Amides, coco, N-[3-(dimethylamino)propyl], N-oxides

Endpoint summary

Description of key information

Skin

An acute dermal study was conducted (OECD 404, GLP) (Stepan, 2000). Three New Zealand White rabbits were exposed to the undiluted test substance. Effects were fully reversible within 7 days. The test substance was concluded to be not irritant.

A study (non-OECD, GLP not specified) in five Kleinrusse rabbits exposed to a diluted solution containing 25% active matter of the test substance was performed (Henkel KGaA, 1987). The following gradings were observed: Erythema 1.87 and oedema 1.13, concluding the substance is a mild irritant to skin.

Eye

A mucous membrane irritation study (OECD 405, GLP not specified) was conducted (Henkel KGaA, 1987). Four Kleinrusse rabbits were exposed to a diluted solution containing 25% active matter of the test substance. Results presented 3/4 animals with irreversible eye lesions after an observation period of 21 days. The test substance is categorized as seriously eye damaging. 

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

Study period:

From October 26, 1999 to November 16, 1999

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD guideline 404 and EU guideline B.4. GLP study.

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

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 to 16 weeks old
- Weight at study initiation: 2.48 to 3.09 kg
- Housing: The animals were individually housed in suspended metal cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: a minimum of five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17-23 ºC
- Humidity (%): 30 to 70 %
- Air changes (per hr): the rate was approximately fifteen changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours continuous light and 12 hours of darkness.

Type of coverage:

semiocclusive

Preparation of test site:

other: clipped

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL of the test material.

Duration of treatment / exposure:

4 hour

Observation period:

14 days

Number of animals:

3 animals

Details on study design:

TEST SITE
- Area of exposure: 2.5 x 2.5 cm
- Type of wrap if used: the trunk of the rabbit was wrapped in an elasticated corset (TUBIGRIP)

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Any residual test material was removed by gentle swabbing with cotton wool soaked in 74% Industrial Methylated Spirits.
- Time after start of exposure: 4 hours after application (when the patch was removed).

SCORING SYSTEM:
according to the OECD guideline nº 404.

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24 h

Score:

0

Irritation parameter:

edema score

Basis:

animal #1

Time point:

48 h

Score:

0

Irritation parameter:

edema score

Basis:

animal #1

Time point:

72 h

Score:

0

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24 h

Score:

0

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

48 h

Score:

0

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

72 h

Score:

0

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 7 days

Irritation parameter:

edema score

Basis:

animal #2

Time point:

48 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 7days

Irritation parameter:

edema score

Basis:

animal #2

Time point:

72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 7 days

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 7 days

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

48 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: Loss of skin elasticity.

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

72 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: Loss of skin elasticity and loss of skin flexibility. At day 7, crust formation was observed. At day 14, slight desquamation was observed.

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 7 days

Irritation parameter:

edema score

Basis:

animal #3

Time point:

48 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 7 days

Irritation parameter:

edema score

Basis:

animal #3

Time point:

72 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 7 days

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 7 days

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

72 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: Loss of skin elasticity and loss of skin flexibility. At day 7, crust formation was observed. At day 14, slight desquamation was observed.

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

48 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: Loss of skin elasticity

Irritant / corrosive response data:

No corrosive effects were noted.

Individual skin reactions following 4-hours exposure period

 

Skin Reaction	Observation Time	Individual Scores -Rabbit Number and Sex (Bodyweight kg)			Total
Erythema/Eschar Formation		1Male (3.09)	198 Male (2.74)	200 Female (2.48)
	1 Hour	0	2	2	( 4 )
	24 Hours	0	2	2	4
	48 Hours	0	2Le	2le	( 4 )
	72 Hours	0	2LeLf	2LeLf	4
	7 Days	-	OCf	OCf	( 0 )
	1 4 Days	-	OD	OD	( 0 )
Oedema Formation	1Hour	0	2	2	( 4 )
	24 Hours	0	1	2	3
	48 Hours	0	1	2	( 3 )
	72Hours	0	1	1	2
	7 Days	-	0	0	( 0 )
	14 Days	-	0	0	( 0 )
Sum of 24 and 72-hour Readings (S)                 13
Primary Irritation Index                               13/6 - 2.2

( ): total values not used for calculation of the primary index

Cf: crust formation

D: slight desquamation

Le: loss of skin elasticity

Lf: loss of skin flexibility

- : observation not required

 

