Registration Dossier

Administrative data

Description of key information

Acute oral toxicity:

In the acute oral toxicity key study in female Crl:WI (SPF) rats, following the acute toxic class method in accordance with the OECD Guideline 423, the LD50 was established to be 500 mg/kg bw on basis of LD50 cut off mg/kg bw, because three animals died in 2000 mg/kg bw at first step (Toxicoop, 2017). The test item was therefore ranked into category 4 based on GHS criteria.

In two acute oral toxicity supporting studies in Sprague-Dawley strain rats according to OECD401, the LD50 of the test material was considered to be approximately 2000 mg/kg bw (K1, Safepharm, 1991) and LD50 > 200 mg/kg bw but < 2000 mg/kg bw (K1, Safepharm, 1992). According to further acute oral toxicity supporting studies in Sprague-Dawley strain rats according to OECD401 conducted by Safepharm in 1992 and 1994, the LD50 was found to be greater than 2000 mg/kg bw.

In conclusion, the test item was ranked into category 4 based on GHS criteria which is in line with the result of the key study by Toxicoop (2017).

Acute inhalation toxicity:

The study concerning the acute inhalation toxicity does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size.

Acute dermal toxicity:

study in progress (05/2018).

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
18 January 1991 - 12 February 1991
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Version / remarks:
1981
Qualifier:
according to guideline
Guideline:
EU Method B.1 (Acute Toxicity (Oral))
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
no
Specific details on test material used for the study:
Description: straw-coloured Iiquid
Container: plastic screw-top container
Storage conditions: room temperature



Species:
rat
Strain:
Sprague-Dawley
Sex:
not specified
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River (UK) Ltd., Manston, Kent, U.K.
- Age at study initiation: 5 to 8 weeks old
- Weight at study initiation: males: 129 - 173 g
females: 124 - 143 g
- Fasting period before study: overnight fast inmediately before dosing and aprox. 2 h after dosing
- Housing: in groups of up to five by sex in solid-floor polypropylene cages with sawdust bedding.
- Diet: Rat and Mouse Expanded Diet No. 1, Special Diet Services Limited, Witham, Essex, U.K.) ad libitum
- Water: ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16 - 21 °C
- Humidity (%): 34 - 56%
- Air changes (per hr): aprox. 15
- Photoperiod : 12 h dark / 12 h light

Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
MAXIMUM DOSE VOLUME APPLIED: 2.6 ml/kg

Doses:
1414, 2000 and 2828 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: observation 1/2, l, 2 and 4 hours after
dosing and subsequently once daily. Weighing: day 0, 7 and 14
- Necropsy of survivors performed: yes, gross pathological examination
- Other examinations performed: clinical signs, body weight, appearance of macroscopic abnormalities
Preliminary study:
HPB 29090: RANGE-FINDING ACUTE ORAL TOXICITY TEST IN THE RAT PROJECT NUMBER: 47/923:
The acute oral median lethal dose (LD50) of the test material in the Sprague-Dawley strain rat was found to be greater than 200 mg/kg bodyweight but less than 2000 mg/kg bodyweight
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
>= 2 000 mg/kg bw
Based on:
test mat.
Mortality:
2000 mg/kg: 20% males, 100% females
2828 mg/kg: 20% males, 80% females
Clinical signs:
Signs of toxicity noted in all dose groups were hunched posture lethargy, ptosis, ataxia and loss of righting reflex. Laboured respiration was also noted in animals treated with l4l4 and 2828 mg/kg. Additional signs of systemic toxicity noted in animals treated with 2000 or 2828 mg/kg were coma and decreased respiratory rate. Isolated incidents of increased salivation and hypothermia were noted in animals treated with 2000 mg/kg. An isolated incident of tiptoe gait was noted in one animal treated with 2828 mg/kg. Surviving animals appeared normal on the day of dosing or one to three days after treatment.
Body weight:
Surviving animals showed expected gain in bodyweight during the study
Gross pathology:
Common abnormalities noted at necropsy of animals that died or were killed in extremis during the study were haemorrhagic or abnormally red
lungs, dark, pale or patchy pallor of the liver, dark or pale kidneys and haemorrhage of the small and large intestlnes. One female treated with 2000 mg/kg and one male treated with 2828 mg/kg also showed pale spleen. The two females treated with 2000 mg/kg that were killed in extremis during the study also showed blood-filled bladder. Haemorrhage of the gastric mucosa was noted in one female treated with 2828 mg/kg. No abnormalities were noted at necropsy of animals killed at the end of the study.
Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
The acute oral median lethal dose (LD50) of the test material, was considered to be approximately 2000 mg/kg bodyweight.
Executive summary:

