29H,31H-phthalocyanine

Endpoint summary

Description of key information

- Skin irritation: not irritating

- Eye irritation: not irritating

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: US Federal Register 38 No 187 § 1500.41

Version / remarks:

24h occlusive exposure, investigation of hypoderm and underlying muscles

GLP compliance:

no

Specific details on test material used for the study:

Heliogengrün 9140

Species:

rabbit

Strain:

other: Albino

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: no data
- Age at study initiation: no data
- Weight at study initiation: 3-4kg
- Housing: single
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least three days

ENVIRONMENTAL CON DITIONS
- Temperature (°C): 21-23°C
- Humidity (%): 34 - 65%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

occlusive

Preparation of test site:

clipped

Remarks:

intact and scarified sites were tested on the same animal

Vehicle:

other: 50% Lutrol 400 (= PEG 400) in water

Remarks:

for moistioning

Controls:

other: yes, untreated skin sites

Amount / concentration applied:

0.5g per site (1g per animal)

Duration of treatment / exposure:

24h

Observation period:

7 days

Number of animals:

6

Details on study design:

TEST SITE
- Area of exposure: 6.25cm2
- Type of wrap if used: PVC foil

REMOVAL OF TEST SUBSTANCE
- Washing (if done): warm vehicle
- Time after start of exposure:24h

OBSERVATION TIME POINTS
24h and 72h for skin findings, 7 days for hypoderm and muscle

SCORING SYSTEM: Draize

Irritation parameter:

edema score

Basis:

mean

Time point:

24 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable, since no findings

Irritation parameter:

edema score

Basis:

mean

Time point:

72 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable, since there were no findings

Irritation parameter:

erythema score

Basis:

other: all animals

Time point:

24 h

Remarks on result:

not determinable

Remarks:

The green test material had stained the skin.

Irritation parameter:

erythema score

Basis:

mean

Time point:

72 h

Score:

0

Max. score:

0

Reversibility:

other: Not applicable, since no findings

Remarks on result:

other: For half of the animals, green staining of the skin interfered with scoring.

Irritant / corrosive response data:

At the end of treatment (24h reading), scoring for erythema was not possible as the green test item had stained the skin.
At the 72h reading, scoring was still not possible for 3 animals. For the other 3 animals, no erythema (score = 0) was observed. Findings were equal for intact and scarified skin.

Edema was not observed in any animal at any time point (Score = 0).

Other effects:

There were no adverse findings on the hypoderm and muscles underlying the treated skin sites.

Interpretation of results:

GHS criteria not met

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

- Synonyms: Copper tri/tetrachlorophthalocyanine pigment
- Substance type: blue powder
- Analytical purity: >85%
- Lot/batch No.: M200058
- Stability under test conditions: guaranteed for 4 hours
- Storage condition of test material: darkness at approx. 20 °C in a fume cupboard
- Other: different CAS No. are existing: 29719-96-8, 68987-63-3, 16040-69-0, 27614-71-7

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland GmbH, Kisslegg, Germany
- Weight at study initiation: 2.4 - 2.7 kg
- Housing: in fully air-conditioned rooms in separate cages arranged in a battery
- Diet: ssniff (R) K-H (V2333), ad libitum and hay (approx. 15 g daily)
- Water: tap water, ad libitum
- Acclimation period: 1 week under study conditions

ENVIRONMENTAL CONDITIONS
- Temperature: 20 +- 3 °C
- Humidity: 50 +- 20 %
- Photoperiod: 12 hrs dark / 12 hrs light

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

other: sesame oil

Controls:

other: untreated shaved skin served as control.

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.5 g test material, pasted with 0.4 ml sesame oil

Duration of treatment / exposure:

The exposure period was 4 hours.

Observation period:

72 hours after removal of the patches.

Number of animals:

3 females

Details on study design:

About 24 h before the start of the study the hair in the dorsal region of the body of the animals was removed with an electric clipper over an area of ca. 25 cm2. Only animals with intact skin were used.
Each animal was treated with the test material, the substance was evenly distributed over the whole surface of a 2.5 cm x 2.5 cm cellulose patch on a piece of surgical plaster (specially produced by Beiersdorf AG). The plaster was fixed to the prepared skin area and then covered with a semi-occlusive bandage.
The exposure period was 4 h, after the exposure all remnants of the test material were carefully removed from the skin with warm tap water.
Examinations of the skin took place 30 - 60 min as well as 24, 48 and 72 h after removal of the patches.
Erythema, eschar formation and edema were evaluated numerically, acc. to the score of Draize. All other changes of the skin were recorded.

Irritation parameter:

erythema score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

No signs of irritations were observed during the study.

Interpretation of results:

GHS criteria not met

Conclusions:

No signs of irritation were seen on the skin of the rabbits during the whole study period under the experimental conditions chosen here. Therefore, the test material has not to be classified according to GHS- or EU criteria.

Reference 3

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Meets generally accepted scientific standards; well documented study.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

; application time: 1, 5, 15 min and 20 h (OECD: 4 h); 2 animals used (OECD: 3); observation period 8 days (OECD: 14); skin changes were recorded on working days; dose ca. 1 g (OECD: 0.5 ml), additionally testing of the ear (not recommended)

Principles of method if other than guideline:

BASF-TEST:
Before OECD guideline 404 was established, skin irritation was tested using an internal method. White Vienna rabbits were used. Usually, 2 animals were treated for 1, 5, 15 min and 20 h using occlusive conditions. An application site of 2.5 cm x 2.5 cm was covered with the test substance. In addition, skin tissue from the ear was tested by wrapping the ear. These results from the ear, however, would not be taken into account for evaluation, as they do not represent testing of the dorsal/lateral flank of the back. After the application time, the skin was washed with water which sometimes contained a mild detergent. The animals were observed for 8 days. Skin reactions were recorded after 24 h and after 8 days of exposure. In the raw data, skin reactions were additionally recorded at reading time points 1 h, 48 h, 72 h, 96 h and 7 days after application.

GLP compliance:

no

Remarks:

GLP was not compulsory at the time the study was conducted.

Specific details on test material used for the study:

- Analytical purity: 100 %

Species:

rabbit

Strain:

Vienna White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Average weight at study initiation: ca. 2.7 kg

Type of coverage:

occlusive

Preparation of test site:

shaved

Vehicle:

water

Controls:

other: Untreated, clipped skin of the same animal served as control.

Amount / concentration applied:

Concentration: 50 %

Duration of treatment / exposure:

Dorsal skin: 1, 5, 15 min and 20 h

Observation period:

The animals were observed following exposure for 7 days on working days. Skin changes were recorded after removal of the bandage as well as after 1h, 24h, 48h, 72h, 96 h and after 7 and 8 days.