Individual daily and mean scores for dermal irritation following 4-hour exposure

 

Skin Reaction	Reading (Hours)	Individual Scores - Rabbit Number and Sex (Bodyweight kg)
		1Male (3.09)	198Male (2.48)	200Female (2.74)
Erythema/Eschar Formation	24	0	2	2
	48	0	2	2
	72	0	2	2
total		0	6	6
Mean Score		0.0	2.2	2.0
Oedema Formation	24	0	1	2
	48	0	1	2
	72	0	1	1
Total		0	3	5
Mean Score		0.0	1.0	1.7

 

Interpretation of results:

GHS criteria not met

Conclusions:

Well-defined erythema was noted at two treated skin sites one hour after patch removal and at the 24, 48 and 72-hour observations. Slight oedema was noted at two treated skin sites one hour after patch removal. Very slight oedema was noted at two treated skin sites at the 72-hour observation. Mean values (all animals mean of 24/48/72h) are 0.9 oedema and 1.3 for erythema. One treated skin site appeared normal throughout the study. Crust formation was noted at two treated skin sites at the 7-day observation with slight desquamation at these two treated skin sites at the 14-day observation. Hence the substance was not classified for irritation according to GHS, EU.

Executive summary:

The Acute Dermal Irritation/corrosion assay for the test substance was performed in New Zealand rabbits. 0.5 mL of the test material was applied to a site of about 2.5 x 2.5 cm of intact skin of 3 rabbits and covered by semi-occlusive dressing. The administration of the test substance was performed in a sequential manner. The first animal was exposed to the test substance stepwise at three sites of the skin: site 1 for three minutes, site 2 for 1 hour and site 3 for 4 hours. Any residual test substance was removed by swabbing with cotton wool soaked in 74% Industrial Methylated Spirits. After considerations of the skin reactions produced on the first animal, the remaining two animals were exposed to the test substance for 4 hours. The treated areas of the animals were examine for erythema/eschar and oedema as well as other local skin alterations approximately 1, 24, 48, and 72 hours after patch removal. Additional observations were made on days 7 and 14 to assess the reversibility of skin reactions. Dermal alterations were scored according to OECD guideline nº 404. Well-defined erythema was noted at two treated skin sites one hour after patch removal and at the 24, 48 and 72-hour observations. Slight oedema was noted at two treated skin sites one hour after patch removal. Very slight oedema was noted at two treated skin sites at the 72-hour observation.One treated skin site appeared normal throughout the study.Crust formation was noted at two treated skin sites at the 7-day observation with slight desquamation at these two treated skin sites at the 14-day observation. No corrosive effects were noted.

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

Oct 1986 - Dec 1986

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Qualifier:

according to guideline

Guideline:

other: European Community

Version / remarks:

Amtsblatt der Europäischen Gemeinschaft L 251, 27. Jg.vom 19.09.1984 S. 106 - 108

Principles of method if other than guideline:

The test subsance was used with an active ingredients of 25% and was tested for its primary skin irritation under occlusive conditions on the shaved back skin of rabbits.

GLP compliance:

not specified

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Purity: 35%
- 25% active matter

Species:

rabbit

Strain:

other: Kleinrusse Chbb:HM/Fa

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Supplier: Firma Thomae, Bilbach, Germany
- Weight at study initiation: 2048 - 2308 g

Type of coverage:

occlusive

Preparation of test site:

shaved

Vehicle:

not specified

Amount / concentration applied:

0.5 ml of 25% active matter were applied

Duration of treatment / exposure:

4 h

Number of animals:

5

Details on study design:

TEST SITE
- Area of exposure: back. The test substance was applied on the left as well as on the right body site
- Coverage: 2.5 cm x 2.5 cm linth cloth
- Type of wrap if used: plastic foil and an acrylic bandage

OBSERVATION TIME POINTS:
1, 24, 48, 72 h and 7, 10, 14, 17, 21 days

SCORING SYSTEM:
- Method of calculation: Draize scoring system (0 - 4 points for erythema and edema)

OTHER:
The test animals have been observed and only animals with an intact skin were used for the study.