1. A study was performed to assess the acute oral toxicity of the test material in the Sprague-Dawley strain rat. The method used followed that described in the OECD Guidelines for Testing of chemicals (1981) No 401 "Acute oral Toxicity" referenced as Method Bl in commission Directive 84/449/EEC (which constitutes Annex V of council Directive 67/548/EEC).

The results may be used as a basis for classification and labelling under Annex VI of council Directi ve 67/548/EEC (as adapted to technical progress by Comrnission Directive 83/467/ EEC) .

2. Three groups, each of ten fasted animals (five males and five females) were given a single oral dose of undiluted test material administered at dose levels of 1414, 2000 and 2828 mg/kg bodyweight. The surviving animals were observed for fourteen days after the day of dosing. All animals were subiected to gross pathologica] examination.

3. Deaths were noted during the day of dosing and one and two days after treatment. Two females treated with 2000 mg/kg were killed in extremis one day after treatment. Connon signs of systemic toxicity noted were hunched posture, lethargy, ptosis, ataxia and loss of righting reflex. Additional signs of systemic toxicity noted were laboured respiration, coma, decreased respiratory rate, increased salivation, hypothermia and tiptoe gait. Surviving animals appeared normal on the day of dosing or one to three days after treatment.

4. Surviving animals showed expected gain in bodyweight during the study.

5. Common abnormalities noted at necropsy of animals that died or were killed in extremis during the study were haemorrhagic or abnormally red lungs, dark, pale or patchy pallor of the 1iver, dark or pale kidneys and haemorrhage of the small and large intestines. Additional abnormalities noted were pale spleen, haemorrhage of the gastric mucosa and blood-filled bladder.

No abnormalities wrere noted at necropsy of animals killed at the end of the study.

6. The acute oral median lethal dose (LD50) of the test material was considered to be approximately 2000 nrg/kg bodyweight. It is reasonable to assume that the symbol "Xn" and risk phrase R 22 'HARMFUL IF SWALLOWED" are required according to EEC labelling regulations.

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1992-02-18 / 1992-04-06
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Version / remarks:
1981
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
yes
Specific details on test material used for the study:
Description: straw coloured liquid
Container: white plastic jar
Storage conditions: room temperature
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River (UK) Ltd., Manston, Kent, U.K.
- Age at study initiation: 5 to 8 weeks old
- Weight at study initiation: males: 120 - 134 g
females: 122 - 139 g
- Fasting period before study: overnight fast inmediately before dosing and aprox. 2 h after dosing
- Housing: in groups of up to five by sex in solid-floor polypropylene cages with sawdust bedding.
- Diet: Rat and Mouse Expanded Diet No. 1, Special Diet Services Limited, Witham, Essex, U.K.) ad libitum
- Water: ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16 - 21 °C
- Humidity (%): 34 - 56%
- Air changes (per hr): aprox. 15
Route of administration:
oral: gavage
Vehicle:
arachis oil
Remarks:
100 mg/ml
Details on oral exposure:
For the purpose of the initial range-finding study the test material was used as supplied.The specific gravity was determined by Safepharm Laboratories Limited and used to calculate the appropriate dose volume for the required dose level.
For the continuation of the range-finding study and the main study the test material was freshly prepared, as required, as a solution/
suspension at the appropriate concentration in arachis oil B.P.

