Disodium phthalate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 5 November 2012 to 27 December 2012.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD guideline and EU method. GLP study.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Disodium phthalate

EC Number:

240-106-6

EC Name:

Disodium phthalate

Cas Number:

15968-01-1

Molecular formula:

C8H6O4.2Na

IUPAC Name:

disodium phthalate

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): Phthalic acid sodium salt (disodium phthalate)
- Physical state: white powder
- Analytical purity: > 95%
- Storage condition of test material: room temperature in the dark

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories UK Ltd., Leicestershire, UK.
- Age at study initiation: twelve to twenty weeks old.
- Weight at study initiation: 2.49 to 2.94 kg
- Housing: The animals were individually housed in suspended cages.
- Diet (e.g. ad libitum): 2930C Teklad Global Rabbit diet, ad libitum.
- Water (e.g. ad libitum): drinking water, ad libitum.
- Acclimation period: At least five days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 to 23°C
- Humidity (%): 30-70%
- Air changes (per hr): at least fifteen changes per hour.
- Photoperiod (hrs dark / hrs light): twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): A volume of 0.1 ml of the test item, which was found to weigh approximately 71 mg (as measured by gently compacting the required volume into an adapted syringe).

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 7 and 14 to assess the reversibility of the ocular effects.

Number of animals or in vitro replicates:

Three males

Details on study design:

SCORING SYSTEM:
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 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 a considerable area around the eye 3

IRIS
(D) Values
Normal 0
Folds above normal, congestion, swelling, circumcorneal injection (any or all of these or combination of any thereof) iris still reacting to light (sluggish reaction is positive) 1
No reaction to light, haemorrhage, gross destruction (any or all of these) 2

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

TOOL USED TO ASSESS SCORE: Examination of the eye was facilitated by the 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:

other: 24-72 h

Score:

0

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

other: 24-72 h

Score:

0

Results 3

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

other: 24-72 h

Score:

0

Results 4

Irritation parameter:

iris score

Basis:

animal #1

Time point:

other: 24-72 h

Score:

0

Results 5

Irritation parameter:

iris score

Basis:

animal #2

Time point:

other: 24-72 h

Score:

0

Results 6

Irritation parameter:

iris score

Basis:

animal #3

Time point:

other: 24-72 h

Score:

0

Results 7

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

other: 24-72 h

Score:

1.67

Max. score:

2

Reversibility:

fully reversible within: 14 days

Results 8

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

other: 24-72 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 14 days

Results 9

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

other: 24-72 h

Score:

1.33

Max. score:

2

Reversibility:

fully reversible within: 14 days

Results 10

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: 24-72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 14 days

Results 11

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: 24-72 h

Score:

1.67

Max. score:

2

Reversibility:

fully reversible within: 14 days

Results 12

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

other: 24-72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 14 days

Irritant / corrosive response data:

No corneal effects were noted during the study.

Iridial inflammation was noted in one treated eye one hour after treatment.

Severe conjunctival irritation was noted in one treated eye and moderate conjunctival irritation was noted in two treated eyes one hour after treatment. Moderate conjunctival irritation was noted in two treated eyes and minimal conjunctival irritation was noted in one treated eye at the 24-Hour observation. Moderate conjunctival irritation was noted in one treated eye and minimal conjunctival irritation was noted in two treated eyes at the 48 and 72-Hour observations with minimal conjunctival irritation noted in all treated eyes at the 7-Day observation.

A small area of petechial haemorrhage on the nictitating membrane was noted in one treated eye at the 24-Hour observation and in one other treated eye at the 48 and 72-Hour observations.

All treated eyes appeared normal at the 14-Day observation.

Other effects:

All animals showed expected gain in bodyweight during the study.

Any other information on results incl. tables

Table 1 Individual Scores and Individual Total Scores for Ocular Irritation

Rabbit Number and Sex	72650 Male						72715 Male						72794 Male
	IPR = 2						IPR = 2						IPR = 2
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	7 Dy	14 Dy
CORNEA
E = Degree of Opacity	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
F = Area of Cornea Involved	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Score (E x F) x 5	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
IRIS
D	1	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Score (D x 5)	5	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
CONJUNCTIVAE
A = Redness	2	2	2Pt	1Pt	1	0	2	2	2	2	1	0	2	2Pt	1	1	1	0
B = Chemosis	2	1	1	1	1	0	3	2	2	1	1	0	2	1	1	1	1	0
C = Discharge	2	1	0	0	0	0	3	1	1	1	0	0	3	0	0	0	0	0
Score (A + B + C) x 2	12	8	6	4	4	0	16	10	10	8	4	0	14	6	4	4	4	0
Total Score	17	8	6	4	4	0	16	10	10	8	4	0	14	6	4	4	4	0

IPR = Initial pain reaction

Hr = Hour(s)

Dy = Days

Pt = Small area of petechial haemorrhage on the nictitating membrane

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test item does not meet the criteria for classification as eye irritant.

Executive summary:

The study was performed to assess the irritancy potential of the test item to the eye of the New Zealand White rabbit. The method was designed to be compatible with OECD Guideline No. 405 “Acute Eye Irritation/Corrosion” and Method B5 Acute Toxicity (Eye Irritation) of Commission Regulation (EC) No. 440/2008. A single application of the test item to the non-irrigated eye of three rabbits

produced iridial inflammation and moderate to severe conjunctival irritation. A small area of petechial haemorrhage on the nictitating membrane was also noted in two treated eyes. All treated eyes appeared normal at the 14-Day observation. The test item does not meet the criteria for classification as eye irritant.