Number of animals:

2 (1 male and 1 female)

Details on study design:

A paste containing the moistened test substance (50 %) was applied to a 2.5 cm x 2.5 cm patch and placed on the clipped and intact skin of two white Vienna rabbits for 1 min, 5 min, 15 min (short term) and 20 h application time under occlusive conditions. The test substance was removed by washing once with 100 % Lutrol and subsequently with 50 % Lutrol in water. The areas exposed for 20 h were not washed. Skin reactions were recorded after 24h and after 7 days of exposure. In the raw data, skin reactions were additionally recorded at reading time points 1 h, 48 h and on days 5 and 7 after application. The application time used for assessment was 20 h, the test material was applied on dorsal skin, the corresponding reading time points used for assessment were 24, 48 and 72 hours after application. Dermal reactions were classified using a BASF-scoring system, which is convertible into the current OECD Draize scheme. For the evaluation the BASF scores given in the raw data were converted to Draize scores.
The results for ear tests are not described in detail here because they do not represent testing of the dorsal/lateral flank of the back and/or are not comparable to the guideline conditions.

Irritation parameter:

erythema score

Basis:

animal: #1, #2

Time point:

24/48 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal: #1, #2

Time point:

24/48 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

No formation of edema or erythema was observed at any time point. Substance residues were seen on the treated sites until day 7.

Interpretation of results:

GHS criteria not met

Reference 4

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: Code of Federal Regulations, Title 16, Section 1500.41

Deviations:

no

Principles of method if other than guideline:

6 male albino New Zealand white rabbits were used for each trial. After clipping, the skin of the left flank of the animals was left intact, the right flank was scarified. The animals were treated for 24 h using occlusive conditions. The application site, treated with the liquid test substance, was covered with a gauze pad and wrapped with adhesive tape. After the application time, the patches/tape were removed. The primary irritation index was evaluated after 25 and 72 hours after start of application of the test material. The animals were observed for one week, at day 7 another reading was made.

GLP compliance:

no

Specific details on test material used for the study:

- Analytical purity: ca. 98 %

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: at the start of the test, the animals weighed between 2.5 and 3.5 kg
- Housing: Rabbits were kept in individual cages (540 x 360 x 315 mm) or in restraining devices
- Diet: 200 g of food per animal per day (Granules Lapin "entretien", Sanders)
- Water: softened and filtrated water ad libitum, automatically given

ENVIRONMENTAL CONDITIONS
- Temperature (°C): minimum 20 °C
- Air changes: ventilation (renewal of air 12 times per hour )
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

occlusive

Preparation of test site:

other: The rabbits were clipped; the skin of the left flank was left intact ; the skin of the right flank was scarified.

Vehicle:

other: olive oil

Controls:

no

Amount / concentration applied:

0.5 g pure test material, applied in paste form (mixed with olive oil)

Duration of treatment / exposure:

24 hours

Observation period:

7 days

Number of animals:

6

Details on study design:

ADMINISTRATION/EXPOSURE
- The right flank was scarified with a sterile scalpel blade, making three parallel superficial incisions 2 cm long and 0.5 cm apart. The incisions were epidermal.
- The test substance was applied to the rabbit skin, using the right, previously scarified flank and the left intact one, at a rate of 0.5 g test substance per area and animal.
- Occlusion: A loaded gauze patch was applied to intact and scarified skin. The test substance and the gauze pads were kept in contact with the skin by a patch. Patches were then fastened to the body with adhesive tape.
- Vehicle: olive oil (0.5 g in 0.65 ml of neutralized sterilized olive oil per zone)
- Removal of the test substance: after 24 hrs
- Post exposure period: 7 days

EXAMINATION
- Observation period: after exposure at 25 h, at 72 h and 7 days
- Scoring system: readings were made of both scarified and no scarified zones, following the scale proposed by Draize. Only cutaneous reactions observed after treatment of intact skin are considered relevant for hazard assessment and are described in detail.

Irritation parameter:

erythema score

Basis:

animal: #1, #3, #5, #6

Time point:

other: 24-72 h

Score:

1.12

Max. score:

4

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: Slight desquamation was seen in 2 animals after 7 days; first reading time point was in fact 25 hours instead of 24 hours.

Irritation parameter:

edema score

Basis:

animal: #1, #3, #4, #5, #6

Time point:

other: 24-72 h

Score:

0.1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: first reading time point was in fact 25 hours instead of 24 hours.

Irritant / corrosive response data:

Treatment with the test substance resulted in only one out of 6 rabbits in very slight edema after 24 h. The evaluation of erythema after 24 hours was impossible in 2 rabbits due to treatment related colouring of the skin. Very slight to well defined erythema were noticed in 4 rabbits where irritation was reversible after 7 days. Slight desquamation was seen on intact skin in 2/6 animals after 7 days.

Table 1: OECD Draize scores for edema after 24 hours exposure to the test substance:

Rabbit No.		1	2	3	4	5	6
Erythema	25 h	0	*	1	*	2	0
	72 h	1	*	2	*	2	1
	7 d	S	§	0	0	0	S
Edema	25 h	0	0	0	0	1	1
	72 h	0	0	0	0	0	0
	7 d	0	§	0	0	0	0
S = slight desquamation;   C = Formation of a scab without underlying necrosis
* = Reading of erythema was impossible on two rabbits, because of green colouration through the test material
§ = Rabbit No. 2 died on day 6 and was not replaced

Interpretation of results:

GHS criteria not met

Reference 5

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

according to guideline

Guideline:

other: Code of Federal Regulations, Title 16, Section 1500, 42

Deviations:

no

Principles of method if other than guideline:

6 male albino New Zealand white rabbits were used for each trial. After clipping, the skin of the left flank of the animals was left intact, the right flank was scarified. The animals were treated for 24 h using occlusive conditions. The application site, treated with the liquid test substance, was covered with a gauze pad and wrapped with adhesive tape. After the application time, the patches/tape were removed. The primary irritation index was evaluated after 25 and 72 hours after start of application of the test material. The animals were observed for one week, at day 7 another reading was made.

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: at the start of the test, the animals weighed ca. 2.5 - 3.5 kg
- Housing: Rabbits were kept in individual cages (540 x 360 x 315 mm) or in restraining devices
- Diet: 200 g of food per animal per day (Granules Lapin "entretien", Sanders)
- Water: softened and filtrated water ad libitum, automatically given

ENVIRONMENTAL CONDITIONS
- Temperature: minimum 20 °C
- Air changes: ventilation (renewal of air 12 times per hour )
- Photoperiod: 12 hrs dark / 12 hrs light

Type of coverage:

occlusive

Preparation of test site:

clipped

Remarks:

the skin of the left flank was left intact ; the skin of the right flank was scarified.

