Registration Dossier

Administrative data

Description of key information

Skin irritation, OECD TG 404: not irritant
Eye irritation, OECD TG 405: Not irritant
Respi

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
Reference
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
12 FEBRUARY 1986 to 13 JUNE 1986
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: The study was carried out according to GLP guidelines and an international testing protocol (OECD 405). Furthermore, the study was assessed in the current risk assessment report under the excisting substance regulation.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
No observation at 1 hr and no wash at 24 hr
GLP compliance:
yes
Species:
rabbit
Strain:
other: Albino, new zealand
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Weight at study initiation: 2-3.5 kg
- Housing: housed singly under standard laboratory conditions
- Diet: 100 grams of purina rabbit chow supplemented by whole oats daily.
- Water: ad libitum

ENVIRONMENTAL CONDITIONS
- Other: no extraneous material, such as sawdust or woodchips was used in the cages. The suspended pans beneath the cages were littered with preprepared bedding (easi litter).

Vehicle:
unchanged (no vehicle)
Controls:
other: Left eye served as a control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL
Duration of treatment / exposure:
7 days
Observation period (in vivo):
At 24, 48, 72 hrs and at 4 and 7 days. The results were re-evaluated on day 10.
Number of animals or in vitro replicates:
6
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): washing was not done.
- Time after start of exposure: Washing was not done.

SCORING SYSTEM: According to the method of Draize.

TOOL USED TO ASSESS SCORE: a hand held ophtalmoscope and fluorescein.
Irritation parameter:
other: mean ocular irritation score
Basis:
mean
Time point:
other: 24 hrs
Score:
ca. 3.5
Max. score:
19
Reversibility:
not specified
Irritation parameter:
other: mean ocular irritation score
Basis:
mean
Time point:
other: 48 hrs
Score:
ca. 1.17
Max. score:
5
Reversibility:
not specified
Irritation parameter:
other: mean ocular irritation score
Basis:
mean
Time point:
other: 72 hrs, 4 and 7 days
Score:
ca. 0
Max. score:
0
Reversibility:
not specified
Irritant / corrosive response data:
One rabbit had a small central opacity (score 2) at 24 hr, which was fainter at 48 hr and cleared at 72 hr. The same animal showed also score 1 for effect on the iris at 24 hr only and slight conjunctival redness and discharge (both score 1) at 24 hr only. Another animal had slight redness of the conjunctivae (score 1) at 24 and 48 hr but no effects at later times. Primary eye irritation score at 24, 48, and 72 hr was 3.5, 1.17 and 0 (average of three time points was 1.6).

Table 1: Score charts primary eye irritation.

24 hrs
Animal # A-1125 A-1108 A-1109 A-1110 A-1111 A-1112
Cornea
A) Opacity 2 0 0 0 0 0
B) Area 1 0 0 0 0 0
Iris 1 0 0 0 0 0
Conjunctivae
A) redness 1 0 0 0 1 0
B) Chemosis 0 0 0 0 0 0
C) Discharge 1 0 0 0 0 0
Control 0 0 0 0 0 0
Weighted score 19 0 0 0 2 0
Total 21
Primary ocular irritation score 3.5
48 hrs
Animal # A-1125 A-1108 A-1109 A-1110 A-1111 A-1112
Cornea
A) Opacity 1 0 0 0 0 0
B) Area 1 0 0 0 0 0
Iris 0 0 0 0 0 0
Conjunctivae
A) redness 0 0 0 0 0 0
B) Chemosis 0 0 0 0 0 0
C) Discharge 0 0 0 0 0 0
Control 0 0 0 0 0 0
Weighted score 5 0 0 0 2 0
Total 7
Primary ocular irritation score 1.17
72 hrs, 4 and 7 days
Animal # A-1125 A-1108 A-1109 A-1110 A-1111 A-1112
Cornea
A) Opacity 0 0 0 0 0 0
B) Area 0 0 0 0 0 0
Iris 0 0 0 0 0 0
Conjunctivae
A) redness 0 0 0 0 0 0
B) Chemosis 0 0 0 0 0 0
C) Discharge 0 0 0 0 0 0
Control 0 0 0 0 0 0
Weighted score 0 0 0 0 0 0
Total 0
Primary ocular irritation score 0
Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: not specified
Conclusions:
Based on the irritation scores the test material is considered as practically nonirritating to the eye.
Executive summary:

Galaxolide (65% HHCB in DEP) was tested in the eyes of 6 rabbits by a procedure essentially equivalent to OECD 405 (no observation at 1 hr and no wash at 24 hr). 0.1 ml of Galaxolide was instilled into the right eye (the left serving as control) of healthy young adult albino rabbits. Both eyes were scored according to the method of Draize at 24, 48, 72, 96 and 168 hr. Four rabbits had no ocular changes at any time. One had a small central opacity (score 2) at 24 hr, which was fainter at 48 hr and cleared at 72 hr. The same animal showed also score 1 for effect on the iris at 24 hr only and slight conjunctival redness and discharge (both score 1) at 24 hr only. Another animal had slight redness of the conjunctivae (score 1) at 24 and 48 hr but no effects at later times. Primary eye irritation score at 24, 48, and 72 hr was 3.5, 1.17 and 0 (average of three time points was 1.6). Based on the irritation scores the test material (Galaxolide) is considered as practically nonirritating to the eye.

