1-amino-4-hydroxy-2-phenoxyanthraquinone

Endpoint summary

Description of key information

Skin irritation: non-irritant
Eye irritation: non-irritant

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:

June 2nd to 9th, 1980

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods

Qualifier:

according to guideline

Guideline:

EPA OPP 81-5 (Acute Dermal Irritation)

Deviations:

not specified

Remarks:

Test performed according to the Proposed Guideline, Federal register, Vol. 43, n. 163, August 22nd, 1978

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: 2 to 3 kg
- Housing: individual housing in metal cages
- Diet: NAFAG, no. 814, Gossau SG, standard rabbit food, ad libitum
- Water: ad libitum
- Acclimation period: 4 days minimum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2
- Humidity (%): 55 ± 10
- Photoperiod (hrs dark / hrs light): 10 hours light cycle day

Type of coverage:

occlusive

Preparation of test site:

other: intact and abraded skin

Vehicle:

other: propylene glycol + saline (70:30 parts)

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g
- Concentration (if solution): 50 %

Duration of treatment / exposure:

24 h

Observation period:

24 h, 48 h, 72 h, 4 d and 7 d after patch removal

Number of animals:

6 animals: 3 male and 3 female

Details on study design:

TEST SITE
- Area of exposure: 2.5 cm × 2.5 cm

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

ca. 0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

ca. 0.33

Max. score:

4

Reversibility:

fully reversible within: 48h

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

ca. 0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #4

Time point:

24/48/72 h

Score:

ca. 0.33

Max. score:

4

Reversibility:

fully reversible within: 48h

Irritation parameter:

erythema score

Basis:

animal #5

Time point:

24/48/72 h

Score:

ca. 0.33

Max. score:

4

Reversibility:

fully reversible within: 48h

Irritation parameter:

erythema score

Basis:

animal #6

Time point:

24/48/72 h

Score:

ca. 0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

ca. 0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

ca. 0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

ca. 0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #4

Time point:

24/48/72 h

Score:

ca. 0.33

Max. score:

4

Reversibility:

fully reversible within: 48h

Irritation parameter:

edema score

Basis:

animal #5

Time point:

24/48/72 h

Score:

ca. 0.33

Max. score:

4

Reversibility:

fully reversible within: 48h

Irritation parameter:

edema score

Basis:

animal #6

Time point:

24/48/72 h

Score:

ca. 0

Max. score:

4

Remarks on result:

no indication of irritation

Assessment

Due to intensive staining by the test substance, erythema was not assessable. For the purpose of calculating the primary irritation index, erythema was assumed to be equal to edema based on existing correlation (Marzulli and Maibach, Fol. Cosm. Tox. 13, 355, 1975).

Skin reaction scores of rabbits

Abraded skin

Animal/sex	Reaction	24 h	48 h	72 h	4 d	7 d	Mean 24, 48 and 72 h
19 M	Erythema	0	0	0	0	0	0
20 M	Erythema	1	0	0	0	0	0.33
21 M	Erythema	0	0	0	0	0	0
22 F	Erythema	1	0	0	0	0	0.33
23 F	Erythema	1	0	0	0	0	0.33
24 F	Erythema	0	0	0	0	0	0
19 M	Oedema	0	0	0	0	0	0
20 M	Oedema	1	0	0	0	0	0.33
21 M	Oedema	0	0	0	0	0	0
22 F	Oedema	1	0	0	0	0	0.33
23 F	Oedema	1	0	0	0	0	0.33
24 F	Oedema	0	0	0	0	0	0

Intact skin

Animal/sex	Reaction	24 h	48 h	72 h	4 d	7 d	Mean 24, 48 and 72 h
19 M	Erythema	0	0	0	0	0	0
20 M	Erythema	0	0	0	0	0	0
21 M	Erythema	0	0	0	0	0	0
22 F	Erythema	0	0	0	0	0	0
23 F	Erythema	0	0	0	0	0	0
24 F	Erythema	0	0	0	0	0	0
19 M	Oedema	0	0	0	0	0	0
20 M	Oedema	0	0	0	0	0	0
21 M	Oedema	0	0	0	0	0	0
22 F	Oedema	0	0	0	0	0	0
23 F	Oedema	0	0	0	0	0	0
24 F	Oedema	0	0	0	0	0	0

Interpretation of results:

other: not classified according to the CLP Regulation (EC 1272/2008)

Conclusions:

Non-skin irritant.

Executive summary:

The substance was tested for skin irritation potential based on the Proposed guidelines of the US EPA 163.81 -5 "Primary dermal irritation study," Federal register, Vol. 43, No. 163, August 22, 1978 (1980). 0.5 g of test substance was applied to 2.5 cm × 2.5 cm intact and abraded skin in an occlusive preparation to 3 male and 3 female New Zealand White rabbits. 50 % solution of propylene glycol + saline (70:30 parts) was the vehicle used. The exposure duration was 24 h. Animal skin reactions were scored at 24 h, 48 h, 72 h, 4 d and 7 d after patch removal.