Vehicle:

water

Controls:

no

Amount / concentration applied:

0.5 g pure test material made into a paste using 0.8 ml water

Duration of treatment / exposure:

24 hours

Observation period:

7 days

Number of animals:

6

Details on study design:

ADMINISTRATION/EXPOSURE
- The right flank was scarified with a sterile scalpel blade, making three parallel superficial incisions 2 cm long and 0.5 cm apart. The incisions were epidermal.
- The test substance was applied to the rabbit skin, using the right, previously scarified flank and the left intact one, at a rate of 0.5 g test substance per area and animal.
- Occlusion: A loaded gauze patch was applied to intact and scarified skin. The test substance and the gauze pads were kept in contact with the skin by a patch. Patches were then fastened to the body with adhesive tape.
- Removal of the test substance patches: after 24 hrs
- Post exposure period: 7 days

EXAMINATION
- Observation period; after exposure at 25 h, at 72 h and 7 days
- Scoring system: readings were made of both scarified and no scarified zones, following the scale proposed by Draize. Only cutaneous reactions observed after treatment of intact skin are considered relevant for hazard assessment and are described in detail.

Irritation parameter:

erythema score

Basis:

animal: #1, #2, #3, #4, #5, #6

Time point:

other: 24/72 h

Max. score:

4

Remarks on result:

not determinable

Remarks:

first reading time point was in fact 25 hours instead of 24 hours; scoring was not possible due to substance-related discolouration of the treated site.

Irritation parameter:

edema score

Basis:

animal: #1, #2, #3, #4, #5, #6

Time point:

other: 24/72 h

Score:

0

Max. score:

4

Remarks on result:

other: first reading time point was in fact 25 hours instead of 24 hours.

Table 1: Skin irritation parameters of rabbits treated on dorsal skin with the test material for 24 hours:

	Animal No.	1	2	3	4	5	6
Erythema	24 h	*	*	*	*	*	*
	72 h	*	*	*	*	*	*
	7 days	0	*	*	0	0	0
Edema	24 h	0	0	0	0	0	0
	72 h	0	0	0	0	0	0
	7 days	0	0	0	0	0	0

The blue colour of the test material made the macroscopic examination of the erythema impossible. Surplus material which ha dnot penetrated the skin was removed with a sterile gauze soaked in warm water. Reading was made 1 hour later. But: * = Exact reading and exact calculation of the irritation score remained impossible due to blue colouration.

Interpretation of results:

GHS criteria not met

Conclusions:

The test substance can be regarded as "non-irritant" to the skin according to GHS or EU criteria under the experimental conditions chosen.

Reference 6

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

according to guideline

Guideline:

other: Code of Federal Regulations, Title 16, Section 1500.41

Principles of method if other than guideline:

3 male albino New Zealand White rabbits were used. The animals were treated for 24 h using occlusive conditions. The application site, treated with the liquid test substance, was covered with a 2.5 cm x 2.5 cm gauze pad. After the application time, the patches were removed. The scores were evaluated after 24, 48 and 72 hours after application of the substance. The animals were observed for one week after application of the test material, at day 8 another reading was made.

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: W. Ostrop / Meerbusch
- Weight at study initiation: ca. 2.2 kg
- Housing: individual; 1 animal per cage
- Diet: mümmel z (ssniff/Soest), ad libitum
- Water: ad libitum
- Acclimation period: 4-6 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16-18
- Humidity (%): 40 %
- Photoperiod: daylight rhythm

Type of coverage:

occlusive

Preparation of test site:

clipped

Remarks:

skin was partially scarified; test substance was applied on intact and scarified skin.

Vehicle:

unchanged (no vehicle)

Controls:

other: Untreated, clipped skin of each animal served as a control.

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.5 ml
- Concentration: undiluted test substance

Duration of treatment / exposure:

24 hours

Observation period:

8 days

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: 2.5 cm x 2.5 cm
- Type of wrap if used: The material to be tested and the gauze pads were kept in contact with the skin by an adhesive tape (LeukoflexR); the torso was wrapped with an elastic bandage (ElastoflexR)
- The animals were immobilized

REMOVAL OF TEST SUBSTANCE
The skin was not washed out, only the patches were removed after 24 hours.

READING TIME POINTS:
24 h, 48 h and 72 h after application

READING TIME POINS USED FOR ASSESSMENT:
24 h and 72 h

SCORING SYSTEM:
Draize scores. Only cutaneous reactions observed after treatment of intact skin are considered relevant for hazard assessment and are described in detail.

Irritation parameter:

erythema score

Basis:

animal: #1, #2, #3

Time point:

other: 24/72h

Max. score:

4

Remarks on result:

not determinable

Remarks:

Scoring was not possible as the test material caused a colouration of the treates sites.

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: 24/72h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #2

Time point:

other: 24/72h

Score:

2

Max. score:

4

Reversibility:

not fully reversible within: 8 days

Irritation parameter:

edema score

Basis:

animal #3

Time point:

other: 24/72h

Score:

0.5

Max. score:

4

Reversibility:

fully reversible within: 8 days

Table 1: Summary of dermal effects in rabbits after dorsal application of    
the test material after 24 hours of application

Rabbit No.		1	2	3
Erythema	24 h	*	*	*
	72 h	*	*	*
	8 d	*	*	*
Edema	24 h	0	2	1
	72 h	0	2	0
	8 d	0	1	0

* = scoring not possible due to substance related discolouration of the trated skin

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of the test, the test substance has not to be classified according to EU or GHS criteria.

Reference 7

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: No detailled information on test material and results (individual score values) were given.

Principles of method if other than guideline:

The test was performed following a method from "Federal Register" § 191.11, 17 Sept 1964.

GLP compliance:

no

Specific details on test material used for the study:

- Analytical purity: no data given in report

Species:

rabbit

Strain:

other: Albino

Type of coverage:

occlusive

Preparation of test site:

other: The skin of the rabbits was clipped and skin was partially scarified; the test substance was applied on intact and scarified skin.

Vehicle:

other: polyethylene glycol

Controls:

not specified

Amount / concentration applied:

TEST MATERIAL
0.5 g of the powdered test material was mixed to a paste with polyethylene glycol before application.

Duration of treatment / exposure:

24 hours

Observation period:

72 hours

Number of animals:

no data given

Details on study design:

Irritation to the skin was measured by a patch test technique on the clipped intact and abraded skin. Ca. 0.5 g of the semi-solid test material was placed under a square patch such as a surgical gauze (ca. 2.5 cm x 2.5 cm, 2 layers thick). The animals were immobilized with patches secured in place by adhesive tape. The entire trunk of each animal was then wrapped with an impervious material for the 24 hours period of exposure. After 24 hours the patches were removed, the skin was not washed and the resulting reactions were evaluated on the basis of scores according to the method of Draize. A second reading was made at 48 hours after the first reading. Only cutaneous reactions observed after treatment of intact skin were considered relevant for hazard assessment.

Irritant / corrosive response data:

According to the author, no signs of irritation were seen during the 72 hours observation period at the intact skin of any animal.

