Salicylonitrile

Endpoint summary

Description of key information

Skin irritation: erythema=1.0, oedema=0.0, mean scores for all animals, in vivo skin irritation, Test No. 404, Fangfang 2012
Eye Irritation: Irreversible effects on the eye, in vivo eye irritation, Test No. 405, Fangfang 2012

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:

September 29 2011 - May 14 2012

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The study was conducted according to a national guideline method, essentially equivalent to a OECD TG 404 (1981 edition). Although not conducted under GLP (OECD), the study was conducted under a similar national quality assurance system. Adequately documented study, purity data available separately for test batch.

Qualifier:

according to guideline

Guideline:

other: Guidelines for the testing of chemicals: Test No. 404: Acute dermal irritation/corrosion test (Minestry of Environmental Protection People's Republic of China)

Deviations:

not specified

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

1981 edition

Deviations:

yes

Remarks:

Scoring at 30/60min not reported.

GLP compliance:

no

Remarks:

The study was conducted in compliance with a national quality assurance system, with similar objectives to GLP (OECD).

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: not reported
- Weight at study initiation: ca. 2kg
- Housing: Suspended, wire bottom, stainless steel, 1 animal per cage
- Diet: ad libitum (Statement that contaminents in feed and water effecting study outcome not expected)
- Water: ad libitum (Municipal water supply, analyzed by City Water Quality Monitoring Center of Zhejiang Province)
- Acclimation period: 5 days
- Sex: male and female, nulliparous & non-pregnant

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20±3°C
- Humidity (%): 30-70%
- Air changes (per hr): 10-12
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: not reported

Type of coverage:

occlusive

Preparation of test site:

shaved

Vehicle:

water

Remarks:

distilled

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount applied: total dose 0.5g
- Concentration: not reported

VEHICLE
- Amount: not reported

Duration of treatment / exposure:

4 hours

Observation period:

Observations were made at 24h, 48h, 72h, or until "reversibility was observed" up to 14 days. The study reported observations to at least 72h (guideline minimum), possibly to 14 days (as tabulated).

Number of animals:

4

Details on study design:

TEST SITE
- Area of exposure: 3x3cm clipped free of hair; 2x3cm 4 ply surgical gauze patch, secured with non-irritating adhesive tape.
- % coverage:
- Type of wrap if used: trunk of each animal wrapped in orthopedic stockinette to prevent possible ingestion of test substance.

REMOVAL OF TEST SUBSTANCE
- Washing: Washed gently with room temperature tap water without altering the existing response, or the integrity of the epidermis.
- Time after start of exposure: 4 hours

SCORING SYSTEM:
- According to the Guidelines for the testing of chemicals: Test No. 404: Acute dermal irritation/corrosion test (Minestry of Environmental Protection People's Republic of China). For each animal, all of the erythema and edema scores were added, and the sum was divided by 4 to obtain an individual irritation score. The primary irritation index was determined by calculating the mean of the irritation scores for all the animals and was used to obtain a rating for the test substance.

Erythema Formation Score
No erythema 0
Very slight erythema (barely perceptible) 1
Well defined erythema 2
Moderate to severe erythema 3
Severe erythema (beef redness) and eschar formation 4

Edema Formation
No edema 0
Very slight edema (barely perceptible) 1
Slight edema (edges of area well defined by definite raising) 2
Moderate edema (raised approximately 1 mm) 3
Severe edema (raised more than 1 mm and extending beyond area of exposure) 4

Classification of Test Substance
Non-irritating 0.0-0.4
Mildly Irritating 0.5-1.9
Moderately Irritating 2.0-5.9
Severely Irritating 6.0-8.0
The primary irritation index is calculated using only the observation scheduled through 72h.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72h

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

48 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72h

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 72h

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72h

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

48 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72h

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 72h

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72h

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

48 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72h

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 72h

Irritation parameter:

erythema score

Basis:

animal #4

Time point:

24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72h

Irritation parameter:

erythema score

Basis:

animal #4

Time point:

48 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72h

Irritation parameter:

erythema score

Basis:

animal #4

Time point:

72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 72h

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #4

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

Signs of dermal irritation or defects are presented in Table 1. Very slight erythema and no edema in the skin of New Zealand white rabbits was observed after the patch removal. Therefore, the maximum primary irritation index of the test substance was calculated to be 1.00.

