Registration Dossier

Administrative data

Description of key information

Acute Oral: Under the conditions of this study, the acute oral LD50 value of the test item CAS 577978-76-8 was found to be greater than 2000 mg/kg bw in female Crl:WI rats.

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:
19 June 2019 to 05 July 2019
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
yes
Remarks:
See other information section for more details
Qualifier:
according to guideline
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
yes
Remarks:
See other information section for more details
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
yes
Specific details on test material used for the study:
No further details specified in the study report
Species:
rat
Strain:
other: Crl:WIWistar rats
Sex:
female
Details on test animals or test system and environmental conditions:
Species and strain: Crl:WIWistar rats
Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7, D-97633 Sulzfeld
Hygienic level at arrival: SPF
Hygienic level during the study: Standard housing conditions
Number of animals: 6 animals, 3 animals / group
Sex: Female, nulliparous and non-pregnant.
Age of animals at dosing: Young healthy adult rats, ~10 -11 weeks old
Body weight at treatment: 224 – 245 g
Acclimation period: at least 27 days

Husbandry
Animal health: Only healthy animals were used for the test. The staff Veterinarian certified their health status.
Number of animal room: 522/5
Housing: 3 animals / cage
Cage type: Type II polypropylene/polycarbonate
Bedding: SAFE 3/4 S certified wooden chips (J. Rettenmaier & Söhne GmbH + CO. KG, 73494 Holzmühle 1, Rosenberg, Germany) was available to animals during the study. The quality of the bedding was guaranteed by the supplier. Details of bedding quality will be archived with the raw data.
Nesting: ARBOCEL nest building materials (J. Rettenmaier & Söhne GmbH + CO. KG, 73494. Holzmühle 1, Rosenberg, Germany) was available to animals during the study. The quality of the nest building material was guaranteed by the supplier. Details of nest building material quality will be archived with the raw data.
Lighting period: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 20.3 – 24.8°C
Relative humidity: 30 – 85%
Ventilation: 15 – 20 air exchanges/hour
Enrichment: Animals were housed by group to allow social interaction and with deep wood sawdust bedding to allow digging and other normal rodent activities.

The temperature and relative humidity were recorded twice daily during the study.

Food and Water Supply
Animals received ssniff SM R/M "Autoclavable complete diet for rats and mice – breeding and maintenance" produced by ssniff Spezialdiäten GmbH, D-59494 Soest, Germany, ad libitum, except for the night before treatment. The food is considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study. Batch numbers and details of the diet are archived with the raw data.
Animals received tap water from the municipal supply from 500 mL bottles, ad libitum. The water was fit for human consumption and was considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study. Water quality control analysis is performed once every three months and microbiological assessment is performed monthly, by Veszprém County Institute of State Public Health and Medical Officer Service (ÁNTSZ, H-8200 Veszprém, József Attila u. 36, Hungary). The quality control results are retained in the archives at Charles River Laboratories Hungary Kft.

Randomisation
At the end of the acclimatisation period, the animals were selected by hand and assigned to the study based on body weight. The mean body weight of all animals used in the study were within 20%. No computer generated randomisation program was used.

Animal Identification
Animals were individually identified using numbers written on the tail with an indelible marker pen. The numbers were given on the basis of Charles River Laboratories Hungary Kft.'s Master File, for each animal allocated to the treatment groups. The cages were identified by cards, with information about study code, sex, dose group, cage number and individual animal numbers.
Route of administration:
oral: gavage
Vehicle:
other: PEG 400
Details on oral exposure:
A single oral treatment was carried out by gavage for each animal after an overnight food withdrawal
Doses:
Doses
Justification of the dose:
The initial dose level was selected by the Study Director to be that which is most likely to produce mortality in some of the dosed animals. A limit dose of 2000 mg/kg bw was selected as a starting dose.
Initially three females (Group 1) were treated at a dose level of 2000 mg/kg bw. No mortality was observed therefore the second group of animals (Group 2) were treated at a dose level of 2000 mg/kg bw. No mortality was observed in the confirmatory group, therefore no further testing was required according to the test guidelines (OECD 423, Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.tris).
No. of animals per sex per dose:
3 females per group
Control animals:
no
Details on study design:
Procedure
A single oral gavage administration using a constant dose volume of 10 mL/kg bw was followed by a fourteen-day observation if no death occurred. On the night before treatment, the animals were fasted. The food, but not water was withheld during an overnight period. Animals were weighed just before treatment. The test item was administered by oral gavage in the morning. The food was returned 3 hours after the treatment.