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Time point:

14 d

Score:

2.87

Max. score:

8

Reversibility:

fully reversible within: 14 d

Remarks on result:

positive indication of irritation

Table 1: Results of the individual test animals

Animal number	Hours after application										Days after application
	2			24			48		72		7		10		14		17		21		P.I.I
	E		Ö	E	Ö		E	Ö	E	Ö	E	Ö	E	Ö	E	Ö	E	Ö	E	Ö
467	1	0		2		1	1	1	1	0	0	0x	0	0x	0	0
466	1	0		2		1	2	1	1	1x	0	0x	0	0x	0	0
468	1	0		2		1	2	0	2	1	1	0x	0	0x	0	0
460	1	0		3		1	3	2	3	2	1	0x	0	0x	0	0
464	1	0		2		2	1	1	1	0	0	0x	0	0x	0	0
V	1.0			2.2			1.8		1.6		0.4		0.0		0.0
V	0.0			1.2			1.0		0.8		0.0		0.0		0.0						2.87

x = scab

V = average of erythema and oedema

P.I.I = primary (dermal) irritation index

Interpretation of results:

Category 3 (mild irritant) based on GHS criteria

Conclusions:

The test substance appears to be slightly irritant to the skin of rabbits. However, a classification according to Regulation (EU) No. 1272/2008 is not warranted.

Executive summary:

In a comparative study 10 anionic surfactants, 4 amphotric surfactants, 8 nioionic surfactants and 3 kationic surfactants with equal active ingredients of 25% were tested for their primary skin irritation under occlusive conditions on the shaved back skin of rabbits (Method: Off. J. Europ. Commun. 27, L 251, 1984 p. 106 - 108 and Guidelines for Testing of Chemicals, Section 4, No 404, OECD).

Two kationic surfactants were tested earlier after the same method. After a contact time of 4 hours the skin reactions of each animal were evaluated. Various strong reactions in kind of erythema and edema were observed. On the skin areas treated with one surfactant nectrotic skin changes and scar formation appeared. The same reactions had been observed after application of two surfactants in a further study.

According to the criteria in Off. J. Europ. Commun. 26, (L 257) 1983, p. 8 + 19 three surfactnats have to be classified as "corrosive", the test substance and eleven other surfactants have to be classified as "irritant". The other surfactants do not need to be classified as "irritant" or "corrosive". For all tested surfactants this classification refers to a content of 25% active ingredient.

For the concrete substance mentioned here the following gradings were observed: Erythema 1.87 and oedema 1.13, concluding the substance is a mild irritant to 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

Study period:

April 1987 - May 1987

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:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Principles of method if other than guideline:

The irritant potential of four amphoteric surfactants was assessed.

GLP compliance:

not specified

Specific details on test material used for the study:

The test substance was used with adjusted 25% active matter.

Species:

rabbit

Strain:

other: chbb:HM

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Fa. Thomae, Biberach, Germany
- Age at study initiation: 2115.0 to 2412.5 g
- Sex: males

Vehicle:

not specified

Controls:

yes

Amount / concentration applied:

0.1 ml of test substance (25% active matter)

Duration of treatment / exposure:

24 h

Observation period (in vivo):

1, 6, 24, 48, 72 h and 7, 10, 14, 17, 21 days

Number of animals or in vitro replicates:

four animals

Details on study design:

REMOVAL OF TEST SUBSTANCE
- not specified

SCORING SYSTEM: The reactions on cornea, iris and conjunctiva were assessed according to the scoring system by Draize.

TOOL USED TO ASSESS SCORE: 0.5% fluorescein solution

Irritation parameter:

other: opacity area

Basis:

animal #3

Time point:

21 d

Score:

1

Max. score:

4

Reversibility:

not reversible

Remarks on result:

positive indication of irritation

Irritation parameter:

other: opacity area

Basis:

animal #4

Time point:

21 d

Score:

1

Max. score:

4

Reversibility:

not reversible

Remarks on result:

positive indication of irritation

Irritation parameter:

other: opacity area

Basis:

animal #1

Time point:

21 d

Score:

1

Max. score:

4

Reversibility:

not reversible

Remarks on result:

positive indication of irritation

Irritant / corrosive response data:

Corneal reactions which persisted for 21 days were observed in three test animals treated with the test substance. One animal treated with the test substance revealed irreversible corneal damage after 21 days. Reactions to the iris were not observed. Conjunctival reactions were severe and persisted for 21 days after application in the majority of test animals (3/4) when rabbits were treated with the test substance.