Other effects:

None reported.

Table 1: Acute dermal irritation in rabbits

Animal number	Start weight (kg)	Final weight (kg)	Effect	Score time point
				24h	48h	72h	4d	5d	6d	7d	8d	9d	10d	11d	12d	13d	14d
1	2.41	2.49	Erythema	1	1	0	0	0	0	0	0	0	0	0	0	0	0
			Edema	0	0	0	0	0	0	0	0	0	0	0	0	0	0
2	2.52	2.60	Erythema	1	1	0	0	0	0	0	0	0	0	0	0	0	0
			Edema	0	0	0	0	0	0	0	0	0	0	0	0	0	0
3	2.80	2.89	Erythema	1	1	0	0	0	0	0	0	0	0	0	0	0	0
			Edema	0	0	0	0	0	0	0	0	0	0	0	0	0	0
4	2.13	2.22	Erythema	1	1	0	0	0	0	0	0	0	0	0	0	0	0
			Edema	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Primary irritation scores				1.0	1.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0

Descriptive rating: mildly irritating

Interpretation of results:

GHS criteria not met

Conclusions:

The current OECD and EU guidelines deviate from the 1981 OECD guideline primarily with respect to numbers of animals, staggered and successive testing, and the preliminary consideration of weight-of-evidence and in vitro data. The report is thus considered to be adequate for classification and risk assessment purposes.

Executive summary:

In a primary dermal irritation study, male or female New Zealand white rabbits (4 animals) were dermally exposed to 0.5g of 2-cyanophenol in distilled water for 4 hours with a 2x3cm patch. Animals were then observed for up to 14 days. Very slight erythema and no edema in the skin of New Zealand white rabbits was observed after the patch removal in all animals, and was fully reversible within 72h. In this study, 2-cyanophenol was found to be mildly irritating according to Guidelines for the testing of chemicals: Test No. 404: Acute skin irritation/corrosion test (Ministry of Environmental Protection of the People's Republic of China, 2004).

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:

29th September 2011 - 14th May 2012

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The study was conducted according to a national guideline method, essentially equivalent to a OECD TG 405 (1987 edition). Although not conducted under GLP (OECD), the study was conducted under a similar national quality assurance system. Adequately documented study, although not all raw data was reported, this did not effect the conclusions, especially considering the detailed description of the observed effects and persistence. Purity data available separately for test batch.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

1987 edition

Deviations:

yes

Remarks:

Only total scores for cornea and conjunctivae reported.

Qualifier:

according to guideline

Guideline:

other: Guidelines for the Testing of Chemicals: Test No. 405: Acute eye irritation/corrosion test (MEP, People's Republic of China, 2004)

Deviations:

not specified

Principles of method if other than guideline:

The methodology followed was equivalent to the 1987 version of OECD TG 405, which deviates from the latest 2002 version, particularly in the number of animals used. The inital test with 4 animals ("unwashed test") was followed by two further tests of 3 animals each, with eye irrigation at 4 seconds after instillation, and irrigation at 30 seconds, respectively. Evaluation was according to Kay&Calandra, although not explicitly referenced.

GLP compliance:

no

Remarks:

The study was conducted in compliance with a national quality assurance system, with similar objectives to GLP (OECD).

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Age at study initiation: not reported
- Weight at study initiation: ca. 2.3kg
- Housing: Suspended, wire bottom, stainless steel, 1 animal per cage
- Diet: ad libitum (Statement that contaminents in feed and water effecting study outcome not expected)
- Water: ad libitum (Municipal water supply, analyzed by City Water Quality Monitoring Center of Zhejiang Province)
- Acclimation period: 5 days
- Sex: male and female, nulliparous & non-pregnant

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20±3°C
- Humidity (%): 30-70%
- Air changes (per hr): 10-12
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: not reported

Vehicle:

unchanged (no vehicle)

Remarks:

Ground to a fine powder and sieved through 0.154mm mesh

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.1g

Duration of treatment / exposure:

- Initial Test: 24 hours
- Second Test: 4 seconds
- Third Test: 30 seconds

Observation period (in vivo):

21 days with observations at 1h, 24h, 48h, 72h, 96h, 7 days, 14 days, 21 days.

