Reaction mass of sodium sulphate and disodium dodecanedioate and Dodecanoic acid, 12-amino-, sodium salt (1:1)

Endpoint summary

Description of key information

-Skin irritation: The test item is slightly irritant for skin but not classified. An in vitro skin corrosion (OECD 431, Episkin model) and a OECD 404 studyare available (in vivo test on rabbits).
-Eye irritation: The test item is hightly irritant for eyes and is classified. A OECD 405 study is available (in vivo test on rabbits).

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:

28 June 2010 - Sept.2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Compliant to GLP and testing guideline (observed deviation was not considered to have compromised the validity of the results and study); adequate coherence between data, comments and conclusions.

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

temperature recorded in the animal room was sometimes outside of the target ranges specified in the study plan (up to 24°C)

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

temperature recorded in the animal room was sometimes outside of the target ranges specified in the study plan (up to 24°C)

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: Breeder: Grimaud Frères Selection S.A.S., La Corbière, Roussay, France
- Age at study initiation: on the day of treatment, the animals were 2 to 4 months old
- Weight at study initiation:on the day of treatment, the animals had a mean body weight ± standard deviation of 2.7 ± 0.2 kg
- Housing: the animals were individually housed in Pajon cages (50 cm x 57 cm x 75 cm)
- Diet (e.g. ad libitum): free access to breeding pelleted diet “type 110”
- Water (e.g. ad libitum): tap water (filtered using a 0.22 µm filter) contained in bottles
- Acclimation period: at least 5 days before the beginning of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 +/- 3°C
- Humidity (%): 50 +/- 20%
- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air
- Photoperiod (hrs dark / hrs light): 12 h / 12 h

IN-LIFE DATES: From: 29 June 2010 To: 26 July 2010.

Type of coverage:

semiocclusive

Preparation of test site:

other: clipped

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 mL of undiluted test item

Duration of treatment / exposure:

3 minutes, 1 hour, 4 hours.

Observation period:

For each exposure period, cutaneous reactions were evaluated approximately 1 hour, 24, 48 and 72 hours after removal of the dressing and then
daily until the reversibility of cutaneous reactions.

Number of animals:

Three males.

Details on study design:

TEST SITE
- Area of exposure: anterior left flank (application for 3 minutes), anterior right flank (application of 1 hour), posterior right flank (application for
4 hours).
- % coverage: 6 cm2
- Type of wrap if used: gauze pad held in place by a non-irritation semi occlusive dressing and a restraining bandage

REMOVAL OF TEST SUBSTANCE
- Washing (if done): any residual test item was wiped off by means of a moistened cotton pad
- Time after start of exposure: the test item was placed on a dry gauze pad, which was then applied to an area of the skin of approximately 6 cm2 of
the animals. The gauze pad was held in place by a non-irritation semi occlusive dressing and a restraining bandage.
For the 4-hour exposure, a dry gauze pad was applied to the opposite flank, which was acted as control.
After required period of contact with the skin, the dressings were removed.

SCORING SYSTEM:
Dermal irritation was evaluated for each animal according to the following scoring scale:

Erythema and eschar formation:
. no erythema 0
. very slight erythema (barely perceptible) 1
. well-defined erythema 2
. moderate to severe erythema 3
. severe erythema (beet redness) to slight eschar formation (injuries in depth) 4

Edema formation
. no edema 0
. very slight edema (barely perceptible) 1
. slight edema (edges of area well-defined by definite raising) 2
. moderate edema (raised approximately 1 millimeter) 3
. severe edema (raised more than 1 millimeter and extending beyond area of exposure) 4

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: 24 h, 48 h, 72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: day 8

Remarks on result:

other: Exposure of 3 minutes

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: 24 h, 48 h, 72 h

Score:

0

Max. score:

4

Remarks on result:

other: Exposure of 3 minutes

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: 24 h, 48 h, 72 h

Score:

1.3

Max. score:

4

Reversibility:

fully reversible within: day 8

Remarks on result:

other: Exposure of 1 hour

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: 24 h, 48 h, 72 h

Score:

0

Max. score:

4

Remarks on result:

other: Exposure of 1 hour

Irritation parameter:

erythema score

Basis:

mean

Remarks:

3 rabbits

Time point:

other: 24 h, 48 h, 72 h

Score:

0.9

Max. score:

4

Reversibility:

fully reversible within: day 9

Remarks on result:

other: Exposure of 4 hours. Individual scores are : 1.0 - 0.0 - 1.7

Irritation parameter:

edema score

Basis:

mean

Remarks:

3 rabbits

Time point:

other: 24 h, 48 h, 72 h

Score:

0

Max. score:

4

Remarks on result:

other: Exposure of 4 hours

Irritant / corrosive response data:

After a 3 minute exposure, a very slight erythema was observed from day 1 until day 7.
After a 1 hour exposure, a very slight to well-defined erythema was observed from day 1 to day 7
After a 4 hours exposure (three animals), a very slight erythema was observed on day 1 only in one animal, and from day 1 to day 4 in a second one. In the third treated animal, a well-defined erythema was observed from day 1 to 3, decreasing to a very slight erythema from day 4 to day 8.

Other effects:

Dryness of the skin was also noted:
After a 3-minute exposure (animal No. 569), from day 5 to 7;
After a 1 hour exposure (animal No. 569) on days 6 and 7
After a 4-hour exposure, from day 2 to day 11 in 1/3 animal.

After a 1-hour exposure (animal No. 569)
A dryness of the skin was noted on days 6 and 7.

After a 4-hour exposure (three animals)
Dryness of the skin was also noted in this last animal, from day 2 to day 11.

Interpretation of results:

slightly irritating

Remarks:

Migrated information but not classified Criteria used for interpretation of results: other: Regulation EC no.1272/2008 (CLP)

Conclusions:

Under the experimental conditions of this study, the test item, Reaction mass of sodium sulfate, sodium amino-12-dodecanoate and sodium dodecanoedioate, (batch No. T710/712) was slightly irritant when applied topically to rabbits.

Executive summary:

The objective of this study was to evaluate the potential of the test item, Reaction mass of sodium sulfate, sodium amino-12-dodecanoate and sodium dodecanoedioate, to induce skin irritation following a single topical application to rabbits according to OECD (No. 404, 24th April 2002) and Commission Regulation (EC) (No. 440/2008, B.4, 30 May 2008) guidelines.

The study was conducted in compliance with the principles of Good Laboratory Practice Regulations.

 

Methods

The test item, Reaction mass of sodium sulfate, sodium amino-12-dodecanoate and sodium dodecanoedioate, was first applied for periods of 3 minutes, 1 hour and 4 hours to a single male New Zealand White rabbit.

Since the test item was not severely irritant or corrosive to the skin on this first animal, it was then applied for 4 hours simultaneously to two other animals.

 

The quantity of the test item administered was 0.5 mL. The test item was placed on a dry gauze pad, which was then applied to an area of the skin of approximately 6 cm² of the animals. The gauze pad was held in place by a non-irritation semi-occlusive dressing and a restraining bandage. After required period of contact with the skin, the dressings were removed.

 

For each exposure period, cutaneous reactions were evaluated approximately 1 hour, 24, 48 and 72 hours after removal of the dressing and then daily until the reversibility of cutaneous reactions.

Body weight was recorded at the beginning and the end of the observation period. On completion of the observation period, the animals were sacrificed then discarded without macroscopic post‑mortem examination.

The mean values of the scores for erythema and edema were calculated for each animal.

 

Results

After a 3 -minute exposure (one animal)

A very slight erythema was observed from day 1 until day 7. Dryness of the skin was also noted from day 5 to 7.

 

After a 1 -hour exposure (one animal)

A very slight to well-defined erythema was observed from day 1 to day 7, associated with a dryness of the skin on days 6 and 7.

 

After a 4-hour exposure (three animals)

A very slight erythema was observed on day 1 only in one animal, and from day 1 to day 4 in a second one. In the third treated animal, a well-defined erythema was observed from day 1 to 3, decreasing to a very slight erythema from day 4 to day 8. Dryness of the skin was also noted in this last animal, from day 2 to day 11.

 

Mean scores over 24, 48 and 72 hours for each animal were 1.0, 0.0 and 1.7 for erythema and 0.0, 0.0 and 0.0 for edema.

Conclusion

Under the experimental conditions of this study, the test item Reaction mass of sodium sulfate, sodium amino-12-dodecanoate and sodium dodecanoedioate (batch No. T7110/712) was slightly irritant when applied topically to rabbits.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (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 07 July 2010 to ...........

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Compliant to GLP and testing guideline; adequate coherence between data, comments and conclusions.

