Cuprate(4-), [C-(aminosulfonyl)-C-[[[2-[[4-chloro-6-[(2,5-disulfophenyl)amino]-1,3,5-triazin-2-yl]amino]ethyl]amino]sulfonyl]-29H,31H-phthalocyanine-C,C-disulfonato(6-)-N29,N30,N31,N32]-, sodium salts

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2 June 1993 to 6 June 1993.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1993

Report date:

1993

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

OECD Guidelines for Testing of Chemicals (1987) No. 405 "Acute Eye Irritation/Corrosion"

Deviations:

not specified

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

Method B5 in Commission Directive 92/69/EEC (which constitutes Annex V of Council Directive 67/ 548/EEC).

Deviations:

not specified

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Cuprate(4-), [C-(aminosulfonyl)-C-[[[2-[[4-chloro-6-[(2,5-disulfophenyl)amino]-1,3,5-triazin-2-yl]amino]ethyl]amino]sulfonyl]-29H,31H-phthalocyanine-C,C-disulfonato(6-)-N29,N30,N31,N32]-, sodium salts

EC Number:

689-188-9

Cas Number:

149343-84-0

Molecular formula:

C32H24N12O8S4Cu to C76H56N28O32S12Cl4Na8Cu

IUPAC Name:

Cuprate(4-), [C-(aminosulfonyl)-C-[[[2-[[4-chloro-6-[(2,5-disulfophenyl)amino]-1,3,5-triazin-2-yl]amino]ethyl]amino]sulfonyl]-29H,31H-phthalocyanine-C,C-disulfonato(6-)-N29,N30,N31,N32]-, sodium salts

Test material form:

solid: particulate/powder

Details on test material:

Identification: JPR Blue 100
Description: Blue powder
Chemical Name: Cuprate(2-),[2-[[4-chloro-6-[[2-[(29H,31H-phthalocyaninylsulphonyl)amino]ethyl]amino]-1,3,5-triazin-2-yl]amino]-1,4-benzenedisulphonato(4-)-N29,N30,N31,N32]-mono(or bis)aminosulphonyl mono (or di)sulpho derivs., sodium salts.
Lot Number: 303001
Purity: 95.2%
Major Impurities: H2o, NaCl, Na2SO4
Date Received: 30 March 1993
Container: Opaque plastic jar x 7
Storage conditions: Room temperature

Specific details on test material used for the study:

No further details specified in the study report.

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Specification
Three New Zealand White rabbits were supplied by David Percival Ltd., Maston, Sandbach, Cheshire, U.K. At the start of the study the animals weighed 2.61 - 3.09 g, and were approximately twelve to sixteen weeks old. After a minimum acclimatisation period of five days each animal was given a number unique within the study which was written with a black indelible marker-pen on the inner surface of the ear and on a cage label.

Husbandry
The animals were individually housed in suspended metal cages. Free access to mains drinking water and food (RABMA Rabbit Diet, Special Diet Services Ltd., Witham, Essex, U.K.) was allowed throughout the study.
The animal room was maintained at a temperature of 19 – 23 °C and relative humidity of 58 - 66 %. The rate of air exchange was approximately 15 changes per hour and the lighting was controlled by a time switch to give 12 hours light and 12 hours darkness.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

A volume of 0.1 ml of the test material, which was found to weigh approx imately 99 mg.

Duration of treatment / exposure:

Single exposure

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

3 animals

Details on study design:

Immediately before the start of the test, both eyes of the provisionally selected test rabbits were examined for evidence of ocular irritation or defect with the aid of a light source from a standard ophthalmoscope.
Animals showing evidence of ocular lesions were rejected and replaced.
One rabbit was initially treated. A volume of 0.1 ml of the test material, which was found to weigh approximately 99 mg (as measured by gently compacting the required volume into an adapted syringe) was placed into the conjunctival sac of the right eye, formed by gently pulling the lower lid away from the eyeball. The upper and lower eyelids were held together for about one second immediately after application, to prevent loss of the test material, and then released.
The left eye remained untreated and was used for control purposes.
Immediately after administration of the test material, an assessment of the initial pain reaction was made.
After consideration of the ocular responses produced in the first treated animal, two additional animals were treated. In order to minimise pain on application of the test material, one drop of local anaesthetic ("Ophthaine", 0.5% proxymetacaine hydrochloride, E.R. Squibb & Sons , Limited Hounslow, Middlesex, U.K.) was instilled into both eyes of these animals 1 - 2 minutes before treatment.
Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment, according to the numerical evaluation, (from Draize J.H. 1959, Association of Food and Drug Officials of the United States, Austin, Texas, "The Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics").
Any other ocular effects were also noted. Examination of the eye was facilitated by use of the light source from a standard ophthalmoscope.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 3

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 4

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 5

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 6

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 7

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 48 Hours

Results 8

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Results 9

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 48 hours

Results 10

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Results 11

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

7 d

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Results 12

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Irritant / corrosive response data:

No corneal or iridial effects were noted during the study.
Moderate conjunctival irritation was noted in all treated eyes one hour after treatment with minimal conjunctival irritation at the 24-hour observation. Minimal conjunctival redness was noted in one treated eye at the 48-hour observation.
No ocular effects were noted 72 hours after treatment.

