Phosphonium, tetraphenyl-, salt with 4,4'-sulfonylbis[phenol], compd. with 4,4'-sulfonylbis[phenol] (2:2:1)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

The study was performed between the 03 February 2004 and 03 March 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2004

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

not specified

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

484-440-2

EC Name:

-

Cas Number:

502157-74-6

Molecular formula:

C84H68O12S3P2

IUPAC Name:

4-(4-hydroxybenzenesulfonyl)phenol bis(tetraphenylphosphanium) bis(4-(4-hydroxybenzenesulfonyl)benzen-1-olate)

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

Description, Identification and Storage Conditions
Sponsor's identification SBCAT-03
Description white powder
Batch number UK030622B
Date received 09 October 2003
Storage conditions room temperature in the dark


Data relating to the identity, purity and stability of the test material are the responsibility of the Sponsor

For the purpose of the study the test material was used as supplied.
The absorption of the test material was not determined.

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Three New Zealand White rabbits were supplied by David Percival Ltd, Moston, Sandbach, Cheshire, UK. At the start of the study the animals were in the weight range of 2.0 to 3.5 kg and were twelve to twenty weeks old. After an acclimatisation period of at least 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 the cage label.
The animals were individually housed in suspended metal cages. Free access to mains drinking water and food (Certified Rabbit Diet (Code 5322) supplied by BCM IPS Limited, London, UK) was allowed throughout the study. The diet and drinking water were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.
The temperature and relative humidity were set to achieve limits of 17 to 23°C and 30 to 70% respectively. Any occasional deviations from these targets were considered not to have affected the purpose or integrity of the study. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.
The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

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 approximately 72 mg (as measured by gently compacting the required volume into an adapted syringe.

Duration of treatment / exposure:

The upper and lower eyelids were held togetlter for about one second immediately after treatment, to prevent loss of the test material, and then released. The left eye remained untreated and was used for control purposes.
Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment.
Additional observations were made on Days 5, 7 and 14 to assess the reversibility of the ocular
effects.

Observation period (in vivo):

Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment. Additional observations were made on Days 5, 7 and 14 to assess the reversibility of the ocular effects.

Number of animals or in vitro replicates:

3

Details on study design:

The study was performed to assess the irritancy potential of the test material following a single
application to the rabbit eye. The method was designed to meet the requirements of the following:
OECD Guidelines for the Testing of Chemicals No. 405 "Acute Eye Irritation/Corrosion"
(adopted 24 April 2002)
Commission Directive 92/69/EEC Method B5 Acute Toxicity (Eye Irritation)
The albino rabbit has been shown to be a suitable model for this type of study and is
recommended in the test method. The results of the study are believed to be of value in predicting
the likely eye irritancy potential of the test material to man.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

48 h

Score:

2

Max. score:

20

Reversibility:

fully reversible within: 14 days

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

48 h

Score:

0

Max. score:

80

Reversibility:

fully reversible

Results 3

Irritation parameter:

iris score

Basis:

animal #1

Time point:

48 h

Score:

0

Max. score:

10

Reversibility:

fully reversible

Results 4

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

48 h

Score:

14

Max. score:

20

Reversibility:

not reversible

Results 5

Irritation parameter:

iris score

Basis:

animal #2

Time point:

48 h

Score:

0

Max. score:

10

Reversibility:

not reversible

Results 6

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

48 h

Score:

0

Max. score:

80

Reversibility:

not reversible

Results 7

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

48 h

Score:

18

Max. score:

20

Reversibility:

not reversible

Results 8

Irritation parameter:

iris score

Basis:

animal #3

Time point:

48 h

Score:

5

Max. score:

10

Reversibility:

not reversible

Results 9

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

48 h

Score:

2

Max. score:

80

Reversibility:

not reversible

Results 10

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible

Results 11

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

48 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Results 12

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Results 13

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24 h

Score:

2

Max. score:

4

Reversibility:

not reversible

Results 14

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

48 h

Score:

2

Max. score:

4

Reversibility:

not reversible

Results 15

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

72 h

Score:

2

Max. score:

4

Reversibility:

not reversible

Results 16

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24 h

Score:

3

Max. score:

4

Reversibility:

not reversible

Results 17

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

48 h

Score:

3

Max. score:

4

Reversibility:

not reversible

Results 18

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

72 h

Score:

2

Max. score:

4

Reversibility:

not reversible

Results 19

Irritation parameter:

conjunctivae score

Basis:

animal #1

Remarks:

7 male

Time point:

24 h

Score:

6

Max. score:

20

Reversibility:

not specified

Results 20

Irritation parameter:

conjunctivae score

Basis:

animal #1

Remarks:

7 male

Time point:

72 h

Score:

2

Max. score:

20

Reversibility:

not specified

Results 21

Irritation parameter:

conjunctivae score

Basis:

animal #2

Remarks:

61 Male K, K = Animal killed for humane reasons immediately after 14-Day observation

Time point:

24 h

Score:

10

Max. score:

20

Reversibility:

not specified

Results 22

Irritation parameter:

conjunctivae score

Basis:

animal #2

Remarks:

61 Male K, K = Animal killed for humane reasons immediately after 14-Day observation

Time point:

72 h

Score:

12

Max. score:

20

Reversibility:

not specified

Results 23

Irritation parameter:

conjunctivae score

Basis:

animal #3

Remarks:

83 Male K*, K* = Animal killed for humane reasons immediately after 5-Day observation

Time point:

24 h

Score:

18

Max. score:

20

Reversibility:

not specified

Results 24

Irritation parameter:

conjunctivae score

Basis:

animal #3

Remarks:

83 Male K*, K* = Animal killed for humane reasons immediately after 5-Day observation

Time point:

72 h

Score:

14

Max. score:

20

Reversibility:

not specified

Results 25

Irritation parameter:

cornea opacity score

Basis:

animal #1

Remarks:

7Male

Time point:

24 h

Score:

0

Max. score:

80

Reversibility:

not specified

Results 26

Irritation parameter:

cornea opacity score

Basis:

animal #1

Remarks:

7Male

Time point:

72 h

Score:

0

Max. score:

80

Reversibility:

not specified

Results 27

Irritation parameter:

cornea opacity score

Basis:

animal #2

Remarks:

61 Male K, K = Animal killed for humane reasons immediately after 14-Day observation

Time point:

24 h

Score:

0

Max. score:

80

Reversibility:

not specified

Results 28

Irritation parameter:

cornea opacity score

Basis:

animal #2

Remarks:

61 Male K, K = Animal killed for humane reasons immediately after 14-Day observation

Time point:

72 h

Score:

15

Max. score:

80

Reversibility:

not specified

Results 29

Irritation parameter:

cornea opacity score

Basis:

animal #3

Remarks:

83 Male K*, K* = Animal killed for humane reasons immediately after 5-Day observation

Time point:

24 h

Score:

20

Max. score:

80

Reversibility:

not specified

Results 30

Irritation parameter:

cornea opacity score

Basis:

animal #3

Remarks:

83 Male K*, K* = Animal killed for humane reasons immediately after 5-Day observation

Time point:

72 h

Score:

30

Max. score:

80

Reversibility:

not specified

Results 31

Irritation parameter:

iris score

Basis:

animal #3

Remarks:

83 male K*, K* = Animal killed for humane reasons immediately after 5-Day observation

Time point:

72 h

Score:

5

Max. score:

10

Reversibility:

not specified

Results 32

Irritation parameter:

iris score

Basis:

animal #3

Remarks:

83 Male K*, K* = Animal killed for humane reasons immediately after 5-Day observation

Time point:

24 h

Score:

5

Max. score:

10

Reversibility:

not specified

Results 33

Irritation parameter:

iris score

Basis:

animal #2

Remarks:

61 male K, K = Animal killed for humane reasons immediately after 14-Day observation

