[8-[(p-aminophenyl)azo]-7-hydroxy-2-naphthyl]trimethylammonium chloride

Endpoint summary

Description of key information

Basic brown 16 were found to be non-irritating to skin and eye. Therefore, Basic brown 16 (CAS no. 26381-41-9) cannot be classified as per the criteria of CLP regulation for skin and eye irritation.

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:

supporting study

Reliability:

2 (reliable with restrictions)

Qualifier:

equivalent or similar to guideline

Guideline:

other: as below

Principles of method if other than guideline:

Skin irritation study of Basic brown 16

GLP compliance:

not specified

Species:

other: no data

Strain:

not specified

Details on test animals or test system and environmental conditions:

No data available

Type of coverage:

not specified

Preparation of test site:

not specified

Vehicle:

not specified

Controls:

not specified

Amount / concentration applied:

Neat (100 % )

Duration of treatment / exposure:

No data available

Observation period:

No data available

Number of animals:

No data available

Irritation parameter:

other: skin irritation

Remarks on result:

other: Non irritating to skin

Concordance of the in vitro results by OD and HPLC/UPLC-spectrophotometry with in vivo Draize skin test.

Chemical name	CAS n°	Conc	In Vivo Class.
Basic brown 16	26381–41-9	Neat	NC

Overflow: Outside linearity range of absorbance (OD); unqualified: Could not be determined (SD N 18%).

$ Test items #1–48were tested in the EpiSkin™ skin irritation test method and test items 49–50were tested in the EpiSkin™ skin corrosion test method; NC: not classified; Cat.2: UNGHS Category 2; Cat.1A/Cat.1B—UN GHS Category 1 A/UN GHS

Interpretation of results:

not irritating

Remarks:

Migrated informationCriteria used for interpretation of results: EU

Conclusions:

Concordance of the in vitro results by OD and HPLC/UPLC-spectrophotometry with in vivo Draize skin test.

Executive summary:

In a Skin irritation study, Basic brown 16 were tested by using Draize skin test at the concentration of 100 %. Basic brown 16 were found to be non-irritating.

Hence, Basic brown 16 were not classified according to the test.

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

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

other: Data is from opinions on health and safety risks (chemical, biological, mechanical and other physical risks)

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

GLP compliance:

not specified

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS- Age at study initiation:young adult

Vehicle:

not specified

Controls:

yes

Amount / concentration applied:

0.1 g

Duration of treatment / exposure:

72 hours

Observation period (in vivo):

14 days

Number of animals or in vitro replicates:

3 (1 male and 2 females)

Details on study design:

REMOVAL OF TEST SUBSTANCE- Washing (if done):By lukewarm tap water- Time after start of exposure:48 hoursSCORING SYSTEM:corneal opacity, iris, redness and chemosis of the conjunctivae.TOOL USED TO ASSESS SCORE: No data

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

other: 14 days

Reversibility:

not reversible

Remarks on result:

other: mild to moderate, early-onset and transient ocular changes, such as reddening of the conjunctivae and sclerae, discharge and chemosis. These effects were reversible and were no longer evident 14 days after treatment

Irritant / corrosive response data:

Slight opacity of the cornea, affecting the whole area, was noted in all animals at the 1- and 24-hour reading due to staining produced by the test item. Slight opacity of the cornea, affecting the whole area, was again visible in one animal 72 hours after treatment.

Score of Irritation parameter
	24 hrs	48 hrs	72hrs
corneal opacity	0.33	0.67	0.33
iris	0.00	0.00	0.00
conjunctivae	1.67	2.00	2.00
redness	1.67	1.33	0.00
chemosis	1.67	0.00	0.00

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of the test, the study considered Basic Brown 16 to be not irritating to rabbit eye.

Executive summary:

The test substance was applied by instillation of 0.1 g into the left eye of each of three young adult New Zealand White rabbits. The treated and untreated eyes were rinsed with lukewarm tap water 48 hours after instillation. Scoring of irritation effects was performed

approximately 1, 24, 48 and 72 hours, as well as 7, 10 and 14 after test item instillation.The mean score was calculated across 3 scoring times (24, 48 and 72 hours after instillation) for each animal for corneal opacity, iris, redness and chemosis of the conjunctivae, separately.

The individual mean scores for corneal opacity were 0.33, 0.67 and 0.33, respectively. The individual mean scores for the iris were 0.00 for all three animals. The individual mean scores for the conjunctivae were 1.67, 2.00 and 2.00 for reddening and 1.67, 1.33 and 1.67 for chemosis, respectively.The instillation of Basic Brown 16 into the eye resulted in mild to moderate, early-onset and transient ocular changes, such as reddening of the conjunctivae and sclerae, discharge and chemosis.These effects were reversible and were no longer evident 14 days after treatment, the end of the observation period for all animals. Slight opacity of the cornea, affecting the whole area, was noted in all animals at the 1- and 24-hour reading due to staining produced by the test item. Slight opacity of the cornea, affecting the whole area, was again visible in one animal 72 hours after treatment. No abnormal findings were observed in the iris of any animal at any of the examinations. No corrosion was observed at any of the measuring intervals. Brown staining of the treated eyes produced by the test item was observed in all animals 1 and 24 hours after treatment. No clinical signs were observed.Thus, the test item did not induce significant or irreversible damage to the rabbit eye.

