1-Ethoxy-2,3-difluoro-4-(trans-4-propylcyclohexyl)-benzene

Endpoint summary

Description of key information

The test item is considered to possess no skin irritating properties.

No eye irritating potential of the test item could be detected.

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:

key study

Study period:

Mar 12 - Apr 09, 1997

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:

July 1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Strain: Albino rabbit, Crl:KBL(NZW)
Source: Charles River Wiga GmbH, Kifßlegg
Age: about 27 weeks
Initial Weight: 3.66 kg (3.15-4.25 kg)

Identification and adaptation
The animal was kept in the experimental room for more than 7 days to allow them to acclimatize. The animal was uniquely identified by a tattoo in the ear displaying the animal number. An individual cage card was affixed to the cage displaying the study number, test material, day of treatment, and animal number.
Housing and diet
The rabbits were housed in an air-conditioned room of about 28 m^2 in the Institute of Toxicology. Lighting was controlled by a timer to provide a 12-hour light and a 12-hour dark regime. They were kept separately in special rabbit cages (manufacturer: Becker; type K99/30 KU, floor area about 5400 cm^2, a shelter with an integrated sitting board of about 1820 cm^2; overall height: 60 cm) with plastic grids placed on mobile racks. The collection pans underneath the cages were cleaned at least three times a week. The cages were cleaned before the start of the study. Temperature and relative humidity were measured using a thermohygrograph. The room temperature varied from 20 to 21 °C and the relative humidity from 59 to 68 %. The rabbits received a commercial diet for rabbits, Altromin Standard Diet ad libitum and fresh tap water from Makrolon drinking bottles ad libitum. According to the specifications given by the manufacturer, the diet had been checked by independent laboratories. Analysis included both qualitative and quantitative evaluation for heavy metals, aflatoxins, pesticides, and antibiotics. The drinking water was periodically analyzed according to the German regulations for human drinking water.

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

other: aqua pro injectione

Amount / concentration applied:

0.5 g

Duration of treatment / exposure:

4 hours

Observation period:

1, 24, 48, 72 hours after removal of patches and then daily up to day 8

Number of animals:

3 females

Details on study design:

Observations for clinical symptoms
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 8.Skin changes at the application sites were evaluated according to the DRAIZE-, OECD-and EEC recommendations.
Grading scale for evaluation:
Erythema and eschar formation
No erythema: 0
Very slight erythema (barely perceptible): 1
Well defined erythema: 2
Moderate to severe erythema: 3
Severe erythema (beet redness) to eschar formation preventing grading of erythema: 4
Maximum possible: 4
Edema formation
No edema: 0
Very slight edema (barely perceptible): 1
Slight edema (edges of area well defined by definite raising): 2
Moderate edema (raised approx. 1 mm): 3
Severe edema (raised more than 1 mm and extending beyond area of exposure): 4
Maximum possible: 4
Total possible irritation score (maximum): 8
Mean score = Mean grading for erythema or edema of all the rabbits per time point
Mean cumulative score = Mean grading of all readings for erythema and edema per time point
Mean score per animal = Mean grading for each animal of erythema or edema (24, 48, and 72 hours after removal of the patches)

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

0.5 g of the test material was administered. After a single application to the intact dorsal skin of a rabbit, for 4 hours under semiocclusive conditions, no signs of irritation were observed.

Interpretation of results:

GHS criteria not met

Conclusions:

The test item is considered to possess no skin irritating properties.

Executive summary:

The purpose of this assay was to identify the skin irritation/corrosion potential of the test material when applied to the intact skin of previously shaven rabbits for a 4 hours period under semiocclusive conditions.

This study should provide a rational basis for risk assessment to the irritating potential of the test item in man and was conducted according to OECD 404.

To test for primary skin irritation, 0.5 g of the test material was mixed with some drops of aqua pro injectione to ensure good contact with the skin. Afterwards the test material was spread onto 6 cm2 patches and applied to the intact skin of three previously shaven rabbits for a 4 hours period under semiocclusive conditions.

The first examination of the treated skin sites followed 1 hour after removal of the patches. Thereafter, examinations were performed daily for another 7 days.

No signs of irritation were observed after administration. Thus, the test item is considered to possess no skin irritating properties.