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:

not specified

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Breeder: Grimaud Frères Sélection S.A.S., La Corbière, Roussay, France
- Age at study initiation: 2 to 4 months
- Weight at study initiation: mean body weight ± standard deviation of 2.7 ± 0.2 kg
- Housing: Pajon cages
- Diet (e.g. ad libitum): 112C pelleted diet
- Water (e.g. ad libitum): drinking water filtered by a FG Millipore membrane
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 +/- 3°C
- Humidity (%): 50 ± 20%
- Air changes (per hr): 12 cycles/hour
- Photoperiod (hrs dark / hrs light): 12 h / 12 h

IN-LIFE DATES: From 20 July 2010 to 31 August 2010

Vehicle:

unchanged (no vehicle)

Controls:

other: For each rabbit, the right eye, which remained untreated, served as control.

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL.

Duration of treatment / exposure:

Single application to the eye.

Observation period (in vivo):

22 days

Number of animals or in vitro replicates:

Three males.

Details on study design:

SCORING SYSTEM: Draize scale.

TOOL USED TO ASSESS SCORE: UV lamp and 0 5% of sodium fluorescein solution.

Irritation parameter:

conjunctivae score

Remarks:

(Chemosis)

Basis:

mean

Remarks:

3 rabbits

Time point:

other: 24, 48 and 72 h

Score:

2.76

Max. score:

3

Reversibility:

fully reversible within: 21 days

Remarks on result:

other: Individual score = 2.3 + 3 + 3

Irritation parameter:

conjunctivae score

Remarks:

(discharge)

Basis:

mean

Remarks:

3 rabbits

Time point:

other: 24, 48 and 72 hours

Score:

1.33

Max. score:

3

Reversibility:

fully reversible within: 21 days

Remarks on result:

other: Individual scores = 1 + 1.7 + 1.3

Irritation parameter:

cornea opacity score

Remarks:

(opacity)

Basis:

mean

Remarks:

3 rabbits

Time point:

other: 24, 48 and 72 hours

Score:

2.77

Max. score:

4

Reversibility:

fully reversible within: 21 days

Remarks on result:

other: Individual scores = 3 + 2.3 + 3

Irritation parameter:

cornea opacity score

Remarks:

(area)

Basis:

mean

Remarks:

3 rabbits

Time point:

other: 24, 48 and 72 hours

Score:

3.04

Max. score:

4

Reversibility:

fully reversible within: 21 days

Remarks on result:

other: Individual scores = 2.7 + 3.7 + 2.7

Irritant / corrosive response data:

First animal (No. 524)
From day 1 until day 13, very slight toa moderate or marked conjunctival reactions (very slight to moderate chemosis (grade 2 or 3) was noted and
the evaluation of redness of the conjunctiva was masked by a black discoloration of the eye. Then, slight chemosis and redness of the conjunctiva
(grade 1) were observed from day 14 until day 20.
A clear discharge was noted on days 2 and 3.
Evaluation of iris lesions was masked by a black discoloration of the eye from day 1 until day 4. Then, iris lesions (grade 1) were noted until day 8.
Evaluation of corneal opacity was masked by a black discoloration on day 1. Then, a slight to severe corneal opacity (grades 1 to 3), which covered the whole cornea between day 5 and day 8) was noted from day 2 until day 12.
A brownish substance was noted from day 2 until day 22, associated with neovascularisation on day 22., very slight or slight redness of the
conjunctiva and clear to whitish purulent discharge) were observed in all animals from day 1: some of these reactions persisted up to day 10 at the
latest.
A slight iritis was noted in all animals on day 2; it persisted up to day 8, 9 or 10.
A slight corneal opacity was recorded in all animals on day 2; it persisted up to day 9 or 17.
Neovascularisation was noted in all animals, from day 3 or day 5 up to day 8, 10 or 12.

Second animal (No. 514)
A moderate or marked chemosis (grade 2 or 3) was noted from day 1 until day 9. Then, a slight chemosis (grade 1) was noted until day 13.
Evaluation of redness of the conjunctiva was masked by a black discoloration of the eye on days 1 and 2. Then, a slight or moderate redness of the
conjunctiva (grade 1 or 2) was noted until day 6
A clear discharge was noted from day 2 until day 4.
Evaluation of iris lesions and corneal opacity were masked by a black discoloration of the eye on day 1. Then, iris lesions (grade 1) were noted until
day 6 and a slight to severe corneal opacity (grades 1 to 3), which covered the whole cornea Evaluation of iris lesions and corneal opacity were
masked by a black discoloration of the eye on day 1. Then, iris lesions (grade 1) were noted until day 6 and a slight to severe corneal opacity
(grades 1 to 3), which covered the whole cornea on days 2 and 3) was noted from day 2 until day 8.
A brownish substance was noted from day 2 until day 6, followed by a neovascularisation on days 8 and 9.

