N-(5-chloro-2-methoxyphenyl)-3-hydroxy-4-[[2-methoxy-5-[(phenylamino)carbonyl]phenyl]azo]naphthalene-2-carboxamide

Endpoint summary

Description of key information

Skin irritation (OECD 439, in vitro): not irritating

Eye irritation (OECD 405, in vivo): not irritating (read-across from analogue substance C.I. Pigment Red 184 [N-(5-chloro-2-methoxyphenyl)-3-hydroxy-4-[[2-methoxy-5-(phenylcarbamoyl)phenyl]diazenyl]naphthalene-2-carboxamide])

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 28 MAR 2012 to 02 APR 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study (OECD 439) and in compliance with GLP.

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Version / remarks:

Reconstructed Human Epidermis Test Method (Original Guideline adopted July 22, 2010)

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

In October 2011 the EU commission published a recommendation (2011/696/EU) on the definition of nanomaterials. From the results of analyses it is concluded that the registered substance C.I.Pigment Red 269 falls within the boundaries of the nanomaterial definition. Hence, studies were performed using the nanomaterial.

Amount / concentration applied:

Approximately 11 mg of the neat test item were applied to three EPISKIN (Skinethic) tissues. Addtitonally, the tissues were wetted with 20 µL of deionised water.

Duration of treatment / exposure:

The skin equivalents were exposed to the test item for 15 minutes. After completion of the treatment the test item was rinsed off and the skin equivalents were incubated for further 42 hours.

Details on study design:

Approximately 11 mg of the neat test item and 10 µL of the negative control (deionised water) or the positive control (5% sodium lauryl sulfate) were applied to each three EPISKIN (Skinehtic) tissues. Additionally, the test item treated skin equivalents were wetted with 20 µL of deionised water. The test item as well as the controls were rinsed off after 15 minutes treatment. After further 42 hours incubation the tissues were treated with the MTT solution for 3 hours following approximately 70 hours extraction of the colorant from the cells. The amount of extracted colorant was determined photometrically at a wavelength of 570 nm.

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

15 min

Value:

106.8

Other effects / acceptance of results:

After treatment with the test item the relative absorbance values did not decrease relevantly (106.8%; threshold for irritancy: ≤ 50%). Therefore, the test item is not considered to possess an irritant potential.

Results after treatment with test substance and controls

 

Dose group	Treatment Interval	Absorbance 570 nm Tissue 1*	Absorbance 570 nm Tissue 2*	Absorbance 570 nm Tissue 3*	Mean Absorbance of 3 Tissues	Relative Absorbance [%] Tissue 1, 2 + 3**	Standard Deviation [%]	Rel. Absorbance [% of Negative Control]***
Negative Control	15 min	1.107	1.040	1.124	1.090	101.5 95.4 103.1	4.0	100.0
Positive Control	15 min	0.321	0.304	0.336	0.320	29.4 27.9 30.8	1.4	29.4
Test Item	15 min	1.190	1.189	1.115	1.165	109.1 109.1 102.3	4.0	106.8

*       Mean of two replicate wells after blank correction

**      relative absorbance per tissue [rounded values]:
***    relative absorbance per treatment group [rounded values]:

After treatment with the test item the relative absorbance values did not decrease (106.8%; threshold for irritancy: ≤ 50%). Consequently the test item was non irritant to skin.

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

In conclusion, it can be stated that in this study and under the experimental conditions reported, the test item is not irritant to skin.

Executive summary:

This in vitro study was performed to assess the irritation potential of test substance by means of the Human Skin Model Test according to OECD 439.

Three tissues of the human skin model EpiSkin™ were treated with the test item, the negative or the positive control for 15 minutes.

Approximately 11 mg of the neat test item were applied to each tissue, spread to match the tissue size.Additionally, the test item tissues were wetted with 20 µL of deionised water.

10 µL of either the negative control (deionised water) or the positive control (5% Sodium lauryl sulfate) were applied to each tissue.

The test item and the positive and negative controls were washed off the skin tissues after 15 minutes treatment. After further incubation for about 42.5 hours the tissues were treated with the MTT solution for 3 hours following 69.5 hours extraction of the colorant from the cells. The amount of extracted colorant was determined photometrically at 570 nm.

After treatment with the negative control the absorbance values were well within the required acceptability criterion of mean OD in the range frm 0.6 to 1.5 for the15 minutes treatment interval thus showing the quality of the tissues.

Treatment with the positive control induced a decrease in the relative absorbance as compared to the negative control to 29.4% thus ensuring the validity of the test system.