Interpretation of results:

GHS criteria not met

Reference 8

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

; [application time: 1, 5, 15 min and 20 h (OECD: 4 h); 4 animals used (OECD: 3); observation period 8 days (OECD: 14); skin changes recorded on working days; dose ~1 ml (OECD: 0.5 ml), additionally testing of the ear (not recomm. by the OECD guideline]

Principles of method if other than guideline:

Before OECD guideline 404 was established, skin irritation was tested using an internal BASF method. White Vienna rabbits were used. Usually, 2 animals were treated for 1, 5, 15 min and 20 h using occlusive conditions. An application site of 2.5 cm x 2.5 cm was covered with the liquid test substance. In addition, skin tissue from the ear was tested by wrapping the ear. These results from the ear, however, would not be taken into account for evaluation as they do not represent testing of the dorsal/lateral flank of the back. After the application time, the skin was washed with water which sometimes contained a mild detergent. The animals were observed for 8 days. Skin reaction was recorded after 1 h, 24 h, 48 h, 72 h, 96 h and after 7 and 8 days.

GLP compliance:

no

Remarks:

GLP was not compulsory at the time the study was conducted.

Specific details on test material used for the study:

- Analytical purity: 100 %

Species:

rabbit

Strain:

Vienna White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Breed of Medical-Biological Reseach Laboratories of BASF
- Average weight at study initiation: ca. 2.75 kg
- Diet: Altromin-MS (Altrogge, Lage/L. Germany), ad libitum
- Water: ad libitum

Type of coverage:

occlusive

Preparation of test site:

shaved

Vehicle:

water

Controls:

no

Amount / concentration applied:

The test substance, moistured with water and mixed to a paste (concentration: 50 %), was applied to a 2.5 cm x 2.5 cm patch (~1 ml)

Duration of treatment / exposure:

20 hours

Observation period:

The animals were observed following exposure for 8 days on working days. Skin changes were recorded after removal of the bandage after 1 hour as well as after 24 h, 48 h, and on day 6 and 8.

Number of animals:

4 animals, 2 males and 2 females

Details on study design:

The test substance, moistured with water and mixed to a paste, was applied to a 2.5 cm x 2.5 cm patch (~1 ml) and placed on the clipped and intact skin of four white Vienna rabbits for 1 min, 5 min, 15 min and 20 h under occlusive conditions. The test substance was removed by washing once with 50 % Lutrol. Skin reactions were recorded after 1 h, 24 h, 48 h, and on day 6 and 8. The application time used for assessment was 20 hours, the test material was applied on dorsal skin. The corresponding reading time points used for assessment were 24 and 48 hours after application. Dermal reactions were classified using a BASF-scoring system, which is convertible into the current OECD Draize scheme. For the evaluation, the BASF scores given in the raw data were converted to Draize scores.

Irritation parameter:

erythema score

Basis:

animal: #1, #2, #3, #4

Time point:

24/48 h

Score:

0

Max. score:

0

Irritation parameter:

edema score

Basis:

animal: #1, #2, #3, #4

Time point:

24/48 h

Score:

0

Max. score:

0

Irritant / corrosive response data:

No formation of erythema or edema were observed (Draize score = 0) at any reading time point and after any application time.

Interpretation of results:

GHS criteria not met

Reference 9

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

- Batch no SF20927
- blue powder
- storage conditions: room temperature in the dark
- no data on purity available

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: David Percival Ltd, Moston, Sandbach, Cheshire, UK
- Age at study initiation: 12-20 weeks
- Weight at study initiation: 2 - 3.5kg
- Housing: individulally
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum):ad libitum
- Acclimation period: 5 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): 12/12

IN-LIFE DATES: From: June 1 2004 To: June 8 2004

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Controls:

not required

Amount / concentration applied:

0.5g

Duration of treatment / exposure:

4h

Observation period:

24, 48 and 72 hours

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: 2.5cm x 2.5 cm
- Type of wrap if used: surgical adhesive tape

REMOVAL OF TEST SUBSTANCE
- Washing (if done): gentle swabbing with cotton wool soaked in 74% Industrial Methylated Spirits
- Time after start of exposure:4h

OBSERVATION TIME POINTS
24, 48 and 72 hours

SCORING SYSTEM: Draize
- Method of calculation: according to OECD 404

Irritation parameter:

erythema score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

edema score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

0

Skin Reaction	Observation Time		IndividualScores-Rabbit Numberand sex			Total
			104 Male	105 Male	106 Male
	1 Hour		1 STA	1 STA	1 STA	( 3 )
Erythema/Eschar	24 Hours		1 STA	1 STA	1 STA	3
Formation	48 Hours		O STA	O STA	O STA	( 0 )
	72 Hours		O STA	O STA	O STA	0
	1 Hour		0	0	0	( 0 )
Oedema Formation	24 Hours		0	0	0	0
	48 Hours		0	0	0	( 0 )
	72 Hours		0	0	0	0
Sum of 24 and 72-hour Readings (S)		:	3
Primary Irritation Index (S/6)		:	3/6=0.5

STA = Blue colored staining of the skin

(  ) =Total values not used for calculation of primary irritation index

Interpretation of results:

GHS criteria not met

Conclusions:

Copper phthalocyanine is not irritating to rabbit skin.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Qualifier:

according to guideline

Guideline:

other: US FEDERAL REGISTER 38 NO. 187, Paragraph 1500.42 page 27019 of 27. SEPT. 1973

GLP compliance:

no

Specific details on test material used for the study:

Heliogengrün 9140

Species:

rabbit

Strain:

other: Albino

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: no data
- Age at study initiation: no data
- Weight at study initiation: 3 - 4 kg
- Housing: single
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 3 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-23°C
- Humidity (%): 45 - 65%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

100 mg

Duration of treatment / exposure:

single application

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

6

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no washing after application


SCORING SYSTEM: Draize
Time points investigated: 1h, 24h, 72h, and longer if needed

TOOL USED TO ASSESS SCORE: not specified

Irritation parameter:

cornea opacity score

Basis:

other: all six animals

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: Not applicable, since no findings

Irritation parameter:

iris score

Basis:

other: all six animals

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: Not applicable since no adverse findings

Irritation parameter:

conjunctivae score

Basis:

other: all six animals

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: Not applicable, since no adverse findings

Irritation parameter:

chemosis score

Basis:

other: all six animals

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: Not applicable since no adverse findings

Irritant / corrosive response data:

One hour after the application, two rabbits showed slight wetness of the treated eyes.