MAXIMUM DOSE VOLUME APPLIED: 10 ml/kg (100mg/ml concentration)
Doses:
2000 mg/kg
No. of animals per sex per dose:
1
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: observation 1/2, l, 2 and 4 hours after
dosing and subsequently once daily. Weighing: day 0, 7 and 14
- Necropsy of survivors performed: no
- Other examinations performed: clinical signs, body weight
Preliminary study:
Range-finding Study: No deaths were noted.
Common signs of systemic toxicity noted were lethargy, ataxia, hunched posture and ptosis. Incidents of decreased respiratory rate, laboured respiration, pallor of the extremities, loss of righting reflex and coma were also noted.
The results of the range-finding study indicated that the acute oral median lethal dose of the test material was greater than 2000 mg/kg bodyweight.
Based on this information, a dose level of 2000 mg/kg bodyweight was selected for the main study.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
Two females were found dead one day after dosing.
Clinical signs:
Common signs of systemic toxicity noted on the day of dosing were hunched posture, lethargy, ataxia and laboured respiration.
Incidents of loss of righting reflex and ptosis were also noted. Isolated incidents of hunched posture and lethargy were noted on day l. Surviving animals appeared normal 1 or 2days after dosing.
Body weight:
Surviving animals showed expected gain in bodyweight during the study.
Gross pathology:
Abnormalities noted at necropsy of the females that died during the study were haemorrhagic lungs, dark liver, dark kidneys, haemorrhage of the gastric mucosa, sloughing of the nonglandular epithelium of the stomach and haemorrhage of the small intestine.
No abnormalities were noted at necropsy of animals that were killed at the end of the study.
Interpretation of results:
Category 5 based on GHS criteria
Conclusions:
The acute oral median lethal dose (LD50) of the test material in the Sprague-Dawley strain rat was found to be greater than 2000 mg/kg bodyweight. No symbol and risk phrase are required according to EEC Iabelling regulations.
Executive summary:

l. A study was performed to assess the acute oral toxicity of the test material in the Sprague-Dawley strain rat. The method used followed that described in the OECD Guidelines for Testing of Chemicals (1981) No. 401 "Acute Oral Toxicity" referenced as Method Bl in Commission Directive 84/449/EEC (which constitutes Annex V of Council Directive 67/548/EEC). The results may be used as a basis for classification and labelling under Annex VI of Council Directive 67/548/EEC (as adapted to technical progress by Commission Directive 83/467/EEC).

2. Following a range-finding study, a group of ten fasted animals (five males and five females) was given a single oral dose of test material, as a solution/suspension in arachis oil B.P. at a dose level of 2000 mg/kg bodyweight. The animals were observed for fourteen days after the day of dosing and were then killed for gross pathological examination.

3. Two females were found dead one day after dosing. Common signs of systemic toxicity noted were hunched posture, lethargy, ataxia, and laboured respiration. Incidents of loss of righting reflex and ptosis were also noted. Surviving animals appeared normal 1 or 2 days after dosing.

4. Surviving animals showed expected gain in bodyweight during the study.

5. Abnormalities noted at necropsy of the females that died during the study were haemorrhagic lungs, dark liver, dark kidneys, haemorrhage of the gastric mucosa, sloughing of the non-glandular epithelium of the

stomach and haemorrhage of the small intestine. No abnormalities were noted at necropsy of animals that were killed at the end of the study.

6. The acute oral median lethal dose (LD50) of the test material in the Sprague-Dawley strain rat was found to be greater than 2000 mg/kg bodyweight. No symbol and risk phrase are required according to EEC labelling regulations .

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
18 February 1992 - 19 March 1992
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Version / remarks:
1981
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
no
Specific details on test material used for the study:
Description: straw coloured liquid
Container: white plastic jar
Storage conditions: room temperature
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River (UK) Ltd., Manston, Kent, U.K
- Age at study initiation: approximately five to eight weeks old
- Weight at study initiation: males: 139 - 175 g
females: 146 - 158 g
- Fasting period before study: overnight fast immediately before dosing
- Housing: in groups of up to five by sex in solid floor polypropylene cages with sawdust bedding
- Diet (e.g. ad libitum): Rat and Mouse Expanded Diet No. 1, Special Diet Services
Limited, Witham, Essex, U.K. ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 21 °C
- Humidity (%): 52 - 53 %
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 hrs light/ 12 hrs dark
Route of administration:
oral: gavage
Vehicle:
arachis oil
Details on oral exposure:
VEHICLE: arachis oil B.P.
- Concentration in vehicle: 200 mg/ml
For the purpose of the initial range-finding study the test material was used as supplied.The specific gravity was determined by Safepharm Laboratories Limited and used to calculate the appropriate dose volume for the required dose level.
For the continuation of the range-finding study and the main study the test material was freshly prepared, as required, as a solution/ suspension at the appropriate concentration in arachis oil B.P.