Thus from overall observation of experiment the test substance Basic Brown 16 was observed to be not irritating to rabbit eye.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Additional information

Skin irritation:

In a study conducted by N. Alepeeet. al.(2016), skin irritation was evaluated by Concordance of the in vitro results by OD and HPLC/UPLC-spectrophotometry with in vivo Draize skin test at the concentration of 100 %. Basic brown 16 were found to be non-irritating. Hence, Basic brown 16 were not classified according to the test.

In a opinion given by Scientific Committee on Consumer Safety for Basic Brown 16 (2012), skin irritation were evaluated in three young adult New Zealand White rabbits by using Basic Brown 16 applied by topical semi-occlusive application of 0.5 g to the intact left flank for four hours. The scoring of skin reactions was performed at 1, 24, 48 and 72 hours after removal of the dressing. The mean score was calculated across 3 scoring times (24, 48 and 72 hours after patch removal) for each animal for erythema/eschar grades and for oedema grades, separately. The mean erythema/eschar score and the mean oedema score was 0 for all three animals. No signs of irritation were observed in treated rabbits. However slight brown staining was observed in all animals at the 1 and 24-hourreading. No corrosive effects at any of the measuring intervals and no clinical signs were observed. No significant or irreversible damage to skin were observed. Thus, based on the experimental observation and result it is concluded that the Basic Brown 16 is not irritating to rabbit skin.

Similar to above reference, in 2003, skin irritation were evaluated in New Zealand white rabbit treated with Basic brown 16 in the concentration of 5 g. Animals were observed for skin reactions after 24 and 72 hours. No skin reactions were observed in treated rabbits. Therefore, Basic brown 16 were considered to be non-irritating.

Similar to above reference, in 2011 for read across Basic Red 76, skin irritation were evaluated on New Zealand White rabbits according to OECD 404 Guidelines to determine the irritation potency of Basic Red 76. Approximately 24 hours prior to the treatment, the dorsal fur was shaved to expose an area of about 150 cm² intact shaved back skin of each animal and were treated with aliquot of 0.5 g of the moistened Basic Red 76. The patch was removed 4 hours after semi-occlusive contact. The skin reactions were assessed approximately 1, 24, 48 and 72 hours after termination of the exposure. There were no visible signs of irritation. However, slight red staining was observed in all animals from the 1-hour reading up to 72 hours after treatment. Staining persisted in one animal up to a 14-day examination. Under the conditions of the study, the undiluted Basic Red 76 was not irritating to rabbit skin.

Similar to above reference, in 2003 for read across Basic Red 76, skin irritation were evaluated on New Zealand White male and female rabbits by using Basic Red 76 applied without any vehicle at the level of 0.5 g/in 2 to the backs of 3 rabbits of each sex with either shorn intact or scarified skin. The sample was occlusively covered and left in place for 24 hours. Readings were made according to Draize upon removal of the test material and daily for 14 days post administration. There were no observable reactions to the dye. Therefore, Basic Red 76 was considered “not irritant” to rabbit skin.

Similar to above reference, in 2003 for read across Basic Red 76, skin irritation were evaluated on New Zealand White rabbits by using Basic Red 76 in the concentration of 0.5 g dampened with 0.5 ml distilled water and applied to an area of 1 in 2 on the backs of 3 rabbits each with either shorn intact or scarified skin. The sample was covered by an impervious material and left in place for 24 hours and skin reactions were recorded after 24 and 72 hours. No reactions were observed in treated skin. Therefore, Basic Red 76 was considered “not irritant” to rabbit skin.

Similar to above reference, in 2008 for another read across (8-((4-Amino-3-nitrophenyl)azo)-7-hydroxy-2-naphthyl)trimethylammonium chloride, skin irritation were evaluated on New Zealand White rabbits by using (8-((4-Amino-3-nitrophenyl)azo)-7-hydroxy-2-naphthyl)trimethylammonium chloride in the concentration of 0.5 grams applied on to clipped skin for 4 hours using a semi-occlusive dressing and observed for 1, 24, 48 and 72 hours after exposure. Very slight erythema and/or very slight to slight oedema were observed in treated rabbits, which had resolved within 48 hours. Yellow-brown staining of the treated skin was observed throughout the observation period, which did not hamper the scoring of the skin reactions. No remnants of the test substance were present on the skin. Thus, based on the above results, (8-((4-Amino-3-nitrophenyl) azo)-7-hydroxy-2-naphthyl) trimethylammonium chloride was considered to be not irritant to rabbit skin.