Interpretation of results:

GHS criteria not met

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Synonyms: Copper tri/tetrachlorophthalocyanine pigment
- Substance type: blue powder
- Analytical purity: >85%
- Lot/batch No.: M200058
- Stability under test conditions: guaranteed for 4 hours
- Storage condition of test material: darkness at approx. 20 °C in a fume cupboard
- Other: different CAS No. are existing: 29719-96-8, 68987-63-3, 16040-69-0, 27614-71-7

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland GmbH, Kisslegg, Germany
- Weight at study initiation: 2.6 - 2.9 kg
- Housing: in fully air-conditioned rooms in separate cages arranged in a battery
- Diet: ssniff (R) K-H (V2333), ad libitum and hay (approx. 15 g daily)
- Water: tap water, ad libitum
- Acclimation period: 1 week under study conditions

ENVIRONMENTAL CONDITIONS
- Temperature: 20 +- 3 °C
- Humidity: 50 +- 20 %
- Photoperiod: 12 hrs dark / 12 hrs light

Vehicle:

unchanged (no vehicle)

Controls:

other: the untreated eye served as control

Amount / concentration applied:

100 mg

Duration of treatment / exposure:

24 h

Observation period (in vivo):

72 h after administration of the test material

Number of animals or in vitro replicates:

3 females

Details on study design:

About 24 h before the start of the study the test eyes of all animals were examined under UV light for corneal lesions after instillation of one drop of a 0.01 % fluorescein-sodium solution. Only animals without ocular abnormalities were used for the study.
100 mg of the test material was administered once to the conjunctival sac of the left eyeof three animals. In each case the untreated eye served as control.
24 h after administration the treated eyes were washed out thoroughly with isotonic saline at approx. 37 °C.
The eyes were examined 1, 24, 48 and 72 h after administration of the test material. At 24 and 72 h, the eyes were further examined for corneal lesions under UV light after instillation of one drop of a 0.01 % fluorescein-sodium solution. Lesions in cornea, iris or conjunctivae were graded numerically acc. to Draize. All other changes or toxic effects were recorded when apparent.

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

chemosis score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

conjunctivae score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0.56

Max. score:

4

Reversibility:

fully reversible within: 72 h

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.67

Max. score:

4

Reversibility:

fully reversible within: 72 h

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 72 h

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.67

Max. score:

4

Reversibility:

fully reversible within: 72 h

One hour after administration the conjunctiva of one animal was very slightly swollen. Furthermore, all animals showed definitely injected blood vessels at one hour up to 48 h after administration. In addition to these observations, serious eye discharge discoloured by compound was noted. 72 h after administration all signs of irritation had disappeared.

Interpretation of results:

GHS criteria not met

Executive summary:

Slight signs of irritation (a mean score of 0.56 for 3 animals at 24 -48 -72 h for conjunctivae redness) were seen in the eyes of the rabbits which were fully reversible within 72 h under the experimental conditions chosen here. Therefore, the test material has not to be classified according to GHS- or EU criteria.

Reference 3

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Well documented and acceptable study which meets generally accepted scientific standards.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

[volume of 50 mg applied (OECD: volume of 0.1 ml); 2 animals were used (OECD: 3); observation period was 8 days (OECD: 21); examination using fluorescein (OECD: no use of fluorescein)]

GLP compliance:

no

Remarks:

GLP was not compulsory at the time the study was conducted.

Specific details on test material used for the study:

- Analytical purity: 100 %

Species:

rabbit

Strain:

Vienna White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 3.18 kg, 3.48 kg

Vehicle:

unchanged (no vehicle)

Controls:

other: the second eye of the same rabbit was treated with 50 mg talcum

Amount / concentration applied:

undiluted
Amount applied: 50 mg

Duration of treatment / exposure:

24 h; the substance was not washed out

Observation period (in vivo):

8 days

Number of animals or in vitro replicates:

2 animals (1 male and 1 female)

Details on study design:

Ca. 50 mg of the undiluted test substance was applied to the right conjuntival sac of one male and one female Vienna White rabbit. The left eye treated with 50 mg talcum served as a control. The substance was not washed out.
Ocular reactions were recorded 10 min, 1, 3, 24, 48, 72 hours, as well as on days 6 and 8 after instillation using a BASF-scoring system, which is convertible into the current OECD Draize scheme. For the evaluation, the BASF scores given in the raw data were converted to Draize scores. The reading time points used for assessment were 24, 48 and 72 hours after application. At day 8, the eyes were additionally examined by using fluorescein.
Observations of the condition of the cornea were made with the naked eye and also with the aid of an ophtalmoscope of Heine.

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

animal: #1, #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

chemosis score

Basis:

animal: #1, #2

Time point:

24/48/72 h

Score:

0

Max. score:

3

Irritation parameter:

conjunctivae score

Basis:

animal: #1, #2

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 8 days

OECD Draize scores for the 2 test animals; the original BASF scores were  
converted into the scores according to the OECD Draize system.  
No further irritating effects were seen compared to the control treatment 
(talcum powder) due to the physical properties (mechanical friction) of the test 
substance.

- Erythema scores (conjunctiva redness):

             /observation after [talcum]                            
             --------------------------------------------- /Mean 24, 48,
Test animal  /1 hour /24 hours /48 hours /72 hours /8 days /and 72 hours
---------------------------------------------------------- -------------
first        /1[1]   /1[1]     /1[1]     /1[1]     /0      /1[1]
second       /1[1]   /1[1]     /1[1]     /1[1]     /0      /1[1]

Mean         /1[1]   /1[1]     /1[1]     /1[1]     /0      /1[1]
------------------------------------------------------------------------

- No changes were observed in cornea opacity, conjunctiva chemosis or on the 
iris.

Interpretation of results:

GHS criteria not met

Reference 4

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Well documented study, sufficient for assessment.

Qualifier:

according to guideline

Guideline:

other: Code of Federal Regulations, Title 16, Section 1500, 42

Principles of method if other than guideline:

This test was used for eye irritation test before OECD guideline 405 was established. It is generally similar to the Draize test, with the following deviations: 100 mg of the test substance was applied to the right eye (OECD recommends a volume of 0.1 ml), 6 animals were used (OECD: 3); observation period was 7 days (OECD: 21).

GLP compliance:

no

Specific details on test material used for the study:

- Analytical purity: ca. 98 %

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Weight at study initiation: at the start of the test, the animals weighed ca. 2.5 kg
- Housing: Rabbits were kept in individual cages (540 x 360 x 315 mm) or in restraining devices
- Diet: 200 g of food per animal per day (Granules Lapin "entretien", Sanders)
- Water: softened and filtrated water ad libitum, automatically given

ENVIRONMENTAL CONDITIONS
- Temperature (°C): minimum 20 °C
- Air changes: ventilation (renewal of air 12 times per hour)
- Photoperiod (hrs dark / hrs light): 12/12

Vehicle:

unchanged (no vehicle)

Controls:

other: The left eye was treated with 50 mg talcum powder and served as control.

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 100 mg
- Concentration: undiluted

Duration of treatment / exposure:

24 h; the substance was not washed out.

