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Description of key information

Oral (OECD 423), rat: LD50 > 2000 mg/kg bw; LD50 cut-off value = 5000 mg/kg bw (limit test)
Dermal (OECD 402), rat: LD50 > 2000 mg/kg bw (limit test)

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:
22 Jul - 08 Aug 2003
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP - Guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
adopted 1996
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Version / remarks:
adopted 1996
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Version / remarks:
adopted June 1996
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF) Nov. 2000
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
other: Wistar strain Crl:(WI) BR (outbred, SPF-Quality)
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: approximately 10 weeks
- Weight at study initiation: female (mean) 230 ± 32 g; male (mean) 358 ± 19 g
- Fasting period before study: over night
- Housing: 3 animals per sex per cage in labelled Makrolon type IV cages containing purified sawdust as bedding material
- Diet: standard pelleted laboratory animal diet (Altromin, code VRF 1), ad libitum
- Water: tap water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18.1 - 24.5
- Humidity (%): 47 - 79
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
propylene glycol
Details on oral exposure:
The dose level of 2000 mg/kg bw was reached by administration of two times 1000 mg/kg bw (10 mL/kg bw) within 24 hours. The first on t=0 and the second on approx. t=1 hour.

VEHICLE
- Amount of vehicle (if gavage): Two administrations of formulation of the test substance at 10 mL/kg bw
- Justification for choice of vehicle: vehicle was selected based on trial formulations performed at the testing laboratory
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
1st step: 3 females
2nd step: 3 males
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Mortality was observed twice daily. Animals were observed at periodic intervals on the day of dosing and once daily thereafter for clinical signs. Weighing was done prior to dosing (day 1) and on days 8 and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, gross pathology
Statistics:
No statistical analysis was performed.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: According to OECD Guideline 423, Annex 2d, a cut-off value of 5000 mg/kg bw was derived based on a limit test with 2000 mg/kg bw, since no mortality occurred in any step.
Mortality:
No mortality occurred.
Clinical signs:
Clinical signs were noted in all animals between days 1 and 3 and included lethargy, hunched posture, uncoordinated movements and piloerection in all animals. In addition, ptosis were noted in all males on day 1.
Body weight:
Body weight gain was normal during the study.
Gross pathology:
No abnormalities were found at macroscopic post mortem examination of the animals.

Table 1. Table for acute oral toxicity

Dose
[mg/kg bw]

Toxicological results*

Duration of clinical signs

Time of death

Mortality (%)

Males

2000

0/3/3

Day1 – Day2

---

0

Females

2000

0/3/3

Day1 – Day3

---

0

LD50 > 2000 mg/kg bw

 Day 1 refers to the day of test substance administration.                                                                                    

* first number = number of dead animals                                 

  second number = number of animals with clinical signs         

  third number = number of animals used

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
DSD: not classified
CLP: not classified
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
The available information comprises an adequate and reliable study (Klimisch score 1), and is thus sufficient to fulfil the standard requirements set out in Annex VII, 8.5, of Regulation (EC) No 1907/2006.

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:
04 Feb - 18 Feb 2004
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP - Guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
adopted 1987
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Version / remarks:
adopted 1992
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Version / remarks:
adopted June 1996
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), November 2000, including most recent partial revisions
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
other: Wistar strain Crl:(WI) BR (outbred, SPF-Quality)
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: approximately 9 weeks
- Weight at study initiation: male (mean) 306 ± 14 g; female (mean) 216 ± 14 g
- Housing: individually in labelled Makrolon type III cages containing purified sawdust as bedding material
- Diet: standard pelleted laboratory animal diet (Altromin, code VRF1), ad libitum
- Water: tap water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 15.4 - 22.9
- Humidity (%): 29 - 80
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12
Type of coverage:
occlusive
Vehicle:
propylene glycol
Details on dermal exposure:
TEST SITE
- Area of exposure: 25 cm² for males and 18 cm² for females
- % coverage: 10
- Type of wrap if used: surgical gauze patch, successively covered with aluminium foil and Coban elastic bandage, for females bandages were fixed using a piece of micropore tape

REMOVAL OF TEST SUBSTANCE
- Washing: The skin was cleaned of residual test substance using water.
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 10 mL/kg bw
- Constant volume or concentration used: yes
- Concentration (if solution): 200 mg/mL (w/w) (calculated from 10 mL/kg bw amount and 2000 mg/kg bw)

VEHICLE
- Amount(s) applied (volume or weight with unit): 10 mL/kg bw
Duration of exposure:
24 h
Doses:
2000 mg/kg bw
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: checks were done twice daily for mortalities, clinical signs were recorded at periodic intervals on the day of dosing and once daily thereafter, body weights were determined before dosing (day 1) and on days 8 and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, gross pathology
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: no mortality was observed during the study
Mortality:
No mortality occurred.
Clinical signs:
Flat/hunched posture and/or chromodacryorrhoea were noted among all animals between days 1 and 5.
Body weight:
Body weight gain was within the normal range during the study.
Gross pathology:
No toxicologically relevant abnormalities were found at macroscopic post mortem examination of the animals.
Other findings:
White staining of the treated skin area was noted among most animals between days 2 and 9. In addition, erythema (days 2 and/or 3) or scales (between days 3 and 12) of grade 1 were noted on the treated skin area of some females (table 2).