Third animal (No. 522)
A moderate or marked chemosis was noted from day 1 until day 8. Then, a slight chemosis (grade 1) was noted until day 22.
Evaluation of redness of the conjunctiva was masked by a black discoloration of the eye on day 1. A moderate or severe redness of the conjunctiva
(grade 2 or 3) was noted from day 2 until day 6. Then, a slight redness of the conjunctiva (grade 1) was noted until day 12.
A clear discharge was noted on days 2 and 3.
Evaluation of iris lesions was masked by a black discoloration of the eye on day 1. Then, iris lesions (grade 1) were noted until day 5.
Evaluation of corneal opacity was masked by a black discoloration on day 1. Then, a slight to severe corneal opacity (grades 1 to 3) was noted from
day 2 until day 7.
A neovascularisation was noted from day 3 until day 6 and a brownish substance was noted from day 4 until day 22.

Mean scores calculated over 24, 48 and 72 hours were:
- for chemosis: 2.3, 3.0 and 3.0 for animals Nos.524, 514 and 522, respectively,
- for corneal opacity: 3.0, 2.3 and 3.0 for each animal Nos.524, 514 and 522, respectively,
- for redness of the conjunctiva: 3.0 for animal No. 522,
- for the iris lesions: 1.0 for animals Nos. 514 and 522.
Due to eye discoloration, the mean scores were not calculable in the animals Nos. 524 and 514 for redness of the conjunctiva and in the animal No. 524 for the iris lesions.

Table : Individual ocular examinations and mean values f the scores recorded for each animal (24, 48/ and 72 hours).

 

Rabbit number	Region of eye	Description of ocular reactions	Scores				Mean score irritation^2	Fully reversible
			1h D1	24h D2	48h D3	72h D4
524	Conj^1	Chemosis	3	3	2	2	2.3	Yes (D21)
		Redness	LC	LC	LC	LC	-	Yes (D21)
		Discharge	E	2	1	0	1.0	Yes (D4)
	Iris		LC	LC	LC	LC	-	Yes (D9)
	Cornea	Intensity	LC	3	3	3	3.0	Yes (D13)
		Area	LC	2	3	3	2.7	Yes (D13)
514	Conj^1	Chemosis	3	3	3	3	3.0	Yes (D14)
		Redness	LC	LC	2	2	-	Yes (D7)
		Discharge	E	3	1	1	1.7	Yes (D5)
	Iris		LC	1	1	1	1.0	Yes (D7)
	Cornea	Intensity	LC	3	2	2	2.3	Yes (D9)
		Area	LC	4	4	3	3.7	Yes (D9)
522	Conj^1	Chemosis	3	3	3	3	3.0	No at D22
		Redness	LC	3	3	3	3.0	Yes (D12)
		Discharge	E	2	2	0	1.3	Yes (D4)
	Iris		LC	1	1	1	1.0	Yes (D6)
	Cornea	Intensity	LC	3	3	3	3.0	Yes (D13)
		Area	LC	3	3	2	2.7	Yes (D13)

 

¹Conj = Conjunctivae

²Mean score on days 2, 3 and 4

E = Scoring masked by residual test item

LC = reading masked by a lack discoloration of the eye

Interpretation of results:

highly irritating

Remarks:

Migrated information Criteria used for interpretation of results: other: Regulation EC n°1272/2008

Conclusions:

The test item, Reaction mass of sodium sulfate, sodium amino-12-dodecanoate and sodium dodecanoedioate, (batch No. T710/712) was severely
irritant when administered by ocular route to rabbits.

Executive summary:

The objective of this study was to evaluate the potential of the test item, Reaction mass of sodium sulfate, sodium amino-12-dodecanoate and sodium dodecanoedioate, to induce irritation following a single ocular administration in rabbitsaccording to OECD (No. 405, 24th April 2002) andCommission Regulation (EC) (No. 440/2008, B.5, 30 May 2008)guidelines.

The study was conducted in compliance with the principles of Good Laboratory Practice Regulations.

