day 15 after treatment

.

5

 

No  findings noted

Scheduled necropsy

.

 

 

 

Day 15 after treatment

.


FEMALES 2000 MG/KG

6

 

No  findings noted

Scheduled necropsy

.

 

 

 

Day 15 after treatment

.

7

General observations

Beginning  autolysis.

Spontaneous death

.

 

 

 

Day 2 after treatment

.

8

 

No  findings noted

Scheduled necropsy

.

 

 

 

Day 15 after treatment

.

9

 

No  findings noted

Scheduled necropsy

.

 

 

 

Day 15 after treatment

.

10

 

No  findings noted

Scheduled necropsy

.

 

 

 

Day 15 after treatment

.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The dermal LD50 value of MEA Polyborate 1:3 in Wistar rats was established to exceed 2000 mg/kg body weight.