4-trans-Methoxy-4-trans-propyl-[1,1-bicyclohexyl]

Endpoint summary

Description of key information

Skin irritation was observed in a study according to OECD TG 404 in rabbits (reference 7.3.1 -1).

No eye irritating potential could be detected in a study according to OECD TG 405 in rabbits (reference 7.3.2 -1).

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

Jan 10 - May 24, 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

2002

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Remarks:

CRL:KBL(NZW)

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River WIGA GmbH, Kisslegg
- Age at study initiation: 27 weeks
- Weight at study initiation: 3.95 kg
- Housing: kept serarately in a special rabbit cage (manufacturer: Becker; type K99/30 KU) on a mobile rack
- Diet: ad libitum, commercial diet for rabbits, Provimi Kliba 3418.0 and ssniff K snack
- Water: ad libitum, fresh community tap water from Makrolon drinking bottles
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 15 to 27
- Humidity (%): 23 to 98
The room temperature and relative atmospheric humidity in the animal room was transiently outside the target range of 16 to 22 °C and > 40 %. These deviations did not affect the integrity or outcome of the study.
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.5 mL

Duration of treatment / exposure:

4 hours

Observation period:

1, 24, 48, 72 hours after removal of patches and then daily up to day 22

Number of animals:

1f

Details on study design:

TEST SITE
- Area of exposure: 0.5 mL of the test material was spread onto a patch of about 6 cm2.
- Type of wrap: The patch was fastened to the left side of the animal’s back and kept in place by a self-adhesive fabric (Fixomull®stretch, Beiersdorf).

REMOVAL OF TEST SUBSTANCE
- Washing: no, residues were wiped off
- Time after start of exposure: 4 hours

OBSERVATION TIME POINTS
The rabbit was examined for skin alterations, behavior, and general condition 1 hour after removal of the patch, after 24,48,72 hours and then daily up to experimental day 22.

SCORING SYSTEM:
- Method of calculation: Skin changes at the application site was evaluated according to the DRAIZE-, OECD- and EEC recommendations.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

not fully reversible within: 22 days

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 8 days

Irritant / corrosive response data:

After a single application to the intact dorsal skin of a rabbit, for 4 hours under semiocclusive conditions, erythema (scores 2 to 4) from day 1 up to the end of the study and edema (scores 1 to 3) from day 1 up to day 7 of the study occurred. Furthermore, small scales from day 13 up to the end of the study and epithelization from day 15 up to the end of the study were observed. Brittle skin from day 8 up to day 14 was seen at the site of application.

Other effects:

- Other adverse systemic effects: No signs of clinical toxicity were detected. The animal survived the observation period. Body weight development of the treated rabbit was inconspicuous.

Pretests:

Prior to testing the pH of the preparation was determined with pH-test strips. The pH was found to be about 5.

The test material was screened for its irritating potential using the hen's egg test on the chorioallantoic membrane (HET-CAM) as described below. Fertile hen eggs, on day 10 of incubation, provided the test system. The eggshell was entered above the air cell by a dentist's rotary and then pared off. The shell with the attached outer membrane and inner egg membrane was carefully opened. After the membrane had been moistened with isotonic sodium chloride solution, it was removed to expose the chorioallantoic membrane (CAM) of the fertilized egg, which lies immediately below. The CAM of fertilized and incubated hen eggs provides a suitable model for membranes with an intact and functional vascular system. The CAM has no nerve fibers in the early stages of development and is, thus, regarded as being insensitive to pain. In this model the cumulative irritation score of the test item was 0.00.

Interpretation of results:

Category 2 (irritant) based on GHS criteria

Conclusions:

The test material caused irritative skin effects.

