Reaction mass of 3a,4,5,6,7,7a-hexahydro-4,7-methanoinden-5-yl acetate and 3a,4,5,6,7,7a-hexahydro-4,7-methanoinden-6-yl acetate

Endpoint summary

Description of key information

- Skin corrosion /irritation (in vivo): not irritating (method similar to OECD 404)

- Eye irritation (in vivo): not irritating (method similar to OECD 405)

- Respiratory irritation: not irritating (based on the non-irritant properties of skin and eyes)

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:

02-11-1976 - 13-12-1976

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

10 animals used; different scoring system; observation < 14 days (Reversibility check); positve and negative control substances used.

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 9-12 weeks

Type of coverage:

semiocclusive

Preparation of test site:

other: clipped

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 ml
- Concentration (if solution): 100%

Controls:
Positive control: Cyclamen Aldehyde
Negative control: Diethyl Phthalate

Duration of treatment / exposure:

4 hours

Observation period:

at 4, 24, 48 and 72 hours

Number of animals:

10

Details on study design:

TEST SITE
- Area of exposure: dorsal
- % coverage: no data
- Type of wrap if used: Patches are prepared by heat-sealing 1" x 1" 24 ply gauze pads on to 1 1/4" x 1 1/4" squares of polythene sheeting which is then attached to 3 1/2" x 1" strips of adhesive tape. Test solutions are applied to a dry patch. The patches are firmly attached to ensure good contact between the skin and test substance.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): wiped clean
- Time after start of exposure:4 hours

SCORING SYSTEM: The sites are scored for erythema, oedema, cracking and scaling and any other feature using an 8-point anchored ordinate scale ranging from “a” (very slight) to “h” (severe). At the end of the test the reaction grades are converted to corresponding numerical scores which are used to calculate the total irritation score per animal, an overall score per treatment group, a mean irritation score per site, and a mean irritation score per site per day for each treatment group. A mean irritation ranking for each treatment group is also given.
Reaction Grades and Scores of erythema, oedema, cracking and scaling:
a marginal/very slight = 1
b slight = 2
c fairly distinct =3
d quite distinct = 4
e becoming well developed = 6
f well developed = 8
g becoming severe = 10
h severe = 12

STATISTICS:
The Wilcoxon Matched-Pairs Signed Ranks Test, which considers the size and direction of differences between reactions is used to test the significance of differences between pairs of treatment groups.

Irritation parameter:

erythema score

Basis:

animal: 1, 2, 4, 5, 6, 7, 8, and 9

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

not fully reversible within: 72 hours

Remarks on result:

other: Animal 1, 2, 4, 5, 6, 7, 8, and 9

Irritation parameter:

erythema score

Basis:

animal: 3

Time point:

24/48/72 h

Score:

1.33

Max. score:

4

Reversibility:

not fully reversible within: 72 hours

Remarks on result:

other: Animal 3

Irritation parameter:

erythema score

Basis:

animal: 10

Time point:

24/48/72 h

Score:

1.67

Max. score:

4

Reversibility:

not fully reversible within: 72 hours

Remarks on result:

other: Animal 10

Irritation parameter:

edema score

Basis:

animal: 6 and 9

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: Animal 6 and 9

Irritation parameter:

edema score

Basis:

animal: 1 and 8

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

other: not fully reversible in 72 hours in 1/2 animals

Remarks on result:

other: Animal 1 and 8

Irritation parameter:

edema score

Basis:

animal: 2, 3, 4, 5, 7, and 10

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

other: not fully reversible in 72 hours in 5/6 animals

Remarks on result:

other: Animal 2, 3, 4, 5, 7, and 10

Irritant / corrosive response data:

Under the conditions of this test the test substance produced slight to moderate irritation consisting of marginal to slight erythema and oedema with variable cracking and scaling ranging from slight to distinct. The test substance was significantly more irritant than the negative control, diethyl phthalate, which produced only a marginal response. The test ingredient was significantly less irritant than the positive control, cyclamen aldehyde, which produced well-developed erythema fairly distinct cracking and slight scaling.