Doses:
Range finding study: 2000 mg/kg
Main study 200 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: 0.5, 1, 2, 4 hours after dosing and subsequently once daily
- Frequency of weighing: prior to dosing on Day 0 and on Days 7 and 14 or at death
- Necropsy of survivors performed: yes
- observation of behavioural and clinical observations, gross lesions, bodyweight changes, mortality and any other toxicological effects.
Preliminary study:
One male treated with 2000 mg/kg was found dead one day after dosing. The other male treated with 2000 mglkg was killed in extremis three days after dosing. Common signs of systemic toxicity noted were hunched posture, lethargy, ataxia, ptosis, decreased respiratory rate and laboured respiration with isolated
incidents of comatose and loss of righting reflex.
The results of the range-finding study indicated that the acute oral median lethal dose of the test material was less than 2000 mglkg bodyweight.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 200 - < 2 000 mg/kg bw
Based on:
test mat.
Mortality:
2/4 animals in Range-finding study
Main study: There were no deaths
Clinical signs:
Common signs of systemic toxicity noted were hunched posture, lethargy, ataxia, ptosis, decreased respiratory rate and laboured respiration with isolated
incidents of comatose and loss of righting reflex.
No signs of system'ic toxic'ity were noted during the main study
Body weight:
All animals showed expected gain in bodyweight during the study.
Gross pathology:
No abnormalities were noted at necropsy.
Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
The acute oral median lethal dose (LD50) of the test material jn the Sprague-Dawley strain rat was found to be greater than 200 mg/kg bodyweight but less than 2000 mg/kg bodyweight. The test material was classified as HARMFUL and the symbol "Xn" and risk phrase R 22 "HARMFUL IF SWALLOWED" are therefore required according to EEC labelling regulations.
Executive summary:

1. A study was performed to assess the acute oral toxicity of the test material in the Sprague-Dawley strain rat. The method used followed that described in the OECD Guidelines for Testing of Chemicals (1981) No. 40l "Acute Oral Toxicity" referenced as Method Bl in Commission Directive 84/449/EEC (which constitutes Annex V of Council Directive 67/548/EEC).

The results may be used as a basis for classification and labelling under Annex VI of Council Directive 57/548/EEC (as adapted to technical progress by Commission Directive 83/467 /EEC) .

2. Following a range-finding study, a group of ten fasted animals (five males and five females) was given a single oral dose of test material, as a solution/suspension in arachis oil B.P. at a dose level of 200 mg/kg bodyweight. The animals were observed for fourteen days after the day of dosing and were then killed for gross pathological examination.

3. There were no deaths. No signs of systemic toxicity were noted during the study.

4. All animals showed expected gain in bodywe'ight during the study.

5. No abnormalities were noted at necropsy.

6. The acute oral median lethal dose (LD50) of the test material in the Sprague-Dawley strain rat was found to be greater than 200 mg/kg bodyweight but less than 2000 mg/kg bodyweight. The test material was classified as HARMFUL and the symbol "Xn" and risk phrase R 22 "HARMFUL IF SWALLOWED" are therefore required according to EEC labelling regulations.

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
18 February 1992 - 19 March 1992
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Version / remarks:
1981
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
yes
Specific details on test material used for the study:
Description: straw coloured Iiquid
Container: white plastic jar
Storage conditions: room temperature
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River (UK) Ltd., Manston, Kent, U.K
- Age at study initiation: approximately five to eight weeks old
- Weight at study initiation: Males: 148 - 158 g
Females: 130 - 148 g
- Fasting period before study: overnight
- Housing:in groups of up to five by sex in solid floor polypropylene cages with sawdust bedding
- Diet (e.g. ad libitum): ad libitum Rat and Mouse Expanded Diet No. l, Special Diet Services
Limited, Wiitham, Essex, U.K
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 21°C
- Humidity (%): 52 - 53 %
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 hrs dark / 12 hrs light
Route of administration:
oral: gavage
Vehicle:
arachis oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 200 mg/ml
For the purpose of the initial range-finding study the test material was used as supplied.The specific gravity was determined by Safepharm Laboratories Limited and used to calculate the appropriate dose volume for the required dose level.
For the continuation of the range-finding study and the main study the test material was freshly prepared, as required, as a solution/ suspension at the appropriate concentration in arachis oil B.P.


