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Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
02-09-2008 to 23-09-2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study performed under GLP. All relevant validity criteria were met; note that for local effects only: the reshaving at day 8 is a deviation is suggestive of effects that are non-test item related, which limits the reliability for assessment.
Cross-reference
Reason / purpose for cross-reference:
reference to same study
Reference
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
02-09-2008 to 23-09-2008
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Guideline study performed under GLP. All relevant validity criteria were met; note that for local effects only: the reshaving at day 8 is a deviation is suggestive of effects that are non-test item related, which limits the reliability for assessment.
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
other: OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
yes
Remarks:
note that for local effects only: the reshaving at day 8 is a deviation is suggestive of effects that are non-test item related, which limits the reliability for assessment.
Qualifier:
according to guideline
Guideline:
other: EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
yes
Remarks:
See above
GLP compliance:
yes
Species:
rat
Strain:
Wistar
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Recognised supplier
- Age at study initiation: 9 - 11 weeks.
- Weight at study initiation: 197.3 - 264.0 g; the weight variation did not exceed ±20% of the mean weight for each sex.
- Fasting period before study: Not applicable
- Housing: during acclimation: group housed by sex; during study: individually housed in Makrolon type-4 cages furnished with softwood bedding.
- Diet (e.g. ad libitum): Certified diet from recognised supplier, provided ad libitum.
- Water (e.g. ad libitum): ad libitum.
- Acclimation period: At least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 30 - 70%
- Air changes (per hr): 10 to 15 air changes per hour
- Photoperiod: 12 h light / 12 h dark

IN-LIFE DATES: From: To: 2008-09-02 to 2008-09-23
Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
other: polyethylene glycol
Remarks:
PEG 300 (details available in full study report)
Controls:
not required
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg
- Concentration (if solution): See below.
- Constant volume or concentration used: 4 mL volume ; at a dose level of 2000 mg/kg bw test item.
Duration of treatment / exposure:
24 hours
Observation period:
Once daily during days 2-15. All abnormalities were recorded.
Number of animals:
5 per sex per dose (5 male/5 female)
Details on study design:
TEST SITE
- Area of exposure: the day before treatment the back and flanks were clipped free of hair. Dorsal area application. Note: that the skin was re-shaved on day 8 to facilitate the reading of local reactions.
- % coverage: Approximately 10% of total body surface
- Type of wrap if used: The area of application was covered by a semi-occlusive dressing and wrapped with a piece of elastic adhesive bandage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The treated skin and surrounding hair wiped with luke warm water and dried with disposable towl to remove any residual test item
- Time after start of exposure: 24 h

OBSERVATION TIME POINTS
(indicate if minutes, hours or days) : Clinical observations and mortality checks were conducted at approximately 0.5, 1, 3, and 5 hours and subsequently twice daily for days 2 to 15. Local effects were examined once daily days 2 to 15 after the completion of the 24-hour exposure period.

SCORING SYSTEM:
- Method of calculation: Full details on the scoring and criteria (appears consistent with Draize for Erythema) are given in the full study report. Individual bodyweights were recorded prior to application of the test item on Day 1 and on Days 8 and 15.
Irritation parameter:
erythema score
Remarks:
mean
Basis:
animal #1
Remarks:
male
Time point:
other: 48/72/96h
Score:
1.33
Reversibility:
fully reversible within: 7 days
Irritation parameter:
erythema score
Remarks:
mean
Basis:
animal #2
Remarks:
male
Time point:
other: 48/72/96h
Score:
1.33
Reversibility:
fully reversible within: 7 days
Irritation parameter:
erythema score
Remarks:
mean
Basis:
animal #3
Remarks:
male
Time point:
other: 48/72/96h
Score:
1.33
Reversibility:
fully reversible within: 7 days
Irritation parameter:
erythema score
Remarks:
mean
Basis:
animal #4
Remarks:
male
Time point:
other: 48/72/96h
Score:
1.33
Reversibility:
fully reversible within: 7 days
Irritation parameter:
erythema score
Remarks:
mean
Basis:
animal #5
Remarks:
male
Time point:
other: 48/72/96h
Score:
1.33
Reversibility:
fully reversible within: 7 days
Irritation parameter:
erythema score
Remarks:
mean
Basis:
animal: #1
Remarks:
female
Time point:
other: 48/72/96h
Score:
1
Reversibility:
fully reversible within: 7 days
Irritation parameter:
erythema score
Remarks:
mean
Basis:
animal: #2
Remarks:
female
Time point:
other: 48/72/96h
Score:
1.33
Reversibility:
fully reversible within: 7 days
Irritation parameter:
erythema score
Basis:
animal: #3
Remarks:
female
Time point:
other: 48/72/96h
Score:
1.33
Reversibility:
fully reversible within: 7 days
Irritation parameter:
erythema score
Remarks:
mean
Basis:
animal: #4
Remarks:
female
Time point:
other: 48/72/96h
Score:
1.33
Reversibility:
fully reversible within: 7 days
Irritation parameter:
erythema score
Remarks:
mean
Basis:
animal: #5
Remarks:
female
Time point:
other: 48/72/96h
Score:
1
Reversibility:
fully reversible within: 7 days
Irritant / corrosive response data:
- Dermal reactions: All indicated slight (score = 1) to moderate (score =2) erythema from day 2 to day 6. The sites were re-shaved on day 8 after which one indication of slight erythema on days 8 to 12 and 9/10 indications of 'scaling'. See 'other information on results incl. tables' for further information.
Other effects:
- Other adverse local effects: None. See 'other information on results incl. tables' for further information.
- Other adverse systemic effects: None.