Table 1: Corneal reactions to rabbit eyes after application of the test substance with 25% active matter

Animal number	Hours after application														Days after application
	1		6			24			48			72			7		10		14		17		21
	A	B	A	B		A	B		A	B		A	B		A	B	A	B	A	B	A	B	A	B
543	0	0	1		2	1		4	1		4	1		3	1	2	1	1	1	1	1	1	1		1
588	1	2	1		2	1		4	1		4	1		4	1	2	1	2	0	0	0	0	0		0
589	1	2	1		2	1		4	1		4	1		4	1	4	1	4	1	4	1	4	1		4
590	1	2	1		4	1		4	1		4	1		4	1	4	1	4	1	4	1	4	1		4
A (mean)	0.75		1.00			1.00			1.00			1.00			1.00		0.75		0.75		0.75		0.75
B (mean)	1.50		2.50			4.00			4.00			3.75			3.00		2.75		2.25		2.25		2.25

A = opacity grade; B = opacity area

Table 2: Conjunctiva reactions to rabbit eyes after application of the undiluted test substance

Animal number	Hours after application															Days after application
	1			6			24			48			72			7			10			14			17			21
	A	B	C	A	B	C	A	B	C	A	B	C	A	B	C	A	B	C	A	B	C	A	B	C	A	B	C	A	B	C
543	1	2	3	1	2	3	2	1	3	3	2	3	3	1	3	2	1	3	1	0	1	1	0	1	0	0	1	0	0	0
588	1	1	3	1	1	3	3	1	3	3	1	3	3	1	3	2	1	3	2	0	3	2	0	1	2	0	1	1	0	3
589	2	1	3	1	1	3	2	1	3	3	1	3	3	1	3	3	1	3	3	1	3	2	0	1	2	0	2	2	0	1
590	1	2	3	2	1	3	3	1	3	2	1	3	2	1	3	2	0	1	2	0	1	2	0	1	2	0	1	1	0	3
A (mean)	0.75			1.00			1.00			1.00			1.00			1.00			0.75			0.75			0.75			0.75
B (mean)	1.50			2.50			4.00			4.00			3.75			3.00			2.75			2.25			2.25			2.25

A = erythema; B = chemosis; C = exudation;  red bold = bloody; bold = bloody infiltrated

Interpretation of results:

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

Conclusions:

The results generated by the performed study concludes the test substance to be severly irritant to the mucous membrane of the eyes with irreversible damages to the cornea and the conjuctiva. Thus, the test substance must be labelled with the R-phrase R-41.

Executive summary:

Three amphonic surfactants and the test substance wre tested in a concentration of 25% active ingredient for their primary eye irritation on rabbits by a single application and permanent contact (Method: Off. J. Europ. Commun. 27, L 251, 1984 p. 109 - 112 and Guidelines for Testing of Chemicals, Section 4, No 405, OECD). In each group were animals with corneal lesions, which had to be regarded as irreversible. All 4 test samples had to be signed with the risk phrases R-41.

Reference 2

Endpoint:

eye irritation: in vitro / ex 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

Principles of method if other than guideline:

HETCAM-Test used for detecting severe eye irritation in hen's eggs.

GLP compliance:

not specified

Species:

other: chorioallantoic membrane of hen's eggs (HET-CAM) Test

Vehicle:

water

Controls:

no

Amount / concentration applied:

0.01, 0.05, 0.1, 0.5, 1, 5, and 10% active substance in water. 300 µl test substance were applied on each egg.

Observation period (in vivo):

300 seconds

Number of animals or in vitro replicates:

Each test concentration was tested on six eggs.

Details on study design:

Evaluation criteria was according to the following scheme:
++ = strong reaction = 3 points
+ = moderate reaction = 2 points
(+) = sllight reaction = 1 point
- = no reaction = 0 points

Evaluated observations were known CAM-reactions, such as haemorrhage, coagulation, and other specific alterations

Calculating the mean of the six tested eggs the results were presented as follows:

Points Evaluation
0 - 3 no irritating potential
4 - 6 slight irritating potential
7 - 12 moderate irritating potential
13 - 18 severe irritating potential

The condition of the eggs were assessed at the end of the testing period, thus, after 300 seconds. The irritating threshold was set as the concentration inducing the highest slightest irritating concentration.