Executive summary:

The liquid test material was spread onto patches and applied to the intact skin of a previously shaven rabbit for 4 hours under semi-occlusive conditions. The study was performed initially with one animal, which showed signs of irritation (erythema, edema,  brittle skin, and small  scales) and therefore no more animals were treated. The first examination of the treated skin followed 1 hour after patch removal. Thereafter, examinations were performed daily for a further 21 days. No signs of pain were observed after administration of the test material. Under the conditions of the present study erythema (scores 2 to 4) from day 1 up to the end of the study and edema (scores 1 to 3) from day 1 up to day 7 of the study occurred. Furthermore, small scales were observed from day 13 up to the end of the study and epithelization from day 15 up to the end of the study. Brittle skin from day 8 up to day 14 was seen at the site of application. The test material caused irritative skin effects.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Apr 12 - Aug 23, 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

2002

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Remarks:

Crl:KBL(NZW)

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River Wiga GmbH, Kifilegg
- Age at study initiation: Approx. 18 weeks
- Weight at study initiation: mean 3.36 kg
- Housing: animals were kept separately in special rabbit cages (manufacturer: Becker; type K99/30 KU)
- Diet: ad libitum, Provimi Kliba 3418.0 and ssniff® K snack
- Water: ad libitum, tap water from Makrolon drinking bottles
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 15 to 21
- Humidity (%): 42 to 56
- Photoperiod (hrs dark / hrs light): 12/12

The room temperature in the animal room was transiently outside the target range of 16 to 22 °C. This minor and short deviation did not influence the integrity or outcome of the study.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.1 mL of the test material was instilled into the conjunctival sac of the left eye of each animal by gently pulling the lower lid away from the eyeball to form a cup into which the test material was placed.

Observation period (in vivo):

1 hour after treatment, after 24, 48, and 72 hours, then daily up to day 5 of the experimental part

Number of animals or in vitro replicates:

3f

Details on study design:

SCORING SYSTEM: Eye changes were evaluated according to DRAIZE, OECD and EEC recommendations.

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 3 days

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.67

Max. score:

3

Reversibility:

fully reversible within: 4 days

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.67

Max. score:

3

Reversibility:

fully reversible within: 4 days

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Remarks on result:

not measured/tested

Irritant / corrosive response data:

No signs of irritation were observed at the cornea or iris. One animal showed redness of the conjunctiva (score 1) on day 2 and the other two animals showed redness (score 1) from the first reading up to day 3 of the experimental part. All animals showed chemosis (score 1) and discharge (score 3) one hour after instillation of the test material. No abnormalities were detected in the untreated eyes.

Other effects:

- Other observations: No signs of clinical toxicity were detected. All animals survived the observation period. Body weight development of the treated rabbits was inconspicuous.

Interpretation of results:

GHS criteria not met

Conclusions:

No eye irritating potential could be detected.

Executive summary:

The liquid test material was tested for its eye irritating properties. Prior to testing, the hen's egg test on the chorioallantoic membrane was performed and no irritating potential could be detected. The test for primary eye irritation was performed initially with one animal and followed by the confirmatory test with two further animals.
For the test of primary eye irritation, the test material was applied into the conjunctival sac of rabbits. The first examination of the eyes followed 1 hour after instillation. The other examinations of the eyes were performed daily for a further 7 days. No signs of irritation were observed at the cornea or iris. One animal showed redness of the conjunctiva (score 1) on day 2 and the other two animals showed redness (score 1) from the first reading up to day 3 of the experimental part. All animals showed chemosis (score 1) and discharge (score 3) one hour after instillation of the test material. No eye irritating potential could be detected.