 

Methods

The test item, Reaction mass of sodium sulfate, sodium amino-12-dodecanoate and sodium dodecanoedioate, was first administered to a single male New Zealand White rabbit.

Since the test item was irritant on this first animal, it was then evaluated in sequential manneron two other animals.

The quantity of the test item administered was 0.1 mL.

For animal Nos. 514 and 522, a local anaesthetic was used prior to treatment.

The test item was introducedinto the conjunctival sac of the left eye. The right eye, which remained untreated, served as control.

The eyes were not rinsed after administration of the test item.

Ocular reactions were observed approximately 1 hour, 24, 48 and 72 hours after the administration and then daily until the reversibility of the ocular reactions (day 14; one animal) or the end of the observation period (two animals).

Body weight was recorded at the beginning and the end of the observation period. On completion of the observation period, the animals were sacrificed then discarded without macroscopicpost‑mortemexamination.

 

The mean values of the scores for chemosis, redness of the conjunctiva, iris lesions and corneal opacity were calculated for each animal.

The interpretation of results was carried out according to the classification criteria laid down in Council Directive 67/548/EEC (and subsequent adaptations).

 

Results

Slight to marked chemosis was noted in all animals from day 1 until day 13, 20 or 22.

Evaluation of redness of the conjunctiva was masked by a black eye discoloration in all animals on day 1 or from day 1 until day 2 or 13. Then slight to severe redness of the conjunctiva was noted until day 6, 12 or 20.

A clear discharge was noted in all animals, from day 2 until 3 or 4.

Evaluation of iris lesion was masked by a black eye discolorationin all animals on day 1 (two animals) or from day 1 until day 4 (one animal). Then, iris lesions were noted until day 5, 6 or 8. The evaluation of corneal opacity was masked by a black eye discoloration on day 1. Then, slight to marked corneal opacity, which covered the whole cornea in two animal during 2 or 4 consecutive days, was noted until day 7, 8 or 12.

Brownish area was noted in all animals from day 2 or 3 until day 6 (one animal) or 22 (two animals). Neovascularisation was observed on days 8 and 9 (one animal) from day 3 until day 6 (one animal) or on day 22 (one animal).

Mean scores calculated over 24, 48 and 72 hours were:

.            for chemosis: 2.3, 3.0 and 3.0 for each animal,

.            for corneal opacity: 3.0, 2.3 and 3.0 for each animal,

.            for redness of the conjunctiva: 3.0 for animal No. 522,

.            for the iris lesions: 1.0 for animals No. 514 and 522.

Due to eye discoloration, the mean scores were not calculable in the animals Nos. 524 and 514 for redness of the conjunctiva and in the animal No. 524 for the iris lesions.

 

Conclusion

The test item, Reaction mass of sodium sulfate, sodium amino-12-dodecanoate and sodium dodecanoedioate, was severely irritant when administered by ocular route to rabbits.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Additional information

In vitro skin corrosion (OECD 431, Episkin model):

The purpose of this test is to evaluate the corrosivity potential of the test item using the EPISKIN^TMin vitro Reconstituted Human Epidermis (RHE) Model after treatment periods of 3, 60 and 240 minutes. This method was designed to meet the requirements of the following OECD Guideline for the Testing of Chemicals No. 431 “In Vitro Skin Corrosion: Human Skin Model Test” (adopted 13 April 2004)

The relative mean viability of the test item treated tissues was: 123.2% after 3 minutes of exposure , 93.9% at 60 minutes of exposure and 68.7% after 240 minutes of exposure. The quality criteria required for acceptance of results in the test were satisfied.

The test item was considered to be Non-Corrosive to the skin according to the regulation EC no.1272/2008 (CLP).

 

Skin irritation study (OECD 404):

The test item was evaluate to induce skin irritation following a single topical application to rabbits according to OECD 404. The test item was first applied for periods of 3 minutes, 1 hour and 4 hours to a single male New Zealand White rabbit. For each exposure period, cutaneous reactions were evaluated approximately 1 hour, 24, 48 and 72 hours after removal of the dressing and then daily until the reversibility of cutaneous reactions.

Body weight was recorded at the beginning and the end of the observation period. On completion of the observation period, the animals were sacrificed then discarded without macroscopicpost‑mortem examination.

The mean values of the scores for erythema and edema were calculated for each animal.

After a 3 -minute exposure (one animal): A very slight erythema was observed from day 1 until day 7. Dryness of the skin was also noted from day 5 to 7.