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

6

Details on study design:

100 mg of the undiluted test substance was applied to the right conjuntival sac of 6 New Zealand White rabbits. The left eye was treated with 50 mg talcum powder and served as control. The substance was not washed out.
Ocular reactions were recorded at 1 h, 24 h, 48 h, 72 h, 96 h and 7 days after instillation using a scoring system, which is convertible into the current OECD Draize scheme. For the evaluation, the scores given in the raw data were converted to Draize scores. The reading time points used for assessment were 24, 48 and 72 hours after application.
Observations of the condition of the cornea were made with the naked eye and also with the aid of an ophtalmoscope of Heine.

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #2, #3, #4, #5, #6

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 72 h

Irritation parameter:

iris score

Basis:

animal: #1, #2, #3, #4, #5, #6

Time point:

24/48/72 h

Score:

0.11

Max. score:

2

Reversibility:

fully reversible within: 72 h

Irritation parameter:

conjunctivae score

Basis:

animal: #1, #2, #3, #4, #5, #6

Time point:

24/48/72 h

Score:

0.11

Max. score:

3

Reversibility:

fully reversible within: 72 h

Irritation parameter:

chemosis score

Basis:

animal: #1, #2, #3, #4, #5, #6

Time point:

24/48/72 h

Score:

0.06

Max. score:

3

Reversibility:

fully reversible within: 72 h

Irritant / corrosive response data:

The following effects were observed, but were completely reversible within 72 hours:
- Test animals showed slight conjunctiva chemosis and redness, which were completely reversible in 5/6 animals within 48 hours;
- Iris congestion was also observed in 3/6 animals and was also completely reversible within 48 hours in 2 of the animals concerned;
- The cornea was slightly affected in 1/6 animals at 24 h only.

Table 1: Eye irritation parameters according to the method of Draize

Reading time point	Animal 1, treated eye (right)
	Conjunctiva Chemosis	Conjunctiva Redness	Iris score	Cornea score	Additional findings
1 h	1	1	1	0
24 h	1	0	0	0
48 h	1	0	0	0
72 h	0	0	0	0
96 h	0	0	0	0
7 d	0	0	0	0
Reading time point	Animal 2, treated eye (right)
	Conjunctiva Chemosis	Conjunctiva Redness	Iris score	Cornea score	Additional findings
1 h	1	0	1	0
24 h	0	0	0	0
48 h	0	0	0	0
72 h	0	0	0	0
96 h	0	0	0	0
7 d	0	0	0	0
Reading time point	Animal 3, treated eye (right)
	Conjunctiva Chemosis	Conjunctiva Redness	Iris score	Cornea score	Additional findings
1 h	1	1	0	0
24 h	2	1	1	1	discharge
48 h	0	1	1	0
72 h	0	0	0	0
96 h	0	0	0	0
7 d	0	0	0	0
Reading time point	Animal 4, treated eye (right)
	Conjunctiva Chemosis	Conjunctiva Redness	Iris score	Cornea score	Additional findings
1 h	1	1	0	0
24 h	0	0	0	0
48 h	0	0	0	0
72 h	0	0	0	0
96 h	0	0	0	0
7 d	0	0	0	0
Reading time point	Animal 5, treated eye (right)
	Conjunctiva Chemosis	Conjunctiva Redness	Iris score	Cornea score	Additional findings
1 h	1	0	1	0
24 h	1	0	0	0
48 h	0	0	0	0
72 h	0	0	0	0
96 h	0	0	0	0
7 d	0	0	0	0
Reading time point	Animal 6, treated eye (right)
	Conjunctiva Chemosis	Conjunctiva Redness	Iris score	Cornea score	Additional findings
1 h	1	0	0	0
24 h	1	0	0	0
48 h	0	0	0	0
72 h	0	0	0	0
96 h	0	0	0	0
7 d	0	0	0	0

Interpretation of results:

GHS criteria not met

Reference 5

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

according to guideline

Guideline:

other: Code of Federal Regulations, Title 16, Section 1500.42

Principles of method if other than guideline:

3 New Zealand White rabbits were used. Approximately 100 µl of the test substance was applied to the conjunctival sac of one eye of each animal. The non- or saline- treated eye served as control. In studies with solids, talcum powder was used as control. The animals were observed after 24 h, 28 h, 72 h and after 8 days following application.
The eyes were not washed out after 24 h as optionally recommended in OECD Guideline 405. However, at the time the study was conducted, this requirement was not compulsory. This fact could be critical for solids which thus remained on the cornea for several days, causing mechanical damage. Probably, this would have been less severe if it had been washed out after 1 day.

GLP compliance:

no

Remarks:

GLP was not compulsory at the time the study was conducted

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: W. Ostrop, Meerbusch
- Weight at study initiation: 2.2 kg average weight
- Housing: individual; 1 animal per cage
- Diet: Mümmel Z (ssniff, Soest), ad libitum
- Water: ad libitum
- Acclimation period: 4-6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16-18 °C
- Humidity (%): 40 %
- Photoperiod: daylight rhythm

Vehicle:

unchanged (no vehicle)

Controls:

other: the second eye of the same rabbit served as control and was not treated

Amount / concentration applied:

undiluted
Amount applied: a volume of 0.1 ml of the crystalline substance

Duration of treatment / exposure:

24 h; the substance was not washed out

Observation period (in vivo):

8 days

Number of animals or in vitro replicates:

3

Details on study design:

100 µl of the undiluted test substance was applied to the conjuntival sac of the right eye of each animal. The left eye was not treated and served as a control. The substance was not rinsed or washed out.
Ocular reactions were recorded 24 h, 48 h, 72 h and 8 days after instillation and were evaluated using Draize scores. The reading time points used for assessment were 24, 48 and 72 hours after application.

Irritation parameter:

iris score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

3

Irritation parameter:

chemosis score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #2, #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

No irritation response was seen an iny animal at any time point.

Table 1: Eye irritation parameters; scores according to the method of Draize
Reading time point	Animal 1, treated eye
	Cornea score	Iris score	Conjunctivae score	Chemosis	Additional findings
24 h	0	0	0	0
48 h	0	0	0	0
72 h	0	0	0	0
8 d	0	0	0	0
Reading time point	Animal 2, treated eye
	Cornea score	Iris score	Conjunctivae score	Chemosis	Additional findings
24 h	0	0	0	0
48 h	0	0	0	0
72 h	0	0	0	0
8 d	0	0	0	0
Reading time point	Animal 3, treated eye
	Cornea score	Iris score	Conjunctivae score	Chemosis	Additional findings
24 h	0	0	0	0
48 h	0	0	0	0
72 h	0	0	0	0
8 d	0	0	0	0

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of the test, the test material has not to be classified according to GHS- or EU criteria.

Reference 6

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1973

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Principles of method if other than guideline:

The procedure employed was that predescribed by the Food and Drug Administration of the U.S.A. in The Federal Register (17 September 1964, § 191.12). The ocular reactions were scored by the method described by Draize in "Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics", p. 51. The scores for Conjunctivae redness and chemosis were combined and expressed as one single value.

