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Administrative data

Description of key information

Acute oral toxicity: Key study: OECD 423 and EU method B.1 tris. GLP study. The LD50 of the test item is higher than 2000 mg/kg body weight by oral route in the rat. The LD50 cut-off of the test item may be considered to be higher than 5000 mg/kg body weight by oral route in the rat.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From June 5th, 2018 to June 20th, 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
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
Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Six Sprague Dawley rats (SPF Caw) supplied by Elevage JANVIER LABS (53940 Le Genest St Isle – France)
- Females nulliparous and non-pregnant: yes
- Age at study initiation: At the beginning of the study, the animals were 8-week old, with a mean body weight of 195 g (SD: 2.9).
- Fasting period before study: yes
- Housing: Healthy female rats were housed by group of three in solid-bottomed clear polycarbonate cages with a stainless steel mesh lid. Each cage contains 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): standard laboratory foodstuff (ENVIGO 2016) ad libitum.
- Water (e.g. ad libitum): Tap water ad libitum.
- Acclimation period: at least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25ºC
- Humidity (%): 30-70%
- Air changes (per hr): at least 10 times/hour
- Photoperiod (hrs dark / hrs light): 12 hours light/12 hours dark.

IN-LIFE DATES: From: 05 June 2018 (step 1) & 06 June 2018 (step 2)
Route of administration:
oral: gavage
Vehicle:
DMSO
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 0.2 g/mL in stock solution.
- Justification for choice of vehicle: Dimethyl sulfoxide (DMSO) was chosen as it produced the most suitable formulation at the requested concentration.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

DOSAGE PREPARATION: In the first and second step of the study, 2.0053 g and 2.0001 g of the test item were weighed and DMSO was added to two 10 mL volumetric flasks. Just before the administration, the preparations were stirred using a vortex to obtain yellowish solutions. Each preparation was administered under a volume of 10 mL/kg body weight using a suitable syringe graduated fitted with an oesophageal metal canula.

CLASS METHOD
- Rationale for the selection of the starting dose: As no information regarding the acute toxicity of the test item was available suggesting toxicity of the test item and in accordance with the principles of animal welfare, the first tested dose was 2000 mg/ kg b.w.
Doses:
Step 1: 2000 mg/kg bw.
Step 2: 2000 mg/kg bw.
No. of animals per sex per dose:
3 female rats were used in step 1 (Rf2644, Rf2658, Rf2659) and 3 female rats in step 2 (Rf2649 to Rf2651)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Systematic examinations were carried out to identify any behavioural or toxic effects on the major physiological functions 30 min, 1h, 3h, 4h, 24h, 48h after administration of the test item and continued daily during 14 days. Weight was determined on day 0 (directly before administration), 2, 7 and 14 before euthanasia.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, gross examinations.
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:
> 5 000 mg/kg bw
Based on:
test mat.
Mortality:
No treatment-related mortalities occurred during the study: one death (1/6) was noted in animals treated at the dose of 2000 mg/kg body weight on day 9. The mortality was preceded by dyspnea from day 2 to day 6 post dose, noisy breathing between days 7 and 9, swollen abdomen and piloerection on day 8 and a loss of body weight (-14% on day 2 and -29% on day 7). Rigor mortis and swollen abdomen were noted before the necropsy. The macroscopic examination of the animal revealed black spots on the corpus and air inside the digestive system. Thus, thiis death was due to a gavage injury.
Clinical signs:
In the surviving animal (5/6), no clinical signs related to the administration of the test item were observed during the study
Body weight:
The body weight evolution of the animals remained normal during the study.
Gross pathology:
The macroscopic examination of the animals at the end of the study did not reveal treatment related changes.