Irritation parameter:

other: irritation threshold

Run / experiment:

1

Value:

0.05

Vehicle controls validity:

not applicable

Negative controls validity:

not applicable

Positive controls validity:

not applicable

Remarks on result:

positive indication of irritation

Other effects / acceptance of results:

The irritation threshold of the test substance was 0.05% of the concentration of the test substance (based on 25% active substance).

Conclusions:

The results of this in vitro assay showed that the test substance did induce irritating reactions at a concentration higher than 0.05%.

Executive summary:

The simplified hen's eggs test was used for the evaluation of the irritating potential of the test substance.

Reference 3

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1980

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

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

GLP compliance:

no

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Purity: 33.5%

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

no data

Vehicle:

water

Controls:

not specified

Amount / concentration applied:

0.1 ml of 15% active matter

Duration of treatment / exposure:

uninterrupted (no washing)

Observation period (in vivo):

21 days in total. Observation times were after 2, 6, 24, 48, 72, and 96 hours, and after 8, 12, 16, and 21 days.

Number of animals or in vitro replicates:

5 male rabbits

Details on study design:

Application: The test substance was applied into the conjunctival sac.

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

21 d

Score:

>= 8 - < 12.5

Max. score:

100

Reversibility:

not fully reversible within: 21d

Remarks on result:

positive indication of irritation

Irritant / corrosive response data:

Slight to moderate irrit11.3ations to conjunctiva, cornea and iris.

Table 1: Results of the eye irritation test on rabbits

Test substance	Conc.(%)	Contact time	Hours after application						Days after application
			2	6	24	48	72	96	8	12	16	21
Cornea	15	constant	12.5	11.3	11.3	25	13.8	16.3	27.5	20	15	12.5**
Iris	15	constant	50	50	50	40	40	30	40	20	10	10*
Conjunctiva	15	constant	50	62	44	42	42	30	22	14	8	8*

* = one of the five rabbits showed irreversible alterations to iris and conjunctiva

** = one of the five rabbits showed irreversible alterations to cornea

Interpretation of results:

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

Conclusions:

The test substance caused irreversible eye damage and is considered to be classified as a category 1 substance.

Executive summary:

Seven different anionic, nonionic and amphoteric surfactants were tested comparatively for their irritant potential to the skin and mucous membranes in diverse test systems, such as the mucous membrane irritation test in rabbits. The individual testing methods resulted in slighty different rankings of the surfactants. Considering the results of all three tests, it can be concluded that the test substance has the most irritating potential.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin:

An acute dermal irritation/corrosion assay for the test substance according to OECD guideline 404, in compliance with GLP, was conducted in three New Zealand rabbits (Stepan, 2000). 0.5 mL of the test material was applied to a site of about 2.5 x 2.5 cm of intact skin of 3 rabbits and covered by semi-occlusive dressing. The administration of the test substance was performed in a sequential manner. The first animal was exposed to the test substance stepwise at three sites of the skin: site 1 for three minutes, site 2 for 1 hour and site 3 for 4 hours. Any residual test substance was removed by swabbing with cotton wool soaked in 74% Industrial Methylated Spirits. After considerations of the skin reactions produced on the first animal, the remaining two animals were exposed to the test substance for 4 hours.The treated areas of the animals were examined for erythema/eschar and oedema as well as other local skin alterations approximately 1, 24, 48, and 72 hours after patch removal. Additional observations were made on days 7 and 14 to assess the reversibility of skin reactions. Dermal alterations were scored according to OECD guideline nº 404.Well-defined erythema (score: 2) was noted at two treated skin sites one hour after patch removal and at the 24, 48 and 72-hour observations. Slight oedema (score: 2) was noted at two treated skin sites one hour after patch removal. Very slight oedema (score: 1) was noted at two treated skin sites at the 72-hour observation. One treated skin site appeared normal throughout the study. Crust formation was noted at two treated skin sites at the 7-day observation with slight desquamation at these two treated skin sites at the 14-day observation. No corrosive effects were noted.

In a comparative study 10 anionic surfactants, 4 amphotric surfactants, 8 nioionic surfactants and 3 kationic surfactants with equal active ingredients of 25% were tested for their primary skin irritation under occlusive conditions on the shaved back skin of rabbits (Method: Off. J. Europ. Commun. 27, L 251, 1984 p. 106 - 108 and Guidelines for Testing of Chemicals, Section 4, No 404, OECD).