Table 1: Skin irritation scoring from test report

	4 h	4 h	24 h	24 h	48 h	48 h	72 h	72 h
Animal	Er	Ed	Er	Ed	Er	Ed	Er	Ed
1	A	-	B	A	B	-	B	-
2	A	B	B	A	B	A	B	A
3	B	B	C	B	A	A	A	A
4	A	-	B	A	A	A	A	A
5	A	B	A	A	A	A	A	A
6	A	-	A	-	A	-	B	-
7	B	B	B	B	B	B	B	A
8	A	-	A	-	A	-	A	A
9	A	-	A	-	B	-	A	-
10	A	A	C	B	C	A	B	-

Er = Erythema; Ed = Edema

Table 2: Recalculated scoring system based on proposed conversion (see any other information on materials and methods).

	4 h	4 h	24 h	24 h	48 h	48 h	72 h	72 h	Mean 24/48/72h	Mean 24/48/72h
Animal	Er	Ed	Er	Ed	Er	Ed	Er	Ed	Er	Ed
1	1	0	1	1	1	0	1	0	1	0.33
2	1	1	1	1	1	1	1	1	1	1
3	1	1	2	1	1	1	1	1	1.33	1
4	1	0	1	1	1	1	1	1	1	1
5	1	1	1	1	1	1	1	1	1	1
6	1	0	1	0	1	0	1	0	1	0
7	1	1	1	1	1	1	1	1	1	1
8	1	0	1	0	1	0	1	1	1	0.33
9	1	0	1	0	1	0	1	0	1	0
10	1	1	2	1	2	1	1	0	1.67	1

Er = Erythema; Ed = Edema 

Interpretation of results:

other: Not a skin irritant

Remarks:

according to EU CLP (EC 1272/2008 and its amendments)

Conclusions:

In the in vivo rabbit skin irritation test according to OECD TG 404, the substance is not a skin irritant.

Executive summary:

The skin irritation/corrosion potential of Cyclacet was investigated in a study similar to OECD Test Guideline 404 (Acute Dermal Irritation / Corrosion). In this Rabbit Covered Patch Skin Irritation Test, ten New Zealand White rabbits were exposed to the test substance for 4 hours under semi-occlusive conditions and reactions were scored after removal, and after 24, 48 and 72 hours. The test substance caused slight to moderate irritation, consisting of marginal to slight erythema and oedema with variable cracking and scaling ranging from slight to distinct. The mean erythema scores over the 24, 48 and 72 hour time period, were 1 (8 animals), 1.33 (1 animal) and 1.67 (1 animal). The mean edema scores over the 24, 48 and 72 hour time period, were 0 (2 animals), 0.33 (2 animals), and 1 (6 animals). As all animals had scorings below 2.3, the test substance is not considered to be a skin irritant.

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:

09-05-1977 - 31-05-1977

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

different scoring system

Qualifier:

equivalent or similar to guideline

Guideline:

other: Modified Federal Hazardous Substances Labelling Act Method

Deviations:

not specified

GLP compliance:

no

Species:

rabbit

Strain:

not specified

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Age at study initiation: 9 weeks

Vehicle:

unchanged (no vehicle)

Controls:

not specified

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml
- Concentration (if solution): 100%

Duration of treatment / exposure:

24 hours

Observation period (in vivo):

at 15 min., 1, 2, and 3 days

Number of animals or in vitro replicates:

3

Details on study design:

TOOL USED TO ASSESS SCORE: hand-slit lamp

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.67

Max. score:

4

Reversibility:

fully reversible within: 3 days

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 2 days

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:

0.33

Max. score:

2

Reversibility:

fully reversible within: 2 days

Irritation parameter:

iris score

Basis:

animal: 2, 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:

0.33

Max. score:

3

Reversibility:

fully reversible within: 2 days

Irritation parameter:

conjunctivae score

Basis:

animal: 2, 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:

0.33

Max. score:

4

Reversibility:

fully reversible within: 2 days

Irritation parameter:

chemosis score

Basis:

animal: 2 and 3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

The test substance caused slight corneal lesions affecting up to half of the corneal surface in two animals. One of these lesions was associated with moderate corneal swelling, slight conjunctivitis and discharge. The lesions had healed within three days. The remaining animal was unaffected. One animal showed signs of discomfort when treated.

