Registration Dossier

Administrative data

Description of key information

Oral LD50 female rats = >2000 mg/kg bw; OECD 420; Sanders, A. (2018)

According to the CLP, the test item does not meet the criteria for acute oral toxicity.

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:
31 May 2018 - 21 Jun 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 (Acute Toxicity (Oral))
Version / remarks:
Commission Regulation (EC) No. 440/2008
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Version / remarks:
2002
Deviations:
no
Qualifier:
according to
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), Testing Guidelines for Toxicology Studies, No. 2-1-1 "Acute oral toxicity studies", 12 Nousan No. 8147
Version / remarks:
24 Nov 2000
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes
Specific details on test material used for the study:
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature in the dark
- Stability under test conditions: Not reported
- Solubility and stability of the test substance in the solvent/vehicle: Not reported
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: Not reported

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Test item formulated in arachis oil BP within 4 hours prior to dosing. Homogeneity was accomplished to a visually acceptable level.
- Preliminary purification step (if any): N/A
- Final dilution of a dissolved solid, stock liquid or gel: Not reported
- Final preparation of a solid: N/A

FORM AS APPLIED IN THE TEST (if different from that of starting material): Light yellow liquid

OTHER SPECIFICS: Homogeneity of formulation confirmed by visual inspection.
Species:
rat
Strain:
Wistar
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 at the start of treatment
- Fasting period before study: yes overnight and for approximately 3 - 4 hours post dosing
- Housing: housed in groups of up to 4 individuals in suspended solid-floor polypropylene cages furnished with woodflakes.
- Diet (e.g. ad libitum): free access to food (2014C Teklad Global Rodent diet supplied by Envigo RMS (UK) Limited, Oxon, UK)
- Water (e.g. ad libitum): free access to mains drinking water
- Acclimation period: minimum of 5 days

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

IN-LIFE DATES: Not clarified in the study report

Route of administration:
oral: gavage
Vehicle:
arachis oil
Remarks:
Arachis oil BP used for 300 mg/kg bw dose only
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 30 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg total
- Justification for choice of vehicle: Arachic oil was used because the test item did not dissolve/suspend in distilled water
- Lot/batch no. (if required): n/d
- Purity: n/d

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw (300 mg/kg bw) and 2.24 mL/kg bw (2000 mg/kg bw)

DOSAGE PREPARATION (if unusual): applied unchanged

CLASS METHOD (if applicable) - a fixed dose procedure was used
- Rationale for the selection of the starting dose: In the absence of data regarding toxicity of the test item, 300 mg/kg bw was chosen as the start dose for one female. In the absence of effects at this dose level a further female was tested at 2000 mg/kg bw. As not effects were found here a further 4 females were tested at the maximum guideline required concentration of 2000 mg/kg bw. This is in accordance with the OECD 420 (2001).
Doses:
300, 2000 mg/kg
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Each animal was given single dose of 10 mL/kg dose volume of 2000 mg/kg of the test item of 30 mg/mL concentration.
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: 0.5, 1, 2 and 4 hours after dosing then daily thereafter until day 14 post-dose.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs and body weight
Statistics:
not required
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
>= 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
not determinable due to absence of adverse toxic effects
Mortality:
No deaths reported
Clinical signs:
There were no signs of systemic toxicity noted during the course of the study with excepttion of hunched posture was noted within four hours post dosing in animals treated at a dose level of 300 mg/kg and four animal treated at 2000 mg/kg.
Body weight:
All animals showed expected weight gain. The animal dosed at 300 mg/kg bw gained 16 g in the first week and 11 g in the second week (27 g total). The animals dosed at 200 mg/kg bw gained 16 g in the first week and 11 g in the second week (27 g total).
Gross pathology:
No abnormality noted at necropsy

Table 1:   Number of animals dead (and with evident toxicity)

Dose

(mg/kg bw)

Mortality

(# dead / total)

Time range of deaths

(hours)

Number with evident toxicity

(# / total)

Male

Female

Combined

Male

Female

Combined

300

-

0 / 5

0 / 5

n/a

 