Number of animals or in vitro replicates:

- Initial Test: 4
- Second Test: 3
- Third Test: 3

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: gently rinsed for 30s with 0.9% saline water
- Initial Test, time after start of exposure: 24h
- Second Test, time after start of exposure: 4s and 24h
- Third Test, time after start of exposure: 30s and 24h

SCORING SYSTEM:
As described in The Guidelines for the Testing of Chemicals: Test No. 405: Acute Eye Irritation/Corrosion Test (Ministry of Environmental Protection of the People's Republic of China.

TOOL USED TO ASSESS SCORE:
Corneas of all treated eyes were examined with sodium fluorescein ophthalmic solution at 24h. Any corneas exhibiting staining were followed at subsequent observation points with fluorescein solution until staining no longer occurred.

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #4

Time point:

24/48/72 h

Score:

40

Reversibility:

not fully reversible within: 21 days

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24 h

Score:

20

Reversibility:

fully reversible within: 21 days

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

48 h

Score:

20

Reversibility:

fully reversible within: 21 days

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

72 h

Score:

15

Reversibility:

fully reversible within: 21 days

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24 h

Score:

40

Reversibility:

not fully reversible within: 21 days

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

48 h

Score:

40

Reversibility:

not fully reversible within: 21 days

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

72 h

Score:

20

Reversibility:

not fully reversible within: 21 days

Irritation parameter:

iris score

Basis:

animal: #1, #3, #4

Time point:

24/48/72 h

Score:

5

Reversibility:

not fully reversible within: 21 days

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

5

Reversibility:

fully reversible within: 21 days

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24 h

Score:

12

Reversibility:

not fully reversible within: 21 days

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

48 h

Score:

12

Reversibility:

not fully reversible within: 21 days

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

72 h

Score:

18

Reversibility:

not fully reversible within: 21 days

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

16

Reversibility:

not fully reversible within: 21 days

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

16

Reversibility:

not reversible

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

18

Reversibility:

not fully reversible within: 21 days

Irritation parameter:

overall irritation score

Remarks:

Sum of cornea, iris and conjunctivae scores

Basis:

animal #1

Time point:

21 d

Score:

41

Reversibility:

not fully reversible within: 21 days

Remarks on result:

other: No eye irrigation; see tables for full data

Irritation parameter:

overall irritation score

Remarks:

Sum of cornea, iris and conjunctivae scores

Basis:

animal #2

Time point:

21 d

Score:

10

Reversibility:

not fully reversible within: 21 days

Remarks on result:

other: No eye irrigation; see tables for full data

Irritation parameter:

overall irritation score

Remarks:

Sum of cornea, iris and conjunctivae scores

Basis:

animal #3

Time point:

21 d

Score:

28

Reversibility:

not fully reversible within: 21 days

Remarks on result:

other: No eye irrigation; see tables for full data

Irritation parameter:

overall irritation score

Remarks:

Sum of cornea, iris and conjunctivae scores

Basis:

animal #4

Time point:

21 d

Score:

22

Reversibility:

not fully reversible within: 21 days

Remarks on result:

other: No eye irrigation; see tables for full data

Irritation parameter:

chemosis score

Basis:

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

Time point:

24/48/72 h

Remarks on result:

other: not reported

Irritant / corrosive response data:

Observation results of eye reaction (unwashed test) after the application were shown as diffuse beefy red, swelling with lids more than half closed, discharge with moistening of the lids and hairs, and a considerable area around the eye in the conjunctivae. The main symptoms affecting the cornea include easily discernible translucent area, details of iris slightly obscured, the damage area covered greater than three quarters, or up to whole area. There was observed fluorescein staining with scales in corneas and congestion in iris. Area of angiogenesis was greater than one quarter, but less than half in cornea surface (moderately. The corneas displayed slight dulling of normal luster. The highest average of total score until 4d after instillation was 55.50. Duration of irritation reaction was the average of total score on day 7>40.