GLP compliance:

no

Remarks:

GLP was not compulsory at the time the study was conducted.

Specific details on test material used for the study:

- Analytical purity: no data given in report

Species:

rabbit

Strain:

other: Albino

Vehicle:

unchanged (no vehicle)

Controls:

other: The second, untreated eye of the animal served as control.

Amount / concentration applied:

Concentration: 40 mg

Duration of treatment / exposure:

24 h; the substance was not washed out

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

6

Details on study design:

40 mg of the test material was placed in the conjuntival sac of each animal. The other eye, untreated, served as control. The substance was not washed out after 24 hours. Ocular reactions were recorded after 1, 2, 3, 4 and 7 days after instillation using Draize scores. The reading time points used for assessment were 24, 48 and 72 hours after application.

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #2, #3, #4, #5, #6

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

animal: #1, #2, #3, #4, #5, #6

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Basis:

animal: #1, #2, #3, #4, #5

Time point:

24/48/72 h

Score:

0

Max. score:

3

Irritation parameter:

conjunctivae score

Basis:

animal #6

Time point:

24/48/72 h

Score:

0.66

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Irritant / corrosive response data:

In 5/6 animals no findings were seen at any time point. One animal showed severe conjunctivae redness 24 hours after application of the test material. After 48 hours, the conjunctivae redness was moderate. After 72 hours, no more findings were seen in this animal until the end of the observation period.

Interpretation of results:

GHS criteria not met

Reference 7

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

; 50 mm3 (one sharp-edged spoon) of test material applied (OECD0.1 ml,< 100 mg); 2 animals were used (OECD: 3); the eyes were not washed out after 24 h (OECD: washing after 24 h); observation period was 3 days (OECD: 2); exam. using fluorescein (OECD no)

Principles of method if other than guideline:

BASF-Test:
Before OECD guideline 405 was established, eye irritation was tested using an internal method. White Vienna rabbits were used. Usually approximately 50 µl or one sharp-edged spoon in case of solids of the test substance were applied to the conjunctival sac of one eye of 2 animals. The non- or saline- treated eye served as control. In studies with solids, talcum powder was alternatively used as control. The animals were observed several times on the treatment day and up to 8 days afterwards, and findings were recorded on working days.
The eyes were not washed out after 24 h as specified in OECD Guideline 405, which could be critical for solids which thus remained on the cornea for several days, causing mechanical damage. Probably, this would have been less severe if it had been washed out after 1 day.

GLP compliance:

no

Remarks:

GLP was not compulsory at the time the study was conducted.

Specific details on test material used for the study:

- Analytical purity: 100 %

Species:

rabbit

Strain:

Vienna White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Average weight at study initiation: ca. 3.0 kg
- Diet: Altromin-MS (Altrogge, Lage/L. Germany), ad libitum
- Water: ad libitum

Vehicle:

unchanged (no vehicle)

Controls:

other: the second eye of the same rabbit was treated with 50 mg talcum powder.

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: one sharp-edged spoon of the test substance, undiluted; ca. 50 mg

Duration of treatment / exposure:

24 h; the substance was not washed out

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

2

Details on study design:

One sharp-edged spoon of the undiluted, solid test substance was applied to the right conjuntival sac of one male and one female Vienna White rabbit. The left eye, treated with physiological saline, served as a control. The substance was not washed out.
Ocular reactions were recorded after 10 min, 1, 3 and 24 hours, as well as 2 and 3 days after instillation using a BASF-scoring system, which is convertible into the current OECD Draize scheme. For the evaluation the BASF scores given in the raw data were converted to Draize scores. The reading time points used for assessment were 24, 48 and 72 hours after application.

Irritation parameter:

conjunctivae score

Basis:

animal: #1, #2

Time point:

24/48/72 h

Score:

0.16

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Remarks on result:

other: finding was also seen in control eye

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

animal: #1, #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

chemosis score

Basis:

animal: #1, #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

Slight conjunctivae redness as well as substance residues were observed in both eyes of all animals after 1 min, 1 hour and 3 hours of test substance instillation. In animal 1, no more findings were seen after 24 hours in both eyes. In animal 2, very slight conjunctivae chemosis was seen only in the left control eye after 1 and 3 hours and slight conjunctivae redness was still seen in both eyes after 24 hours. No more findings were seen after 48 hours in both eyes.

Table 1: Eye irritation parameters of 2 rabbits after application of the test material; scores were determined according to the method of Draize

Reading time point	Animal 1, treated eye (right)
	Cornea	Iris	Conjunctivae redness	Conjunctivae chemosis	Additional findings
10 min	0	0	1	0	substance residues
1 h	0	0	1	0	substance residues
3 h	0	0	1	0	substance residues
24 h	0	0	0	0
48 h	0	0	0	0
72 h	0	0	0	0
Reading time point	Animal 1, control eye (left)
	Cornea	Iris	Conjunctivae redness	Conjunctivae chemosis	Additional findings
10 min	0	0	1	0	substance residues
1 h	0	0	1	0	substance residues
3 h	0	0	1	0	substance residues
24 h	0	0	0	0
48 h	0	0	0	0
72 h	0	0	0	0
Reading time point	Animal 2, treated eye (right)
	Cornea	Iris	Conjunctivae redness	Conjunctivae chemosis	Additional findings
10 min	0	0	1	0	substance residues
1 h	0	0	1	0	substance residues
3 h	0	0	1	0	substance residues
24 h	0	0	1	0
48 h	0	0	0	0
72 h	0	0	0	0
Reading time point	Animal 2, control eye (left)
	Cornea	Iris	Conjunctivae redness	Conjunctivae chemosis	Additional findings
10 min	0	0	1	0	substance residues
1 h	0	0	1	1	substance residues
3 h	0	0	1	1	substance residues
24 h	0	0	1	0
48 h	0	0	0	0
72 h	0	0	0	0

Interpretation of results:

GHS criteria not met

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

All substances in this category share a similar structure with a molecular weight > 500 g/mol. Based on the high molecular weight and insolubility, the substances in this category will not be able to penetrate biological membranes. All substances are solids which do not melt at temperatures below 400 °C. Skin irritation studies are available for four substances in this category: CAS 147-14-8, CAS 1328-53-6, CAS 574-93-6 and CAS 27614-71-7. Eye irritation studies are available for five substances in this category: CAS 147-14-8, CAS 1328-53-6, CAS 574-93-6, CAS 27614-71-7 and CAS 68512-13-0. The skin and eye irritation data can be used for read-across to all substances in this category.