Day	1(1 hour)	2(24 hours)	3(48 hours)	7(72 hours)
edema	0	0	0	0
erythma	0	0	0	0

 

According to EU Regulation No. 1272/2008 and CLP the test material must not be classified as an irritant to the skin.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Jul 11 - Sep 24, 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:

2002-04-24

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Strain: Albino rabbit, Crl:KBL(NZW)
Source: Charles River Wiga GmbH, Kifßlegg
Age: about 31-32 weeks
Mean weight: 4.3 kg

- Identification and adaptation: Healthy rabbits were allocated to the study group. The animals were kept in the experimental room for more than 7 days to allow for acclimatization. Each animal was uniquely identified by a tattoo in the ear displaying the animal number. Individual cage cards were affixed to each cage displaying the study number, test material, day of treatment, and animal number.
- Housing and diet: All rabbits were housed in an air-conditioned room of about 28 m^2 in the test facility. Lighting was controlled by a timer to provide a 12-hour light and a 12-hour dark regime. They were kept separately in special rabbit cages (manufacturer: Becker; type K99/30 KU, floor area about 5400 cm^2, a shelter with an integrated sitting board of about 1820 cm^2; overall height: 60 cm) with plastic grids placed on mobile racks. The collection pans underneath the cages were cleaned at least three times a week. The cages were cleaned before the start of the study. Temperature and relative humidity were measured using a thermohygrograph. The room temperature varied from 20 to 22 °C and the relative humidity from 50 to 64 %. The rabbits received diet for rabbits Provimi Kliba 3418.0 and sniff K snack ad libitum, and fresh tap water from Makrolon drinking bottles at least three times a week. The diet is checked periodically according to the specifications of the manufacturer by an independent laboratory, approved by the German government. Analysis includes both qualitative and quantitative evaluation for heavy metals, aflatoxins, pesticides, and antibiotics. The drinking water is periodically analyzed according to the German regulations for human drinking water.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1 g

Observation period (in vivo):

1 hour after treatment, after 24, 48, and 72 hours, then daily up to day 8 of the experimental part.

Number of animals or in vitro replicates:

3 female rabbits

Details on study design:

- Preparation: Before the application, the test material was ground in a mortar using a pestle.
- Administration and dose level: To ensure that only rabbits with normal eyes were included in the study, approximately 24 hours before treatment ophthalmological examinations were performed after instillation of a 0.15 % fluorescein solution (Dr. Thilo & Co.) using an ophthalmoscope. Animals with eye defects, injury or irritation were excluded. 0.1 g 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. The right eye remained untreated and served as control. After instillation, the eyelids were closed for about 30 seconds.
- Observation for clinical symptoms: The rabbits were examined for eye irritation and for changes in behavior and general condition 1 hour after treatment, after 24, 48, and 72 hours, then daily up to day 8 of the experimental part. Eye changes were evaluated according to the DRAIZE- , OECD- and EEC recommendations.
- Evaluation of eye reactions:
- Cornea
A) Opacity-degree of density (area most dense taken for reading)
No ulceration or opacity: 0
Scattered or diffuse areas of opacity (other than slight dullingof normal lustre), details of iris clearly visible: 1
Easily discernible translucent area, details of iris slightly obscured: 2
Nacrous area, no details of iris visible, size of pupil barely discernible: 3
Opaque cornea, iris not discernible through the opacity: 4
B) Area of cornea involved not evaluated (Rating according to the DRAIZE method)
- Iris
Normal: 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperemia, or injection, any of these or combination of any thereof, iris still reacting to light (sluggish reaction is positive): 1
No reaction to light, hemorrhage, gross destruction (any or all of these): 2
- Conjunctivae
A) Redness (refers to palpebral and bulbar conjunctivae, cornea and iris)
Blood vessels normal: 0
Some blood vessels definitely hyperemic (injected): 1
Diffuse, crimson color, individual vessels not easily discernible: 2
Diffuse beefy red: 3
B) Chemosis: lids and/or nictating membranes
No swelling: 0
Any swelling above normal (includes nictating membranes): 1
Obvious swelling with partial eversion of lids: 2
Swelling with lids about half closed: 3
Swelling with lids more than half closed: 4
C) Discharge (Rating according to the DRAIZE method)
No discharge: 0
Any amount different from normal (does not include small amounts observed in inner canthus of normal animals): 1
Discharge with moistening of the lids and hairs just adjacent to lids: 2
Discharge with moistening of the lids and hairs, and of a considerable area around the eye: 3
Mean score of all animals = Mean grading for irritations of cornea, iris, and conjunctivae per time point
Mean score per animal = Mean grading for each animal of irritations of cornea, iris, and conjunctivae (1, 24, 48, and 72 hours after application)
Maximum value = Maximum grading of a sign of irritation within a period

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:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

3

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

3

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

3

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

All animals survived the observation period. Body weight development of the treated rabbits was inconspicuous.