Doses:
2000 mg/kg
No. of animals per sex per dose:
5
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations: 0.5, 1, 2, 4 hours after dosing and subsequently once daily
- Frequency of weighing: prior to dosing on Day 0 and on Days 7 and 14 or at death
- Necropsy of survivors performed: yes
- observation of behavioural and clinical observations, gross lesions, bodyweight changes, mortality and any other toxicological effects.
Preliminary study:
Range-finding Study: There were no deaths. Common signs of systemjc toxicity noted were hunched posture, lethargy, ataxia, ptosis, laboured respiration and
decreased respiratory rate with isolated incidents of loss of righting reflex and comatose.
The results of the range-finding study indicated that the acute oral median lethal dose of the test material was greater than 2000 mglkg bodyweight.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
Two animals (one male and one female) were found dead one day after dosing
Clinical signs:
Common signs of systemic toxicity noted were hunched posture, lethargy and ataxia with additional signs of decreased respiratory rate, ptosis and loss of righting reflex. Incidents of systemic toxicity noted in one female were pilo-erection and laboured respiration. Surviving animals appeared normal one or two days after dosing.
Body weight:
Surviving animals showed expected gain in bodyweight during the study.
Gross pathology:
Abnormalities noted at necropsy of animals that died during the study were haemorrhagic lungs, dark liver, dark kidneys and slight haemorrhage of the gastric mucosa. No abnormalities were noted at necropsy of animals that were killed at the end of the study.
Interpretation of results:
Category 5 based on GHS criteria
Conclusions:
The acute oral median lethal dose (LD50) of the test material, in the Sprague-Dawley strain rat was found to be greater than 2000 mg/kg bodyweight. No symbol and risk phrase are required according to EEC labelling regulations.
Executive summary:

l. A study was performed to assess the acute oral toxicity of the test material in the Sprague-Dawley strain rat. The method used followed that described in the OECD Guidelines for Testing of Chemjcals (1981) No. 401 "Acute Oral Toxicity" referenced as Method Bl in Commission Directive 84/449/EEC (which constitutes Annex V of Council Directive 67/548/EEC).

The results may be used as a basis for classification and 1abelling under Annex VI of Council Directive 67/548/EEC (as adapted to technical progress by Commission Directi ve 83/467 /EEC).

2. Following a range-finding study, a group of ten fasted animals (five males and five females) was given a single oral dose of test material, as a suspension in arachis oil B.P. at a dose level of 2000 mg/kg bodyweight. The animals were observed for fourteen days after the day of dosing and were then killed for gross pathological examination.

3. Two animals (one male and one female) were found dead one day after dosing. Common signs of systemic toxicity noted were hunched posture, lethargy and ataxia with additional signs of decreased respiratory rate, ptosis and loss of righting reflex. Isolated incidents of systemic toxicity noted were pilo-erection and laboured respiration. Surviving animals appeared normal one or two days after dosing.

4. Surviving animals showed expected gain in bodyweight during the study.

5. Abnormalities noted at necropsy of animals that died during the study were haemorrhagic lungs, dark liver, dark kidneys and slight haemorrhage of the gastric mucosa. No abnormalities were noted at necropsy of animals that were killed at the end of the study.

6. The acute oral median lethal dose (LD50) of the test material in the Sprague-Dawley strain rat was found to be greater than 2000 mg/kg bodyweight. No symbol and risk phrase are required according to EEC labelling regulations .