Other effects:

Residual test material was noted around the treated eye of all animals throughout the study.
Blue-coloured staining was noted in all treated eyes throughout the study.

Any other information on results incl. tables

JPR BLUE 100: ACUTE EYE IRRITATION TEST IN THE RABBIT

INDIVIDUAL SCORES AND INDIVIDUAL TOTAL SCORES FOR OCULAR IRRITATION

Rabbit Number and Sex (Bodyweight Kg)	IPR = 3				IPR = 0                                  +				IPR = 0                                  +
	151 Female (3.09)				154 Male (2.61)				165 Female (2.83)
Time After Treatment	1 hr	24 hr	48 hr	72 hr	1 hr	24 hr	48 hr	72 hr	0 hr	24 hr	48 hr	72 hr
CORNEA E = Degree of Opacity F = Area of Opacity	0S 0	0S 0	0S 0	0S 0	0S 0	0S 0	0S 0	0S 0	0S 0	0S 0	0S 0	0S 0
Score (E x F) x 5	0	0	0	0	0	0	0	0	0	0	0	0
IRIS D	0S	0S	0S	0S	0S	0S	0S	0S	0S	0S	0S	0S
Score (D x 5)	0	0	0	0	0	0	0	0	0	0	0	0
CONJUNCTIVAE A = Redness B = Chemosis C = Discharge	2S 2 2Re	1S 1 0Re	0S 0 0Re	0S 0 0Re	2S 2 3Re	2S 1 0Re	1S 0 0Re	0S 0 0Re	2S 2 2Re	1S 1 0Re	0S 0 0Re	0S 0 0Re
Score (A + B + C) x 2	12	4	0	0	14	6	2	0	12	4	0	0
Total Score	12	4	0	0	14	6	2	0	12	4	0	0

Key:       hr = hour(s)          IPR = initial pain reaction  Re = residual test material around treated eye

               + = one drop of Ophthaine instilled into each eye 1 – 2 minutes before dosing   S = blue coloured staining

 

JPR BLUE 100: ACUTE EYE IRRITATION TEST IN THE RABBIT

INDIVIDUAL TOTAL SCORES AND GROUP MEAN SCORE FOR OCULAR IRRITATION

Rabbit Number and Sex	Individual Total Scores At:
	1 Hour	24 Hours	48 Hours	72 Hours
151 Female 154 Male 165 Female	12 14 12	4 6 4	0 2 0	0 0 0
Group Total	38	14	2	0
Group Mean Score	12.7 1 Hour	4.7 24 Hours	0.7 48 Hours	0.0 72 Hours

 

JPR BLUE 100: ACUTE EYE IRRITATION TEST IN THE RABBIT

INDIVIDUAL & MEAN SCORES FOR CORNEA, IRIS & CONJUNCTIVAE REQUIRED FOR EEC LABELLING REGULATIONS

Rabbit Number & Sex (Bodyweight Kg)	Time After Treatment	Corneal Opacity	Iridial Inflammation	Conjunctival Redness	Conjunctival Chemosis
151 Female (3.09)	24 Hours 48 Hours 72 Hours	0 0 0	0 0 0	1 0 0	1 0 0
Total		0	0	1	1
Mean		0.0	0.0	0.3	0.3
154 Male (2.61)	24 Hours 48 Hours 72 Hours	0 0 0	0 0 0	2 1 0	1 0 0
Total		0	0	3	1
Mean		0.0	0.0	1.0	0.3
165 Female (2.83)	24 Hours 48 Hours 72 Hours	0 0 0	0 0 0	1 0 0	1 0 0
Total		0	0	1	1
Mean		0.0	0.0	0.3	0.3

 

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The test material, JPR BLUE 100, produced a maximum group mean score of 12.7 and was classified as a MILD IRRITANT (CLASS 4 ON A 1 TO 8 SCALE) to the rabbit eye according to a modified Kay and Calandra classification system.
The test material did not produce positive criteria in any rabbit according to the EEC labelling regulations and was classified as NON IRRITANT to the rabbit eye. No symbol and risk phrase are therefore required.

Executive summary:

A study was performed to assess the irritancy potential of the test material to the eye of the New Zealand White rabbit. The method used followed that described in the OECD Guidelines for Testing of Chemicals (1987) No. 405 "Acute Eye Irritation/Corrosion" referenced as Method B5 in Commission Directive 92/69/EEC (which constitutes Annex V of Council Directive 67/ 548/EEC).

 

The results may be used as a basis for classification and labelling under Annex VI of Council Directive 67/548/EEC (as adapted to technical progress by Commission Directive 91/325/EEC).

 

A single application of the test material to the non-irrigated eye of three rabbits produced moderate conjunctival irritation. No ocular effects were noted 72 hours after treatment.

 

The test material produced a maximum group mean score of 12.7 and was classified as a mild irritant (Class 4 on a 1 to 8 scale) to the rabbit eye according to the Kay and Calandra classification system.

 

The test material was also classified as non-irritant according to EEC labelling regulations. No symbol and risk phrase are required.