Time point:

72 h

Score:

5

Max. score:

10

Reversibility:

not specified

Results 34

Irritation parameter:

iris score

Basis:

animal #2

Remarks:

61 Male K, K = Animal killed for humane reasons immediately after 14-Day observation

Time point:

24 h

Score:

0

Max. score:

10

Reversibility:

not specified

Results 35

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

7 male

Time point:

72 h

Score:

0

Max. score:

10

Reversibility:

not specified

Results 36

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

7Male

Time point:

24 h

Score:

0

Max. score:

10

Reversibility:

not specified

Results 37

Irritation parameter:

maximum mean total score (MMTS)

Basis:

mean

Time point:

14 d

Score:

40.5

Max. score:

40.5

Reversibility:

not specified

Irritant / corrosive response data:

Individual and group mean scores for ocular irritation are given in "Any other information on results".
White residual test material was noted in all treated eyes during the study.
Diffuse corneal opacity was noted in one treated eye at the 24-hour observation with translucent
corneal opacity at the 48, 72-hour and 5-day observations. Diffuse corneal opacity was noted in
one other treated eye at the 72-hour observation with translucent corneal opacity at the 7-day
observation and opalescent corneal opacity at the 14-day observation. Vascularisation,
generalised ingrowth of vessels for 3 to 4 mm in length, was also noted in this treated eye at the
14-day observation.
Iridial inflammation was noted in one treated eye at the 24, 48, 72-hour and 5-day observations.
Iridial inflammation was noted in one other treated eye at the 72-hour, 7 and 14-day observations.
Moderate conjunctival irritation was noted in all treated eyes one hour after treatment. Severe
conjunctival irritation was noted in one treated eye, moderate conjunctival irritation was noted in
one other treated eye and minimal conjunctival irritation was noted in the remaining treated eye at
the 24 and 48-hour observations. Moderate conjunctival irritation was noted in two treated eyes
with minimal conjunctival irritation in the remaining treated eye at the 72-hour observation.
Moderate conjunctival irritation was noted in one treated eye at the 5-day observation. Severe
conjunctival irritation was noted in one treated eye with minimal conjunctival irritation in the
remaining treated eye at the 7-day observation. Severe conjunctival irritation persisted in one
treated eye at the 14-day observation.
Areas of haemorrhage along the edge of the nictitating membrane and the lower conjunctival
membranes was noted in one treated eye at the 24, 48, 72-hour and 5-day observations and in one
other treated eye at the 7 and 14-day observations. Pale appearance of the nictitating membrane
was noted in one treated eye at the 48, 72-hour and 5-day observations and in one other treated
eye at the 7 and 14-day observations. Blood stained discharge was noted in one treated eye at the
5-day observation.
One treated eye appeared normal at the 14-day observation.
Due to blood stained discharge and signs of pain and discomfort one animal was killed for
humane reasons in accordance with current UK Home Office guidelines on Day 5. Due to signs
of pain and discomfort one other animal was killed for humane reasons in accordance with current UK Home Office guidelines on Day 14.

Any other information on results incl. tables

Individual Scores and Individual Total Scores for Ocular Irritation

Rabbit Number and Sex	7Male						61 Male K						83 Male K*
	IPR=3						IPR=0 +						PR=0 +
Time After Treatment	1 Hr	24 Hr	48 Hr	72 Hr	7 Dy	14 Dy	1 Hr	24 Hr	48 Hr	72 Hr	7 Dy	14 Dy	1 Hr	24 Hr	48 Hr	72 Hr	5 Dy
CORNEA E = Degree of Opacity F = Area of Cornea Involved	0 0	0 0	0 0	0 0	0 0	0 0	0 0	0 0	0 0	1 3	2 4	3V 4	0 0	1 4	2 3	2 3	2 4
Score (Ex F) x 5	0	0	0	0	0	0	0	0	0	15	40	60	0	20	30	30	40
IRIS D	0	0	0	0	0	0	0	0	0	1	1	1	0	1	1	1	1
Score (D x 5)	0	0	0	0	0	0	0	0	0	5	5	5	0	5	5	5	5
CONJUNCTIV AE A= Redness B = Chemosis C = Discharge	2Re 1 1	1 1 1	1 0 0	1 0 0	1 0 0	0 0 0	1Re 2 1	1 2 2	2 2 3	2 2 2	2HP 3 3	2HP 3 3	2Re 2 1	3ReH 3 3	3HP 3 3	2HP 2 3	2H°P 2 3Bs
Score (A + B + C) x 2	8	6	2	2	2	0	8	10	14	12	16	16	10	18	18	14	14
Total Score	8	6	2	2	2	0	8	10	14	32	61	81	10	43	53	49	59

