Registration Dossier

Administrative data

Description of key information

Acute toxicity (oral) LD50 Females = >2000 mg/kg bw; OECD 420; Pooles, A. (2018)

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:
23 October - 08 November 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Study was conducted in accordance with international guidelines and in accordance with GLP. All guideline validity criteria were met.
Qualifier:
according to
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Version / remarks:
2001
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)
Version / remarks:
Commission Regulation (EC) No. 440/2008
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes
Species:
rat
Strain:
Wistar
Remarks:
RccHan:WIST
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Envigo RMS (UK) Limited, Oxon, UK.
- Females (if applicable) nulliparous and non-pregnant: Yes
- Age at study initiation:
8-12 weeks
- Weight at study initiation:
Body weight variation did not exceed ±20% of the mean body weight.
- Fasting period before study:
Yes, overnight fast prior to dosing.
- Housing: The animals were housed in groups of up to four in suspended solid-floor polypropylene cages furnished with softwood flake bedding.
- Diet (e.g. ad libitum):
Free access to feed (apart from during fasting period); 2014C Teklad Global Rodent diet supplied by Envigo RMS (UK) Limited, Oxon, UK.
- Water (e.g. ad libitum):
Free access to mains drinking water throughout the study.
- Acclimation period:
At least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):
19-25 ºC
- Humidity (%):
30-70 % RH
- Air changes (per hr):
≥15 air changes per hour.
- Photoperiod (hrs dark / hrs light):
12:12 h

IN-LIFE DATES: Not reported.
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
VEHICLE
- Concentration in vehicle: N/A - test item applied undiluted and as supplied.
- Amount of vehicle (if gavage): N/A
- Justification for choice of vehicle: N/A
- Lot/batch no. (if required): N/A
- Purity: N/A

MAXIMUM DOSE VOLUME APPLIED: 2.16 mL/kg.

DOSAGE PREPARATION (if unusual): N/A - test item applied undiluted and as supplied.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: N/A - fixed dose limit test.
Doses:
2000 mg/kg
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical observations were made 30 minutes, 1, 2, and 4 hours after dosing and then daily for up to 14 days. Morbidity and mortality checks were made twice daily, early and late during normal working days, and once daily at weekends and public holidays. Individual body weights were recorded on Day 0 (the day of dosing) and on Days 7 and 14 or at death.
- Necropsy of survivors performed: Yes
- Other examinations performed: clinical signs, body weight, gross necropsy (consisting of an external examination and opening of the abdominal and thoracic cavities. Macroscopic abnormalities were recorded)
Statistics:
N/A
Preliminary study:
A single animal was treated at 2000 mg/kg as the starting dose. In the absence of mortality at a dose level of 2000 mg/kg, an additional group of 4 animals was treaed at 2000 mg/kg.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
One animal was killed for humane reasons, 8 days after dosing, due to the occurrence of clinical signs of toxicity that approached the severity limit set forth in the UK Home Office Project Licence.
Clinical signs:
Hunched posture was noted in all animals. Staining around ano-genital region was also noted in three animals. Ataxia, lethargy and pilo-erection were confined to one animal.
Body weight:
Two animals showed body weight loss during the first week with expected gain in body weight during the second week and two animals showed expected gains in body weight.
Gross pathology:
Abnormalities noted at necropsy of the animal that was humanely killed during the study were small left kidney (in comparison to the right), epithelial sloughing and thickened non-glandular epithelium of the stomach, gaseous distension of the small and large intestines and light brown fluid filled abdominal cavity. No abnormalities were noted at necropsy of animals that were killed at the end of the study.

Table 2       Individual clinical observations and mortality data

 

Dose level

(mg/kg)

Animal number and sex

Effects noted after dosing (hours)

Effects noted during period after dosing (days)

0.5

1

2

4

1

2

3

4

5

6

7

8

9

10

11

12

13

14

2000

1-0 F

0

0

0

0

0

0

0

0

0

0

HSa

HSa

H

H

H

H

0

0

2-0 F

0

H

H

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-1 F

H

H

H

0

0

0

0

0

HSa

HSa

H

H

H

H

0

0

0

0

2-2 F

H

H

H

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-3 F

0

H

H

0

0

0

0

0

HSa

HSa

HPA

HLAX

 

 

 

 

 

 

0: no sign of systemic toxicity

A: ataxia

H: hunched posture

L: lethargy

P: pilo-erection

Sa: staining around ano-genital region

X: animal humanely killed due to occurrence of clinical signs of toxicity that approached the severity limit set forth in the UK Home Office Project Licence

 

Table 3       Individual body weight and body weight changes

 