The standard deviations between the % variabilities of the test item, the positive and negative controls were below 4% (threshold of the "OECD TG 439 Guideline for the Testing of Chemicals 439: In vitro Skin Irritation: Reconstructed Human Epidermis Test Method”: 18%).thus ensuring the validity of the study.

After treatment with the test item the relative absorbance values did not decrease (106.8%; threshold for irritancy:≤50%). Therefore, the test item is not considered to possess a skin irritant potential.

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:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

From 06 Jun 2011 to 16 JUN 2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study (OECD 405), in compliance with GLP

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Robinson Services, Inc.
- Age at study initiation: young adult
- Weight at study initiation: no data
- Housing: individually
- Diet: Harlan Teklad Global High Fiber Rabbit Diet 2031 ad libitum
- Water: tap water ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-23
- Humidity (%): 63-83
- Air changes (per hr): 12
- Photoperiod (hrs dark / hrs light): 12/12

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

0.1 mL (0.03 g)

Duration of treatment / exposure:

24 h

Observation period (in vivo):

72 h

Number of animals or in vitro replicates:

2 females, 1 male

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): with physiological saline
- Time after start of exposure: 24 h

SCORING SYSTEM: in accordance with OECD TG 405.
Scores were read 1, 24, 48 and 72 h after test substance application.

TOOL USED TO ASSESS SCORE: hand-slit lamp / fluorescein at 24 h reading

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility not relevant since no effect

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: reversibility not relevant since no effect

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

other: reversibility not relevant since no effects

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility not relevant since no effect

Irritant / corrosive response data:

Iris, and cornea scores were 0 for all three animals at all time points (1, 24, 48 and 72 h). Conjunctiva redness and chemosis scores were also 0 for all three animals at 24, 48 and 72 h reading. Only one female showed slight chemosis at the 1 h reading (score 1, animal 2) and two animals (one male and one female, animal 1 and 2, respectively) showed slight redness (score 1 and 2, respectively) at the 1 hour reading. Slight discharge (score 2, 2, 1 in animal #1, 2, 3, respectively) was observed in all animals at the 1 h reading. All effects were fully reversible withing 24 hours.

Other effects:

All animals appeared active and healthy during the study. There were no other signs of gross toxicity, adverse pharmacologic effects or abnormal behavior.

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test material is not irritating to the eyes under the test conditions of OECD 405.

Executive summary:

Test material was subject to a test of eye irritancy according to OECD guideline 405. 30 mg (0.1 mL) of substance were applied to one eye of three animals. The eyes were washed 24 h after application and eye responses were noted for 72 h after start of the exposure. There was no corneal opacity or iritis observed in any treated eye during the study. Within one hour after test substance instillation, all three treated eyes exhibited conjunctivitis. All animals were free of ocular irritation by 24 hours. No other effects could be observed in any animal. Therefore, it is concluded that the test item is not irritating to eyes.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation

The skin irritating potential of the registered substance was investigated in an in vitro study by means of the Human Skin Model Test according to OECD guideline 439 under GLP conditions (Harlan_1474204). Three tissues of the human skin model EpiSkin™ were treated with the test item, the negative or the positive control for 15 minutes. Approximately 11 mg of the neat test item was applied to each tissue and wetted with 20 µL of deionised water. Deionised water and 5% sodium lauryl sulphate (SLS) served as negative and positive controls, respectively. After treatment with the test item the relative absorbance values did not decrease (mean value: 106.8%; threshold for irritancy: ≤ 50%). Therefore, the test item is not considered to possess a skin irritant potential.

Eye irritation

The analogue substance C.I. Pigment Red 184 (N-(5-chloro-2-methoxyphenyl)-3-hydroxy-4-[[2-methoxy-5-(phenylcarbamoyl)phenyl]diazenyl]naphthalene-2-carboxamide) was subject to a test of eye irritancy according to OECD guideline 405 under GLP conditions (Eurofins_32401). 30 mg (0.1 mL) of the test substance was applied to one eye of three animals. The eyes were washed 24 h after application and eye irritation responses were noted for 72 h after start of the exposure. There was no corneal opacity or iritis observed in any treated eye during the study. Within one hour after test substance instillation, all three treated eyes exhibited conjunctivitis. All animals were free of ocular irritation by 24 hours. No other effects could be observed in any animal. Therefore, it is concluded that the test item is not irritating to eyes.

Justification for classification or non-classification

The available data on skin irritation obtained with the registered substance and on eye irritation derived from an analogue substance do not meet the criteria for classification according to Regulation (EC) No. 1272/2008 (CLP) and are therefore conclusive but not sufficient for classification.