Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
13 August 2018 - 03 December 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report Date:
2018

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
OECD Guidelines for Testing of Chemicals, Section 4, No. 402, “Acute Dermal Toxicity” adopted 09
Oct, 2017
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid
Details on test material:
Colour: light yellow
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: SUQIAN UNITECH CO., LTD; 2018041002
- Purity: 99.29%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature, protected from light

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: On sponsor’s request the dose was calculated based on the purity of the active component and a correction factor of 1.01 was applied.

Test animals

Species:
rat
Strain:
other: WISTAR Crl: WI(Han)
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River, 97633 Sulzfeld, Germany
- Females (if applicable) nulliparous and non-pregnant: [yes]
- Age at study initiation: 7–10 weeks
- Weight at study initiation: 201–220 g
- Diet: Free access to Altromin 1324 maintenance diet for rats and mice
- Water: 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)
- Acclimation period: controlled full-barrier maintained breeding system (SPF)

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 55 ± 10%
- Air changes (per hr): 10 x / hour
- Photoperiod (hrs dark / hrs light): Artificial light, sequence being 12 hours light, 12 hours dark

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
water
Remarks:
aqua ad injectionem (sterile water)
Details on dermal exposure:
The test item was applied at a single dose, uniformly over an area which was approximately 10% of the total body surface. 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. The test item was held in contact with the skin by a dressing throughout a 24-hour period. This consisted of a semi-occlusive dressing made of a porous gauze and non-irritating tape and was fixed with an additional dressing in a suitable manner. At the end of the exposure period the residual test item was removed using aqua ad injectionem (sterile water).
Duration of exposure:
24 hrs
Doses:
Dose range finding : 2000 mg/kg bw.
Main test: 2000 mg/kg bw
No. of animals per sex per dose:
DRF: 1 female
Main test: 2 females
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Observation during the whole observation period. The animals were weighed on day 1 (prior to the application), on days 8 and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: Signs of erythema and oedema were assessed using the scoring system laid down in OECD Guideline 404 24, 48 and 72 hours after patch removal.
A 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 6 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.
All animals were subjected to gross necropsy and examined macroscopically for gross pathological changes. In absence of gross pathological changes no tissues were preserved for a possible histopathological evaluation.

Results and discussion

Preliminary study:
The test item showed neither mortality nor signs of acute dermal toxicity but signs of dermal irritation after a single dose application (erythema grade 1, oedema grade 1 and desquamation. All signs of irritation were reversible within up to day 10.). A slight weight loss was recorded during the first week, but the animal showed weight gain during the second week. The effects on weight development might be secondary to the dressing, and toxicological relevance of this finding cannot clearly be concluded. The abdominal region had a yellow solid mass upon necropsy. (Tables 2,4 6,7). The dose of 2000 mg/kg bw was used in the main study.
Effect levels
Key result
Sex:
female
Dose descriptor:
approximate LD50
Effect level:
> 2 000 - <= 5 000 mg/kg bw
Mortality:
The test item showed neither mortality nor signs of acute dermal toxicity.
Clinical signs:
No signs of toxicity were observed througout the entire observation period (Table 3).

Erythema grade 2 in animal no. 2 (main study) and grade 1 in animal no. 3 (main study) was observed. Besides erythema, animal no. 2 (main study) showed oedema grade 1 and desquamation and animal no. 3 (main study) showed scratches. All signs of irritation were reversible within up to day 10. (Table 5)
Body weight:
A slight weight loss was recorded for 1 out of 2 female animals during the first week, but all of the female animals showed weight gain during the second week. The effects on weight development might be secondary to the dressing, and toxicological relevance of this finding cannot clearly be concluded (Table 6).
Gross pathology:
No specific gross pathological changes were recorded for any other animal in the main study (Table 7).

Any other information on results incl. tables

Table 2:  Clinical Signs of Systemic Toxicity - Individual Data – Dose Range Finding Study

Animal
No. / Sex / Dose

Time of Observation

Observation

1 / female /
2000 mg/kg bw

during the whole observation period

no signs of toxicity

bw = body weight

Table 3:  Clinical Signs of Systemic Toxicity - Individual Data – Main Study

Animal
No. / Sex / Dose

Time of Observation

Observations

2 / female /
2000 mg/kg bw

during the whole observation period

no signs of toxicity

3 / female /
2000 mg/kg bw

during the whole observation period

no signs of toxicity

bw = body weight

Table 4:  Skin Irritation at Application Site – Individual Data – Dose Range Finding Study  

Study Day (Time after Patch Removal)