Table 1. Table for acute dermal toxicity

Dose
[mg/kg bw]

Toxicological results*

Duration of clinical signs

Time of death

Mortality (%)

Males

2000

0/5/5

Day1 – Day5

---

0

Females

2000

0/5/5

Day1 – Day5

---

0

LD50 > 2000 mg/kg bw

 Day 1 refers to the day of test substance administration.                                                                                         

* first number = number of dead animals                                 

 second number = number of animals with clinical signs         

  third number = number of animals used                                                              

Table 2. Presence of skin effects

 

Female rats

Male rats

Animal no.

1

2

3

4

5

1 - 5

White staining

Day2 – Day8

Day2

Day2 – Day8

Day2 – Day9

---

Day2 - Day8

Day2 - Day8

Day2 - Day8

Day2 - Day8

Day2 - Day8

Erythema

(grade

1 - 4)

---

---

Day2 – Day3

(grade1)

---

Day2 (grade 1)

 ---

 ---

 ---

 ---

 ---

Scales (grade

1 - 3)

Day3 – Day12 (grade 1)

---

---

---

---

---

---

---

---

---

Day 1 refers to the day of test substance administration.     

---: sign not observed

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
DSD: not classified
CLP: not classified
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
The available information comprises an adequate and reliable study (Klimisch score 1), and is thus sufficient to fulfil the standard requirements set out in Annex VIII, 8.5, of Regulation (EC) No 1907/2006.

Additional information

The acute toxicity of 2-Hexyl-2 ‘-hydroxy-5 ‘methyldecananilide (AF-654) was assessed in an oral toxicity study and a dermal toxicity study. No inhalation toxicity study is available.

In an acute oral toxicity study performed according to OECD Guideline 423, the limit dose of 2000 mg/kg bw AF-654 was administered by gavage to 3 female Wistar rats in a first step and to 3 male Wistar rats in a second step (Hooiveld, 2003a). Animals were observed daily for clinical signs and body weight was determined weekly. Macroscopic examination was performed at the end of the 14-day observation period at terminal sacrifice. There were no mortalities during the observation period and body weight gain of all animals was normal. Clinical signs were noted between Days 1 and 3 and included lethargy, hunched posture, uncoordinated movements and piloerection in all animals. Moreover, all males showed ptosis on Day 1. Those effects are known to be stress related and are therefore not considered to be due to test substance treatment. Additionally, no treatment-related abnormalities were found during the macroscopic post mortem examination of the animals. Based on the results of this study, the LD50 value was determined to be > 2000 mg/kg bw in rats. In accordance with OECD Guideline 423, Annex 2d, a cut-off value of 5000 mg/kg bw was derived, since no mortality occurred in any step at the limit dose of 2000 mg/kg bw.

The acute dermal toxicity of AF-654 was assessed in a limit test performed in 5 male and 5 female Wistar rats according to OECD Guideline 402 (van Otterdijk, 2004a). A single dose of 2000 mg/kg bw of the test substance in propylene glycol was applied to the clipped skin of rats under occlusive conditions for 24 hours. There was no mortality and no effects on body weight during the 14-day observation period. Flat/hunched posture and/or chromodacryorrhoea were noted among all animals between Days 1 and 5. The necropsy and gross pathological examination did not show any treatment-related effects. White staining of the treated skin area was noted among most animals between Days 2 and 9. In addition, general erythema (Days 2 and/or 3) or scales (between Days 3 and 12) were noted on the treated skin area of some females. The LD50 value for systemic toxicity is considered to be > 2000 mg/kg bw.


Justification for selection of acute toxicity – oral endpoint
There is only one study available.

Justification for selection of acute toxicity – inhalation endpoint
No study required since the substance has a low potential for exposure via the inhalation route taking into account the physico-chemical properties and the conditions of use.

Justification for selection of acute toxicity – dermal endpoint
There is only one study available.

Justification for classification or non-classification

The available data on acute oral and dermal toxicity of the test substance do not meet the criteria for classification according to Regulation (EC) 1272/2008 or Directive 67/548/EEC, and are therefore conclusive but not sufficient for classification.