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Description of key information

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:
14/02/2018 - 07/03/2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Chemrign / batch 02/16
- Expiration date of the lot/batch: 01/01/2021
- Purity test date: 100%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature in the dark
- Stability under test conditions: The test item was formulated within 2 hours ogf being applied to the test system. It is assumed that the formulation was stable for this duration.
- Solubility and stability of the test substance in the solvent/vehicle: The test item was freshly prepared as a solution in arachis oil BP. Arachis oil BP was used beacause the test item did not dissolve/suspend in distilled water.
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test item was freshly prepared as a solution in arachis oil BP.
- Preliminary purification step (if any):
- Final dilution of a dissolved solid, stock liquid or gel:
- Final preparation of a solid:

FORM AS APPLIED IN THE TEST (if different from that of starting material)

TYPE OF BIOCIDE/PESTICIDE FORMULATION (if applicable)

OTHER SPECIFICS:
Species:
rat
Strain:
Wistar
Remarks:
RccHan : WIST from Envigo RMS(US)
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source:
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 8-12 weeks
- Weight at study initiation: The body weight variataion did not exceed +/- 20% of the mean body weight of any previously treated animals
- Fasting period before study: an overnight fast immediately before dosing and for approximately 3 to 4 hours after dosing
- Housing: suspended solid-floor polypropylene cages furnished with wood flakes
- Diet (e.g. ad libitum) and Water (e.g. ad libitum): yes with the exception of an overnight fast immediately before dosing and for approximately 3 to 4 hours after dosing
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25°C
- Humidity (%): 30 to 70% relative humidity
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: To:
Route of administration:
oral: gavage
Vehicle:
arachis oil
Details on oral exposure:
VEHICLE : Arachis oil

- Justification for choice of vehicle: Arachis oil BP was used beacause the test item did not dissolve/suspend in distilled water.

All animals were dosed once only by gavade, using a metal canula attached to graduated syringe.

Rationale for the selection of the starting dose :
In the absence of data regarding the toxicity of the test item, 300 mg/kg was chosen as the starting dose.
Doses:
300 mg/kg, 2000 mg/kg
No. of animals per sex per dose:
1 female rat for the Dose Level 2000 mg/kg
5 females rat for the Dose Level 300 mg/kg
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of clinical observations: 30 minutes, 1, 2 and 4 hours after dosing and then daily for up to 14 Days.
- Frequency of mobidity and mortality : twice daily, early and late during normal working days, and once daily at weekends and public holidays.
- Frequency of body weights : recorded on Day 0 and on Day 7 and 14 or at death.
- Necropsy of survivors performed: yes, gross necropsy : external examination and opening of the abbominal and thoracic cavities. No tissues were retained. Only the appareance of any macroscopic abnormalities was recorded.
- Euthanasia : cervical dislocation
Statistics:
NA
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - <= 2 000 mg/kg bw
Based on:
test mat.
Mortality:
Dose level - 300mg/kg
There were no deaths.
Dose level - 2000mg/kg
The animal was killed for human reasons, due to the occurence of clinical signs of toxicity that approached the severity limit set forth in the UK Home Office Project License.
Clinical signs:
Dose level - 300mg/kg
Signs of systemic toxicity noted in four animals during the day of dosing were hunched posture, ataxia and pilo-erection.
One animal appeared normal throughout the observation period.
Dose level - 2000mg/kg
Signs of systemic toxicity noted were hunched posture, lethargy, ataxia, occasional body tremos, prostration, decreasded respiratory rate, hyperthermia and ptosis.
Body weight:
Dose level - 300mg/kg
All animals showed expected gains in body weight over the observation period.

Gross pathology:
Dose level - 300mg/kg
No abnormalities were noted at necropsy
Dose level - 2000mg/kg
Abnormalities noted at necropsy were dark liver, dark kidney and a pale stomach.
Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
The LD50 of the test item in the female Wistar strain rat was estimated to be in the range of 300-2000 mg/kg bw.
Executive summary:

An acute oral toxicity in the Wistar strain female rat (A. Sanders, 2018, Envigo) has been performed, according the 420 OECD guideline and in compliance with GLP, in order to assess the LD50 of the test item.

Following a sighting test with 1 animal at dose levels of 300 and 2000 mg/kg, a further group of 4 fasted female was given a single oral dose by gavage of test item, as a dissolution in arachis oil BP, at a dose level of 300 mg/kg. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy.