 

Skin irritation

CAS No. 147-14-8:

An OECD TG 404 and GLP compliant study was performed to assess the irritancy potential of Pigment Blue 15 to the skin of the New Zealand White rabbit (Sun Chemicals 2004, K1). A single 4-hour, semi-occluded application of 0.5 g the blue powder (moistened with 0.5 mL of distilled water) to the intact skin of three rabbits produced very slight erythema. No evidence of skin irritation was noted at the 48-hour observation. The test material produced a primary irritation index of 0.5. No corrosive effects were noted. Blue-coloured staining was noted at all treated skin sites throughout the study. This did not affect evaluation of skin reactions.

A Draize test was performed with four White Vienna rabbits, comparable to OECD guideline 404 (BASF 1971, K2). A 20-hour occlusive exposure to ca. 1 ml of the test substance (moistured with water and mixed to a paste) produced no effects (overall scores of 0 for all readings 24, 48 and 72 h).

In a third report, rabbits were exposed to 0.5 g of the powdered test material, mixed to a paste with polyethylene glycol before application, for 24 h under occlusive conditions (Huntingdon Research Centre 1973, K2). According to the author, no signs of irritation were seen during the 72 hours observation period at the intact skin of any animal.

Both results should be considered as overpredictive due to the harsh test conditions if compared to actual guideline recommendations (4 h under semiocclusive conditions).

 

CAS No. 1328-53-6:

A Draize test was performed with six White Vienna rabbits, comparable to OECD TG 404 (IFREB 1980, K2). A 24-hour occlusive exposure to ca. 0.5 g of the test substance in 0.65 mL of neutralized sterilized olive oil per produced the following effects: Treatment with the test substance resulted in only one out of 6 rabbits in very slight edema after 24 h. The evaluation of erythema after 24 hours was impossible in 2 rabbits due to treatment related colouring of the skin. Very slight to well defined erythema were noticed in 4 rabbits where irritation was reversible after 7 days. Slight desquamation was seen on intact skin in 2/6 animals after 7 days.

In a second report also comparable to OECD TG 404, rabbits were exposed to a paste containing the moistened test substance (50 %) for 24 h under occlusive conditions (BASF 1971, K2). No formation of edema or erythema was observed at any time point. Substance residues were seen on the treated sites until day 7.

Both results should be considered as overpredictive due to the harsh test conditions if compared to actual guideline recommendations (4 h under semiocclusive conditions).

 

CAS No. 574-93-6:

A Draize test was performed with three New Zealand White rabbits, comparable to OECD TG 404 (BASF 1979, K2). A 24-hour occlusive exposure to ca. 0.5 mL of the undiluted test material produced the following effects: The evaluation of erythema was impossible in all rabbits due to treatment related colouring of the skin. No to moderate edema was seen in 3 animals, the mean score for edema was 0.83. In one animal the effect was not fully reversible within 8 days.

In another study (IFREB 1979, K2), the test substance can be regarded as "non-irritant" to the skin of rabbits. 

 

CAS No. 27614-71-7:

A test was performed with three New Zealand White rabbits, acc. to OECD TG 404 (Aventis Pharma 2001, K1). A 4-hour semi-occlusive exposure to 0.5 g test material, pasted with 0.4 ml sesame oil produced no signs of irritation on the clipped skin of any rabbit during the whole study period.

 

CAS No. 68512-13-0:

A test was performed with six Albino rabbits, acc. to US Federal Register 38 No 187 § 1500.41 (BASF 1978, K2). A 24-hour occlusive exposure to 0.5 g test material, produced no signs of irritation on the skin of any rabbit during the whole study period.

 

Eye irritation

CAS No. 147-14-8:

A Draize test was performed with two White rabbits, comparable to OECD TG 405 (BASF 1971, K2). Eyes were left unwashed after application of one sharp-edged spoon of the undiluted, solid test substance and were observed for 72 h. The mean score for conjunctivae redness was 0.16 in the treated eye, but was reversible within 72 h. In the control eyes, the mean conjunctivae redness score was also 0.16, reversible within 72 h. Scores for cornea, iris and chemosis were 0 at any reading time point in the treated eyes.

No irritating effects were also observed in a second study, comparable to OECD TG 405 (Huntingdon Research Centre 1973, K2). Eyes were left unwashed after application and were observed for 7 days. In 5/6 animals no findings were seen at any time point. The scores for Conjunctivae redness and chemosis were combined and expressed as one single value. One animal showed severe conjunctivae effects 24 hours after application of the test material. After 48 hours, the conjunctivae score was moderate. After 72 hours, no more findings were seen in this animal until the end of the observation period (7 days).

 

CAS No. 1328-53-6:

No irritating effects were observed in a Draize test, comparable to OECD TG 405 (IFREB 1980, K2). Eyes were left unwashed after application of ca. 100 mg of the undiluted, solid test substance and were observed for 7 days. The following effects observed were observed, but were completely reversible within 72 hours: Test animals showed slight conjunctiva chemosis and redness, which were completely reversible in 5/6 animals within 48 hours; iris congestion was also observed in 3/6 animals and was also completely reversible within 48 hours in 2 of the animals concerned; the cornea was slightly affected in 1/6 animals at 24 h only.

A second Draize test was performed with two White Vienna rabbits, comparable to OECD TG 405 (BASF 1971, K2). Eyes were left unwashed after application of ca. 50 mg of the undiluted, solid test substance into the left eye as well as of ca. 50 mg of talcum (control substance) into the right eye of each animal. Observation period was 8 days. The mean score for conjunctivae redness was 1.0 in the treated eye, but was fully reversible within 8 days. In the control eyes, the mean conjunctivae redness score was also 1.0, and was also fully reversible within 8 days. Scores for cornea, iris and chemosis were 0 at any reading time point in the treated eyes.

 

CAS No. 574-93-6:

No irritating effects were observed in a Draize test, comparable to OECD TG 405 (BASF 1979, K2). Eyes were left unwashed after application of ca. 100 mg of the undiluted, solid test substance and were observed for 8 days. No effects were observed in all 3 animals at any time point.

 

CAS No. 27614-71-7:

No irritating effects were observed in a eye irritation test, acc. to OECD TG 405 (Aventis Pharma 2001, K1). Eyes were washed 24 h after application of ca. 100 mg of the undiluted, solid test substance and were observed for 72 hours after administration of the test material. One hour after administration the conjunctiva of one animal was very slightly swollen. Furthermore, all animals showed definitely injected blood vessels at one hour up to 48 h after administration. In addition to these observations, serous eye discharge discoloured by compound was noted. 72 h after administration, all signs of irritation had disappeared.

 

CAS No. 68512-13-0:

No irritating effects were observed in a Draize test, comparable to OECD TG 405 (InBiFo 1978, K2). Eyes were left unwashed after application of ca. 100 mg of the test substance and were observed for 7 days. No effects were observed in all 6 animals at any time point.

Justification for classification or non-classification

Based on the available information, classification for skin and eye irritation is not warranted in accordance with EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation No. (EC) 1272/2008.