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Toxicological information

Eye irritation

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Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2003-11-03 to 2003-12-03
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2004
Report Date:
2004

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
2002
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.2400 (Acute Eye Irritation)
Version / remarks:
1998
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Version / remarks:
1992
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
impurity

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
Species
Albino Rabbit, New Zealand White, (SPF-Quality)
Recognised by international guidelines as the recommended test system (e.g. EC, OECD)
Source: Charles River Deutschland, Kisslegg, Germany

Number of animals: 3 Males.
Age and body weight: Animals used within the study were at least 6 weeks old and body weights were at least 1.0 kg.

Identification
Earmark.

Conditions
Animals were housed in a controlled environment, in which optimal conditions were considered to be approximately 15 air changes per hour, a temperature of 21.0 ± 3.0°C (actual range: 18.1 - 22.3°C), a relative humidity of 30-70% (actual range: 37 - 74%) and 12 hours artificial fluorescent light and 12 hours darkness per day. Cleaning procedures in the room might have caused the temporary fluctuations above the optimal maximum level of 70% for relative humidity. Based on laboratory historical data, these fluctuations were considered not to have affected the study integrity.

Accommodation
Individually in labelled cages with perforated floors (Scanbur, Denmark, dimensions 56x44x37.5 cm). Acclimatisation period was at least 5 days before start of treatment under laboratory conditions.

Diet
Standard laboratory rabbit diet (Charles River Breeding and Maintenance Diet for Rabbits, Altromin, Lage, Germany) approx. 100 g. per day. Certificates of analysis were examined and retained in the NOTOX archives. In addition, pressed hay (BMI, Helmond, the Netherlands) was provided twice a week.

Water
Free access to tap-water. Certificates of quarterly analysis were examined and retained in the NOTOX archives.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
Each animal was treated by instillation of 39.7 (39.0 - 40.7) mg of the test substance (a volume of approximately 0.1 ml) in the conjunctival sac of one of the eyes after gently pulling the lower lid away from the eyeball.
Duration of treatment / exposure:
The lids were then gently held together for about one second to prevent loss of the test substance. The other eye remained untreated and served as the reference control.
Observation period (in vivo):
The eyes of each animal were examined approximately 1, 24, 48 and 72 hours and 7 and 14 days after instillation of the test substance.
Number of animals or in vitro replicates:
This in-vivo eye irritation study was performed and was started by treatment of a single rabbit (sentinel). The two other animals were treated in a similar manner 7 days later, after considering the degree of eye irritation observed in the first animal.
Details on study design:
All available data relevant to the potential eye irritation/corrosivity of the substance indicated that no severe effects were to be expected. No severe reactions were noted in the skin irritation study (Notox Project 395663). An in-vitro test was considered, but a negative test result was anticipated that still would have to be confirmed in an in-vivo study. Since no severe harm for the animals was to be expected, this in-vivo eye irritation study was performed and was started by treatment of a single rabbit (sentinel). The two other animals were treated in a similar manner 7 days later, after considering the degree of eye irritation observed in the first animal.
A health inspection was performed prior to commencement of treatment, to ensure that the animals were in a good state of health. Special attention was paid to the eyes, which were free from any abnormality.
Each animal was treated by instillation of 39.7 (39.0 - 40.7) mg of the test substance (a volume of approximately 0.1 ml) in the conjunctival sac of one of the eyes after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of the test substance. The other eye remained untreated and served as the reference control.

Immediately after the 24-hour observation, a solution of 2% fluorescein in water (adjusted to pH 7.0) was instilled into both eyes of each animal to quantitatively determine corneal epithelial damage. Any bright green stained area, indicating epithelial damage, was estimated as a percentage of the total corneal area.

Immediately after fluorescein examination on day 2, the treated eye was rinsed with approx 50 ml tepid tap-water, using a velocity of flow which did not affect the eye, to remove any residual test substance. For reference control the other eye was also rinsed.

Observations

Mortality Viability
Twice daily.

Toxicity
At least once daily.

Body Weight
Day of treatment (prior to instillation) and at termination.

Irritation
The eyes of each animal were examined approximately 1, 24, 48 and 72 hours and 7 and 14 days after instillation of the test substance. The irritation scores and a description of all other (local) effects were recorded.

The irritation was assessed according to the following numerical scoring system. At each observation, the highest scores given were recorded:CORNEAL IRRITATlON
Opacity: degree of density (area most dense taken for reading)
No ulceration or opacity (may include slight dulling of normal lustre) ............................................ 0
Scattered or diffuse areas of opacity, details of iris clearly visible ................................................. 1
Easily discernible translucent area, details of iris slightly obscured ............................................... 2
Nacreous area, no details of iris visible, size of pupil barely discernible ........................................ 3
Opaque cornea, iris not discernible through the opacity ............................................................... 4
Area of cornea involved:
No ulceration or opacity ................................................................................................................ 0
One quarter or less but not zero .................................................................................................. 1
Greater than one quarter, but less than half ................................................................................ 2
Greater than half, but less than three quarters .............................................................................. 3
Greater than three quarters, up to whole area .............................................................................. 4
IRIS
Normal ........................................................................................................................................ 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia,
or injection, any of these or combination thereof, iris still reacting to light
(sluggish reaction is positive) ........................................................................................................ 1
No reaction to light, hemorrhage, gross destruction (any or al! of these) ....................................... 2
CONJUNCTlVAL IRRITATION
Redness (refers to palpebrae and sclera, excluding cornea and iris):
Blood vessels normal .................................................................................................................. 0
Some blood vessels definitely hyperaemic (injected) .................................................................... 1
Diffuse, crimson color, individual vessels not easily discernible .................................................... 2
Diffuse beefy red .......................................................................................................................... 3
Chemosis (refers to lids and/or nictitating membranes):
No swelling ......................................................................................................................... 0
Any swelling above normal (includes nictitating 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