One male and two female rabbits showed slight edema and slight erythema on abraded skin at 24 h only. These skin reactions were reversible within 48 h. No skin reactions were observed on intact skin.

Abraded skin of all six animals had the following mean reaction scores (24, 48 and 72 h): erythema score < 1 and oedema score < 1.

Intact skin of all six animals had the following mean reaction scores (24, 48 and 72 h): erythema score = 0 and oedema score = 0.

Therefore, the test substance is a non-skin irritant.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From June 2nd, 1980 to June 9th, 1980

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OPP 81-4 (Acute Eye Irritation)

Deviations:

no

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: New Zealand White breed
- Weight at study initiation: 2 - 3 kg
- Housing: individual housing in metal cages
- Diet (e.g. ad libitum): standard rabbit food, NAFAG, no. 814, Gossau SG
- Water (e.g. ad libitum): Water ad libitum
- Acclimation period: 4 days minimum
- Other: Only rabbits showing normal ophthalmic findings were included in the test

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2
- Humidity (%): 55 ± 10
- Photoperiod (hrs dark / hrs light): 10 hour light cycle day

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

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

Duration of treatment / exposure:

30 seconds

Observation period (in vivo):

1, 2, 3, 4 and 7 days

Number of animals or in vitro replicates:

6 animals: 3 males and 3 females

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The treated eyes of three rabbits (animals n. 22, 23 and 24) were each flushed with 10 ml physiological saline approximately 30 seconds after treatment

TOOL USED TO ASSESS SCORE:
- The reactions were appraised with a slit-lamp after 1, 2, 3, 4 and 7 days

Irritation parameter:

cornea opacity score

Basis:

animal: 6/6

Time point:

other: mean 24, 48 and 72 h

Score:

ca. 0

Max. score:

4

Reversibility:

other: No ocular effect oserved

Irritation parameter:

iris score

Basis:

animal: 6/6

Time point:

other: mean 24, 48 and 72 h

Score:

ca. 0

Max. score:

2

Reversibility:

other: No ocular effect oserved

Irritation parameter:

other: conjunctivae score (redness and chemosis)

Basis:

animal: 6/6

Time point:

other: mean 24, 48 and 2 h

Score:

ca. 0

Max. score:

4

Reversibility:

other: No ocular effect oserved

RABBIT EYE IRRITATION SCORES

n = not rinsed      

r = rinsed

p = purulent

Time point	24 hours						48 hours						72 hours
Rabbit no.	19	20	21	22	23	24	19	20	21	22	23	24	19	20	21	22	23	24
Eye	n	n	n	r	r	r	n	n	n	r	r	r	n	n	n	r	r	r
I. Cornea   A Opacity B Area involved   a = A × B × 5 (max. 80)	0 0   0	0 0   0	0 0   0	0 0   0	0 0   0	0 0   0	0 0   0	0 0   0	0 0   0	0 0   0	0 0   0	0 0   0	0 0   0	0 0   0	0 0   0	0 0   0	0 0   0	0 0   0
II. Iris   b = Score × 5 (max. 10)	0 0	0 0	0 0	0 0	0 0	0 0	0 0	0 0	0 0	0 0	0 0	0 0	0 0	0 0	0 0	0 0	0 0	0 0
III. Conjunctiva   A Redness B Chemosis C Discharge   c = A + B + C × 2 (max. 20)	0 0 0   0	0 0 0   0	0 0 0   0	0 0 0   0	0 0 0   0	0 0 0   0	0 0 0   0	0 0 0   0	0 0 0   0	0 0 0   0	0 0 0   0	0 0 0   0	0 0 0   0	0 0 0   0	0 0 0   0	0 0 0   0	0 0 0   0	0 0 0   0
Total = a + b + c (max. 110)	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Group mean	0			0			0			0			0			0

Time point	4 days						7 days
Rabbit no.	19	20	21	22	23	24	19	20	21	22	23	24
Eye	n	n	n	r	r	r	n	n	n	r	r	r
I. Cornea   A Opacity B Area involved   a = A × B × 5 (max. 80)	0 0   0	0 0   0	0 0   0	0 0   0	0 0   0	0 0   0	0 0   0	0 0   0	0 0   0	0 0   0	0 0   0	0 0   0
II. Iris   b = Score × 5 (max. 10)	0 0	0 0	0 0	0 0	0 0	0 0	0 0	0 0	0 0	0 0	0 0	0 0
III. Conjunctiva   A Redness B Chemosis C Discharge   c = A + B + C × 2 (max. 20)	0 0 0   0	0 0 0   0	0 0 0   0	0 0 0   0	0 0 0   0	0 0 0   0	0 0 0   0	0 0 0   0	0 0 0   0	0 0 0   0	0 0 0   0	0 0 0   0
Total = a + b + c (max. 110)	0	0	0	0	0	0	0	0	0	0	0	0
Group mean	0			0			0			0

EFFECT OF RINSING

Primary irritation index in unrinsed eyes, A = 0

Primary irritation index in rinsed eyes, B = 0

Effect of rinsing, R = A/B = 0/0

Interpretation of results:

other: not classified according to the CLP Regulation (EC 1272/2008)

Conclusions:

Non-eye irritant.

