Registration Dossier

Toxicological information

Acute Toxicity: oral

Currently viewing:

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
09 May 2018 - 29 May 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)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid

Test animals

Species:
rat
Strain:
Sprague-Dawley
Remarks:
(SPF Caw)
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Elevage JANVIER LABS (53940 Le Genest St Isle – France)
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 8 or 9 weeks
- Weight at study initiation: 214.0 g (SD = 18.9 g)
- Fasting period before study: Food was removed on D-1 and then redistributed 4 hours after the test item administration.
- Housing: Rats were housed by group of three in solid-bottomed clear polycarbonate cages with a stainless steel mesh lid. Each cage contained sawdust bedding which was changed at least 2 times a week. Each cage was installed in conventional air conditioned animal husbandry.
- Diet (e.g. ad libitum): Ad libitum. Teklad Global 16% Protein Rodent Diet (ENVIGO 2016).
- Water (e.g. ad libitum): Ad libitum. Drinking water (tap-water from public distribution system). Microbiological and chemical analyses of the water were carried out once every six months by Bureau Veritas – Eurofins (FRANCE).
- Acclimation period: the animals were acclimatized for at least 5 days before treatment.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19ºC to 25ºC
- Humidity (%): 30 to 70%
- Air changes (per hr): >10 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours light (07.00 to 19.00) and 12 hours dark cycle.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
MAXIMUM DOSE VOLUME APPLIED: 2.26 mL/kg body weight (corresponding to 2 g/kg, according to the calculated density)

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Available information suggests that mortality is unlikely at the highest starting dose level (2000 mg/kg body weight). Hence, the study is initiated with the starting dose of 2000 mg/kg body weight.
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
6 (3 female rats per step)
Control animals:
yes
Remarks:
(study performed on three animals receiving distilled water under requirements of OECD Guideline 423)
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: At each step, the animals were observed four times on test day 0 (day of administration), i.e. at T0+30 min, T0+1h, T0+3h and T0+4h, and once daily during days 1 to 14 post administration. The body weights were recorded on test day 0 (just before administration) then on D2, D7, and D14.
- Necropsy of survivors performed: yes. At termination, gross pathological findings were recorded and reported.
- Other examinations performed:
Clinical observations included: spontaneous activity, Preyer’s reflex (noise), respiratory rate, convulsions, tremors, body temperature, muscle tone, palpebral opening, pupil appearance, salivation, lachrymation, righting reflex, back hair appearance.

Results and discussion

Effect levelsopen allclose all
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Key result
Sex:
female
Dose descriptor:
LD50 cut-off
Effect level:
2 500 mg/kg bw
Based on:
test mat.
Mortality:
Two mortalities were noted in animals treated at the dose of 2000 mg/kg body weight at 24 hours post dose (one in each step).
Clinical signs:
The mortalities were preceded by a decrease or an absence of spontaneous activity (2/2), Preyer’s reflex (2/2), muscle tones (2/2), righting reflex (2/2), associated with eyes partly closed (1/2). Rigor mortis (2/2) was noted before the necropsy.
In surviving animals (4/6), a decrease of spontaneous activity (4/4), Preyer’s reflex (2/4), muscle tones (4/4), righting reflex (4/4), associated with eyes partly closed (2/4) were noted in the first hours of the test. The animals recovered a normal activity at 24 hours post-dose.
Body weight:
The body weight evolution of the surviving animals remained normal during the study.
Gross pathology:
The macroscopic examination of the animals found dead revealed a thinning of corpus (1/2) and forestomach (1/2) with red spots (1/2).
The macroscopic examination of the surviving animals at the end of the study did not reveal treatment related changes.

Any other information on results incl. tables

Table 1: Body weight and weight gain in grams

Females

D0

D2

D2-D0

D7

D7-D0

D14

D14-D0

Rf 2491

195

225

30

239

44

269

74

Rf 2492

197

218

21

247

50

269

72

Rf 2493

202

 

 

 

 

 

Rf 2535

229

229

0

252

23

271

42

Rf 2536

241

246

5

290

49

316

75

Rf 2537

220

 

 

 

 

 

MEAN

214.0

229.5

14.0

257.0

41.5

281.3

65.8

SD

18.9

11.9

13.9

22.6

12.6

23.2

15.9

Note: †: Rf2493 and Rf2537 found dead on D1

Applicant's summary and conclusion

Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The LD50 of the test item is higher than 2000 mg/kg body weight by oral route in the rat. The LD50 cut-off may be considered to be 2500 mg/kg body weight by oral route in the rat.
Executive summary:

The acute oral toxicity of the test item was tested in accordance with OECD Test Guideline 423, following GLP. 6 female Sprague-Dawley rats divided in 2 groups were administered sequentially with test item by oral gavage. The first set of 3 female rats was given a single dose of 2000 mg/kg body weight. One mortality was observed after 24 h of treatment, so the second set of 3 female rats was treated at the same dose level. One mortality was also observed at this dose level and at 24 hours post dose. In surviving animals (4/6), a decrease of spontaneous activity (4/4), Preyer’s reflex (2/4), muscle tones (4/4), righting reflex (4/4), associated with eyes partly closed (2/4) were noted in the first hours of the test. The animals recovered a normal activity at 24 hours post-dose. The body weight evolution of the animals remained normal during the study. The macroscopic examination of the animals found dead revealed a thinning of corpus (1/2) and forestomach (1/2) with red spots (1/2), while the animals which survived until the end of the study did not reveal treatment related changes. Based on these results, the LD50 of the test item was determined to be higher than 2000 mg/kg bw and, as per OECD Test Guideline 423, the LD50 cut-off of the test item may be considered to be 2500 mg/kg bw.