OBSERVATIONS

Clinical Observations
Clinical observations were performed on all animals at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. Individual observations were performed on the skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

Body Weight Measurement
The body weights were recorded on Days -1 (prior to removal of food), 0 (prior to administration), 7 and 14 with a precision of 1 g. Terminal body weight of animals were recorded prior to euthanasia.

NECROPSY
Macroscopic examination was performed on all animals. The animals were sacrificed by exsanguination under pentobarbital anaesthesia (Euthanimal 40%; Lot No.: 1811347-03, Expiry Date: 31 December 2021, Produced by: Alfasan Nederland BV, Kuipersweg 9, Woerden, The Netherlands). After examination of the external appearance, the cranial, thoracic and the abdominal cavities were opened and the organs and the tissues were observed. Macroscopic abnormalities were recorded.
Statistics:
Not specified
Preliminary study:
Not specified
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
At a dose level of 2000 mg/kg bw no mortality occurred.
Clinical signs:
After treatment at 2000 mg/kg bw, decreased activity (1/6 animals), hunched back (6/6 animals) and piloerection (6/6 animals) were observed. Symptoms were present only on Day 0. From Day 1, the surviving animals were symptom-free.
Body weight:
There were no effects on body weight or body weight gains.
Gross pathology:
There was no evidence of any macroscopic observations in the animals at dose level 2000 mg/kg bw.
Other findings:
Not specified

INDIVIDUAL CLINICAL OBSERVATIONS

DOSE LEVEL: 2000 mg/kg bw, Treatment on Day 0SEX: FEMALE

Cage No.

Animal Number

Observations

Observation days

Frequency

0

1

2

3

4

5

6

7-14

30’

1h

2h

3h

4h

6h

 

 

 

 

1

 

 

 

 

 

 

8033

Symptom free

+

+

-

-

-

+

+

+

+

+

+

+

+

17/20

Hunched back

-

-

+

+

+

-

-

-

-

-

-

-

-

3/20

Piloerection

-

-

-

+

+

-

-

-

-

-

-

-

-

2/20

 

8034

Symptom free

+

+

-

-

-

+

+

+

+

+

+

+

+

17/20

Hunched back

-

-

+

+

-

-

-

-

-

-

-

-

-

3/20

Piloerection

-

-

-

+

+

-

-

-

-

-

-

-

-

2/20

 

8035

Symptom free

+

+

-

-

-

-

+

+

+

+

+

+

+

16/20

Activity decreased

-

-

-

SI

-

-

-

-

-

-

-

-

-

1/20

Hunched back

-

-

+

+

+

+

-

-

-

-

-

-

-

4/20

Piloerection

-

-

-

+

+

-

-

-

-

-

-

-

-

2/20

 

 

 

2

 

 

 

 

 

 

8036

Symptom free

+

+

-

-

-

+

+

+

+

+

+

+

+

17/20

Hunched back

-

-

+

+

+

-

-

-

-

-

-

-

-

3/20

Piloerection

-

-

-

+

-

-

-

-

-

-

-

-

-

1/20

 

8037

Symptom free

+

+

-

-

+

+

+

+

+

+

+

+

+

18/20

Hunched back

-

-

+

+

-

-

-

-

-

-

-

-

-

2/20

Piloerection

-

-

-

+

-

-

-

-

-

-

-

-

-

1/20

 

8038

Symptom free

+

+

-

-

-

+

+

+

+

+

+

+

+

17/20

Hunched back

-

-

+

+

+

-

-

-

-

-

-

-

-

3/20

Piloerection

-

-

-

+

-

-

-

-

-

-

-

-

-

1/20

 Remarks: + = present      h = hour (s)

                  - = Absent      ‘ = minute

                  Frequency of observation = number of occurrence of observation / total number of observations

 

Severities: Sl = Slight/Small/Few/Small amount

    Mo = Moderate/Several/Moderate amount

    Ex = Severe/Large/Many/Large/Extreme amount

INDIVIDUAL BODY WEIGHT AND BODY WEIGHT GAIN

DOSE LEVEL: 2000 mg/kg bw, Treatment on Day 0  SEX: FEMALE 

Cage No.