Observation results of eye reaction (washed test, 4s after instillation) were shown as diffuse beefy red, swelling with lids about half closed, discharge with moistening of the lids and hairs, and a considerable area around the eye in the conjunctivae. The main symptoms affecting the cornea include easily discernible translucent area, details of iris slightly obscured, the damage area covered greater than three quarters. There was observed fluoresdcein staining within the corneal opacity area and congestion in iris. Area of angiogenesis was greater than ten percent, but less than one quarter in cornea surface (mildly). The corneas displayed slight dulling of normal luster. The highest average of total score until 4d after instillation was 40.67. Duration of irritation reaction was the average of total score on day 7>20.

Observation results of eye reaction (washed test, 30s after instillation) were shown as diffuse beefy red, swelling with lids about half closed, discharge with moistening of the lids and hairs, and a considerable area around the eye in the conjunctivae. The main symptoms affecting the cornea include easily discernible translucent area, details of iris slightly obscured, the damage area covered greater than three quarters. There was observed fluorescein staining within the corneal opacity area and congestion in iris. Area of angiogenesis was greater than ten percent, but less than one quarter in cornea surface (mildly) . The corneas were slight dulling of normal luster. The highest average of total score until 4d after instillation was 53.00. Duration of irritation reaction was the average of total score on day 7<40, more than half animals>30, but no animals>60.

Other effects:

None reported.

Table 1: Initial and final body weights for initial test (“unwashed”).

	Animal 1	Animal 2	Animal 3	Animal 4
Start weight	2.40	2.42	2.30	2.13
Finish weight	2.46	2.50	2.42	2.21

 

Table 2: Ocular reactions for the initial test (“unwashed”).

Time after treatment	Animal number	Score			Total score	Average of total score
		Cornea	Iris	Conjunctivae
1h	1	-	0	6	6	6.5
	2	-	0	6	6
	3	-	0	6	6
	4	-	0	8	8
24h	1	40	5	12	57	55.50
	2	20	5	16	41
	3	40	5	16	61
	4	40	5	18	63
48h	1	40	5	12	57	55.50
	2	20	5	16	41
	3	40	5	16	61
	4	40	5	18	63
72h	1	40	5	18	63	50.75
	2	15	5	16	36
	3	20	5	16	41
	4	40	5	18	63
96h	1	40	5	18	63	50.75
	2	15	5	16	36
	3	20	5	16	41
	4	40	5	18	63
7 day	1	40	5	18	63	44.75
	2	15	5	16	36
	3	20	5	14	39
	4	20	5	16	41
14 day	1	20	5	18	43	30.5
	2	5	5	12	22
	3	10	5	18	33
	4	5	5	14	24
21 day	1	20	5	16	41	25.25
	2	0	0	10	10
	3	5	5	18	28
	4	5	5	12	22

The highest average total score up to 4 days after instillation: 55.50

Duration of irritation reaction: the average total score on day 7: >40

Evaluation: “very severely irritating” (rating 7)

Table 3: Ocular reactions at 21 days for the post-instillation irrigation satellite groups (second and third tests), with eye irrigation at 4s and 30s after instillation respectively.

 

Time after treatment	Animal number	Score			Total score	Average of total score
		Cornea	Iris	Conjunctivae
Second test (4s), 21 days	1	0	0	0	0	0
	2	0	0	0	0
	3	0	0	0	0
Evaluation	The highest average total score up to 4 days after instillation: 40.67 Duration of irritation reaction: the average total score on day 7: >20 Evaluation: “severely irritating” (rating 6)
Third test (30s), 21 days	1	5	5	10	20
	2	5	5	8	18
	3	0	0	8	8
Evaluation	The highest average total score up to 4 days after instillation: 53.00 Duration of irritation reaction: the average total score on day 7: <40, more than half animals >30, but no animals >60 Evaluation: “severely irritating” (rating 6)

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

The current OECD and EU guidelines deviate from the 1987 OECD guideline primarily with respect to numbers of animals and use of satellite groups. Despite only the total scores for cornea and conjunctivae being reported, the descriptive summary allows reconstruction of the maximum scores for corneal opacity (2), the area involved (4), and iris (1). A combination of the descriptive summary of the observations and the effects being observed to 21 days, means the report is adequate for classification and risk assessment purposes.