Executive summary:

The eye irritation potential of the test substance was examined by the slit-lamp procedure on the basis of the US EPA guideline 163.81–4 "Primary Eye Irritation," Federal Register, Vol 43, No. 163, August 22, 1978.

0.1 g of unchanged test substance was applied to the left eye of 3 male and 3 female New Zealand white rabbits for 30 seconds. The untreated right eye served as controls. Treated eyes of three rabbits were each flushed with 10 ml physiological saline approximately 30 seconds after treatment. Eye reactions were scored at 24 h, 48 h, 72 h, 4 d and 7 d after exposure.

The mean irritation indexes (at 24, 48 and 72 h) for 6/6 animals were as follows: Cornea = 0, Iris = 0 and Conjunctivae = 0.

Therefore, the test substance is considered as non-eye irritant.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

The skin irritation potential of the test substance has been examined in five tests (1980, 1978, 1977, 1991 and 1977, in order of relevance).

In the guideline key study (1980), the test substance was examined for irritation potential on intact and abraded skin in an occlusive preparation for 24 h. Skin reactions were scored at 24 h, 48 h, 72 h, 4 d and 7 d after patch removal. Abraded skin of 6/6 animals had the following mean reaction scores (24, 48 and 72 h): erythema score < 1 and oedema score < 1 while intact skin of 6/6 animals had the following mean reaction scores (24, 48 and 72 h): erythema score = 0 and oedema score = 0. All skin reactions were fully reversible within 48 h.

A supporting guideline study (1978) examined irritation potential of the test substance on intact and abraded skin in an occlusive preparation for 24 h. Skin reactions were scored at 24 h, 48 h, 72 h, 4 d and 7 d after patch removal. Intact skin of 6/6 rabbits had the following mean reaction scores (24, 48 and 72 h): erythema = 0 and oedema = 0. Abraded skin of 6/6 rabbits had the following mean reaction scores (24, 48 and 72 h): erythema ≤ 1 and oedema ≤ 1. All skin reactions were fully reversible within 48 h.

Another supporting guideline study (1977) investigated skin irritation potential on abraded and non-abraded skin in an occlusive preparation for a 24 h exposure. Skin reactions were scored at 0 h and 48 h after patch removal. For the purpose of calculating erythema and edema scores at 24, 48 and 72 h after patch removal, the following method was adopted: scores at 0 h were taken as scores at 24 h and scores at 48 h were also taken as scores at 72 h. For all animals, the calculation of mean skin reaction scores (24, 48 and 72 h) after patch removal gave the following values for intact skin: erythema score = 0 and oedema score = 0; and for abraded skin: erythema score = 0 and oedema score < 1. All skin reactions were fully reversible within 48 h.

4 h exposure to the test substance to intact, shaven skin in an occlusive preparation was used to evaluate the skin irritation potential in a supporting guideline line study (1991). Skin reactions were scored at 1 h, 24 h, 48 h and 72 h after patch removal. 4 sites on intact skin of each animal were examined. In calculation of erythema and edema scores, the highest skin reaction score of all four test sites is taken as the skin reaction score for that animal at that time point. For 3/3 animals, the mean skin reaction scores (at 24, 48 and 72 h) were as follows: erythema score ≤ 1 and edema score = 0. All skin reactions resolved within 48 h. Systemic signs or toxic effects on body weight caused by the test substance were not observed. Measurements showed decreases in bodyweight of 0.0, 0.1 and 0.16 kg in animals 1, 2 and 3 respectively. This effect is not considered a result of test substance administration but is attributed to the stress caused by the Elizabethan collar fitted at the rabbits' necks.

Another supporting study (1977) examined skin irritation potential according to a specific national testing guideline. Abraded skin was exposed to the test substance for 24 h in an occlusive preparation and monitored at 0, 24 and 48 h after removal. However, the report for this test is not robust. Rating of erythema and oedema at 24, 48 and 72 h after patch removal are not available in the report.

The primary irritation index resulted to be 0 in the test, as given in the report.

The results of all five tests are coherent and in agreement. The test substance was not irritating to the skin in all tests. Therefore, the test substance is considered as a non-skin irritant.