 

IPR= Initial pain reaction

+ = One drop of local anaesthetic instilled into both eyes I to 2 minutes before treatment

Hr = Hour Dy = Day

Re= White residual test material in treated eye V = Vascularisation, generalised ingrowth of vessels for 3 to 4 mm in length

Bs =Bloodstained discharge

H = Areas of haemorrhage along the edge of the nictitating membrane and on the lower conjunctiva] membrane

P = Pale appearance of the nictitating membrane

H 0 = Larger more numerous areas of haemorrhage scattered over the nictitating membrane and lower conjunctiva] membrane

K* = Animal killed for humane reasons immediate] after 5-Da observation

K = Animal killed for humane reasons immediate] after 14-Da observation

 

Individual Total Scores and Group Mean Scores for Ocular Irritation

Rabbit Number and Sex	Individual Total Scores At:
	1 Hour	24 Hours	48 Hours	72 Hours	5 Days	7 Days	14 Days
7 Male	8	6	2	2	-	2	0
61 Male	8	10	14	32	-	61	81
83 Male	10	43	53	49	59	*	*
Group Total	26	59	69	83	59	63	81
Group Mean Score	8.7	19.7	23.0	27.7	-	31.5	40.5

 

- - Not applicable

* - Animal dead

 

Modified Kay and Calandra Interpretation of Eye Irritation Test

MAXIMUM MEAN SCORE		PERSISTENCE OF SCORE	DESCRIPTION RATING (AND SCORE)
0 to 0.5	Group mean total score at 24 hours= 0   Group mean total score at 24 hours > 0		Non-irritant (1)   Practically non-irritant (2)
0.5 to 2.5	Group mean total score at 24 hours = 0   Group mean total score at 24 hours > 0		Practically non-irritant (2)   Minimal irritant (3)
2.5 to 15	Group mean total score at 48 hours= 0   Group mean total score at 48 hours > 0		Minimal irritant (3)   Mild irritant (4)
15 to 25	Group mean total score at 72 hours= 0   Group mean total score at 72 hours> 0		Mild irritant (4)   Moderate irritant (5)
25 to 50	Group mean total score at 7 days 20 or less                     Group mean total score >20	More than half of the individual total scores at 7 days 10 or less   More than half of the individual total scores less at 7 days > 10 but no individual total score at 7 days >30   More than half of the individual total scores at 7 days > 10 and any individual score at 7 days >30	Moderate irritant (5)             Severe irritant (6)       Severe irritant (6)
50 to 80	Group mean total score at 7 days 40 or less             Group mean total score at 7 days > 40	More than half of the individual total scores at 7 days 30 or less   More than half of the individual total scores at 7 days > 30 but no individual total scores at 7 days > 60.   More than half of the individual total scores at 7 days > 30 and individual total score at > 60	Severe irritant (6)         Severe irritant (6)       Very severe irritant (7)     Very severe irritant (7)
80 to 100	Group mean total score at 7 days 80 or less               Group mean total score at 7 days > 80	More than half of the individual scores at 7 days 60 or less   More than half of the individual scores at 7 days > 60 but no individual total score at 7 days > 100   More than half the individual scores at 7 days > 60 and individual total score at 7 days > 100	Very severe irritant (7)     Very severe irritant (7)           Extremely severe irritant (8)       Extremely severe irritant (8)
100 to 110	Group mean total score at 7 days 80 or less   Group mean total score at 7 davs > 80		Very severe irritant (7)     Extremely severe irritant (8)