Animal No. 1

E/O

C

2 (0±2 h)

0/0

nsf

3 (24±2 h)

1/0

nsf

4 (48±2 h)

1/0

nsf

5 (72±2 h)

1/0

des

6

1/1

des

7

0/0

des

8

0/0

des

9

0/0

nsf

10

0/0

nsf

11

0/0

nsf

12

0/0

nsf

13

0/0

nsf

14

0/0

nsf

15

0/0

nsf

h = hours, d = day(s): study day 1 = day of test item application;

C = Comments; E = erythema; O = oedema;nsf = no specific findings;
0, 1, 2, 3, 4 = scoring system laid down in OECD Guideline 404;
des = desquamation

Table 5:  Skin Irritation at Application Site – Individual Data – Main Study

Study Day (Time after Patch Removal)

Animal No. 2

Animal No. 3

E/O

C

E/O

C

2 (0±2 h)

1/0

nsf

1/0

nsf

3 (24±2 h)

1/0

nsf

1/0

s

4 (48±2 h)

1/0

nsf

1/0

s

5 (72±2 h)

2/1

des

1/0

s

6

2/1

des

0/0

s

7

2/1

des

0/0

s

8

1/0

nsf

0/0

nsf

9

1/0

nsf

0/0

nsf

10

0/0

nsf

0/0

nsf

11

0/0

nsf

0/0

nsf

12

0/0

nsf

0/0

nsf

13

0/0

nsf

0/0

nsf

14

0/0

nsf

0/0

nsf

15

0/0

nsf

0/0

nsf

h = hours, d = day(s): study day 1 = day of test item application;

C = Comments; E = erythema; O = oedema;nsf = no specific findings;
0, 1, 2, 3, 4 = scoring system laid down in OECD Guideline 404;
des = desquamation; s = scratches

Table 6:  Absolute Body Weights in g and Body Weight Change in % - DRF and Main Study

Dose: 2000 mg/kg body weight

Animal No. / Sex

g
Day 1

g
Day 8

g
Day 15

%
Day 1-15

1 / female

201

199

207

3

2 / female

220

221

222

1

3 / female

209

207

208

0

bw = body weight


Table 7:  Macroscopic Findings - Individual Data – DRF and Main Study

Dose: 2000 mg/kg bw

Animal No. / Sex

Organ with Macroscopic Findings

Macroscopic Findings

1 / female

Abdominal region

Yellowish solid mass

2 / female

-

nsf

3 / female

-

nsf

nsf = no specific findings



Applicant's summary and conclusion

Interpretation of results:
Category 5 based on GHS criteria
Remarks:
Not classified under CLP
Conclusions:
Under the conditions of the present study, single dermal application of the test item 4,6-dichloro-N-(1,1,3,3-tetramethylbutyl)-1,3,5-triazin-2-amine to rats at a dose of 2000 mg/kg body weight was associated with neither mortality nor signs of toxicity but signs of irritation. The LD50 was >2000 mg/kg bw.
Executive summary:

In an acute dermal toxicity study (185459), 3 Wistar Crl: WI(Han) female rats were dermally exposed (10% total body area; semi-occlusive) to the test item 4,6-dichloro-N-(1,1,3,3-tetramethylbutyl)-1,3,5-triazin-2-amine (99.29%) in sterile water for 24 hours at doses of  2000 mg/kg bw. At the end of the exposure period the residual test item was removed using sterile water.

The LD50 (female) was >2000 mg/kg bw.

In the dose-range finding study with 1 female, the test item showed neither mortality nor signs of acute dermal toxicity but signs of dermal irritation after a single dose application (erythema grade 1, oedema grade 1 and desquamation; all signs of irritation were reversible within up to day 10). A slight weight loss was recorded during the first week, but the animal showed weight gain during the second week. The effects on weight development might be secondary to the dressing, and toxicological relevance of this finding cannot clearly be concluded. The abdominal region had a yellow solid mass upon necropsy. In the main study with 2 females; the test item showed neither mortality nor signs of acute dermal toxicity. No clinical signs of toxicity were observed throughout the entire observation period. Erythema grade 2 in animal no. 2 and grade 1 in animal no. 3 was observed. Besides erythema, animal no. 2 showed oedema grade 1 and desquamation and animal no. 3 showed scratches. All signs of irritation were reversible within up to day 10. A slight weight loss was recorded for 1 out of 2 female animals during the first week, but all of the female animals showed weight gain during the second week. The effects on weight development might be secondary to the dressing, and toxicological relevance of this finding cannot clearly be concluded. No specific gross pathological changes were recorded for any other animal in the main study.