 

Reference 2

Endpoint:

eye irritation: in vitro / ex vivo

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

an in vitro eye irritation study does not need to be conducted because adequate data from an in vivo eye irritation study are available

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation

A WoE Approach was conducted to investigate the skin irritating properties of the substance:

A study according OECD TG 404 was conducted. The liquid test material was spread onto patches and applied to the intact skin of a previously shaven rabbit for 4 hours under semi-occlusive conditions. The study was performed initially with one animal, which showed signs of irritation (erythema, edema, brittle skin, and small scales) and therefore no more animals were treated. The first examination of the treated skin followed 1 hour after patch removal. Thereafter, examinations were performed daily for a further 21 days. No signs of pain were observed after administration of the test material. Under the conditions of the present study erythema (scores 2 to 4) from day 1 up to the end of the study and edema (scores 1 to 3) from day 1 up to day 7 of the study occurred. Furthermore, small scales were observed from day 13 up to the end of the study and epithelization from day 15 up to the end of the study. Brittle skin from day 8 up to day 14 was seen at the site of application. The test material caused irritative skin effects.

In addition, results for two in vitro tests are available. In a study performed comparable to OECD TG 439 the irritation potential of the test material was assessed by means of the Human Skin Model Test. Three tissues of the human skin model EPISKIN-SM were treated with either the test item, the negative or the positive control for 15 minutes. 15 µL of the liquid test item were applied to each tissue, spread to match the tissue size. 15 µL of either the negative control (deionised water) or the positive control (5% Sodium lauryl sulfate) were applied to each tissue. After treatment with the negative control, the absorbance values were well above the required acceptability criterion of mean OD 0.8 for the 15 minutes treatment interval thus showing the quality of the tissues. Treatment with the positive control induced a sufficient decrease in the relative absorbance as compared to the negative control for the 15 minutes treatment interval thus ensuring the validity of the test system. After treatment with the test item the relative absorbance values were not decreased (117.3%). Therefore, the test item is not considered to possess an irritant potential.

In an in vitro study performed according OECD TG 431 the corrosive potential of the test material was assessed by means of the Human Skin Model Test. Two tissues of the human skin model EPIDERM were treated with either the test item, the negative or the positive control for 3 minutes and 1 hour, respectively. 50 µL of the liquid test item were applied to each tissue, spread to match the tissue size. 50 µL of either the negative control (deionised water) or the positive control (8.0 N KOH) were applied to each tissue. After treatment with the negative control the absorbance values were well above the required acceptability criterion of mean OD ≥ 0.8 for both treatment intervals thus showing the quality of the tissues. Treatment with the positive control induced a decrease in the relative absorbance as compared to the negative control to 19.5% for the 3 minutes treatment interval and 8.2% for the 1 hour treatment interval thus ensuring the validity of the test system. After treatment with the test item the relative absorbance values were not decreased after 3 minutes treatment. After 1 hour treatment relative absorbance values were not reduced relevantly. Therefore, the test item is considered non-corrosive.

In conclusion, the substance was identified as skin irritant based on the available in vivo data.

Eye irritation

A study according OECD TG 405 was conducted. The liquid test material was tested for its eye irritating properties. Prior to testing, the hen's egg test on the chorioallantoic membrane was performed and no irritating potential could be detected. The test for primary eye irritation was performed initially with one animal and followed by the confirmatory test with two further animals. For the test of primary eye irritation, the test material was applied into the conjunctival sac of rabbits. The first examination of the eyes followed 1 hour after instillation. The other examinations of the eyes were performed daily for a further 7 days. No signs of irritation were observed at the cornea or iris. One animal showed redness of the conjunctiva (score 1) on day 2 and the other two animals showed redness (score 1) from the first reading up to day 3 of the experimental part. All animals showed chemosis (score 1) and discharge (score 3) one hour after instillation of the test material. All observations were fully reversible within the test period.No eye irritating potential could be detected.

 

Justification for classification or non-classification

Classification, Labeling, and Packaging Regulation (EC) No 1272/2008

The available data for skin and eye irritation are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. Based on this data, the substance is considered to be classified for skin irritation Cat.2 (H315) under Regulation (EC) No 1272/2008, as amended for the twelfth time in Regulation (EU) 2019/521. No classification for eye irritation is required under Regulation (EC) No 1272/2008, as amended for the twelfth time in Regulation (EU) 2019/521.