Registration Dossier

Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2001
Reliability:
1 (reliable without restriction)

Data source

Reference
Reference Type:
other company data
Title:
Unnamed
Year:
2001
Report Date:
2001

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Diphenylsulfon (DPS)
- Substance type: Organic
- Physical state: Solid, white powder (determined at NOTOX)
- Analytical purity: >99%
- Lot/batch No.: 1712SA
- Purity test date: 15/05/2001
- Expiration date of the lot/batch: 15/05/2002
- Stability under test conditions: not indicated
- Storage condition of test material: at room temperature in the dark
- Other: bulk density: 0.7-0.8

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Kissleg, Germany
- Age at study initiation: at least 6 weeks old
- Weight at study initiation: body weights between 1.0 and 3.5 kg
- Housing: Individually in labelled cages with perforated floors (Scanbur, Denmark, dimensions 53.5*63*38.5 cm).
- Diet (e.g. ad libitum): Standard laboratory rabbit diet (Charles River Breeding and Maintenance Diet for Rabbits, Altromin, Germany) approx. 100g per day. Certificates of analysis were examined and retained in the NOTOX archives. In addition, hay (BMI, Helmond, the Netherlands) was provided twice a week.
- Water (e.g. ad libitum): Free acces to tap-water. Certificates of quarterly analysis were examined and retained in the NOTOX archives.
- Acclimation period: At least 5 days before start of treatment under laboratory conditions.


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 +- 3°c
- Humidity (%): 30-70%
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light and 12 hours dark per day


Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: The other eye remained untreated and served as the reference control.
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit):71.4 +- 0.6 mg (a volume of approximately 0.1 ml).
- Concentration (if solution):


VEHICLE
- Amount(s) applied (volume or weight with unit):
- Concentration (if solution):
- Lot/batch no. (if required):
- Purity:
Duration of treatment / exposure:
24 h
Observation period (in vivo):
The eyes of each animal were examined approximately 1, 24, 48 and 72 hours after instillation of the test substance.
Number of animals or in vitro replicates:
3
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): 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.
- Time after start of exposure: 24h


SCORING SYSTEM:
The irritation will be assessed according to the following numerical scoring system. At each observation, the highest scores given will be recorded:

CORNEAL IRRITATION:
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 quarers, up the 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 all of these)................................................2

CONJUNCTIVAL 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 vesses 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


TOOL USED TO ASSESS SCORE: fluorescein
Where standard lighting is considered inadequate for observing minor effects, eye examinations will be performed using an ophthalmic examination lamp.
In cases of equivocal results when comparing the threated and untreated eyes, the illustrated guide from the Consumer Product Safety Commission, Washington, D.C. 20207 will be used for additional control purposes.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
other: 48h
Max. score:
1
Reversibility:
fully reversible within: 48 h
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: 24 h
Max. score:
0
Reversibility:
fully reversible
Irritation parameter:
iris score
Basis:
mean
Time point:
other: 24h
Max. score:
1
Reversibility:
fully reversible within: 24 h
Irritant / corrosive response data:
IRRITATION:
Instillation of approximately 71 mg of the test substance into one eye of each of three rabbits resulted in effects on the iris and conjunctivae. One eye of each of three rabbits resulted in effects on the iris and conjunctivae. Iridial irritation (grade 1) was observed in all animals and had resolved within 24 hours. Irritation of the conjunctivae was seen as redness, chemosis and discarge, which had completely resolved within 48 hours in all animals.

No corneal opacity was observed, and treatment of the eyes with 2% fluorescein, 24 hours after test substance instillation revealed no corneal epithelial damage in any of the animals.

CORROSION
There was no evidence of ocular corrosion

COLOURATION:
No staining of (peri) ocular tissues by the test substance was observed. Remnants of the test substance were present in the eyes of all animals on day 1.

TOXICITY / MORTALITY
No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred

Any other information on results incl. tables

Individual eye irritation scores:

 Time after dosing Opacity (cornea)  area (cornea)  fluor area (%) (cornea)  Iris  redness (conjunctivae)  chemosis (conjunctivae)  discharge (conjunctivae)  Comments 
892                 
1 h     b
24 h   1  0  0  /
48h   0   0  0  0  0  /
72 h   0   0  0  0  0  /
893                 
1 h   0  0    1  1  1  1  b
24 h   0  0  0  0  1  0  0  /
48 h   0  0    0  0  0  0  /
72 h   0  0    0  0  0  0  /
  894                
 1 h  0  0    1  1  1  1  b
 24 h  0  0  0  0  1  0  0  /
 48 h  0  0    0  0  0  0  /
 72 h  0  0    0  0  0  0  /

(Fluor area (%): green staining (percentage of total corneal area) after fluorescein treatment.)

Comments: b: remnants of the test substance in the eye.

mean value eye irritation scores

( # animal, mean 24 -72 hours)

animal# Corneal opacity  Iris  redness (conjunctivae)  chemosis (conjunctivae) 
892  0.3 
893  0.3 
894  0.3 

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Based on these results and according to the EC criteria for classification and labelling requirements for dangerous substances and preparations (Guidelines in Commission Directive 93/21/EEC), diphenylsulfon (DPS) does not have to be classified and has no obligatory labelling requirement for eye irritation.