Pentasodium bis{7-[4-(1-butyl-5-cyano-1,2-dihydro-2-hydroxy-4-methyl-6-oxo-3-pyridylazo)phenylsulfonylamino]-5'-nitro-3,3'-disulfonatonaphthalene-2-azobenzene-1,2'-diolato} chromate (III)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

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:

Adopted February 24, 1987

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

July 31, 1992

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Pentasodium bis{7-[4-(1-butyl-5-cyano-1,2-dihydro-2-hydroxy-4-methyl-6-oxo-3-pyridylazo)phenylsulfonylamino]-5'-nitro-3,3'-disulfonatonaphthalene-2-azobenzene-1,2'-diolato} chromate (III)

EC Number:

419-210-2

EC Name:

Pentasodium bis{7-[4-(1-butyl-5-cyano-1,2-dihydro-2-hydroxy-4-methyl-6-oxo-3-pyridylazo)phenylsulfonylamino]-5'-nitro-3,3'-disulfonatonaphthalene-2-azobenzene-1,2'-diolato} chromate (III)

Cas Number:

178452-71-6

Molecular formula:

C66H48CrN16Na5O28S6

IUPAC Name:

chromium(3+) pentasodium bis(7-{4-[2-(1-butyl-5-cyano-2-hydroxy-4-methyl-6-oxo-1,6-dihydropyridin-3-yl)diazen-1-yl]benzenesulfonamido}-2-[2-(5-nitro-2-oxido-3-sulfonatophenyl)diazen-1-yl]-3-sulfonatonaphthalen-1-olate)

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Remarks:

CRL : KBL

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
-Source: Charles River Stolzenseeweg 32-36 D-88353 Kisslegg
-Age at study initiation: 15 weeks
-Weight at study initiation:
male: 2.7 kg
females: 2.7 - 2.8 kg
-Housing: Individually in stainless steel cages with an automatic cleaning system, equipped with feed hoppers, drinking water bowls and wood for gnawing.
-Diet: pelleted standard Kliba 341, Batch no. 87/94 rabbit maintenance diet ("Kliba", Klingentalmuehle AG, CH-4303 Kaiseraugst), ad libitum.
-Water: community tap water from Itingen, ad libitum.
-Acclimation period: five days

ENVIRONMENTAL CONDITIONS
-Temperature: 20 ± 3 °C
-Humidity: 40-70 %
-Air changes: 10-15 air changes per hour
-Photoperiod: 12 hours cycle dark/light
-Other: music during the light period

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes

Amount / concentration applied:

TEST MATERIAL
-Amount: 0.1 g

Duration of treatment / exposure:

single injection

Observation period (in vivo):

1, 24, 48 and 72 hours and 7, 14 and 21 days after administration

Number of animals or in vitro replicates:

1 male and 2 females

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: no

OBSERVATIONS
The eyes of each animal were examined for ocular irritation 1, 24, 48 and 72 hours, as well as 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. Moreover, Viability/Mortality and Clinical Signs were observed daily during the observation period. Also, body weights were observed the start of acclimatization, on day 1 of test (application day) and at termination of observation.

SCORING SYSTEM:
CORNEA
- Opacity: degree of density (area most dense taken for reading)
No ulceration or opacity: 0
Scattered or diffuse areas of opacity (other than slight dulling of normal lustre), details of iris clearly visible: 1
Easily discernible translucent area, details of iris slightly obscured: 2
Nacreous areas, no details of iris visible, size of pupil barely discernible: 3
Opaque cornea, iris not discernible through the opacity: 4
- Area of cornea affected
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 any combination thereof, iris still reacting to light (sluggish reaction is positive): 1
No reaction to light, haemorrhage, gross destruction (any or all of these): 2
CONJUNCTIVAE
-Redness: (refers to palpebral and bulbar conjunctivae, excluding cornea and iris).
Blood vessels normal
Some blood vessels definitely hyperaemic (injected): 1
Diffuse, crimson colour, individual vessels not easily discernible: 2
Diffuse beefy red: 3
-Chemosis: lids and/or nictating membrane
No swelling: 0
Any swelling above normal (includes nictating membrana): 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: 0
Any amount different from normal (does not include small amounts observed in inner canthus of
normal animals): 1
Discharge with moistening of the lids and hairs, just adjacent to lids: 2
Discharge with moistening of the lids and hairs, in a considerable area around the eye: 3.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Results 2

Irritation parameter:

iris score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Results 4

Irritation parameter:

chemosis score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Irritant / corrosive response data:

CORROSION
No corrosion of the cornea was observed at any of the reading times.

Other effects:

COLORATION
Green staining of the conjunctivae by the test substance persisted in both surviving animals until termination. Staining of the sclera was reversible in both animals within the 21-day observation period, whereas staining of the cornea persisted in one of the two animals up to 21 days.

VIABILITY / MORTALITY AND CLINICAL SIGNS
No clinical signs of systemic toxicity were observed in the animals during the test and observation period. Male rabbit no. 4 was found dead in its cage on the morning of test day 7. This finding was considered to be a spontaneous death unrelated to the treatment with the test substance.

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

NECROPSY
Male rabbit no. 4 was found dead on the morning of test day 7 and was subsequently necropsied. Dark red discoloration of the lung lobes was noted, but this finding was considered to be unrelated to the treatment with the test substance.

Applicant's summary and conclusion

Interpretation of results:

other: Classified according to the CLP Regulation (EC n. 1272/2008) Eye Dam. 1

Conclusions:

The test item induced irreversible damage to the rabbit eye, as the green coloured staining of the conjunctivae by the test item persisted throughout the observation period.

Executive summary:

The eye irritation test was performed to the rabbit eye according to OECD Guidelines 405 (1987) and the method B.5 of EEC-Directive 92/69 EEC. 0.1 g of test item were installed into the left eye of 3 young adult New Zealand White rabbits (1 male and 2 females). The treated eyes were not rinsed after the application. Scoring of irritation effects was performed after test item application at 1, 24, 48 and 72 hour, as well as 7, 10, 14, 17 and 21 day.

Animal #1 was found dead on day 7, but the death was considered to be not treatment-related. Mean values at 24, 48 and 72 hours for iris, corneal opacity and the conjunctivae (redness and chemosis) were 0.00 in all animals. No corrosion was observed at any of the measuring intervals. The test item caused dark green staining of cornea, conjunctivae and sclera in all animals at the 1-hour reading. Light green staining was then observed in the following readings. For animal #1 staining of cornea, conjunctivae and sclera persisted until its death. For animals #2 and #3 staining of the conjunctivae persisted up to 21 days. Staining of the sclera was reversible within 14 days (animal #2) and 24 hours (animal #3). Staining of the cornea was still present in animal #2 after 21 days, whereas in animal #3 was reversible on day 21.