Hexasodium 2,2'-[vinylenebis[(3-sulphonato-4,1-phenylene)imino[6-morpholino-1,3,5-triazine-4,2-diyl]imino]]bis(benzene-1,4-disulphonate)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1975

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1975

Report date:

1975

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Hazardous Substances Regulations" under the U.S.A. Federal Hazardous Substances Labelling Act Sect. 191.12 (February 1965), with slight modification.

Deviations:

no

Principles of method if other than guideline:

None

GLP compliance:

no

Test material

Test material information

Constituent 1

Reference substance name:

Hexasodium 2,2'-[vinylenebis[(3-sulphonato-4,1-phenylene)imino[6-morpholino-1,3,5-triazine-4,2-diyl]imino]]bis(benzene-1,4-disulphonate)

EC Number:

257-827-7

EC Name:

Hexasodium 2,2'-[vinylenebis[(3-sulphonato-4,1-phenylene)imino[6-morpholino-1,3,5-triazine-4,2-diyl]imino]]bis(benzene-1,4-disulphonate)

Cas Number:

52301-70-9

Molecular formula:

C40H40N12O20S6.6Na

IUPAC Name:

hexasodium 2,2'-[vinylenebis[(3-sulphonato-4,1-phenylene)imino[6-morpholino-1,3,5-triazine-4,2-diyl]imino]]bis(benzene-1,4-disulphonate)

Test animals / tissue source

Species:

rabbit

Strain:

other: Porton strain

Details on test animals or tissues and environmental conditions:

Animals:
Healthy Neu Zealand White rabbits (Porton strain) aged 11-12 weeks with average body weights of 2.43 kgs. (M) and 2.20 kgs. (F), bred on the premises and maintained under a semi-barrier system. Six rabbits (3 M and 3 F) were acclimatised in the test area for one week prior to the start of the trial.

Husbandry:
The rabbits were caged singly in an experimental room maintained at a temperature of 20 °C. (+/- 1 °C) and a relative humidity of 50-70 % Animals were exposed to artificial light for 10 hours daily from 08.00 -10 .00 hours. A commercial irradiated diet (Styles-Oxoid) was fed ad lib. Sterile filtered water was available at all times.

Test system

Vehicle:

not specified

Amount / concentration applied:

100 mg

Duration of treatment / exposure:

None

Observation period (in vivo):

The rabbits were examined 1, 24, 48 and 72 hours after application of the test compound and for any further period that was considered necessary.

Number of animals or in vitro replicates:

6 animals

Details on study design:

The test was carried out in accordance with the procedure set out in the "Hazardous Substances Regulations" under the U.S.A. Federal Hazardous Substances Labelling Act Sect. 191.12 (February 1965), with slight modification.

The eyes of the experimental animals were examined and found normal prior to the test. 100 mg of the test compound were instilled into the conjunctival sac of the left eye. The eyelids were then held closed for 1 second. The right eye served as a control. After 30 seconds the compound was, as far as possible, flushed out of the eyes of three of the rabbits with warm water for 1 minute. The rabbits were examined 1, >24, 48 and 72 hours after application of the test compound and for any further period that was considered necessary. The ocular reactions were scored by the method described in "Appraisal of the Safety of Chemicals in Food Drugs and Cosmetics" page 51, published by the Association of Food and Drug Officials of the U.S.A.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0.55

Max. score:

4

Reversibility:

fully reversible within: 6 days

Results 2

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

1.66

Max. score:

2

Reversibility:

not fully reversible within: 10 days

Results 3

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

1.3

Max. score:

3

Reversibility:

fully reversible within: 10 days

Results 4

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

A mild to severe conjunctival reaction was seen in all treated eyes one hour after application of the compound. This had improved slightly by 24 hours. The unwashed eyes did not show very much change after this until day 10 when small areas of necrosis were seen in the nictitating membranes. These were still present at termination (day 14). The washed eyes however, had returned to normal by day 6 (2/3) and day 10 (1/3). A corneal reaction consisting of slight opacity and damage to the superficial epithelium was seen in all treated eyes at one hour. This was still present at 24 hours and thereafter, the eyes returned to normal by 48 hours (2/3 washed eyes), 72 hours (2/3 unwashed eyes) and 6 days (1/3 washed, 1/3 unwashed eyes). Increased folding of the iris was seen in all unwashed eyes at 24 hours. These had returned to normal by 48 hours (1/3 and 72 hours (2/3)).

Other effects:

None

Any other information on results incl. tables

None

Applicant's summary and conclusion

Interpretation of results:

other: Not classified as irritant according to the CLP Regulation (EC) No.1272/2008

Conclusions:

the substance is classified as not an eye irritant.

Executive summary:

A key study was performed to determine the eye irritation for the substance according to Hazardous Substances Regulations" under the U.S.A. Federal Hazardous Substances Labelling Act Sect. 191.12 (February 1965), with slight modification. 100 mg of the test compound were instilled into the conjunctival sac of the left eye. The eyelids were then held closed for 1 second. The right eye served as a control. After 30 seconds the compound was, as far as possible, flushed out of the eyes of three of the rabbits with warm water for 1 minute. The rabbits were examined and the scored for the effects on eye 1, 24, 48 and 72 hours after application of the test compound and for any further period that was considered necessary.

 

A mild to severe conjunctival reaction was seen in all treated eyes one hour after application of the compound. This had improved slightly by 24 hours. The unwashed eyes did not show very much change after this until day 10 when small areas of necrosis were seen in the nictitating membranes. These were still present at termination (day 14). The washed eyes however, had returned to normal by day 6 (2/3) and day 10 (1/3).

 

A corneal reaction consisting of slight opacity and damage to the superficial epithelium was seen in all treated eyes at one hour. This was still present at 24 hours and thereafter, the eyes returned to normal by 48 hours (2/3 washed eyes), 72 hours (2/3 unwashed eyes) and 6 days (1/3 washed, 1/3 unwashed eyes). Increased folding of the iris was seen in all unwashed eyes at 24 hours. These had returned to normal by 48 hours (1/3) and 72 hours (2/3).

 

The testing has been carried out with eyes of 3 rabbits washed after 30 seconds, while remaining 3 rabbits never received any washing. However, the modern guidelines correctly recommend "If the solid test substance has not been removed from the eye of the test animal by physiological mechanisms at the first observation time point of 1 hour after treatment, the eye may be rinsed with saline or distilled water." Further the washing/flushing the eyes after exposure is a more realistic scenario for workers, as such an exposure will lead to an attempt to flush out the substance entered in the eye as soon as possible. Hence, not washing the eyes at all is considered to be an elevated testing condition, and the results obtained with such an exposure are not considered for hazard assessment.

 

In line with the above argument, the findings of necrosis of the nictitating membranes of unwashed eyes seen in this study, is not considered for the hazard conclusion. The scoring for washed eyes is below the threshold of the classification, hence substance is classified as not an eye irritant.

 

Comment:

Since the study compared also a cohort that was instilled with the solid test material remaining in the eye and a cohort that was instilled with the test material for about 1 minute followed by a flush out procedure (as recommended by OECD guideline 405) results form animals of the first cohort have not been taken into account for hazard assessment since this test regimen represent an elevated testing condition while noone is able to judge if the scorings and effects observed are due to the intrinic properties of the test item or due to a mechanical effect of the crystals remaining in the conjunctival sac of the eye. In addition the wash-out procedure represents a more realistic scenario to assess risk of eye damage to workers, professionals and consumers (in case applicable at all).