Applicant assessment indicates: within local effects the test item caused only mild transient irritation (score = 2) on day 2 which was then slight (score 1) on days 3, 4 and fully reversed by day 7. When the sites were re-shaved on day 8 this led to score = 1 slight irritation and scaling that persisted to the end of the observation period. This was clearly not a test item related effect, as the effects had already reversed on day 7 in all males/females (information provided in the full study report). The OECD TG 404 test guideline for skin irritation specifies that clipping of the fur is done at the start of the test and to attempts are to minimise abrasion of the skin which can interfere with assessment. It appears that the effects post day 8 were the result of a deviation for the reading of local effects which made fully reversed effects re-appear at the treated sites. Expert judgement indicates these were non-test item related effects.

Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of this study the dermal LD50 was established to exceed 2000 mg/kg bw in male/female Wistar rat. Under the conditions of this study and under the Globally Harmonized Classification System of Classification and Labelling of Chemicals (GHS), the LD50 cut-off value was considered to be greater than 5000 mg/kg body weight. The test item caused only mild transient irritation (score = 2) on day 2 which was then slight (score 1) on days 3, 4 and fully reversed by day 7. The test item was not considered to be skin irritating.
Executive summary:

The study was performed according to OECD TG 402 and EU Method B.3 Acute Toxicity (Dermal) and in accordance with GLP to assess the acute dermal toxicity of the test substance in the Wistar HanRcc: WIST (SPF) strain rat. A group of ten animals (five males and five females) was given a single, 24 hour, semi-occluded dermal application of the diluted test item in PEG 300 vehicle to intact skin at a dose level of 2000 mg/kg body weight. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy. There was no mortality during the study. There were no signs of system toxicity or abnormalities on necropsy. Animals showed expected gains in body weight. Minimal signs of dermal irritation were noted (score 2) at day 2 and (score 1) days 4 to 5 or 6, which had fully reversed at day 7. The dermal LD50 was established to exceed 2000 mg/kg bw in male/female Wistar HanRcc: WIST (SPF) rat. Under the conditions of this study, and according to the GHS criteria, the LD50 cut-off value was considered to be greater than 5000 mg/kg body weight. The test item was not considered to be skin irritating.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2008
Report date:
2008

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
-
EC Number:
482-160-5
EC Name:
-
Cas Number:
130786-09-3
Molecular formula:
C12H13N
IUPAC Name:
(2Z)-2-phenylhex-2-enenitrile
Test material form:
liquid
Details on test material:
- Physical state: liquid
- Storage condition of test material: Refrigerator (range of 5 ± 3 °C), in the dark under nitrogen
- Other: colourless

Test animals

Species:
rat
Strain:
Wistar
Remarks:
HanRcc: WIST (SPF) strain
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Recognised supplier
- Age at study initiation: 9 - 11 weeks.
- Weight at study initiation: 197.3 - 264.0 g; the weight variation did not exceed ±20% of the mean weight for each sex.
- Fasting period before study: Not applicable
- Housing: during acclimation: group housed by sex; during study: individually housed in Makrolon type-4 cages furnished with softwood bedding.
- Diet (e.g. ad libitum): Certified diet from recognised supplier, provided ad libitum.
- Water (e.g. ad libitum): ad libitum.
- Acclimation period: At least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 30 - 70%
- Air changes (per hr): 10 to 15 air changes per hour
- Photoperiod: 12 h light / 12 h dark