Interpretation of results:

other: Not an eye irritant

Remarks:

according to EU CLP (EC 1272/2008 and its amendments)

Conclusions:

The test substance is not eye irritating.

Executive summary:

The primary eye irritation potential of Cyclacet was investigated in a study similar to OECD Test Guideline 405 (Acute Eye Irritation / Corrosion). A single administration of Cyclacet in the eye of 3 rabbits caused slight corneal lesions affecting up to half of the corneal surface in two animals. One of these lesions was associated with moderate corneal swelling, slight conjunctivitis and discharge. The lesions had healed within three days. The remaining animal was unaffected. One animal showed signs of discomfort when treated. Mean chemosis, conjunctivae and iris scores at the 24, 48, and 72 hour time point were 0.33 in 1 animals and 0 in the other two animals. The effects seen in the one animal was reversible within one day. Corneal opacity scores were 0.67, 0.33, and 0 for the three animals respectively. The corneal opacity was absent within three days. It is concluded that Cyclacet is not an eye iritant.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Additional information

Skin irritation: The skin irritation/corrosion potential of Cyclacet was investigated in a study similar to OECD Test Guideline 404 (Acute Dermal Irritation / Corrosion). In this Rabbit Covered Patch Skin Irritation Test, ten New Zealand White rabbits were exposed to the test substance for 4 hours under semi-occlusive conditions and reactions were scored after removal, and after 24, 48 and 72 hours. The test substance caused slight to moderate irritation, consisting of marginal to slight erythema and oedema with variable cracking and scaling ranging from slight to distinct. The mean erythema scores over the 24, 48 and 72 hour time period, were 1 (8 animals), 1.33 (1 animal) and 1.67 (1 animal). The mean edema scores over the 24, 48 and 72 hour time period, were 0 (2 animals), 0.33 (2 animals), and 1 (6 animals). As all animals had scorings below 2.3, the test substance is not considered to be a skin irritant.

Eye irritation: The primary eye irritation potential of Cyclacet was investigated in a study similar to OECD Test Guideline 405 (Acute Eye Irritation / Corrosion). A single administration of Cyclacet in the eye of 3 rabbits caused slight corneal lesions affecting up to half of the corneal surface in two animals. One of these lesions was associated with moderate corneal swelling, slight conjunctivitis and discharge. The lesions had healed within three days. The remaining animal was unaffected. One animal showed signs of discomfort when treated. Mean chemosis, conjunctivae and iris scores at the 24, 48, and 72 hour time point were 0.33 in 1 animals and 0 in the other two animals. The effects seen in the one animal was reversible within one day. Corneal opacity scores were 0.67, 0.33, and 0 for the three animals respectively. The corneal opacity was absent within three days. It is concluded that Cyclacet is not an eye irritant.

Respiratory irritation: For respiratory irritation mostly human data are used for the assessment because no suitable in vitro or in vivo tests are available that can identify respiratory irritation (ECHA guidance R.7.2.3 (2008) and ECHA guidance, 2017, page 240). There are no human data available that indicate respiratory reactions of the substance e.g. from consumer experience or occupational exposure. In addition the substance is not corrosive or severely irritating and therefore respiratory irritation is not considered a concern.

Justification for classification or non-classification

Based on the available corrosion and irritation information the substance does not need to be classified for skin, eye and irritation is not warranted according to EU CLP (EC 1272/2008 and its amendments).