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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:
31 October 2011 - 21 November 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report date:
2012

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
- the relative humidity recorded in the animal room was sometimes outside of the target ranges specified in the study plan,no pH measurement was performed as the test item was insoluble in water.
Qualifier:
according to guideline
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
idem above
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
Bis(piperidinothiocarbonyl) hexasulphide
EC Number:
213-537-2
EC Name:
Bis(piperidinothiocarbonyl) hexasulphide
Cas Number:
971-15-3
Molecular formula:
C12H20N2S8
IUPAC Name:
1,1'-(hexasulfane-1,6-diyldicarbonothioyl)dipiperidine
Constituent 2
Reference substance name:
Dipentamethylenethiuram hexasulfide
IUPAC Name:
Dipentamethylenethiuram hexasulfide
Test material form:
solid: particulate/powder

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Breeder: Hypharm, La Corbière, Roussay, France
- Age at study initiation: at the beginning of the study, the animals were 2 to 4 months old
- Mean body weight at study initiation: the animals had a mean body weight ± standard deviation of 3.323 kg ± 0.129 kg
- Fasting period before study: no
- Housing: individually housed in noryl cages
- Diet: free access to pelleted breeding diet “type 110C”
- Water: tap water filtered with a 0.22 µm filter (free access)
- Acclimation period: at least 5 days before the beginning of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 ± 3°C
- Humidity (%): 50 ± 20%
- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air
- Photoperiod (hrs dark / hrs light): 12 h/12 h (7:00 - 19:00).

IN-LIFE DATES: 09 November 2011 to 21 November 2011

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: untreated right eye served as a control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied: 0.1 g/animal.
Duration of treatment / exposure:
Not applicable: single application not followed by rinsing.
Observation period (in vivo):
1, 24, 48 and 72 h; if relevant, daily until reversibility of reactions
Number of animals or in vitro replicates:
3 males.
Details on study design:
REMOVAL OF TEST SUBSTANCE: No

SCORING SYSTEM: Draize scale.

- Conjunctival chemosis (lids and/or nictitating membranes):
0 no swelling
1 any swelling above normal (includes nictitating membranes)
2 obvious swelling with partial eversion of lids
3 swelling with lids about half-closed
4 swelling with lids more than half-closed

- Conjunctival redness (palpebral and bulbar conjunctivae, cornea and iris):
0 blood vessels normal
1 a number of blood vessels definitely hyperemic (injected)
2 diffuse, crimson colour, individual vessels not easily discernible
3 diffuse, beefy red

- Discharge:
0 absence of discharge
1 slight discharge (does not include small amounts normally found in inner canthus)
2 discharge with moistening of lids and hairs adjacent to lids
3 discharge with moistening of lids and hairs on wide area around the eye

- Iris lesions
0 normal
1 markedly deepened rugae, congestion, swelling, moderate circum-corneal hyperemia,or injection, any of these or combination of any thereof, iris still reacting to light (sluggish reaction is positive)
2 no reaction to light, haemorrhage, gross destruction (any or all of these)

- Cornea intensity of opacity (direct examination and, if necessary, with an UV lamp)
0 no ulceration or opacity
1 scattered or diffuse areas of opacity (other than slight dulling or normal lustre), details of iris clearly visible
2 easily discernible translucent area, details of iris slightly obscured
3 nacreous areas, no details of iris visible, size of pupil barely discernible
4 opaque cornea, iris not discernible through the opacity

- Cornea area of opacity (direct examination and, if necessary, with an UV lamp)
1 one quarter (or less) but not zero
2 greater than one quarter but less than a half
3 greater than one half but less than three quarters
4 greater than three quarters up to whole area

- Any other lesions observed were noted

TOOL USED TO ASSESS SCORE: UV lamp after instillation of 0.5% sodium fluorescein solution

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
3 rabbits
Time point:
24/48/72 h
Score:
0.53
Max. score:
4
Reversibility:
fully reversible within: 6 days
Remarks on result:
other: Individual scores : 0.3 -1.3 -0.0 showing no significant eye irritation
Irritation parameter:
conjunctivae score
Remarks:
(redness)
Basis:
mean
Remarks:
3 rabbits
Time point:
24/48/72 h
Score:
0.77
Max. score:
3
Reversibility:
fully reversible within: 72 hours
Remarks on result:
other: Individual scores: 0.7 -1.3 -0.3 showing no significant eye irritation.
Irritation parameter:
iris score
Basis:
mean
Remarks:
3 rabbits
Time point:
24/48/72 h
Score:
0.23
Max. score:
2
Reversibility:
fully reversible within: 72 hours
Remarks on result:
other: Individual scores: 0.0 -0.7-0.0 showing no significant eye irritation
Irritation parameter:
cornea opacity score
Remarks:
(opacity)
Basis:
mean
Remarks:
3 rabbits
Time point:
24/48/72 h
Score:
0.33
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Remarks on result:
other: Individual scores: 0.0 -1.0 -0.0 showing eye irritation in 1/3 animals.
Irritant / corrosive response data:
First animal (No. 57): A moderate chemosis (grade 2) was observed at the 1-hour observation, became slight 24 hours after treatment and had disappeared on day 3. A severe redness of the conjunctiva (grade 3) was observed 1 hour after treatment, then became slight on days 2 and 3, and had disappeared on day 4.
No iris or corneal lesions were observed.

