reaction product of: C.I. Leuco Sulfur Black 1 with (3-chloro-2-hydroxypropyl)trimethylammonium chloride

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 15. Aug. to 05. Sep. 1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1995

Report date:

1995

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

1987

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

424-510-1

EC Name:

-

Cas Number:

220150-59-4

Molecular formula:

not applicable for UVCB substance

IUPAC Name:

Reaction products of Phenol, 2,4-dinitro-, sulfurized, leuco derivatives and (3-chloro-2-hydroxypropyl)trimethylammonium chloride

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Remarks:

CRL:KBL(NZW)BR

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River, Stolzenseeweg 32 - 36, 88353 Kisslegg, Germany
- Age at study initiation: 16 to 21 weeks
- Weight at study initiation: males: 3.1 kg; females: 2.9 to 3.5 kg
- Housing: individually in stainless steel cages with an automatic cleaning system, equipped with feed hoppers, drinking water bowls and wood for gnawing
- Diet: ad libitum ; pelleted standard Kliba 341, batch no. 90/95 rabbit maintenance diet ("Kliba" Klingentalmühle AG, 4303 Kaiseraugst, CH) (analytical test report demonstrates suitability)
- Water: ad libitum; community tap water from Itingen (bacterial, chemical and contaminant test reports demonstrate suitability)
- Acclimation period: 5 days under laboratory conditions after health examination

ENVIRONMENTAL CONDITIONS
- Temperature: 20 °C ± 3
- Humidity: 30 to 70 %
- Air changes: 10 to 15 per hr
- Photoperiod (hrs dark / hrs light): 12/12

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.1 g
- Concentration: undiluted; solid

Duration of treatment / exposure:

One single application

Observation period (in vivo):

21 days

Number of animals or in vitro replicates:

Males: 1
Females: 2
Total: 3

Details on study design:

TREATMENT
- On test day 1, 0.1 g of undiluted test item was placed in the conjunctival sac of the left eye of each animal after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent the loss of the test item. The right eye remained untreated and served as the reference control.
- Animals were monitored for irritation, corrosivity, viability/mortality, body weight and colouration.

REMOVAL OF TEST SUBSTANCE
- Washing: no washing or removal

SCORING SYSTEM: irritation
- The eyes of each animal were examined for ocular irritation approximately 1, 24, 48 and 72 hours and 7, 14 and 21 days after administration. Any findings ascertained during examination were recorded and the numerical irritation scores were assigned thereafter based upon the severity of the individual findings.
- The following numerical system was used to grade the eye lesions:

CORNEAL IRRITATION: opacity; degree of density (densest area used for assessment)
0 No ulceration or opacity
1* Scattered or diffuse areas of opacity (other than slight dulling of normal luster), details of iris clearly visible
2* Easily discernible translucent area, details of iris slightly obscured
3* Nacrous area, no details of iris visible, size of pupil barely discernible
4* Opaque cornea, iris not discernible through the opacity

IRIDIC IRRITATION:
0 Normal
1* 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)
2* No reaction to Iight, haemorrhage, gross destruction (any or all of these)

CONJUNCTIVAL IRRITATION: redness (refers to palpebral and bulbar conjunctivae, cornea and iris)
0 Blood vessels normal
1 Some blood vessels definitely hyperaemic (injected)
2* Diffuse, crimson colour, individual vessels not easily discernible
3* Diffuse beefy red

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

* Starred figures indicate positive effect

TOOL USED TO ASSESS SCORE: eye examinations were made with a Varta Cliptrix diagnostic-lamp (A. Riegger, Basel, CH)

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #2 and #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 2

Irritation parameter:

iris score

Basis:

animal: #1, #2 and #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal: #1, #2 and #3

Time point:

24/48/72 h

Score:

0

Max. score:

3

Results 4

Irritation parameter:

chemosis score

Basis:

animal: #1, #2 and #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

IRRITATION
- Irritation scores of 0.0 were observed when applied to the conjunctival sac of the rabbit eye. Slight swelling and slight reddening of the conjunctivae were noted in two of three animals after 1 hour. These findings were reversible within 24 hours.

CORROSION
- No corrosive effects were observed at any of the reading times.

Other effects:

BODY WEIGHT
- The body weight of the animals was within the normal range of variability.

COLOURATION
- Black or light black staining of the cornea noted in all animals was reversible in one animal after 72 hours and in a second animal after 21 days. The third animal showed light black corneal staining until the end of the observation period of 21 days.
- Black or light black staining of the ocular adnexa (conjunctivae and/or nictitating membrane) was noted in all animals. Black or light black staining of the nictitating membrane persisted in all animals until the end of the observation period at 21 days.

VIABILITY / MORTALITY
- No clinical signs of systemic toxicity were observed in the animals during the test and observation periods, and no mortality occurred.

PATHOLOGY
- All animals were sacrificed by an intravenous injection of NARCOREN (Rhone Merieux GmbH, 88471 Laupheim, Germany) into the ear vein at a dose of at least 1.0 mL/kg bw.
- No necropsy was performed after animals were sacrificed at termination of the study.

Applicant's summary and conclusion

Interpretation of results:

other: Category 1 (Serious Eye Damage) based on CLP criteria (EC 1272/2008)

Conclusions:

The test item was considered to have eye damage potential.

Executive summary:

Eye irritation potential was evaluated in an experimental study according to the OECD Guideline 405 (1987) and EU method B.5 (1992). 0.1 g of undiluted, solid test item was administered to the left conjunctival sac of three rabbits (one male and two female) and was not rinsed. Observations and irritation scores were recorded at 1, 24, 48 and 72 hours and 7, 14 and 21 days after application of the test item.

Irritation scores for all parametres were calculated as means at 24, 48 and 72 hours. All irritation scores were 0.0 at these timepoints. Black or light black staining of the cornea was reversible in one test animal after 72 hours and in a second test animal after 21 days. The third test animal showed light black corneal staining until the end of the study period (21 days). Black or light black stainging of the nictitating membrane was observed in all three test animals until the end of the study period (21 days). No corrosive effect was observed.