Executive summary:

In a primary eye irritation study, 0.1g of 2-cyanophenol was instilled into the conjunctival sac of the right eye of male or female New Zealand white rabbits (4) for 24 hours. Treated eyes were gently washed with 0.9% saline solution after 24 hours. Animals then were observed for 21 days.  Irritation was scored by the method of Draize. Ocular reactions after instillation were diffuse beefy red conjunctivae, swelling with lids more than half closed, discharge with moistening of the lids and hairs, and a considerable area around the eye. The cornea showed discernible translucent areas, details of iris slightly obscured, and the damaged area covered greater than three quarters, or up to whole area. There was congestion evident in the iris. Effects were persistent to 21 days. In this study, 2 -cyanophenol was found to be very severely irritating (rating 7) according to Guidelines for the testing of chemicals: Test No. 405: Acute eye irritation/corrosion test (Ministry of Environmental Protection of the People's Republic of China, 2004).

Two satellite groups, each of 3 animals, were instilled with 0.1g of 2-cyanophenol into the conjunctival sac of the right eye for 24 hours. Treated eyes were gently washed with 0.9% saline solution after 4 and 30 seconds respectively. Animals then were observed for 21 days.  Irritation was scored by the method of Draize.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation/corrosion:

A single in-vitro study conducted according to guideline OECD TG 431 and under GLP is available. The study is considered to be relevant, and reliable for the purposes of risk assessment, but not adequate for full sub-category classification and labelling. The Epiderm Skin Corrosivity Test (Model EPI-200) gave a 99.4% mean percentage cell viability after 3 minutes, but only 11.7% after one hour, which is below the threshold of 15% for classification as corrosive.

 

Access to further relevant information has been obtained, which became available after the initial submission of this dossier in 2010. The new information is the result of in vivo testing conducted by a non-Registrant in accordance with the requirements of a national registration scheme outside of the European Union.

 

In a primary dermal irritation study, male or female New Zealand white rabbits (4 animals) were dermally exposed to 0.5g of the test material in distilled water for 4 hours with a 2x3cm patch. Animals were then observed for up to 14 days. Very slight erythema and no edema was observed after patch removal in all animals, and was fully reversible within 72h. In this study, the test material was found to be mildly irritating according to Guidelines for the testing of chemicals: Test No. 404: Acute dermal irritation/corrosion test (Ministry of Environmental Protection of the People's Republic of China, 2004). The guideline is equivalent to OECD TG 404 (1981).

 

The current OECD and EU Test Methods deviate from the 1981 OECD guideline primarily with respect to numbers of animals, staggered and successive testing, and the preliminary consideration of weight-of-evidence and in vitro data, and thus the study by Fangfang(2012) is still considered to follow valid methodology. The study is considered to be reliable with restrictions because it was not conducted under harmonised GLP (OECD Mutual Acceptance of Data), although it was conducted under a similar national quality control scheme. The report is considered to be adequate for classification purposes.

 

No acute dermal toxicity studies are available to provide additional relevant observations. However, in a skin sensitisation local lymph node assay (Weber, 2008), a range finding study with 2 animals / concentration (64.8% and 50% w/w, applied 3 times on consecutive days) was reported, with no overt systemic toxicity, no excessive local skin irritation at the application site, nor important increase in ear thickness observed. Similarly, in the main study with 5 animals / concentration (64.8%, 50% and 25% w/w, applied 3 times on consecutive days) no systemic toxicity, no excessive local skin irritation at the application site, nor an important increase in ear thickness was observed.

