Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
26 June 2018 - 14 Aug 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 423 (Acute Oral toxicity - Acute Toxic Class Method)
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes

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: The test item was weighed out into a tared plastic vial on a precision balance and finely ground with the help of a mortar and pestle. The test item was then suspended with the vehicle corn oil to gain a final volume of 10 mL and to achieve a dose of 2000mg/kg body weight at a dose volume of 10 mL/kg body weight. Homogeneity of the test item in the vehicle was maintained by vortexing the prepared suspension thoroughly before each dose administration.

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: 8–10 weeks
- Weight at study initiation: Step 1: 173–181 g;Step 2: 178–183 g
- Fasting period before study: 16 to 19 hours
- Housing: Full barrier in an air-conditioned room
- 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: at least five days

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

Route of administration:
oral: gavage
Vehicle:
other: corn oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 0.2 g/mL
- Amount of vehicle (if gavage): 10 mL/kg
- Justification for choice of vehicle: The test item does not make a solution or suspension with water, so the corn oil has to be used.
- Lot/batch no. (if required): MKCD1821

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: The dose level of 2000 mg/kg bw was used as the starting dose, according to the test guideline, No compound-related mortality was recorded for any other animals of step 1 or 2.
Doses:
Steps 1 & 2: 2000 mg/kg body weight
No. of animals per sex per dose:
3 per step
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: 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 4 hours post-dose). Thereafter, the animals were observed for clinical signs once daily until the end of the observation period. The animals were weighed on day 1 (prior to the administration) and on days 8 and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, gross necropsy.

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50 cut-off
Effect level:
2 500 mg/kg bw
Mortality:
For one animal (-Step 1, Animal No.1) compound-related mortality occurred. No compound-related mortality was recorded for any other animals of step 1 or 2.
Clinical signs:
The most relevant clinical findings were reduced spontaneous activity, hunched posture, piloerection, eyes half closed, diarrhoea and slight weight loss during the first study week. All animals recovered within up to 8 days post-dose (Table 1).
Body weight:
One day after test item administration a slight weight loss was observed in 5 out of 6 animals. By the end of the first week, 1 of 5 surviving animals still showed a slight weight loss. All of the surviving animals showed an overall weight gain during the 14-day study period (for individual data see Table 2).
Gross pathology:
Detailed pathological findings of the study are presented in Table 3.
No specific gross pathological changes were recorded for any animal.

Any other information on results incl. tables

Table 1: Clinical Signs - Individual Data

Step Animal No. / Sex Starting Dose (mg/kg bw) Time Point  Observations

1

1 / Female 2000 0 min – 30 min nsf
30 min – 120 min Hunched posture,
slight piloerection
120 min – d 2 Slightly reduced spontaneous activity, hunched posture, moderate piloerection, moderate diarrhoea
d 2 Slightly reduced spontaneous activity, hunched posture, moderate piloerection, slight diarrhoea,
slight weight loss (2%)
d 3 found dead
2 / Female 2000 0 min – 30 min nsf
30 min – 120 min Hunched posture,
slight piloerection
120 min – d 2 Slightly reduced spontaneous activity, hunched posture, moderate piloerection, moderate diarrhoea
d 2 – d 3 Slightly reduced spontaneous activity, hunched posture, moderate piloerection, moderate diarrhoea, slight weight loss
(3% by d 3)
d 4 Moderately reduced spontaneous activity, hunched posture, moderate piloerection
d 5 – d 8 Slight piloerection
d 9 – d 15 nsf
3 / Female 2000 0 min – 30 min nsf
30 min – 120 min Hunched posture,
slight piloerection
120 min – d 2 Slightly reduced spontaneous activity, hunched posture, moderate piloerection, diarrhoea moderate
d 2 Slightly reduced spontaneous activity, hunched posture, moderate piloerection, moderate diarrhoea, slight weight loss (<1%)
d 3 Slightly reduced spontaneous activity, hunched posture, moderate piloerection, slight diarrhoea
d 4 Moderately reduced spontaneous activity, hunched posture,
moderate piloerection
d 5 Slight piloerection
d 6 Moderate piloerection
d 7 – d 8 Slight piloerection
d 9 – d 15 nsf