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
February 23 - March 30, 2017
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)
Version / remarks:
17th December 2001
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
Species and strain: Crl:WI rats
Source: TOXI COOP ZRT. Cserkesz u. 90.
1103 Budapest, Hungary
Hygienic level at arrival: SPF
Hygienic level during the study: Good conventional
Justification of strain: The Wistar rats as a rodent is one of the standard species of acute toxicity studies
Number of animals: 3 animals/group
Sex: Female, nulliparous and non pregnant animals
Age of animals: Young adult rat, 8 weeks old in first, second and third step
Body weight range
at starting (first step): 165 - 170 g
Body weight range
at starting (second step): 166 - 178 g
Body weight range
at starting (third step): 167 - 171 g
Acclimatization time: 5 days in first step, 6 days in second step and 7 days in third step

Housing: Group caging (3 animals/cage)
Cage type: Type II polypropylene/polycarbonate.
Bedding: Laboratory bedding.
Light: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 22 ± 3 °C
Relative humidity: 30 - 70 %
Ventilation: above 10 air exchanges/hour by central air-condition system.

Animals received ssniff® SM R/M-Z+H complete diet for rats and mice produced by ssniff Spezialdiäten GmbH, D-59494
Soest Germany and tap water from municipal supply, as for human consumption from bottle ad libitum.

In life phase: February 28- March 17, 2017


Route of administration:
oral: gavage
Vehicle:
other: Helianthi annui oleum raffinatum
Details on oral exposure:
All doses were formulated in the vehicle. Concentration of formulations was adjusted to maintain a treatment volume
of 10 mL/kg bw. The test item was applied in a concentration of 200 and 30 mg/mL. Formulations were prepared just
before the administration and were stirred continuously during the treatment.

Vehicle:
Name: Helianthi annui oleum raffinatum
Batch number: 1607-4569
Date of expiration: 30.11.2017
Produced by: Parma Produkt Kft.

The acute toxic class method was carried out involving a stepwise procedure with the use of 2000 mg/kg bw as the starting
dose in three female rats. All females died, so the test was continued at 300 mg/kg bw dose level on further three female rats.
No animal died in the second step at 300 mg/kg bw dose level, three further female rats were treated with the same (300 mg/kg bw) dose.
No animal died in the third step, too, so the test was finished, the stopping criteria of Annex 2d of OECD Guideline No. 423. was met.
Doses:
2000 mg/kg bw
300 mg/kg bw
No. of animals per sex per dose:
2000 mg/kg bw: 3 animals
300 mg/kg bw: 6 animals
Control animals:
no
Details on study design:
5 days in first step, 6 days in second step and 7 days in third step of acclimatization, treatment’s day,
14 days post-treatment observation period, necropsy on Day 15.

Frequency of observations:
Animals were observed individually after dosing at least once during the first 30 minutes, then 1 h, 2 h, 3 h, 4 h after the treatment
and once each day for 14 days thereafter.

Body weight measurement:
The body weights were recorded on day 0 (just before the treatment), on day 1, on day 7 and on day 15 with a precision of 1 g.

Necropsy:
All animals treated with 2000 mg/kg bw dose spontaneously died during the study and were necropsied on Day 1.
All animals treated with 300 mg/kg bw dose survived until the scheduled necropsy on Day 15.
Statistics:
No statistics was used in the study.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - < 2 000 mg/kg bw
Based on:
test mat.
Mortality:
All rats dosed at 2000 mg/kg bw died on Day 1.
No death occurred at 300 mg/kg bw single oral dose of the test item.
Clinical signs:
2000 mg/kg bw: CNS - and emotion symptoms (decreased activity, bedding digging, closed eyes), disturbances of coordination
(incoordination, prone position), decreased righting reflex, decreased muscular tension (grip- and limb tone, body tone, abdominal tone)
and disturbances of the autonomic functions (dyspnea, redness skin and mucous membrane) were observed in animals between treatment day and Day 1.

2000 mg/kg bw: no clinical symptoms were observed on the day of the treatment and during the 14-day observation period.
Body weight:
The body weight development was undisturbed in all survivor animals.
Gross pathology:
All animals treated with 2000 mg/kg bw dose died spontaneously during the study and were necropsied on the Day 1.
All animals treated with 300 mg/kg bw dose survived until the scheduled autopsy on Day 15.
Autopsy revealed treatment related internal changes as haemorrhaged mucous membrane in the stomach, urinary bladder full of urine,
stomach full of gas and spotted liver in 2000 mg/kg bw dose group.
All organs of the animals treated with 300 mg/kg bw proved to be free of treatment related gross pathological changes.