Second animal (No. 58): A moderate chemosis (grade 2) was noted on days 1 and 2 and was then graded slight from day 3 to day 5. A severe redness of the conjunctiva (grade 3) was observed on days 1 and 2, was slight on day 3 and was no longer observed from day 4. Iris lesions (grade 1) were noted on days 2 and 3. A slight corneal opacity (intensity: grade 2 to 1 and area: grade 1) was recorded on days 2 and 3 and had completely recovered on day 4.
A whitish purulent discharge was also observed on days 2 to 4.

Third animal (No. 59): A moderate chemosis (grade 2) was observed at the 1-hour observation only, together with a severe redness of the conjunctiva (grade 3). The redness became slight on day 2 and was no longer observed from day 3. No iris or corneal lesions were observed.
Other effects:
No unscheduled deaths occurred during the study.
No clinical signs other than ocular reactions were noted in any animals.
The body weight of the animals was unaffected by the test item-treatment.

Any other information on results incl. tables

First animal

 

 

 

 

 

 

Irritation parameter

Basis

Time point

Score

Max. score

Reversibility

Remarks

chemosis

mean

24, 48 and 72 h

0.3

1

fully reversible within: 48h

 

conj

mean

24, 48 and 72 h

0.7

1

fully reversible within: 72h

 

iris

mean

24, 48 and 72 h

0

0

other: not applicable

 

cornea

mean

24, 48 and 72 h

0

0

other: not applicable

 

 

 

 

 

 

 

 

 second animal

 

 

 

 

 

 

Irritation parameter

Basis

Time point

Score

Max. score

Reversibility

Remarks

chemosis

mean

24, 48 and 72 h

1.3

2

fully reversible within: 6 days

 

conj

mean

24, 48 and 72 h

1.3

3

fully reversible within: 72h

 

iris

mean

24, 48 and 72 h

0.7

1

fully reversible within: 72h

 

cornea

mean

24, 48 and 72 h

1

2

fully reversible within: 72h

 

 

 

 

 

 

 

 

Third animal

 

 

 

 

 

 

Irritation parameter

Basis

Time point

Score

Max. score

Reversibility

Remarks

chemosis

mean

24, 48 and 72 h

0

0

other: not applicable

 

conj

mean

24, 48 and 72 h

3

1

fully reversible within: 48h

 

iris

mean

24, 48 and 72 h

0

0

other: not applicable

 

cornea

mean

24, 48 and 72 h

0

0

other: not applicable

 

 

 

 

 

 

 

 

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Remarks:
not eye irritating
Conclusions:
The test item was slightly irritant when administered by the ocular route to rabbits.
Executive summary:

The objective of this study was to evaluate the potential eye irritant properties of the test item, Dipentamethylenethiuram hexasulfide, following a single administration to rabbits.

This study was conducted in compliance with OECD guideline (OECD 404) and with the principles of Good Laboratory Practice.

Methods

The test item Dipentamethylenethiuram hexasulfidewas first administered to a single male New Zealand White rabbit.

As mean value from grading at 24, 48 and 72 hours after instillation was < 2 for conjunctival edema (chemosis) or for conjunctival redness and/or < 1 for iris lesion or for corneal opacity, the test item was administered in the left eye of two other animals.

 

The test item was administered inthe conjunctival sac of the left eye. The right eye remained untreated and served as control.A quantity of 0.1 g/animal was used. Just after the 1-hour scoring, both eyes were rinsed with a sterile isotonic saline solution (0.9% NaCl).

 

Each animal was observed at least once a day for mortality and clinical signs.Ocular reactions were observed approximately 1 hour, 24, 48 and 72 hours after the administration and then daily until the reversibility of the ocular reactions (daytwo animals, daythe other one). The mean values of the scores for chemosis, redness of the conjunctiva, iris lesions and corneal opacity were calculated for each animal. Body weight was recorded on the day of treatment and the end of the observation period.

On completion of the observation period, the animals were sacrificed then discarded without macroscopic post-mortem examination.

Results

Moderate chemosis and severe redness of the conjunctiva were observed in all animals on day 1 and/or 2. Then slight conjunctival reactions persisted up to day 5 at the latest.

Iris lesions and a moderate then slight corneal opacity were noted in 1/3 animals on days 2 andaddition, whitish purulent discharge was observed from day 2 until day 4.

 

Mean scores calculated for each animal over 24, 48 and 72 hours were as follows:

.       chemosis: 0.3, 1.3 and 0.0; showing no significant eye irritation,

.       redness of the conjunctiva: 0.7, 1.3 and 0.3; showing no significant eye irritation,

.       iris lesions: 0.0, 0.7 and 0.0; showing no significant eye irritation,

.       corneal opacity: 0.0, 1.0 and 0.0; showing eye irritation in 1/3 animals.

 

No ocular reactions were observed in the right eye of all animals.

 

Conclusion

The test item was slightly irritant when administered by the ocular route to rabbits.

However, the test item is not classified as irritating to the eyes according to the criteria of CLP Regulation.