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

Key studies: Three skin irritation studies have been performed according to directive 79/831/EEC and under GLP conditions. Groups of either 3 or 4 New Zealand White female rabbits were used.

In the first study, 0.5 ml of Galaxolide (65% HHCB in DEP) was applied over an area of approximately six cm2for 4 hr under semi-occlusive lint patches (held in place with Elastoplast plastic adhesive bandage 10 cm wide) on the dorsal skin (clipped free of fur) of three rabbits for a period of 4 hrs. Undiluted DEP and benzyl benzoate (BB) were similarly applied to groups of three rabbits. Scores for erythema and oedema per animal were given after 1, 24, 48, 72 and 168 hrs and the average scores per animal for erythema and oedema over 24, 48 and 72 hrs were calculated. The results for Galaxolide were an erythema score of 1.3 (on all three) and an oedema score of 0.4 (highest score 1). After 168 hrs, erythema was still observed in two animals, while slight desquamation of the skin surface at the treated site was seen in all three rabbits. For DEP and for BB, the scores were zero for erythema and oedema on all three rabbits. (Haynes, 1984)

In the second study 0.5 ml of Galaxolide 50 (65% HHCB in DEP), was placed evenly over a 2.5 cm2 of surgical lint, which was then placed on the skin of each of 4 rabbits and held by an Elastoplast adhesive bandage 10 cm wide for a period of 1 hr. Scores of skin results were given after 1, 24, 48, 72, and 168 hrs. An average score (average of all 4 animals over 24, 48, and 72 hrs) for erythema of 2.1 and for oedema of 1.5 was calculated for Galaxolide. After 168 hrs erythema and oedema were still observed in 4/4 and 3/4 animals, respectively. The average score for DEP was 0.2 for erythema and zero for oedema.(Haynes, 1985)

In the third study, 0.5 ml of either undiluted or 50% solutions of Galaxolide 50 DEP (65% HHCB in DEP) or Galaxolide 50 BB (65% HHCB in benzyl benzoate) were placed evenly over a 2.5 cm2patches of surgical lint. These patches were then placed on the skin of 4 rabbits and held by an adhesive bandage 10 cm wide for a period of 1 hr. The average erythema/oedema scores calculated over 24, 48 and 72 hours were 1.8/1.3 for 100% Galaxolide 50 DEP (reversible after 168 hrs), 1.3/0.3 for 50% Galaxolide 50 DEP in DEP (reversible in 3/4 animals after 168 hrs), 1.8/0.8 for 100% Galaxolide 50 BB (reversible in 3/4 animals after 168 hrs), and 1.3/0.7 for 50% Galaxolide 50 BB in BB (reversible after 168 hrs), respectively. In this study, there were no solvent controls.

 

Additional studies: Two additional skin irritation studies (not performed according to GLP) are available which support the findings of the key studies.

The first test was conducted with undiluted Galaxolide-50 (65% HHCB in DEP) on 3 albino rabbits. A single application on clipped skin (abraded and non-abraded) resulted in average scores (Draize) of 1 for erythema at both 24 and 72 hr (both abraded and non-abraded skin); no erythema or oedema was seen with Galaxolide-50. It was concluded that Galaxolide-50 was a moderate irritant. (Levenstein, 1973a).

The second test was performed under identical conditions with 25% or 50% Galaxolide-50 (equivalent to 16% or 33% HHCB) in alcohol, as well as undiluted Galaxolide 50 (65% HHCB in DEP) for 24 or 72 hr. The 25% solution produced no erythema or oedema at either abraded or unabraded sites. The 50% solution produced a score of 1 for erythema at 24 hr on abraded skin but no erythema or oedema at any other site or time period. It was concluded that this solution could be considered a very mild irritant. The undiluted Galaxolide 50 produced scores of 1 for erythema at 24 hr on both abraded and unabraded sited but no erythema at 72 hr and no oedema at any time on either site. (Levenstein, 1975a)

 

Photoirritation: There are some indications from animal studies that HHCB could be a photoirritant. The results in human tests do not indicate a photoirritating effect in humans. Also, an in vitro phototoxicity test (in compliance with test guideline B.41 (EU/COLIPA Test)) was negative. No criteria on classification of photoirritating substances are available in Annex VI.