A single oral administration - followed by a fourteen-day observation period - was performed by gavage.

The day before treatment the animals were fasted. The food but not water was withheld overnight.

Animals were weighed before the application and the food was given back 3 hours after the treatment.

Starting dose was selected on the basis of the available information about the test item.

The acute toxic class method was carried out involving a stepwise procedure with the use of 2000 mg/kg bw

as the starting dose in three female rats. All females died, so the test was continued at 300 mg/kg bw dose level

on further three female rats. No animal died in the second step at 300 mg/kg bw dose level, three further female rats

were treated with the same (300 mg/kg bw) dose.

No animal died in the third step, too, so the test was finished, the stopping criteria of Annex 2d

of OECD Guideline No. 423. was met.

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
The test item was ranked into classes of Globally Harmonized Classification System (GHS)
described in the OECD Guideline No. 423 as below:

Dose (mg/kg bw) 2000 300
Mortality (dead/treated) 3/3 0/6
LD50 (mg/kg bw) between 300 and 2000
GHS category 4

The estimated LD50 value is 500 mg/kg bw on basis of LD50 cut off mg/kg bw, because three animals died in 2000 mg/kg bw at 1st step.
Executive summary:

A single oral administration - followed by a fourteen-day observation period - was performed by gavage.

The day before treatment the animals were fasted. The food but not water was withheld overnight.

Animals were weighed before the application and the food was given back 3 hours after the treatment.

Starting dose was selected on the basis of the available information about the test item.

The acute toxic class method was carried out involving a stepwise procedure with the use of 2000 mg/kg bw

as the starting dose in three female rats. All females died, so the test was continued at 300 mg/kg bw dose level

on further three female rats. No animal died in the second step at 300 mg/kg bw dose level, three further female rats

were treated with the same (300 mg/kg bw) dose.

No animal died in the third step, too, so the test was finished, the stopping criteria of Annex 2d

of OECD Guideline No. 423. was met.

GHS category: 4

The estimated LD50value is 500 mg/kg bw on basis of LD50cut off mg/kg bw, because three animals died in 2000 mg/kg bw at 1ststep.

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
Between 22 February 1994 and 16 March 1994
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Version / remarks:
1987
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
yes
Specific details on test material used for the study:
Date received: 11 February 1994
Description: brown slightly viscous liquid
Container: opaque plastic jar
Storage conditions: room temperature
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Harlan U.K. Ltd., Blackthorn, Bicester, Oxon, U.K
- Age at study initiation: approx. 8 weeks old
- Weight at study initiation: males: 140 - 164 g
females: 130 - 145 g
- Fasting period before study: overnight fast immediately before dosing and for approximately two hours after dosing
- Housing: housed in groups of up to five by sex in solid floor polypropylene cages furnished with woodflakes
- Diet (e.g. ad libitum): ad libitum Rat and Mouse Expanded Diet No. 1, Special Diets Services Limited, Witham, Essex, U.K.
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 -23 °C
- Humidity (%): 38 - 59%
- Air changes (per hr): approx. 15
- Photoperiod (hrs dark / hrs light): 12h dark / 12 h light

Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 200 mg/ml
- Amount of vehicle (if gavage): 10 ml/kg