 

A supplementary literature search for skin and eye irritation data related to the positional isomers 3-hydroxybenzonitrile and 4-hydroxybenzonitrile returned no relevant data (SciFinder, keyword and synonyms; eChemPortal, CAS number; 21.6.2012), in addition to the original literature search.

 

The results of a valid in vivo study conducted according to a recognised guideline method and under a suitable quality assurance system, are regarded as being more relevant than those from in vitro test systems. For the purposes of formally meeting data submission requirements under Regulation (EC) No. 1907/2006, Article 13(4), studies must be conducted in accordance with GLP and the OECD MAD scheme (see REACH FAQ (31.5.2012), 11.7). As a result, although Fangfang (2012) is considered to be a scientifically valid study, the data requirements for this endpoint must be considered on the basis of Annex XI, 1.1.2, concerning the use of existing data on properties from experiments not carried out according to GLP or recognised guidelines. The study is 1) adequate for the purposes of classification, labelling and risk assessment, 2) the study adequately and reliably covers the parameters foreseen in the equivalent OECD TG 404 and EU Test Method B.4, 3) the exposure durations and observations were as required in those guidelines, and 4) the study was adequately documented. As a result, the study is considered to meet the data requirements for Regulation (EC) No. 1907/2006, Annex VIII, 8.1.1, and inter alia Annex VII, 8.1., and are carried forward for risk assessment and classification purposes. The available data is considered to be conclusive for skin irritation / corrosion.

 

Eye irritation/corrosion:

Because the substance was initially classified as corrosive to skin in a validated and accepted in-vitro test (OECD TG 431), its eye corrosivity was not tested in animals under REACH, since similar severe effects were to be expected in an in-vitro or in-vivo eye test (see stepwise testing strategy in OECD TG 405).Access to further relevant information has been obtained, which became available after the initial submission of this dossier in 2010. The new information is the result of in vivo testing conducted by a non-Registrant in accordance with the requirements of a national registration scheme outside of the European Union. 

 

In a primary eye irritation study, 0.1g of test material was instilled into the conjunctival sac of the right eye of male or female New Zealand white rabbits (4 animals) for 24 hours. Treated eyes were gently washed with 0.9% saline solution after 24 hours. Animals then were observed for 21 days. Irritation was scored by the method of Draize, and evaluation appeared to be according to Kay and Calandra (not explicitly referenced). Ocular reactions after instillation were diffuse beefy red conjunctivae, swelling with lids more than half closed, discharge with moistening of the lids and hairs, and a considerable area around the eye. The cornea showed discernible translucent areas, details of iris slightly obscured, and the damaged area covered greater than three quarters, or up to whole area. There was congestion evident in the iris. Effects were persistent to 21 days.

 

Two satellite groups, each of 3 animals, were instilled with 0.1g of test material into the conjunctival sac of the right eye for 24 hours. Treated eyes were gently washed with 0.9% saline solution after 4 and 30 seconds respectively. Animals then were observed for 21 days. Irritation was scored by the method of Draize. Effects were fully reversible within 21 days for the satellite group with the washing after 4s. Effects were persistent to 21 days for the satellite group with washing at 30s.In the main study, the test material was found to be very severely irritating according to Guidelines for the testing of chemicals: Test No. 405: Acute eye irritation/corrosion test (Ministry of Environmental Protection of the People's Republic of China, 2004). The guideline is equivalent to OECD TG 405 (1987).

 

The current OECD and EU Test Method guidelines deviate from the 1987 edition of OECD Guideline 405 primarily with respect to numbers of animals, and use of satellite groups, and thus the study is considered to follow valid methodology. Despite only the total scores for cornea and conjunctivae being reported, the descriptive summary allows reconstruction of the maximum scores for corneal opacity (2), the area involved (4), and iris (1). The study is considered to be reliable with restrictions due to lack of harmonised GLP (OECD MAD), and above mentioned completeness of the raw data reporting. The report is considered to be adequate for classification purposes on the basis of the raw data presented, the descriptive summaries of the observations, and the clear non-reversal of ocular damage within 21 days.