After a 1 -hour exposure (one animal): A very slight to well-defined erythema was observed from day 1 to day 7, associated with a dryness of the skin on days 6 and 7.

After a 4-hour exposure (three animals): A very slight erythema was observed on day 1 only in one animal, and from day 1 to day a second one. In the third treated animal, a well-defined erythema was observed from day 1 to 3, decreasing to a very slight erythema from day 4 to day 8. Dryness of the skin was also noted in this last animal, from day 2 to day 11.

Mean scores over 24, 48 and 72 hours for each animal were 1.0, 0.0 and 1.7 for erythema and 0.0, 0.0 and 0.0 for edema.

Under the experimental conditions of this study, the test item Reaction mass of sodium sulfate, sodium amino-12-dodecanoate and sodium dodecanoedioate was slightly irritant when applied topically to rabbits, but it’s not classified according to the regulation EC no.1272/2008.

 

 

Eye irritation study (OECD 405):

The test item,Reaction mass of sodium sulfate, sodium amino-12-dodecanoate and sodium dodecanoedioate,was first administered to a single male New Zealand White rabbit. The test item was introduced into the conjunctival sac of the left eye. The right eye, which remained untreated, served as control. The eyes were not rinsed after administration of the test item. Ocular reactions were observed approximately 1 hour, 24, 48 and 72 hours after the administration and then daily until the reversibility of the ocular reactions (day 14; one animal) or the end of the observation period (two animals). Body weight was recorded at the beginning and the end of the observation period. On completion of the observation period, the animals were sacrificed then discarded without macroscopic post‑mortem examination.

The mean values of the scores for chemosis, redness of the conjunctiva, iris lesions and corneal opacity were calculated for each animal.

 Slight to marked chemosis was noted in all animals from day 1 until day 13, 20 or 22. Evaluation of redness of the conjunctiva was masked by a black eye discoloration in all animals on day 1 or from day 1 until day 2 or 13. Then slight to severe redness of the conjunctiva was noted until day 6, 12 or 20. A clear discharge was noted in all animals, from day 2 until 3 or 4.

Evaluation of iris lesion was masked by a black eye discolorationin all animals on day 1 (two animals) or from day 1 until day 4 (one animal). Then, iris lesions were noted until day 5, 6 or 8. The evaluation of corneal opacity was masked by a black eye discoloration on day 1. Then, slight to marked corneal opacity, which covered the whole cornea in two animal during 2 or 4 consecutive days, was noted until day 7, 8 or 12.

Brownish area was noted in all animals from day 2 or 3 until day 6 (one animal) or 22 (two animals). Neovascularisation was observed on days 8 and 9 (one animal) from day 3 until day 6 (one animal) or on day 22 (one animal).

Mean scores calculated over 24, 48 and 72 hours were:

.            for chemosis: 2.3, 3.0 and 3.0 for each animal,

.            for corneal opacity: 3.0, 2.3 and 3.0 for each animal,

.            for redness of the conjunctiva: 3.0 for animal No. 522,

.            for the iris lesions: 1.0 for animals No. 514 and 522.

Due to eye discoloration, the mean scores were not calculable in the animals Nos. 524 and 514 for redness of the conjunctiva and in the animal No. 524 for the iris lesions.

 To sum up, the test item, Reaction mass of sodium sulfate, sodium amino-12-dodecanoate and sodium dodecanoedioate, was severely irritant when administered by ocular route to rabbits.

Effects on skin irritation/corrosion: slightly irritating

Effects on eye irritation: highly irritating

Justification for classification or non-classification

Skin irritation:

According to EU Directive 67/584/EEC and EU regulation (EC) No 1272/2008 (CLP), the Reaction mass of sodium sulfate, sodium amino-12-dodecanoate and sodium dodecanoedioate is not classified for skin irritation.

Justification (CLP): Mean score of erythema (and/or edema) is not equal or higher to 2.3 (maximal score = 4 for each score).

Eye irritation:

The Reaction mass of sodium sulfate, sodium amino-12-dodecanoate and sodium dodecanoedioate is classified as highlty irritating in Category 1 (H318, causes serious eye damage) according to EU regulation (EC) No 1272/2008 (CLP), and R41 "Causes of serious damage to eyes" according to EU Directive 67/584/EEC.

Justification (CLP): At least in 2 of 3 tested animals, a positive response of cornea opacity higher or equal to 3 is calculated as the mean scores following grading at 24, 48 and 72 hours after the installation of the test material.