 

Conclusion and discussion (skin irritation): All key studies showed very slight to well-defined erythema and very slight oedema. In only one of the tests did the mean erythema score for Galaxolide 50 DEP exceed 2.0 (the calculated score was 2.1). The solvent in this study, DEP, scored 0.2 for erythema and zero for oedema. The irritating effect was not reversible in 7 out of in total 15 animals in these three studies during an observation period of 7 days, as at that time point still some erythema and/or oedema was seen.

Unfortunately, the observation period was not sufficiently long (14 days, according to the test method guideline in Annex V) to evaluate fully the reversibility of the effects. However, the test method guideline also states that the irritation scores should be evaluated in conjunction with the nature and severity of lesions and their reversibility or lack of reversibility. Taking that also into account, the results of the animal studies do not indicate that HHCB is a skin irritant. There is a difficulty though, because according to Annex VI of Directive 67/548, inflammation of the skin is also significant if it persists in at least 2 animals at the end of the observation period [without specifying the length of the observation period and the severity of the effects]. If this guidance is followed strictly, the animal studies would warrant classification as a skin irritant. This issue was discussed at the TC-C&L meeting of November 2005, where it was concluded that it is not warranted to classify HHCB as a skin irritant. (Haynes, 1984, Haynes, 1985, Haynes, 1986)

 

Eye irritation

Key study: Galaxolide (65% HHCB in DEP) was tested in the eyes of 6 rabbits by a procedure essentially equivalent to OECD 405 (no observation at 1 hr and no wash at 24 hr). 0.1 ml of Galaxolide was instilled into the right eye (the left serving as control) of healthy young adult albino rabbits. Both eyes were scored according to the method of Draize at 24, 48, 72, 96 and 168 hr. Four rabbits had no ocular changes at any time. One had a small central opacity (score 2) at 24 hr, which was fainter at 48 hr and cleared at 72 hr. The same animal showed also score 1 for effect on the iris at 24 hr only and slight conjunctival redness and discharge (both score 1) at 24 hr only. Another animal had slight redness of the conjunctivae (score 1) at 24 and 48 hr but no effects at later times. Primary eye irritation score at 24, 48, and 72 hr was 3.5, 1.17 and 0 (average of three time points was 1.6). Based on the irritation scores the test material (Galaxolide) is considered as practically nonirritating to the eye.(Sauer, 1980)

 

Additional information: In an additional study, Galaxolide 50 (65% in DEP) was tested in the eyes of three rabbits with an observation time of 168 hr according to method that seemed similar to the key study. No irritation was seen in all three rabbits at any observation time. (Levenstein, 1975b).

Furthermore, two studies were available that were considered to be not relevant for the evaluation of the eye irritating potential of HHCB, as they were both performed with a solution of HHCB in ethanol, which possesses eye irritating potential and is therefore considered to be an inappropriate solvent for eye irritation tests. (Levenstein, 1973b, Wolven and Levenstein, 1963a).

 

Conclusion and discussion (eye irritation): HHCB has been tested for ocular irritation in rabbits in several studies. Some studies used ethanol, a known eye-irritant as solvent, and are not used. In other relevant studies, some ocular irritation was found. However, the effects were not severe enough to require classification according to EU guidelines.

 

Respiratory tract 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 (REACH guidance R.7.2.3). There are no human data such as indicated in R7.2.3 the ECHA guidance that indicate respiratory reactions of the substance e.g. from consumer experience or occupational exposure and there are no human data on file in the RIFM database, which is a database of the fragrance association, in which all available human and animal data of fragrance substances are presented (RIFM, http://www.rifm.org/). In addition, the substance is not corrosive or severely irritating (absence of classification and labeling for skin and eye irritation) which further minimizes the respiratory irritation hazard ((REACH guidance 7.2.1.2).

No further studies are deemed necessary.


Justification for selection of skin irritation / corrosion endpoint:
Three key studies were identified for skin irritation, all have been performed according to directive 79/831/EEC and under GLP conditions.

Justification for selection of eye irritation endpoint:
The key study for eye irritation was performed according to a method similar to OECD405 and under GLP conditions

Justification for classification or non-classification

Based on the available skin and eye irritation studies, there is no need to classify HHCB for skin or eye irritation in accordance with the criteria outlined in Annex VI of 67/548/EEC (DSD) and Annex I of 1272/2008/EC (CLP).

HHCB does not need to be classified as a respiratory irritant in accordance with the criteria outlined in Annex VI of 67/548/EEC and Annex I of 1272/2008/EC.