Registration Dossier

Administrative data

Description of key information

EYE

Key study:non-irritant; rabbit; equivalent to EU Method B5; Potokar (1984b)

Key value for chemical safety assessment

Skin irritation / corrosion

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Referenceopen allclose all

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
24.09.1984-28.09.1984
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, equivalent to a valid guidelines and the study was conducted under GLP conditions. The study was performed with test material being used to support the substance on the basis of read-across.
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Thomae, Biberach (Germany)
- Weight at study initiation: 2.8-2.9 kg
- Housing: Individually
- Diet: Altromin standard diet 2023 ad libitum
- Water: tap water ad libitum
- Acclimation period: one month

ENVIRONMENTAL CONDITIONS
- Temperature: 20-21 °C
- Humidity: approximately 45-50 %
- Photoperiod: 12 hrs dark / 12 hrs light
Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL was instilled into the conjunctival sac of the right eye. The left eye remained untreated as a control.
Observation period (in vivo):
96 hours after exposure
Number of animals or in vitro replicates:
6
Details on study design:
SCORING SYSTEM:

CORNEA
No ulceration or opacity - 0
Diffuse areas of opacity (except the normal brightness appears slightly cloudy), details of the iris are clearly visible - 1
Easily identifiable translucent area, details of the iris are somewhat shadowed - 2
Nacreous areas, no details of the iris are visible, size of the pupil not identifiable - 3
Opaque cornea, iris not identifiable due to opacity - 4

IRIS
Normal - 0
Markedly opened folds, congestion, enlargement, slight circumcorneal hyperaemia or injection; one of these symptoms or a combination of the various symptoms; the iris still reacts to light (a sluggish reaction is positive) - 1
No reaction to light, haemorrhage, severe damage (one or all of these symptoms) - 2

CONJUNCTIVAE
Redness:
Blood vessels normal - 0
Some blood vessels exhibit a distinct hyperemia (injection) - 1
Diffuse, crimson colour, difficult to identify individual blood vessels - 2
Diffuse, vibrant red - 3

Chemosis:
Eyelids and/or nictitating membrane, no swelling - 0
Each above the normal swelling (including nictitating membrane) - 1
Unambiguous swelling with ectropion of the eyelids - 2
Swelling with approximately half-closed eyelids - 3
Swelling with more than half-closed eyelids - 4
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
all animals
Time point:
other: Average of 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
fully reversible
Irritation parameter:
iris score
Basis:
mean
Remarks:
all animals
Time point:
other: Average of 24, 48 and 72 hours
Score:
0
Max. score:
2
Reversibility:
fully reversible
Irritation parameter:
conjunctivae score
Remarks:
Redness
Basis:
animal: 67 + 70 + 72
Time point:
other: Average of 24, 48 and 72 hours
Score:
0.33
Max. score:
3
Reversibility:
fully reversible within: 48 hours
Irritation parameter:
conjunctivae score
Remarks:
Redness
Basis:
animal: 68 + 69 + 71
Time point:
other: Average of 24, 48 and 72 hours
Score:
0
Max. score:
3
Reversibility:
fully reversible
Irritation parameter:
chemosis score
Basis:
animal: 67 + 68 + 70 + 71 + 72
Time point:
other: Average of 24, 48 and 72 hours
Score:
0.33
Max. score:
4
Reversibility:
fully reversible within: 48 hours
Irritation parameter:
chemosis score
Basis:
animal: 69
Time point:
other: Average of 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
fully reversible
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
other: 72 h
Score:
0.7
Irritant / corrosive response data:
One hour after instillation, all of the animals exhibited minor redness of the conjunctivae and three animals also exhibited minor localised swelling of the conjunctivae in the upper eye area. After 24 hours, three animals exhibited minor redness and five animals exhibited minor swelling. 48 and 72 hours after treatment, all animals were without any symptoms.

This implies an irritation index according to the following formula:

Total reactions after 1, 24, 48 and 72 hours / (Number of animals x number of reading events)

i.e. 17 / (6 x 4) = 0.70

Table 1: Assessment of the reactions of the cornea and iris

Animal number   67  68  69  70  71  72
   Cornea  Iris  Cornea  Iris  Cornea  Iris  Cornea  Iris  Cornea  Iris  Cornea  Iris
 1 hour  0  0  0  0  0  0  0  0  0  0  0  0
 24 hours  0  0  0  0  0  0  0  0  0  0  0  0
 48 hours  0  0  0  0  0  0  0  0  0  0  0  0
 72 hours  0  0  0  0  0  0  0  0  0  0  0  0

Table 2: Assessment of the conjunctival reactions

 Animal number  67  68  69  70  71  72
   A  B  A  B  A  B  A  B  A  B  A  B
 1 hour  1  1  1  0  1  0  1  1  1  1  1  0
 24 hours  1  1  0  1  0  0  1  1  0  1  1  1
 48 hours  0  0  0  0  0  0  0  0  0  0  0  0
 72 hours  0  0  0  0  0  0  0  0  0  0  0  0

A = Redness

B = Chemosis

Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of the test, a single application of the test material is tolerated by the mucous membrances of the rabbit eye with minor, rapidly reversible reactions of the conjunctivae.
Executive summary:

The eye irritation of the test material was determined under GLP conditions performed in line with the standardised guideline EU Method B5. During the study 6 rabbits were instilled with 0.1 mL of the test material into to the conjunctival sac of the right eye of each animal. Animals were were observed for a period of 96 hours with observations made at 1, 24, 48, 72 and 96 hours after application. The test material was tolerated by the mucous membranes of the eye with minor, rapidly reversible reactions of the conjuctivae.