 

A supplementary literature search for skin and eye irritation data related to the positional isomers 3-hydroxybenzonitrile and 4-hydroxybenzonitrile returned no relevant data (SciFinder, keyword and synonyms; eChemPortal, CAS number; 21.6.2012), in addition to the original literature search.

 

For the purposes of formally meeting data submission requirements under Regulation (EC) No. 1907/2006, Article 13(4), studies must be conducted in accordance with GLP and the OECD MAD scheme (see REACH FAQ (31.5.2012), 11.7). As a result, although Fangfang (2012) is considered to be a scientifically valid study, the data requirements for this endpoint must be considered on the basis of Annex XI, 1.1.2, concerning the use of existing data on properties from experiments not carried out according to GLP or recognised guidelines. The study is 1) adequate for the purposes of classification, labelling and risk assessment, 2) the study adequately and reliably covers the parameters foreseen in the equivalent OECD TG 405 and EU Test Method B.5, 3) the exposure durations and observations were as required in those guidelines, and 4) the study was documented sufficiently for classification and risk assessment. As a result, the study is considered to meet the data requirements for Regulation (EC) No. 1907/2006, Annex VIII, 8.2.1, and inter alia Annex VII, 8.2., and are carried forward for risk assessment and classification purposes. The available data is considered to be conclusive for eye irritation / corrosion.

Justification for selection of skin irritation / corrosion endpoint:
The study was conducted according to a national guideline method, essentially equivalent to a OECD TG 404 (1981 edition). Although not conducted under GLP (OECD), the study was conducted under a similar national quality assurance system. Adequately documented study, purity data available separately for test batch.

Justification for selection of eye irritation endpoint:
The study was conducted according to a national guideline method, essentially equivalent to a OECD TG 405 (1987 edition). Although not conducted under GLP (OECD), the study was conducted under a similar national quality assurance system. Adequately documented study, although not all raw data was reported, this did not effect the conclusions, especially considering the detailed description of the observed effects and persistence. Purity data available separately for test batch.

Justification for classification or non-classification

Skin irritation/corrosion:

The substance met the criteria for classification as "corrosive" as defined in the EpiDerm Skin Corrosivity Test (EU Method B.40 bis) (Farmer, 2008). This method can only discriminate between "corrosive" and "non-corrosive" chemicals, but not between the three Skin Corrosive Category 1 subcategories: 1A, 1B, and 1C – despite the reported cell viability at 3 min (not validated). In the absence of further information, the substance was classified with the more conservative classification Skin Corrosion Category 1A (H314: Causes severe skin burns and eye damage), according to Regulation (EC) 1272/2008, Annex I, Part 3, 3.2.2. However, the results of the in vitro skin corrosivity study are overridden by the recent in vivo data for skin irritation (Fangfang, 2012), indicating that the substance does not meet the criteria for classification set out in Regulation (EC) 1272/2008, Annex I, Part 3, 3.2.2.7. Mean scores for erythema and oedema in all animals were <2 (1.0 and 0.0 for all animals, respectively), and fully reversible within 72h. As a consequence, classification of the substance for skin irritation/corrosion is not required.

 

Eye irritation/corrosion:

The substance met the criteria for classification as "corrosive" as defined in the EpiDerm Skin Corrosivity Test (EU Method B.40 bis) (Farmer, 2008). This method can only discriminate between "corrosive" and "non-corrosive" chemicals, but not between the three Skin Corrosive Category 1 subcategories: 1A, 1B, and 1C. In the absence of further information, the substance would be classified with the more conservative classification Skin Corrosion Category 1A (H314: Causes severe skin burns and eye damage), according to Regulation (EC) 1272/2008, Annex I, Part 3, 3.2.2. However, the results of the in vitro skin corrosivity study are overridden by the recent in vivo data for both skin and eye irritation (Fangfang, 2012), indicating no classification for skin irritation, and classification in accordance with Regulation (EC) 1272/2008, Annex I, Part 3, 3.3.2.6 for having Irreversible effects on the eye (Category 1) (H318: Causes serious eye damage).