The animal treated at the dose level of 2000 mg/kg was killed for human reasons due to the occurenc of clinical signs of toxicity.

At the dose level of 300 mg/kg, there were no deaths, all animals showed expected gains in body weight and no abnormalities were noted at necropsy. Animals showed signs of systemic toxicity as hunched posture, ataxia and pilo-erection. One animal appeared normal throughout the observation period.

Therefore the LD50 ot the test item in the female Wistar strain rat is estimated to be in the range of 300-2000 mg/kg bw.

The test item is classified as Category 4 according to GHS criteria.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
300 mg/kg bw
Quality of whole database:
Correct

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
20/09/1991 - 31/12/1991
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
test procedure in accordance with national standard methods with acceptable restrictions
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 434 (Acute Dermal Toxicity - Fixed Dose Procedure)
GLP compliance:
yes
Test type:
fixed dose procedure
Limit test:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material:
- Expiration date of the lot/batch:
- Purity test date:
- Appearance : yellow liquid


STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material:
- Stability under test conditions: There was no apparent change in the physical appearance of the test articles during administration.
- Solubility and stability of the test substance in the solvent/vehicle:
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: All test articles were dosed as received from the sponsor
- Preliminary purification step (if any):
- Final dilution of a dissolved solid, stock liquid or gel:
- Final preparation of a solid:

FORM AS APPLIED IN THE TEST (if different from that of starting material)

TYPE OF BIOCIDE/PESTICIDE FORMULATION (if applicable)

OTHER SPECIFICS:
Species:
rabbit
Strain:
New Zealand White
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Supplier : CAMM Research Lab Animals
- Females (if applicable) nulliparous and non-pregnant: NA
- Age at study initiation: 8-12 weeks
- Weight at study initiation: 1.814 to 3.188 kg
- Fasting period before study: No
- Housing: Rabbits were housed individually in cages sized in accordance with the "Guide for the Care and Use of Laboratory Animals" of the Institute of Laboratory Animal Resources, National Research Council.
- Diet : ad libitum
- Water : ad libitum
- Acclimation period: Minimum of 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20°C ± 3°C
- Humidity (%): relative humidity of 30 to 70%
- Air changes (per hr): NA
- Photoperiod (hrs dark / hrs light): 12/12

Rationale for Test System: The albino rabbit is preferred because of its size, skin permeability and extensive database.
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
Approximately 24 hours before testing, fur was Performance: clipped from the dorsal area of the trunk of the test animals.

The test article was applied directly onto the exposed intact skin of the animals taking care to spread the substance evenly over the entire area. A square gauze patch was placed on the animals to cover the dosed area. The animal.s were wrapped with rubber dam and an elastic bandage to retard evaporation.

The test article was held in contact with the skin for twenty-four hours.

Following the twenty-four hour period of exposure, the wrappings were removed and the skin sites were wiped with water and gauze to remove any residual test article.

Duration of exposure:
24 hours
Doses:
2000 mg/kg
No. of animals per sex per dose:
10 : 5 males and 5 females
Control animals:
no
Details on study design:
Rationale for Dose Selection: As required by the regulatory agencies

Observations were recorded daily through Day 14.
Body weights were recorded at initiation and on Days 7 and 14.
All rabbits were sacrificed by a lethal injection on Day 14 and a gross necropsy was performed.
Statistics:
NA
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortalities were recorded.
Clinical signs:
No clinical signs observed
Body weight:
No changes in body weight gain.
Gross pathology:
No visible lesions were observed.
Interpretation of results:
Category 5 based on GHS criteria
Conclusions:
The LD50 of the test item is greated than 2000 mg/kg bw.
Executive summary:

In a Dermal Limit Test (Vincent B. Ciafalo, 1992), performed according to national standard methods and in compliance with GLP, one group of 10 rabbits (5 males and 5 females) per study was exposed to the test article at 2000 mg/kg during 24 hours in a semi-occlusive system.

Animals were observed for clinical signs and mortality once daily for fourteen days.

No clinical signs, no mortalities and no lesions at gross necropsy were observed.

Therefore the LD50 of the test item is greater than 2000 mg/kg bw.

Butyl-NENA is classified as Category 5 accordign to the GHS criteria.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
Correct

Additional information

Justification for classification or non-classification

Acute Oral Toxicity : Category 4 under GHS

Acute Dermal Toxicity : Category 5 under GHS