Animal Number

Body weight (g)

 

Body Weight Gain (g)

Days

-1

0

7

14

-1-0

0-7

7-14

-1- 14

 

1

8033

246

227

247

249

-19

20

2

3

8034

243

229

254

263

-14

25

9

20

8035

243

231

249

255

-12

18

6

12

 

2

8036

268

245

270

277

-23

25

7

9

8037

258

243

260

270

-15

17

10

12

8038

236

224

239

241

-12

15

2

5

Mean

249.0

233.2

253.2

259.2

-15.8

20.0

6.0

10.2

Standard deviation

11.8

8.7

10.8

13.4

4.4

4.0

3.4

6.0

INDIVIDUAL INTERNAL AND EXTERNAL MACROSCOPIC OBSERVATIONS

 

DOSE LEVEL: 2000 mg/kg bw, Treatment on Day 0        SEX: FEMALE 

Cage No.

Animal Number

Necropsy Date/

Necropsy Day

 

External Observations

 

Internal Observations

 

Organ/Tissue

 

1

8033

03 July 2019

Day 14

No external observations recorded

No internal observations recorded

Not applicable

8034

03 July 2019

Day 14

No external observations recorded

No internal observations recorded

Not applicable

8035

03 July 2019

Day 14

No external observations recorded

No internal observations recorded

Not applicable

 

2

8036

05 July 2019

Day 14

No external observations recorded

No internal observations recorded

Not applicable

8037

05 July 2019

Day 14

No external observations recorded

No internal observations recorded

Not applicable

8038

05 July 2019

Day 14

No external observations recorded

No internal observations recorded

Not applicable

Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of this study, the acute oral LD50 value of the test item CAS 577978-76-8 was found to be greater than 2000 mg/kg bw in female Crl:WI rats.
Executive summary:

The single-dose oral toxicity study of CAS 577978-76-8 in rats was performed according to the acute toxic class method (OECD 423 and Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.Tris) in Crl:WIWistar rats.

Two groups of 3 female Crl:WI rats were treated with the test item at dose levels of 2000 mg/kg body weight (bw).

A single oral treatment was carried out by gavage for each animal after an overnight food withdrawal. Food was made available again 3 hours after the treatment. The test item was formulated in PEG 400 at concentrations of 200 mg/mL at a dose volume of 10 mL/kg bw.

Initially three females (Group 1) were treated at a dose level of 2000 mg/kg bw. No mortality was observed therefore the second group of animals (Group 2) were treated at a dose level of 2000 mg/kg bw as well. No mortality was observed in the confirmatory group, therefore no further testing was required according to the test guidelines (OECD 423, Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.tris).

Clinical observations were performed at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. Body weight was measured on Days -1, 0, 7 and 14 (before necropsy). All animals were subjected to a necropsy and a macroscopic examination.

The results of the study were summarized as follows:

Mortality

At a dose level of 2000 mg/kg bw no mortality occurred.

Clinical Observations

After treatment at 2000 mg/kg bw, decreased activity (1/6 animals), hunched back (6/6 animals) and piloerection (6/6 animals) were observed. Symptoms were present only on Day 0. From Day 1 all animals were symptom-free.

Body Weight and Body Weight Gain

There were no effects on body weight or body weight gains.

Necropsy

There was no evidence of the macroscopic observations at a dose level of 2000 mg/kg bw.

Conclusion:

Under the conditions of this study, the acute oral LD50 value of the test item CAS 577978-76-8 was found to be greater than 2000 mg/kg bw in female Crl:WI rats.

According to the GHS criteria, CAS 577978-76-8 can be ranked as "Category 5" for acute oral exposure.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
K1 – study performed to current international/national guidelines in accordance with GLP

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

According to the GHS criteria, CAS 577978-76-8 can be ranked as "Category 5" for acute oral exposure.