Doses:
2000 mg/kg
No. of animals per sex per dose:
5 male
5 female
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: 0.5,1, 2 and 4 hours after dosing and subsequently once daily for 14 days
- Necropsy of survivors performed: gross pathological examination. External examination and opening of the abdominal and thoracic cavities
- Other examinations performed: clinical signs, body weight, organ weights, histopathology
Preliminary study:
Range finding study : dose level 2000 mg/kg. 1 dose by gavage. 1 female, 1 male.
Results: There were no deaths. Common signs of systemic toxicity noted were ataxia, hunched posture and lethargy with an isolated incident of tiptoe gait. No necropsies were performed.
Based on this information, a dose level of 2000 mglkg bodyweight was selected for the main study.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
One male was found dead one day after dosing.
Clinical signs:
Common signs of systemic toxicity noted were ataxia, hunched posture, lethargy, ptosis, decreased respiratory rate and laboured respiration with additional signs of tiptoe gait. Isolated jncidents of systemic toxicity noted were coma, loss of righting reflex and splayed gait. Surviving animals recovered two to five days after dosing.
Body weight:
Surviving animals showed expected gain in bodyweight during the study except for one female which showed no gain in bodyweight during the first week and expected gain in bodyweight during the second week.
Gross pathology:
Abnormalities noted at necropsy of the male that died during the study were haemorrhagic Iungs, dark Iiver and dark kidneys. No abnormalities were noted at necropsy of animals that were killed at the end of the study.
Interpretation of results:
Category 5 based on GHS criteria
Conclusions:
The acute oral median lethal dose (LD50) of the test material, in the Sprague-Dawley strain rat was found to be greater than 2000 mg/kg bodyweight. No symbol and risk phrase are required according to EC labelling regulations.
Executive summary:

1. A study was performed to assess the acute oral toxicity of the test material in the Sprague-Dawley strain rat. The method used followed that described in the OECD Guidelines for Testing of Chemicals (1987) No. 401 "Acute Oral Toxicity" referenced as Method Bl of Commission Directive 84/449/EEC (which constitutes Annex V of Council Directive

67/548/EEc) .

The results may be used as a basis for classification and label1ing under Annex VI of Council Directive 67/548/EEC (as adapted to technical progress by Commission Directive 9l/325/EEC) .

2. Following a range-finding study, a group of ten fasted animals (five males and five females) was given a single oral dose of test material, as a solution in distilled water at a dose level of 2000 mg/kg bodyweight. The animals were observed for fourteen days after the day of dosing and were then killed for gross pathological examination.

3. One male was found dead one day after dosing. Common signs of systemic toxicity noted were ataxia, hunched posture, lethargy, ptosis, decreased respiratory rate and laboured respiration with additional signs of tiptoe gait. Isolated incidents of systemic toxicity noted were coma, loss of righting reflex and splayed gait. Surviving animals recovered two to five days after dosing.

4. Surviving animals showed expected gain in bodyweight during the study except for one female which showed no gain in bodyweight during the first week and expected gain in bodyweight during the second week.

5. Abnormalities noted at necropsy of the male that died during the study were haemorrhagic 1ungs, dark liver and dark kidneys. No abnormalities were noted at necropsy of animals that were killed at the end of the study.

6. The acute oral median lethal dose (LD50) of the test material in the Sprague-Dawley strain rat was found to be greater than 2000 mg/kg bodyweight. No symbol and risk phrase are required according to EC labelling regulations.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
500 mg/kg bw

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 oral toxicity:

An acute oral toxicity study with the test item according to the acute toxic class method in female Crl:WI (SPF) rats (OECD guideline 423) was performed (Toxicoop, 2017).

A single oral administration - followed by a fourteen-day observation period - was performed by gavage. The acute toxic class method was carried out involving a stepwise procedure with the use of 2000 mg/kg bw as the starting dose in three female rats. All females died, so the test was continued at 300 mg/kg bw dose level on further three female rats. No animal died in the second step at 300 mg/kg bw dose level, three further female rats were treated with the same (300 mg/kg bw) dose. No animal died in the third step, too, so the test was finished, the stopping criteria of Annex 2d of OECD Guideline No. 423. was met.

The test item was ranked into classes of Globally Harmonized Classification System (GHS) described in the OECD Guideline No. 423 as below:

Dose (mg/kg bw): 2000 / 300

Mortality (dead/treated): 3/3 / 0/6

LD50 (mg/kg bw): between 300 and 2000

GHS category: 4

The estimated LD50 value is 500 mg/kg bw on basis of LD50 cut off mg/kg bw, because three animals died in 2000 mg/kg bw at first step.

Justification for classification or non-classification

The median lethal dose of the test substance after single oral administration to female rats, observed over a period of 14 days is: LD50 (female rat): 500 mg/kg bw. The test item is ranked into Category 4 based on GHS criteria.

No studies were available for acute toxicity via the inhalation and dermal route so no classification could be derived for acute toxicity after exposure to the test item via those exposure routes.