IN-LIFE DATES: From: To: 2008-09-02 to 2008-09-23

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
polyethylene glycol
Remarks:
PEG 300 (details available in full study report)
Details on dermal exposure:
TEST SITE
- Area of exposure: the day before treatment the back and flanks were clipped free of hair. Dorsal area application. Note: that the skin was re-shaved on day 8 to facilitate the reading of local reactions.
- % coverage: Approximately 10% of total body surface
- Type of wrap if used: The area of application was covered by a semi-occlusive dressing and wrapped with a piece of elastic adhesive bandage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The treated skin and surrounding hair wiped with luke warm water and dried with disposable towl to remove any residual test item
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg
- Concentration (if solution): See below.
- Constant volume or concentration used: 4 mL volume ; at a dose level of 2000 mg/kg bw test item.
Duration of exposure:
24 hours
Doses:
2000 mg/kg
No. of animals per sex per dose:
5 per sex per dose (5 male/5 female)
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical observations and mortality checks were conducted at approximately 0.5, 1, 3, and 5 hours and subsequently twice daily for days 2 to 15. Local effects were examined once daily days 2 to 15 after the completion of the 24-hour exposure period. Full details on the scoring and criteria (appears consistent with Draize for Erythema) are given in the full study report. Individual bodyweights were recorded prior to application of the test item on Day 1 and on Days 8 and 15.
- Necropsy of survivors performed: yes
Statistics:
No statistical analyses were performed.

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality was observed.
Clinical signs:
- Clinical observations: No signs of systemic toxicity were noted during the observation period.
- Dermal reactions: All indicated slight (score = 1) to moderate (score =2) erythema from day 2 to day 6. The sites were re-shaved on day 8 after which one indication of slight erythema on days 8 to 12 and 9/10 indications of 'scaling'. See 'other information on results incl. tables' for further information.
Body weight:
Animals showed expected gains in body weight.
Gross pathology:
No abnormalities were noted at necropsy.

Any other information on results incl. tables

Applicant assessment indicates: within local effects the test item caused only mild transient irritation (score = 2) on day 2 which was then slight (score 1) on days 3, 4 and fully reversed by day 7. When the sites were re-shaved on day 8 this led to score = 1 slight irritation and scaling that persisted to the end of the observation period. This was clearly not a test item related effect, as the effects had already reversed on day 7 in all males/females (information provided in the full study report). The OECD TG 404 test guideline for skin irritation specifies that clipping of the fur is done at the start of the test and to attempts are to minimise abrasion of the skin which can interfere with assessment. It appears that the effects post day 8 were the result of a deviation for the reading of local effects which made fully reversed effects re-appear at the treated sites. Expert judgement indicates these were non-test item related effects.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of this study the dermal LD50 was established to exceed 2000 mg/kg bw in male/female Wistar rat. Under the conditions of this study and under the Globally Harmonized Classification System of Classification and Labelling of Chemicals (GHS), the LD50 cut-off value was considered to be greater than 5000 mg/kg body weight.
Executive summary:

The study was performed according to OECD TG 402 and EU Method B.3 Acute Toxicity (Dermal) and in accordance with GLP to assess the acute dermal toxicity of the test substance in the Wistar HanRcc: WIST (SPF) strain rat. A group of ten animals (five males and five females) was given a single, 24 hour, semi-occluded dermal application of the diluted test item in PEG 300 vehicle to intact skin at a dose level of 2000 mg/kg body weight. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy. There was no mortality during the study. There were no signs of system toxicity or abnormalities on necropsy. Animals showed expected gains in body weight. Minimal signs of dermal irritation were noted (score 2) at day 2 and (score 1) days 4 to 5 or 6, which had fully reversed at day 7. The dermal LD50 was established to exceed 2000 mg/kg bw in male/female Wistar HanRcc: WIST (SPF) rat. Under the conditions of this study, and according to the GHS criteria, the LD50 cut-off value was considered to be greater than 5000 mg/kg body weight.

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