Table 1. Body weights and weight gain of the animals (grams)

FEMALES

D0

D2

D2-D0

D7

D7-D0

D14

D14-D0

Rf 2644

Rf 2658

Rf 2659

208

195

188

226

212

160

18

17

-28

248

224

133

40

29

-55

273

243

65

48

 

Rf 2649

Rf 2650

Rf 2651

208

201

189

203

222

238

-5

21

49

219

240

250

11

39

61

236

267

278

28

66

89

MEAN

Standard deviation

198.2

8.9

210.2

27.3

12.0

26.1

219.0

44.0

20.8

40.6

259.4

18.7

59.2

22.7

†: Animal Rf2659 found dead on D9 

Table 2. Clinical signs, overall list. 

Dose

(mg/kg bw)

Observation

time

Mortality

Step 1. / Animal Rf.

Step 2. / Animal Rf.

2644

2658

2659*

2649

2650

2651

2000

30min

0

N

N

N

N

N

N

1h

0

N

N

N

N

N

N

3h

0

N

N

N

N

N

N

4h

0

N

N

N

N

N

N

D1

0

N

N

N

N

N

N

D2-6

0

N

N

Dy

N

N

N

D7

0

N

N

N

N

N

N

D8

0

N

N

Pi

N

N

N

D9

1

N

N

-

N

N

N

D10-14

0

N

N

-

N

N

N

*Rf2659: wheezing on D2, noisy breathing on D7, noisy breathing and swollen stomach on D8, found dead on D9 (due to gavage injury).

 

Table 3. Necropsy findings, overall list.

Observations

Step 1. / Animal Rf.

Step 2. / Animal Rf.

2644

2658

2659*

2649

2650

2651

General Appearance

N

N

Rigor mortis,

swollen abdomen

N

N

N

Oesophagus

N

N

N

N

N

N

Stomach

N

N

Black spot on the corpus

N

N

N

Duodenum

N

N

presence of air inside the

digestive system

N

N

N

Jejunum

N

N

N

N

N

N

Ileon

N

N

N

N

N

N

Caecum

N

N

N

N

N

N

Colon

N

N

N

N

N

N

Rectum

N

N

N

N

N

N

Spleen

N

N

N

N

N

N

Liver

N

N

N

N

N

N

Thymus

N

N

N

N

N

N

Trachea

N

N

N

N

N

N

Lungs

N

N

N

N

N

N

Heart

N

N

N

N

N

N

Kidneys

N

N

N

N

N

N

Urinary bladder

N

N

N

N

N

N

Ovaries

N

N

N

N

N

N

Uterus

N

N

N

N

N

N

Treatment area

N

N

N

N

N

N

Adrenals

N

N

N

N

N

N

Pancreas

N

N

N

N

N

N

Particulars

 

 

bw: 118 g

 

 

 

 *Animal found dead on D9 due to gavage injury; N: Nothing to report.

Interpretation of results:
GHS criteria not met
Remarks:
(EU criteria)
Conclusions:
The oral LD50 of the test item in female rats was greater thatn 2000 mg/kg bw. The LD50 cut-off of the test item may be considered to be higher than 5000 mg/kg bw.
Executive summary:

The potential acute toxicity of the test item was studied on female Sprague-Dawley rats, according to OECD TG 423 and EU method B.1 tris, under GLP conditions. Since no data indicating acute toxicity was available, a first step was performed by administering a single dose of 2000 mg/kg bw test item to three animals by gavage. As no mortality was observed, a second step was performed by dosing three additional animals with the same dose. One death (1/6) was noted in the treated animals on day 9. However, the macroscopic examination of the animal revealed that it was due to a gavage injury. Therefore, no treatment-related mortality was observed in the study. In the surviving animals (5/6), no clinical signs related to the administration of the test item were observed during the study. The body weight evolution of the animals remained normal during the study. The macroscopic examination of the animals at the end of the study did not reveal treatment related changes. Based on the results, the oral LD50 was found to be > 2000 mg/kg bw. In accordance with the OECD TG 423, the LD50 cut-off of the test item may be considered to be higher than 5000 mg/ kg body weight by oral route in the rat.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
5 000 mg/kg bw
Quality of whole database:
Klimish score 1.

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for classification or non-classification

Based on the available information (oral LD50 5000 mg/kg bw), the substance is not classified for acute toxicity according to CLP Regulation (EC) No. 1272/2008.