The test material is considered sufficiently similar to (z)-3-hexenyl salicylate on the basis of read-across such that the results from this study can be used to address the eye irritation potential of (z)-3-hexenyl salicylate.

Endpoint:
eye irritation: in vivo
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
1. HYPOTHESIS FOR THE ANALOGUE APPROACH
Please see 'Justification for read-across to support the REACH registration of (z)-3-hexenyl salicylate' document attached (section 13) for full details.

It is proposed to read-across to another salicylate substance in order to fulfil data requirements.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Please see ''Justification for read-across to support the REACH registration of (z)-3-hexenyl salicylate' document attached for full details.

Source substance: cyclohexyl salicylate (EC 400-410-3, CAS 25485 88-5).
Target substance: (z)-3-hexenyl salicylate (EC 265-745-8, CAS 65405-77-8)

3. ANALOGUE APPROACH JUSTIFICATION
Please see 'Justification for read-across to support the REACH registration of (z)-3-hexenyl salicylate' document attached for full details.

(z)-3-hexenyl salicylate (the target substance) and the read-across substance cyclohexyl salicylate (source substance) have been characterised using the categories and databases present in the OECD QSAR Toolbox. From the profiling, it can be seen that the two substances share structural similarities and also ‘mechanistic action’ similarities which are both general and endpoint specific. The main difference observed is structural. The target substance ((z)-3-hexenyl salicylate) contains an acyclic hexenyl chain and the source substance (cyclohexyl salicylate) contains a cyclic hexyl group.
The output from the OECD QSAR Toolbox shows that the profiles of (z)-3-hexenyl salicylate and cyclohexyl salicylate are sufficiently similar such that any available data from the source substance can be used to address the following endpoints in the REACH registration dossier for (z)-3-hexenyl salicylate:

4. DATA MATRIX
Please see 'Justification for read-across to support the REACH registration of (z)-3-hexenyl salicylate' document attached for full details.
Reason / purpose:
read-across source
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
all animals
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
fully reversible
Irritation parameter:
iris score
Basis:
mean
Remarks:
all animals
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
fully reversible
Irritation parameter:
conjunctivae score
Remarks:
Redness
Basis:
animal: 67 + 70 + 72
Time point:
24/48/72 h
Score:
0.33
Max. score:
3
Reversibility:
fully reversible within: 48 hours
Irritation parameter:
conjunctivae score
Remarks:
Redness
Basis:
animal: 68 + 69 + 71
Time point:
24/48/72 h
Score:
0
Max. score:
3
Reversibility:
fully reversible
Irritation parameter:
chemosis score
Basis:
animal: 67 + 68 + 70 + 71 + 72
Time point:
24/48/72 h
Score:
0.33
Max. score:
4
Reversibility:
fully reversible within: 48 hours
Irritation parameter:
chemosis score
Basis:
animal: 69
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
fully reversible
Irritant / corrosive response data:
One hour after instillation, all of the animals exhibited minor redness of the conjunctivae and three animals also exhibited minor localised swelling of the conjunctivae in the upper eye area. After 24 hours, three animals exhibited minor redness and five animals exhibited minor swelling. 48 and 72 hours after treatment, all animals were without any symptoms.
Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of the test, a single application of the test material is tolerated by the mucous membrances of the rabbit eye with minor, rapidly reversible reactions of the conjunctivae.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Eye irritation

The eye irritation of the test material was determined under GLP conditions using a method equivalent to the standardised guideline EU Method B5. During the study 6 rabbits were with 0.1 mL of the test material added to the conjunctival sac of the right eye of each animal. Animals were observed for a period of 96 hours with observations made at 1, 24, 48, 72 and 96 hours after application. The test material was tolerated by the mucous membranes of the eye with minor, rapidly reversible reactions of the conjuctivae.

The test material is considered sufficiently similar to (z)-3-hexenyl salicylate on the basis of read-across such that the results from this study can be used to address (z)-3-hexenyl salicylate.

Skin irritation

A waiver was submitted to address the skin irritation/corrosion data requirement. The endpoint was considered suitably addressed with the use of alternative data, the conclusions of which were considered suitable for classification and labelling and risk assessment purposes.

Justification for classification or non-classification

According to Regulation EC 1272/2008, the substance does not meet the classification criteria for skin or eye irritation.