0 / 5

0 / 5

2000

-

0 / 5

0 / 5

n/a

-

0 / 5

0 / 5

Table 2:   Number of animals hunched at observations time x

Dose

(mg/kg bw)

Hunched

(# hunched / total)

Time (h)

Male

Female

Combined

300

0.5

-

0 / 1

0 / 1

1

 

1 / 1

1 / 1

2

 

1 / 1

1 / 1

3

 

1 / 1

1 / 1

4

 

1 / 1

1 / 1

1-14 (days)

 

0 / 1

0 / 1

2000

0.5

 

4 / 5

4 /5

1

 

4 / 5

4 /5

2

 

4 / 5

4 /5

3

 

4 / 5

4 /5

4

 

4 / 5

4 /5

1-14

-

0 / 5

0 / 5

Table 3:   Body weight gain (g)

Body weight gain (g)

Animal #

Week 1

Week 2

Female 1-0

16

11

Female 2-0

19

16

Female 3-0

23

16

Female 3-1

23

27

Female 3-2

27

9

Female 3-3

27

14

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 (Globally Harmonized Classification System - Unclassified).
Executive summary:

OECD 420 (2018) - In an acute oral toxicity study, a group of fasted, 8-12 week old female Wistar rats were given a single oral dose of AD-464 at a single dose rate of 2000 mg/kg bw (limit test) and observed for 14 days. A sighting study in line with the guideline with 1 animal dosed at 300 mg/kg bw was also conducted.

In the absence of mortality during the observation period, the oral LD50 was estimated to be greater than 2000 mg/kg bw.

In addition, there were no treatment related clinical signs, necropsy findings or changes in body weight observed in any of the individuals.

In conclusion, the test item, AD-464 did not meet the criteria for classification according to Regulation (EC) No. 1272/2008 on the Classification, Labelling and Packaging of Substances and Mixture.

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 2,000 mg/kg.

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available
Quality of whole database:
An acute toxicity study via the inhalation route is not required for this 1-10 tonnage band registration. However, consideration of the hazard classification was carried out since the test item contains phenol (that has a harmonised classification of Cat 3 for this hazard) at a weight percent level greater than the generic cut-off value of 0.1 %.
In accordance with CLP the acute toxicity estimate should be derived by the equation below, subsequently the classification will be derived from this estimate.
100/ATEmixture = sum concentration ingredient/ATE ingredient
The ATE of mixture containing phenol at 1.85% therefore = 162 mg/m3
Hence, the mixture is NOT classified for acute toxicity via the inhalation route.

Acute toxicity: via dermal route

Endpoint conclusion
Endpoint conclusion:
no study available
Quality of whole database:
An acute toxicity study via the idermal route is not required for this 1-10 tonnage band registration. However, consideration of the hazard classification was carried out since the test item contains phenol (that has a harmonised classification of Cat 3 for this hazard) at a weight percent level greater than the generic cut-off value of 0.1%.
In accordance with CLP the acute toxicity estimate should be derived by the equation below, subsequently the classification will be derived from this estimate.
100/ATEmixture = sum concentration ingredient/ATE ingredient
The ATE of mixture containing phenol at 1.85% therefore = 16216 mg/kg bw
Hence, the mixture is NOT classified for acute toxicity via the dermal route.

Additional information

Acute oral toxicity

OECD 420 (2018) - In an acute oral toxicity study (OECD 420), groups of fasted, 8 -12 week old, female Wistar (RccHan™:WIST) rats were given a single oral dose of AD-464 at doses 2000 and 300 mg/kg bw and observed for 14 days.

Oral LD50 female rats = >2000 mg/kg bw.

According to the UN GHS classification and Regulation (EC) No. 1272/2008 (relarting to the classification, labeling and packageing of subtances and mixtures), the test item does not meet the criteria for acute oral toxicity.

Justification for classification or non-classification

Based on the OECD 420 study conducted on the test item, the substance does not meet the criteria for classification under acute oral toxicity in accordance with GHS and Regulation (EC) No 1272/2008 (CLP).

Based on CLP mixture principles and ATE calculations, where required and possible (i.e. for phenol), the substance does not meet the criteria for classification under acute inhalation and acute dermal toxicity.