1,7-Naphthalenedisulfonic acid, 4-amino-3-[(1E)-2-(2,4-disulfophenyl)diazenyl]-5-hydroxy-6-[(1E)-2-(4-nitro-2-sulfophenyl)diazenyl]-, sodium salt (1:5)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

13 July 2015 - 22 September 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study was performed at a GLP laboratory in accordance with prescribed OECD, EU, US and Japanese test guidelines. A minor deviation from the daily mean relative humidity occurred however this deviation was determined not to affect the validity of the study.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

Deviations from the maximum level of daily mean relative humidity occurred. Laboratory historical data do not indicate an effect of the deviation. The study integrity was not adversely affected by the deviation.

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

Deviations from the maximum level of daily mean relative humidity occurred. Laboratory historical data do not indicate an effect of the deviation. The study integrity was not adversely affected by the deviation.

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

yes

Remarks:

Deviations from the maximum level of daily mean relative humidity occurred. Laboratory historical data do not indicate an effect of the deviation. The study integrity was not adversely affected by the deviation.

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: JMAFF Guidelines (2000), including the most recent revisions.

Deviations:

yes

Remarks:

Deviations from the maximum level of daily mean relative humidity occurred. Laboratory historical data do not indicate an effect of the deviation. The study integrity was not adversely affected by the deviation.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

pentasodium 4-amino-3-[(1E)-2-(2,4-disulfonatophenyl)diazen-1-yl]-5-hydroxy-6-[(1E)-2-(4-nitro-2-sulfonatophenyl)diazen-1-yl]naphthalene-1,7-disulfonate

EC Number:

811-858-8

Cas Number:

2149571-68-4

Molecular formula:

C22 H11 N6 O18 S5 .5Na

IUPAC Name:

pentasodium 4-amino-3-[(1E)-2-(2,4-disulfonatophenyl)diazen-1-yl]-5-hydroxy-6-[(1E)-2-(4-nitro-2-sulfonatophenyl)diazen-1-yl]naphthalene-1,7-disulfonate

Constituent 2

Reference substance name:

pentasodium 4-amino-3-[(2,4- disulfonatophenyl)diazenyl]-5-hydroxy-6-[(4-nitro-2- sulfonatophenyl)diazenyl]naphthalene-1,7-disulfonate

IUPAC Name:

pentasodium 4-amino-3-[(2,4- disulfonatophenyl)diazenyl]-5-hydroxy-6-[(4-nitro-2- sulfonatophenyl)diazenyl]naphthalene-1,7-disulfonate

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): 206474/A and K1600 BLACK DYE
- Physical state: Powder

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River France, L’Arbresle Cedex, France
- Age at study initiation: 12 and 24 weeks old
- Weight at study initiation: at least 1.5 kg.
- Housing: individually housed in labeled cages with perforated floors and shelters
- Diet (e.g. ad libitum): Pelleted diet for rabbits, approximately 100 grams per day, Hay and wooden sticks were available during the study period.
- Water (e.g. ad libitum): free access to tap water
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 24°C
- Humidity (%): 40 to 70%
- Air changes (per hr): 10 air changes per hour
- Photoperiod (hrs dark / hrs light): 12-hour light/12-hour dark cycle

Test system

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: on average, 81.9 mg (range 81.8 – 82.0 mg)

Duration of treatment / exposure:

1 hour

Observation period (in vivo):

24 hours

Number of animals or in vitro replicates:

3 males

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: he treated eye was rinsed with approximately 50 mL tepid tap water
- Time after start of exposure: 1 hour

SCORING SYSTEM:
CORNEAL IRRITATION

Opacity: degree of density (area most dense taken for reading)
No ulceration or opacity (may include slight dulling of normal luster): 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 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 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

TOOL USED TO ASSESS SCORE: fluorescein

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

overall irritation score

Remarks:

Corneal opacity

Basis:

mean

Remarks:

Animal 944

Time point:

other: 24, 48 and 72 hours

Score:

ca. 0

Results 2

Irritation parameter:

overall irritation score

Remarks:

Corneal opacity

Basis:

mean

Remarks:

Animal 933

Time point:

other: 24, 48 and 72 hours

Score:

ca. 0

Results 3

Irritation parameter:

overall irritation score

Remarks:

Corneal opacity

Basis:

mean

Remarks:

Animal 940

Time point:

other: 24, 48 and 72 hours

Score:

ca. 0

Results 4

Irritation parameter:

overall irritation score

Remarks:

Iris

Basis:

mean

Remarks:

Animal 944

Time point:

other: 24, 48 and 72 hours

Score:

ca. 0

Results 5

Irritation parameter:

overall irritation score

Remarks:

Iris

Basis:

mean

Remarks:

Animal 933

Time point:

other: 24, 48 and 72 hours

Score:

ca. 0

Results 6

Irritation parameter:

overall irritation score

Remarks:

Iris

Basis:

mean

Remarks:

Animal 940

Time point:

other: 24, 48 and 72 hours

Score:

ca. 0

Results 7

Irritation parameter:

overall irritation score

Remarks:

Conjunctivae - Redness

Basis:

mean

Remarks:

Animal 944

Time point:

other: 24, 48 and 72 hours

Score:

ca. 1.7

Reversibility:

fully reversible within: 7 days

Results 8

Irritation parameter:

overall irritation score

Remarks:

Conjunctivae - Redness

Basis:

mean

Remarks:

Animal 933

Time point:

other: 24, 48 and 72 hours

Score:

ca. 1.3

Reversibility:

fully reversible within: 7 days

Results 9

Irritation parameter:

overall irritation score

Remarks:

Conjunctivae - Redness

Basis:

mean

Remarks:

Animal 940

Time point:

other: 24, 48 and 72 hours

Score:

ca. 1

Reversibility:

fully reversible within: 72 hours

Results 10

Irritation parameter:

overall irritation score

Remarks:

Conjunctivae - Chemosis

Basis:

mean

Remarks:

Animal 944

Time point:

other: 24, 48 and 72 hours

Score:

ca. 0.3

Reversibility:

fully reversible within: 24 hours

Results 11

Irritation parameter:

overall irritation score

Remarks:

Conjunctivae - Chemosis

Basis:

mean

Remarks:

Animal 933

Time point:

other: 24, 48 and 72 hours

Score:

ca. 0

Reversibility:

fully reversible within: 24 hours

Results 12

Irritation parameter:

overall irritation score

Remarks:

Conjunctivae - Chemosis

Basis:

mean

Remarks:

Animal 940

Time point:

other: 24, 48 and 72 hours

Score:

ca. 0.3

Reversibility:

fully reversible within: 24 hours

Irritant / corrosive response data:

Some irritation was observed during the study. The irritation of the conjunctivae consisted of redness, chemosis and discharge. This irritation had completely resolved within 72 hours in one animal and within 7 days in the other two animals. Irritation of the iris was found in one animal and had resolved within 24 hours.

Any other information on results incl. tables

No signs of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Blue staining of test substance remnants was observed on the cornea, iris and conjunctivae on Day 1.

The staining had not completely resolved after rinsing the eye on Day 1, but had completely resolved

during the second observation, 24 hours after instillation.

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Based on these results, the test substance does not have to be classified and has no obligatory labelling
requirement for eye irritation according to the:
- Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United
Nations (2011) (including all amendments),
- Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures
(including all amendments).

Executive summary:

Study was performed at a GLP laboratory in accordance with prescribed OECD, EU, US and Japanese test guidelines. A minor deviation from the daily mean relative humidity occurred however this deviation was determined not to affect the validity of the study.

Single samples of approximately 82 mg of the test substance were instilled into one eye of each of three rabbits. Observations were made 1, 24, 48 and 72 hours and/or 7 days after instillation.

Instillation of the test substance resulted in irritation of the conjunctivae and iris. The irritation of the

conjunctivae consisted of redness, chemosis and discharge. The irritation had completely resolved

within 72 hours in one animal and within 7 days in the other two animals. Irritation of the iris was found in one animal and had resolved within 24 hours.

 

Blue staining of test substance remnants was observed on the cornea, iris and conjunctivae on Day 1.

The staining had not completely resolved after rinsing the eye on Day 1, but had completely resolved

during the second observation, 24 hours after instillation. The staining hampered scoring of the cornea (opacity and area) and conjunctivae (redness) on Day 1. For the iris, only the light reaction could be scored during the first observation. Blue staining of the fur on the head and paws, caused by the test substance, was noted during the observation period.

 

Based on these results, the test substance does not have to be classified and has no obligatory labelling requirement for eye irritation according to the:

- Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United

Nations (2011) (including all amendments),

- Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures

(including all amendments).