The eye irritation potential of the test substance was examined in four guideline tests (1980, 1978, 1977, 1977).

The key guideline study (1980) examined eye irritation potential by the slit-lamp procedure. Unchanged test substance was applied to the left eye of 3 male and 3 female New Zealand white rabbits for 30 s. Treated eyes of three rabbits were each flushed with 10 ml physiological saline approximately 30 s after treatment. At 24 h, 48 h, 72 h, 4 d and 7 d after exposure, eye reactions were scored.

The mean irritation indexes (at 24, 48 and 72 h) for 6/6 animals were as follows: Cornea = 0, Iris = 0 and Conjunctivae = 0.

In a supporting guideline study (1978), eye irritation potential was investigated by the slit-lamp procedure. Unchanged test substance was also applied to the left eye of 3 male and 3 female New Zealand white rabbits for 30 s. Treated eyes of female rabbits were each flushed with 10 ml lukewarm water approximately 30 s after treatment and eye reactions were scored at 1 d, 2 d, 3 d, 4 d and 7 d after exposure. The mean irritation indexes (at 1, 2 and 3 d) for 6 out of 6 (6/6) animals were the following: corneal score = 0 and iris score = 0. 5/6 animals had a mean conjunctivae score (at 1, 2 and 3 d) of 0. 1 female animal had a mean conjunctivae score (at 1, 2 and 3 d) = 2, in a rinsed eye and this reaction was reversible within 2 d.

Another supporting guideline study (1977) examined eye irritation potential of the test substance by the slit-lamp procedure. Unchanged test substance was applied to the left eye of 3 male and 3 female New Zealand white rabbits for 30 s. Treated eyes of female rabbits were each flushed with 10 ml lukewarm water approximately 30 s after treatment and eye reactions were scored at 1 d, 2 d, 3 d, 4 d and 7 d after exposure. The mean irritation indexes (at 1, 2 and 3 d) for 6 out of 6 (6/6) animals were the following: corneal score = 0 and iris score = 0. 5/6 animals had a mean conjunctivae score (at 1, 2 and 3 d) of 0. 1 male animal had a mean conjunctivae score (at 1, 2 and 3 d) = 2, in an unrinsed eye and this reaction was reversible within 2 d.

Another study (1977) examined the eye irritation potential of the test substance according to a specific national testing guideline. 100 mg of the test substance was applied to one eye of 6 SPF-Albino-Himalayan rabbits for 30 s. After application of the test substance, the highest irritation index was determined to be 23 after 7 hours, as given in the test report. However, this result is unsuitable for a risk assessment as corneal, iris and conjunctivae scores at 24 h, 48 h and 72 h were not reported. Therefore, this result is not taken into consideration in evaluating the eye irritation potential of the test substance.

The results of the other three guideline tests (1980, 1978 and 1977) for which irritation scores are available are used for the evaluation of eye irritation potential. These results are in agreement. The test substance was not irritating to the eyes in these three tests. Therefore, the test substance is a non-eye irritant.

Justification for classification or non-classification

For skin irritation and eye irritation, the CLP Regulation (EC 1272/2008) was used for overall judgment.

In the CLP Regulation (EC 1272/2008), skin irritation is defined as "the production of reversible damage to the skin following the application of a test substance for up to 4 hours." The same regulation, defines a Category 2 skin irritant as "Mean value of ≥ 2,3 - ≤ 4,0 for erythema/ eschar or for oedema in at least 2 of 3 tested animals from gradings at 24, 48 and 72 hours after patch removal."

In the four comprehensive tests, edema and erythema scores on intact skin were equal to 0.

Therefore, the test substance is a non-skin irritant.

Moreover the CLP Regulation (EC 1272/2008), defines eye irritation as follows: "Eye irritation means the production of changes in the eye following the application of test substance to the anterior surface of the eye, which are fully reversible within 21 days of application." The same regulation defines a Category 2 eye irritant as, "if, when applied to the eye of an animal, a substance produces: — at least in 2 of 3 tested animals, a positive response of: — corneal opacity ≥ 1 and/or — iritis ≥ 1, and/or — conjunctival redness ≥ 2 and/or — conjunctival oedema (chemosis) ≥ 2 — calculated as the mean scores following gradings at 24, 48 and 72 hours after installation of the test material, and which fully reverses within an observation period of 21 days."

Mean corneal, iris and conjunctivae scores (at 24, 48 and 72 h) were equal to 0 for 5/6 animals in the three comprehensive tests. For 1/6 animals in conjunctivae scores were equal to 2, in rinsed and unrinsed eyes (1978 and 1977 respectively). These eye reactions were fully reversible within 2 d. The fourth test (1977) does not provide details on conjunctivae score, iris score and corneal score, therefore, it was not taken into account in the evaluation of eye irritation potential.

Therefore, based on these results, the test material is considered as a non-eye irritant.