Disodium phthalate

Endpoint summary

Description of key information

Key study: Skin irritation. OECD guideline 404 and EU method B.4. GLP study.
Based on the available results, the test substance is not classified as skin irritating.
Key study: Eye irritation OECD guideline 405 and EU method B.5. GLP study.
Based on the available results, the test substance is not classified as eye irritating.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 23 October 2012 to 02 November 2012.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD guideline and EU method. GLP study.

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system 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: 2.27 or 2.51 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.

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

other: moistened sufficiently with 0.5 ml of distilled water to achieve a paste

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): At each test site a quantity of 0.5 g of the test item.

Duration of treatment / exposure:

4 hours

Observation period:

1, 24, 48 and 72 hours

Number of animals:

Two males.

Details on study design:

TEST SITE
- Area of exposure: 2.5 x 2.5 cm
- Type of wrap if used: Each cotton gauze patch was secured in position with a strip of surgical adhesive tape. To prevent the animal interfering with the patches, the trunk of the rabbit was wrapped in an elasticated corset for the duration of the exposure period.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Any residual test item was removed by gentle swabbing with cotton wool soaked in distilled water.
- Time after start of exposure: 4 hours

SCORING SYSTEM:
Erythema and Eschar Formation Value
No erythema .............................................................................................................. ............... 0
Very slight erythema (barely perceptible) ..............................................................................1
Well-defined erythema ............................................................................................................. 2
Moderate to severe erythema ................................................................................................. 3
Severe erythema (beef redness) to eschar formation preventing grading of erythema 4

Oedema Formation Value
No oedema ............................................................................................................................................ 0
Very slight oedema (barely perceptible) ........................................................................................... 1
Slight oedema (edges of area well-defined by definite raising) .................................................... 2
Moderate oedema (raised approximately 1 millimetre) ...................................................................3
Severe oedema (raised more than 1 millimetre and extending beyond the area of exposure) 4

Any other skin reactions and clinical signs of toxicity, if present, were also recorded.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: 24-72 h

Score:

0

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

other: 24-72 h

Score:

0

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: 24-72 h

Score:

0

Irritation parameter:

edema score

Basis:

animal #2

Time point:

other: 24-72 h

Score:

0

Irritant / corrosive response data:

No evidence of skin irritation was noted during the study.

Other effects:

Both animals showed expected gain in bodyweight during the study.

Table 2 Individual Skin Reactions Following 4-Hour Exposure

Skin Reaction	Observation Time (following patch removal)	Individual Scores – Rabbit Number and Sex		Total
		72599 Male	72631 Male
Erythema/Eschar Formation	Immediately	0	0	( 0 )
	1 Hour	0	0	( 0 )
	24 Hours	0	0	0
	48 Hours	0	0	( 0 )
	72 Hours	0	0	0
Oedema Formation	Immediately	0	0	( 0 )
	1 Hour	0	0	( 0 )
	24 Hours	0	0	0
	48 Hours	0	0	( 0 )
	72 Hours	0	0	0

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test item produced a primary irritation index of 0.0 and was classified as non-irritant to rabbit skin according to the Draize classification scheme. No corrosive effects were noted.

Executive summary:

The study was performed to assess the irritancy potential of the test item to the skin of the New Zealand White rabbit. The method was designed to be compatible with OECD Guideline No. 404 “Acute Dermal Irritation/Corrosion” and Method B4 Acute Toxicity (Skin Irritation) of Commission Regulation (EC) No. 440/2008. 3-minute and 1-hour semi-occluded applications of the test item to the intact

skin of one rabbit produced no evidence of skin irritation. A single 4-hour, semi-occluded application of the test item to the intact skin of two rabbits produced no evidence of skin irritation. The test item produced a primary irritation index of 0.0 and was classified

as non-irritant to rabbit skin according to the Draize classification scheme. No corrosive effects were noted.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

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.

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

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.

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.

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

other: 24-72 h

Score:

0

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

other: 24-72 h

Score:

0

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

other: 24-72 h

Score:

0

Irritation parameter:

iris score

Basis:

animal #1

Time point:

other: 24-72 h

Score:

0

Irritation parameter:

iris score

Basis:

animal #2

Time point:

other: 24-72 h

Score:

0

Irritation parameter:

iris score

Basis:

animal #3

Time point:

other: 24-72 h

Score:

0

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

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

other: 24-72 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 14 days

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

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: 24-72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 14 days

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

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.

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

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.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Key study: Skin irritation. OECD guideline 404 and EU method B.4. GLP study.

Based on the available results, the test substance is not classified as skin irritating.

Key study: Eye irritation OECD guideline 405 and EU method B.5. GLP study.

Based on the available results, the test substance is not classified as eye irritating.

Justification for selection of skin irritation / corrosion endpoint:
Only one study available. Klimisch 1 and the study was carried out in accordance with internationally valid GLP principles.

Justification for selection of eye irritation endpoint:
Only one study available. Klimisch 1 and the study was carried out in accordance with internationally valid GLP principles.

Justification for classification or non-classification

Based on the available data, the substance is not classified as skin or eye irritating.