No signs of clinical toxicity were detected. No signs of irritation were observed at the cornea or iris. All animals showed discharge (score 1) one hour after instillation of the test material. Furthermore, a yellowish discoloration of the hairs around the eyes was seen in all treated animals up to day 6 and 7 of the experimental part. No abnormalities were detected in the untreated eyes.

Eve irritation of all animals (mean score):

	24 h	48 h	72 h
Cornea (mean /max)	0/0	0/0	0/0
Iris (mean / max)	0/0	0/0	0/0
Conjunctivae (mean / max)	0/0	0/0	0/0

Interpretation of results:

GHS criteria not met

Conclusions:

No eye irritating potential of the test item could be detected.

Executive summary:

The purpose of this primary eye irritation assay was to provide information on possible health hazards in case of acute eye contact with a test material and serve as a rational basis for risk assessment to the eye irritating potential of the test item in man. To test for eye irritation a test according to the OECD-Guidelines for Testing of Chemicals No. 405, the annex to Directive 92/69 EEC, and the recommendations of DRAIZE (1959) was performed.

All animals survived the observation period. Body weight development of the treated rabbits was inconspicuous. No signs of clinical toxicity were detected. No signs of irritation were observed at the cornea or iris. All animals showed discharge (score 1) one hour after instillation of the test material. Furthermore, a yellowish discoloration of the hairs around the eyes was seen in all treated animals up to day 6 and 7 of the experimental part. No abnormalities were detected in the untreated eyes.

No eye irritating potential of the test item could be detected, thus the test material should not be classified as eye irritant according to EU Regulation No. 1272/2008 (CLP).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation

The purpose of this assay was to identify the skin irritation/corrosion potential of the test material when applied to the intact skin of previously shaven rabbits for a 4 hours period under semiocclusive conditions.

This study should provide a rational basis for risk assessment to the irritating potential of the test item in man and was conducted according to OECD 404.

To test for primary skin irritation, 0.5 g of the test material was mixed with some drops of aqua pro injectione to ensure good contact with the skin. Afterwards the test material was spread onto 6 cm2 patches and applied to the intact skin of three previously shaven rabbits for a 4 hours period under semiocclusive conditions.

The first examination of the treated skin sites followed 1 hour after removal of the patches. Thereafter, examinations were performed daily for another 7 days.

No signs of irritation were observed after administration. Thus, the test item is considered to possess no skin irritating properties.

Day	1(1 hour)	2(24 hours)	3(48 hours)	7(72 hours)
edema	0	0	0	0
erythma	0	0	0	0

According to EU Regulation No. 1272/2008 and CLP the test material must not be classified as an irritant to the skin.

 

Eye irritation

The purpose of this primary eye irritation assay was to provide information on possible health hazards in case of acute eye contact with a test material and serve as a rational basis for risk assessment to the eye irritating potential of the test item in man. To test for eye irritation a test according to the OECD-Guidelines for Testing of Chemicals No. 405, the annex to Directive 92/69 EEC, and the recommendations of DRAIZE (1959) was performed.

All animals survived the observation period. Body weight development of the treated rabbits was inconspicuous. No signs of clinical toxicity were detected. No signs of irritation were observed at the cornea or iris. All animals showed discharge (score 1) one hour after instillation of the test material. Furthermore, a yellowish discoloration of the hairs around the eyes was seen in all treated animals up to day 6 and 7 of the experimental part. No abnormalities were detected in the untreated eyes.

No eye irritating potential of the test item could be detected, thus the test material should not be classified as eye irritant according to EU Regulation No. 1272/2008 (CLP).

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008
The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008.

Based on available data on skin irritation/corrosion, the test item does not require classification for causing skin irritation or corrosion according to Regulation (EC) No 1272/2008 (CLP).

Based on available data on eye irritation/corrosion, the test item does not require classification according to Regulation (EC) No 1272/2008 (CLP).