 

 

Draize Scale for Scoring Ocular Irritation

 

CONJUNCTIVAE
(A) Redness (refers to palpebral and bulbar conjunctivae excluding cornea and iris)
Vessels normal										0
Vessels definitely injected above normal										1
More diffuse, deeper crimson red, individual vessels not easily discernible										2
Diffuse beefy red										3
(B) Chemosis
No swelling										0
Any swelling above normal (includes nictitating membrane)										1
Obvious swelling with partial eversion of lids										2
Swelling with lids about half closed										3
Swelling with lids half closed to completely closed										4
(C) Discharge
No discharge										0
Any amount different from normal (does not include small amounts observed										1
in inner can thus of normal animals)
Discharge with moistening of the lids and hairs just adjacent to lids										2
Discharge with moistening of the lids and hairs a considerable area around the eye										3
THE TOTAL SCORE= (A+ B + C) x 2										MAXIMUM TOTAL= 20
lRIS
(D) Values
Normal										0
Folds above normal, congestion, swelling, circumcomeal injection
(any or all of these or combination of any thereof) iris still reacting to light										1
(sluggish reaction is positive)
No reaction to light, haemorrhage, gross destruction (any or all of these)										2
THE TOTAL SCORE= D x 5										MAXIMUM TOTAL= 10
CORNEA
(E) Degree of Opacity (most dense area used)
No opacity										0
Scattered or diffuse areas, details of iris clearly visible										1
Easily discernible translucent areas, details of iris slightly obscured										2
Opalescent areas, no details of iris visible, size of pupil barely discernible										3
Opaque, iris not discernible through the opacity										4
(F) Area of Cornea Involved
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
THE TOTAL SCORE= (Ex F) x 5										MAXIMUM TOTAL= 80
MAXIMUM TOTAL SCORE POSSIBLE= 110

 

Initial Pain Reaction

 

Class	Reaction by Animal	Descriptive Rating
0	No response	No initial pain
1	A few blinks only, normal within one or two minutes	Practically no initial pain
2	Rabbit blinks and tries to open eye, but reflex closes it	Slight initial pain
3	Rabbit holds eye shut and puts pressure on lids, may rub eye with paw	Moderate initial pain
4	Rabbit holds eye shut vigorously, may squeal	Severe initial pain
5	Rabbit holds eye shut vigorously, may squeal, claw at eye, jump and try to escape	Very severe initial pain

 

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

The test material produced a maximum group mean score of 40.5 and was classified as at least a SEVERE IRRITANT (CLASS 6 ON A 1 TO 8 SCALE) to the rabbit eye according to a modified Kay and Calandra classification system.

Executive summary:

The study was performed to assess the irritancy potential of the test material to the eye of the New Zealand White rabbit. The method was designed to meet the requirements of the following:
• OECD Guidelines for the Testing of Chemicals No. 405 "Acute Eye Irritation/Corrosion"
(adopted 24 April 2002)
• Commission Directive 92/69/EEC Method BS Acute Toxicity (Eye Irritation)

A single application of the test material to the non-irrigated eye of three rabbits produced translucent to opalescent corneal opacity, iridial inflammation and moderate to severe conjunctival irritation. Other ocular effects noted were vascularisation, with a generalised ingrowth of vessels for 3 to 4 mm in length, areas of haemorrhage on the nictitating and lower conjunctival membranes, pale appearance of the nictitating membrane and blood stained discharge. One treated eye appeared normal at the 14-day observation. Two animals were killed for humane reasons on Day 5 or 14, due to the severity of the reactions, in accordance with current UK Home Office guidelines.

The test material produced a maximum group mean score of 40.5 and was classified as at least a severe irritant (Class 6 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system.