Dose level

(mg/kg)

Animal number and sex

Body weight (g) at day

Body weight (g) at death

Body weight gain(g) during week

0

7

14

1

2

2000

1-0 F

152

149

167

-

-3

18

2-0 F

158

169

180

-

11

11

2-1 F

158

149

170

-

-9

21

2-2 F

178

186

205

-

8

19

2-3 F

165

136

-

129

-29

-

 

Table 4       Individual necropsy findings

 

Dose level

(mg/kg)

Animal number and sex

Time of death

Macroscopic observations

2000

1-0 F

Killed Day 14

No abnormalities detected

2-0 F

Killed Day 14

No abnormalities detected

2-1 F

Killed Day 14

No abnormalities detected

2-2 F

Killed Day 14

No abnormalities detected

2-3 F

Humanely killed on Day 8

Kidneys: small left kidney, compared to right.

Non-glandular epithelium of the stomach: epithelial sloughing; thickened.

Abdominal cavity: fluid filled (light brown colour).

Small intestine: gaseous distention.

Large intestine: gaseous distention.

Interpretation of results:
GHS criteria not met
Conclusions:
The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was estimated to be greater than 2000 mg/kg body weight. Test item does not meet the GHS criteria for classification in accordance with CLP Regulation (EC) No. 1272/2008.
Executive summary:

OECD 420 (2018) - In an acute oral toxicity study (OECD 420), five fasted, 8-12 week old, female Wistar strain rats were given a single oral dose of test item at dose level of 2000 mg/kg bw (limit test). Clinical observations were made 30 minutes, 1, 2, and 4 hours after dosing and then daily for up to 14 days. Morbidity and mortality checks were made twice daily, early and late during normal working days, and once daily at weekends and public holidays.

Individual body weights were recorded on Day 0 (the day of dosing) and on Days 7 and 14 or at death.  After the 14 days observation, all surviving animals underwent necropsy.

Mortality

 

One animal was humaely killed for humane reasons on Day 8 due to occurrence of toxicity that approached the severity limit set forth in the UK Home Office Project Licence. No mortality was observed in the other 4 animals.

 

Clinical observations

 

Hunched posture was noted in all animals. Staining around the ano-genital region was noted in 3/5 animals. Ataxia, lethargy and pilo-erection were confined to the one animal that was humanely killed on Day 8.

 

Body weight

 

Two animals showed body weight loss during the first week with expected body weight gain in the second. Two animals showed expected body weight gain.

 

Necropsy

 

Abnormalities were confined to the animal that was humanly killed on Day 8. Abnormalities included; small left kidney (compared to the right); epithelial sloughing and thickened non-glandular epithelium of the stomach; gaseous distention of the large and small intestines an light-brown filled abdominal cavity. No abnormalities were noted in the other 4 animals.

 

The acute oral median dose level (LD50) of the test item was estimate to be greater than 2000 mg/kg bw. The test item does not meet the GHS criteria for classification in accordance with CLP Regulation (EC) No. 1272/2008.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
The endpoint is concluded based on a single key study with a Klimisch rating of 1. No effects were observed up to the limit dose of 2000 mg/kg bw/day.

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

Acute toxicity (oral);

OECD 420 (2018) - In an acute oral toxicity study (OECD 420), five fasted, 8-12 week old, female Wistar strain rats were given a single oral dose of test item at dose level of 2000 mg/kg bw (limit test) and observed for 14 days.

One animal was humanely killed for humane reasons on Day 8 due to occurrence of toxicity that approached the severity limit set forth in the UK Home Office Project Licence. No mortality was observed in the other 4 animals. Hunched posture was noted in all animals. Staining around the ano-genital region was noted in 3/5 animals. Ataxia, lethargy and pilo-erection were confined to the one animal that was humanely killed on Day 8. Two animals showed body weight loss during the first week with expected body weight gain in the second. Two animals showed expected body weight gain. At necropsy, abnormalities were confined to the animal that was humanly killed on Day 8. Abnormalities included; small left kidney (compared to the right); epithelial sloughing and thickened non-glandular epithelium of the stomach; gaseous distention of the large and small intestines and light-brown filled abdominal cavity. No abnormalities were noted in the other 4 animals.

 

The acute oral median dose level (LD50) of the test item was estimate to be greater than 2000 mg/kg bw. The test item does not meet the GHS criteria for classification in accordance with CLP Regulation (EC) No. 1272/2008.

Justification for classification or non-classification

Acute oral toxicity - Oral LD50 Females = >2000 mg/kg bw. According to the CLP Regulation (EC) No. 1272/2008, the substance does not meet the classification criteria for acute oral toxicity.