Two kationic surfactants were tested earlier after the same method. Five Kleinrusse Chbb:HM/Fa rabbits were used and the left and right body sites were shaved in preparation of the application. The amount of 0.5 ml of a diluted solution containing 25% active matter of the test substance was applied on the skin. The area of exposure was wrapped in with a 2.5 cm x 2.5 cm linth cloth and covered occlusive. The exposure duration was 4 h. Observation time points were 1, 24, 48, 72 h and 7, 10, 14, 17, 21 days post-treatment. The scoring system for edema and erythema reactions was the scoring system proposed by Draize. The primary dermal irritation index was 2.87 out of 8. Mean values for edema and erythema for animal 1, animal 2, animal 3, animal 4, and animal 5 were 0.6 and 1.3, 1.0 and 1.6, 0.6 and 2.0, 1.6 and 3.0, 1.0 and 1.3, respectively. No mortality was observed. A positive indication of irritation was reported.

A study was conducted according to OECD guideline 404, in compliance with GLP, in order to determine the skin irritation potential of the test substance (Cognis, 1987). Five New Zealand White rabbits were exposed under occlusive conditions to a diluted solution containing 25% active matter of the test substance. The test site of the animal has been shaved in preparation to the exposure. Treatment duration was 4 h. Observation time points after the patch removal were 24, 48, and 72 h. Erythema as well as edema of each animal were assessed.The score for erythema and edema was 1.86 and 1.0, respectively. The substance was rated as being irritant.

All three available studies concluded an irritating or non-corrosive effect of the test substance. However, the results do not trigger a classification for skin irritation applying the criteria set forth in the Regulation (EC) No. 1272/2008.

The registrant of the target substance decides in a worst case approach to classify the substance as skin irritant cat 2 as only dilutions are tested and various results received, also from the read across substance, that make a more irritating effect for the pure substance possible.

As the substance is normally produced as a 35% solution and none of the studies show a skin irritant effect cat 2 a specific concentration limit (SCL) is established at 35% for non skin classification.

Eye:

A study was performed according to OECD guideline 405 in 1987 (without specifying GLP compliance) in order to determine the eye irritation potential of the test substance (Henkel KGaA, 1987). Four Kleinrusse Chbb:HM rabbits were exposed to a diluted solution containing 25% active matter of the test substance using the untreated eye as control. The amount of 0.1 ml were applied and left in the eye for 24 h. Observation time points were 1, 6, 24, 48, 72 h and 7, 10, 14, 17, 21 days. Corneal reactions which persisted for 21 days were observed in three test animals, one animal revealed irreversible corneal damage after 21 days, and conjunctival reactions were severe and persisted for 21 days after application in three animals. Thus, the test substance was categorized as seriously eye damaging.

Further a HET-CAM conducted test with no information regarding GLP compliance and the use of an OECD guideline in 1986 (BASF, 1986). Chorioallantoic membrane of hen's eggs was exposed to concentrations of 0.01, 0.05, 0.1, 0.5, 1, 5, and 10% active substance in water. 300 µl were applied on each egg. The observation period was 300 seconds and the membranes were evaluated according to a point scheme; no reaction (0 points), slight reaction (1 point), moderate reaction (2 points), and strong reaction (3 points). Results demonstrate that the test substance induced irritating reactions at a concentration higher than 0.05%.

A mucous membrane irritation test with no information regarding GLP compliance and the use of an OECD guideline has been performed in 1980. Five male New Zealand White rabbits were exposed to a diluted solution containing 15% active matter in water. The amount of 0.1 ml was applied into the conjunctival sac for an uninterrupted duration (no washing). Observation time points were after 2, 6, 24, 48, 72, and 96 hours, and after 8, 12, 16, and 21 days. Slight to moderate irritations (overall irritation score between 17.0 and 43.0 as means from 24/48/72 h) to conjunctiva, cornea and iris were observed. Results demonstrated that at least one animal experienced irreversible effects to iris, conjunctiva, and cornea, thus, categorizing the test substance as seriously eye damaging.

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No. 1272/2008

The available and relevant experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008.

As a result the substance is considered to be classified for irritation under Regulation (EC) No. 1272/2008 as skin irritant category 2 (H315) in a worst case approach but establishing an SCL of 35% for not irritancy and seriously eye damaging category 1 (H318).