Similar to above reference, in 2008 for another read across (8-((4-Amino-3-nitrophenyl)azo)-7-hydroxy-2-naphthyl)trimethylammonium chloride, skin irritation were evaluated on New Zealand White male and female rabbits by using (8-((4-Amino-3-nitrophenyl)azo)-7-hydroxy-2-naphthyl)trimethylammonium chloride in the concentration of 0.5 g per square inch either to the shorn intact or to scarified skin on the back. The sample was covered by a linen cloth and plastic foil which were fixed by an elastic bandage and left in place for 24 hours. Readings were made according to Draize upon removal of the test material and then daily for the following 14 days. No irritations were observed on intact or to scarified skin on the back of treated animal. Therefore, (8-((4-Amino-3-nitrophenyl) azo)-7-hydroxy-2-naphthyl)trimethylammonium chloride was considered to be not irritant to rabbit skin.

Thus, on the basis of available values for target Basic Brown 16 and two read across Basic Red 76 and Basic brown 17, it can be concluded that the substance cannot be classified as per the criteria of CLP regulation for skin irritation.

 

Eye irritation:

In a opinion given by Scientific Committee on Consumer Safety for Basic Brown 16 (2008), eye irritation were evaluated in New Zealand White rabbits by using Basic Brown 16 applied I 0.1 g into the left eye of each of three young adult rabbits. Mild to moderate, early-onset and transient ocular changes, such as reddening of the conjunctivae and sclerae, discharge and chemosis were observed in treated rabbits. These effects were reversible and were no longer evident on 14 days after treatment. Slight opacity of the cornea, affecting the whole area, was noted in all animals at the 1- and 24-hour reading due to staining produced by the Basic Brown 16. Slight opacity of the cornea, affecting the whole area, was again visible in one animal after 72 hours. No abnormal findings were observed in the iris of any animal. No corrosion was observed at any of the measuring intervals. Brown staining of the treated eyes was observed in all animals at 1 and 24 hours after treatment. No clinical signs were observed. Thus, the Basic Brown 16 did not induce significant or irreversible damage to the rabbit eye. Therefore, Basic Brown 16 was considered to be not irritating to rabbit eye.

Similar to above reference, in 2003 for read across Basic Red 76, eye irritation were evaluated in New Zealand White rabbits in the concentration of 0.1 ml of 0.5% solution instilled into the conjunctival sac of the left eye of three rabbits. No effects on the cornea or iris in any of the test animals, however, there was a discoloration of the conjunctivae were observed. Therefore, Basic Red 76 was considered to be not irritating to rabbit eyes.

Similar to above reference, in 2011 for read across, eye irritation were evaluated in New Zealand White rabbits according to OECD 405 Guidelines by using Basic Red 76 in the concentration of 100 mg instilled into the conjunctival sac of the left eye animals. Undiluted Basic Red 76 resulted in mild, early-onset and transient ocular changes, such as reddening of the conjunctivae and sclerae, discharge and chemosis. The effects were reversible and were no longer evident after 72 hours of treatment. No abnormal findings were observed in the cornea or iris and at any of the measuring intervals. Slight red staining of the treated eyes was observed in all animals at the 1- and 24- hour reading and persisted in two animals up to 48 hours and in one animal up to 72 hours after treatment. Therefore, undiluted Basic Red 76 was slightly irritating to rabbit eyes.

Similar to above reference, in 2003 for another read across Basic Brown 17, eye irritation were evaluated in New Zealand White rabbits in the concentration of 0.1 ml of a 0.5 % (w/v) in 0.9% saline solution) was instilled into the conjunctival sac of the left eye of each of three rabbits. No washing of test material solution was carried out. Apart from a discoloration of the conjunctivae, no effects were noted on the cornea and iris in treated rabbits. Thus, Basic Brown 17 was considered to be not irritating to rabbit eyes.

Similar to above reference, in 2003 for another read across Basic Brown 17, eye irritation were evaluated in New Zealand White rabbits in the concentration of 45 mg (a volume of approximately 0.1 ml) were instilled into one eye of each of three rabbits. Irritation was noted on the iris in two test animals and on the conjunctivae in all animals. lridial irritation grade 1 was observed and had resolved within 24 or 72 hours. The irritation of the conjunctivae consisted of redness, chemosis and discharge and had completely resolved within 7 days in all treated rabbits. No corneal opacity was observed, and treatment of the eyes with 2% fluorescein, 24 hours after test substance instillation revealed no corneal epithelial damage. Thus, Thus, Basic Brown 17 was considered to be not irritating to rabbit eyes.

Based on the available data for target Basic Brown 16 and two read across Basic Red 76 and Basic brown 17, it can be concluded that the substance cannot be classified as per the criteria of CLP regulation for skin irritation.

Justification for selection of skin irritation / corrosion endpoint:
Basic brown 16 were found to be non-irritating to skin

Justification for selection of eye irritation endpoint:
Basic brown 16 were found to be non-irritating to eye

Justification for classification or non-classification

It can be concluded that the substance Basic Brown 16 (CAS no. 26381 -41 -9) and its two read across Basic Red 76 (CAs no. 68391 -30 -0) and Basic brown 17 (CAS no. 68391 -32 -2) cannot be classified as per the criteria of CLP regulation for skin irritation and eye irritation.