Discharge:
No discharge (may include small amounts observed in inner canthus of normal animals) ............ 0
Any amount different from normal and/or lacrimation .................................................................... 1
Discharge with moistening of the lids and hairs just adjacent to lids ............................................. 2
Discharge with moistening of the lids and hairs (considerable area around the eye) .................... 3

Where standard lighting was considered inadequate for observing minor effects, eye examinations were performed using an ophthalmic examination lamp.
In cases of equivocal results when comparing the treated and untreated eyes, the illustrated guide from the Consumer Product Safety Commission. Washington, D.C. 20207 was used for additional control purposes.
Interpretation
The results will be evaluated according to the OECD Harmonized Integrated Hazard Classification System for Human Health and Environmental Effects of Chemical Substances (OECD, 1998) and the EC criteria for classification and labelling of dangerous substances and preparations (Council Directive 67/548/EEC and all adaptations to technical progress and amendments of this Directive published in the Official Journal of the European Communities).

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
animal: 697-712-715
Time point:
24/48/72 h
Score:
0
Irritation parameter:
iris score
Basis:
animal: 697-712-715
Time point:
24/48/72 h
Score:
0
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal: 697
Time point:
24/48/72 h
Score:
3
Reversibility:
fully reversible within: 14 days
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal: 712
Time point:
24/48/72 h
Score:
2.7
Reversibility:
fully reversible within: 14 days
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal: 715
Time point:
24/48/72 h
Score:
2.7
Reversibility:
fully reversible within: 14 days
Irritation parameter:
chemosis score
Basis:
animal: 697
Time point:
24/48/72 h
Score:
1.7
Reversibility:
fully reversible within: 14 days
Irritation parameter:
chemosis score
Basis:
animal: 712-715
Time point:
24/48/72 h
Score:
1
Reversibility:
fully reversible within: 14 days
Irritant / corrosive response data:
Irritation and Corrosion (Table 1)
Instillation of approximately 40 mg of naphthalene-1,5-diol (a volume of approximately 0.1 ml) into one eye of each of three rabbits resulted in irritation of the conjunctivae, which consisted of redness, chemosis and discharge. The irritation had completely resolved within 14 days.
No iridial irritation or cornea opacity was observed, and treatment of the eyes with 2% fluorescein, 24 hours after test substance instillation revealed no corneal epithelial damage.

Corrosion
There was no evidence of ocular corrosion.
Colouration / Remnants
Remnants of the test substance were present in the eye of all animals on day 1 .

Other effects:
Toxicity / Mortality
No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Applicant's summary and conclusion

Interpretation of results:
Category 2 (irritating to eyes) based on GHS criteria
Conclusions:
Based on these results and according to the:
OECD Harmonized Integrated Hazard Classification System for Human Health and Environmental Effects of Chemical Substances (OECD, 1998), naphthalene-1,5-diol should be classified as: irritating to eyes (Class 2).
EC criteria for classification and labelling requirements for dangerous substances and preparations (Council Directive 67/548/EEC), naphthalene-1,5-diol should be labelled as: irritating to eyes (R 36).
Executive summary:

Acute eye irritation/corrosion study with naphthalene-1,5 -diol in the rabbit.

The study was carried out based on the guidelines described in: "Acute Toxicity - Eye irritation", OECD No.405 (2002); EC Commission Directive 92/69/EEC, 8.5, "Acute Eye Irritation/Corrosion" (1992); US EPA, OPPTS 870.2400, Acute Eye Irritation, (1998) and JMAFF guidelines (2000).

Single samples of approximately 40 mg of naphthalene-1,5-diol (a volume of approximately 0.1 ml) were instilled into one eye of each of three rabbits. Observations were made 1, 24, 48 and 72 hours and 7 and 14 days after instillation.

Instillation of the test substance resulted in irritation of the conjunctivae, which consisted of redness, chemosis and discharge. The irritation had completely resolved within 14 days.

Remnants of the test substance were present in the eye of all animals on day 1.

Based on these results and according to the:

- OECD Harmonized Integrated Hazard Classification System for Human Health and Environmental Effects of Chemical Substances (OECD, 1998), naphthalene-1,5-diol should be classified as: irritating to eyes (Class 2).

- EC criteria for classification and labelling requirements for dangerous substances and preparations (Council Directive 67/548/EEC), naphthalene-1,5-diol should be labelled as: irritating to eyes (R 36).