2

4 / Female 2000 0 min – 30 min nsf
30 min – 60 min Slight piloerection
60 min – 120 min Slightly reduced spontaneous activity, hunched posture,
slight piloerection
120 min – 180 min Slightly reduced spontaneous activity, hunched posture, moderate piloerection, half eyelid-closure, moderate diarrhoea
180 min – 240 min Spontaneous activity moderately reduced, hunched posture, moderate piloerection,
moderate diarrhoea
240 min – d 2 Spontaneous activity moderately reduced, hunched posture, moderate piloerection, half eyelid-closure, moderate diarrhoea
d 2 Slight piloerection, severe diarrhoea, slight weight loss (1%)
d 3 Slight diarrhoea
d 4 – d 15 nsf
5 / Female 2000 0 min – 30 min nsf
30 min – 60 min Slight piloerection
60 min – 120 min Slightly reduced spontaneous activity, hunched posture,
slight piloerection
120 min – 180 min Slightly reduced spontaneous activity, hunched posture, moderate piloerection, half eyelid-closure, moderate diarrhoea
180 min – 240 min Moderately reduced spontaneous activity, hunched posture, moderate piloerection,
moderate diarrhoea
240 min – d 2 Moderately reduced spontaneous activity, hunched posture, severe piloerection, half eyelid-closure, moderate diarrhoea
d 2 Slight piloerection, severe diarrhoea, slight weight loss (7%)
d 3 Slight diarrhoea
d 4 – d 15 nsf
6 / Female 2000 0 min – 30 min nsf
30 min – 60 min Slight piloerection
60 min – 120 min Spontaneous activity slightly reduced, hunched posture,
slight piloerection
120 min – 180 min Slightly reduced spontaneous activity, hunched posture, moderate piloerection, half eyelid-closure, diarrhoea moderate
180 min – 240 min Moderately reduced spontaneous activity, hunched posture, moderate piloerection,
moderate diarrhoea
240 min – d 2 Moderately reduced spontaneous activity, hunched posture, moderate piloerection, half eyelid-closure, moderate diarrhoea
d 2 Slight piloerection,
severe diarrhoea
d 3 Slight diarrhoea
d 4 – d 15 nsf

bw = body weight; d = day (day 1 = day of administration); min = minute(s); nsf = no specific findings

Table 2:Absolute Body Weights in g and Body Weight Change in %

Step Animal No. / Sex Starting Dose (mg/kg bw) BW (g) Body Weight Change in Comparison to Day 1 (%)
Day 1 Day 8 Day 15 Day 15
1 1 / Female 2000 181 Found dead on study day 3
2 / Female 2000 175 194 225 29
3 / Female 2000 173 205 208 20
2 4 / Female 2000 183 196 203 11
5 / Female 2000 179 177 197 10
6 / Female 2000 178 184 210 18

bw= body weight

Table 3:Findings of the Necropsy - Individual Data

Step Animal No. / Sex Starting Dose (mg/kg bw) Organ Macroscopic Findings
1 1 / Female 2000  -  nsf
2 / Female  -  nsf
3 / Female  -  nsf
2 4 / Female  -  nsf
5 / Female  -  nsf
6 / Female  -  nsf

bw = body weight;nsf = no specific findings

Applicant's summary and conclusion

Interpretation of results:
Category 5 based on GHS criteria
Remarks:
Not classified according to CLP
Conclusions:
The median lethal dose of 4,6-dichloro-N-(1,1,3,3-tetramethylbutyl)-1,3,5-triazin-2-amine after a single oral administration to female rats, observed over a period of 14 days is: LD50 cut-off (rat): 2500 mg/ kg bw
Executive summary:

In an acute oral toxicity test (183811), 2 groups of female Wistar Crl: WI(Han) rats (3/group) were administered 4,6-dichloro-N-(1,1,3,3-tetramethylbutyl)-1,3,5-triazin-2-amine (99.29%) in corn oil by oral gavage at a dose of 2000 mg/kg bw.

The LD50 (cut-off) was 2500 mg/kg bw.

One animal of step 1 was found dead 2 days post-application. All remaining animals survived until the end of the study showing signs of toxicity. The most relevant clinical findings were reduced spontaneous activity, hunched posture, piloerection, eyes half closed, diarrhoea and slight weight loss during the first study week. All animals recovered within up to 8 days post-dose. Throughout the 14-day observation period, the weight gain of the surviving animals was within the normal range of variation for this strain. At necropsy, no treatment-related macroscopic findings were observed in any animal of any step.