Registration Dossier

Administrative data

Description of key information

When tested for skin irritation in vivo, Cocamidopropyl hydroxysultaine as a 16% solids or 41.5% solution, was not irritating.
When tested for eye irritation in vivo at concentrations ranging from 10% solids to 41.5%, Cocamidopropyl hydroxysultaine induced ocular damage that was persistent at the end of the observation period (7 to 21 days) in some animals. Cocamidopropyl hydroxysultaine is therefore considered a severe eye irritant. Furthermore, an in vitro test on chorioallantoic membrane of Hen's egg performed on the commercial product containing the registered substance at ca. 42% (commercial product diluted at 1.25% in the in vitro test) confirmed the severe eye irritating potential of the registered substance.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vivo
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Study period:
27 February 1989 - 2 March 1989
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
GLP guideline (US CFR) compliant study with acceptable restrictions. The study was performed on an analogue substance (for justification of read-across between Lauramidopropylhydroxysultaine and cocamidopropylhydroxysultaine, please refer to corresponding assessment report in Section 13).
Reference:
Composition 0
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
yes
Remarks:
Abraded and nonabraded skin instead of intact skin used, exposure period of 24 hours instead of 4 hours, occlusive dressing instead of semi-occlusive, observations conducted up to 72 hours instead of 14 days, not conducted at the 48 h timepoint
Qualifier:
according to
Guideline:
other: modification of Section 1500.41 of the Code of Federal Regulations
Deviations:
no
GLP compliance:
yes
Test material information:
Composition 1
Species:
rabbit
Strain:
New Zealand White
Type of coverage:
occlusive
Preparation of test site:
other: clipped, then abraded and nonabraded sites used
Vehicle:
unchanged (no vehicle)
Controls:
no
Duration of treatment / exposure:
24 hours
Observation period:
72 hours
Number of animals:
3 (2M + 1F)
Irritation parameter:
primary dermal irritation index (PDII)
Remarks:
intact skin - erythema and eschar
Basis:
mean
Time point:
other: 24 hours
Score:
1.33
Max. score:
2
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
primary dermal irritation index (PDII)
Remarks:
intact skin - erythema and eschar
Basis:
mean
Time point:
other: 72 hours
Score:
0
Max. score:
0
Irritation parameter:
primary dermal irritation index (PDII)
Remarks:
abraded skin - erythema and eschar
Basis:
mean
Time point:
other: 24 hours
Score:
1.33
Max. score:
2
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
primary dermal irritation index (PDII)
Remarks:
abraded skin - erythema and eschar
Basis:
mean
Time point:
other: 72 hours
Score:
0
Max. score:
0
Irritation parameter:
primary dermal irritation index (PDII)
Remarks:
intact skin - edema
Basis:
mean
Time point:
other: 24 hours
Score:
0.33
Max. score:
1
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
primary dermal irritation index (PDII)
Remarks:
intact skin - edema
Basis:
mean
Time point:
other: 72 hours
Score:
0
Max. score:
0
Irritation parameter:
primary dermal irritation index (PDII)
Remarks:
abraded skin - edema
Basis:
mean
Time point:
other: 24 hours
Score:
0.33
Max. score:
1
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
primary dermal irritation index (PDII)
Remarks:
abraded skin - edema
Basis:
mean
Time point:
other: 72 hours
Score:
0
Max. score:
0

Animal number

Erythema / Eschar

Edema

24 h

72 h

Individual score

24 h

72 h

Individual score

Intact skin

8173M

1

0

0.5

0

0

0

8174M

1

0

0.5

0

0

0

8175F

2

0

1

1

0

0.5

 

Mean score

0.7

Mean score

0.2

Abraded skin

8173M

1

0

0.5

0

0

0

8174M

1

0

0.5

0

0

0

8175F

2

0

1

1

0

0.5

 

Mean score

0.7

Mean score

0.2

Details of skin irritation scores (according to Draize scale)

Interpretation of results:
slightly irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Cocamidopropyl hydroxysultaine, as a 16% solids solution, is not considered a skin irritant.
Executive summary:

Cocamidopropyl hydroxysultaine, as a 16% solids aqueous solution, has been tested for acute skin irritation on 2 male and one female New Zealand White rabbits, in accordance with procedures of section 1500.41 of US Code of Federal Regulations, and in compliance with Good Laboratory Practice.

The test article was applied as such in a single dermal dose of 0.5 mL to a clipped area of the abraded and non-abraded skin (appr. 10% of the skin surface), for 24 hours under an occlusive dressing. Cutaneous examinations were performed at 24 and 72 hours after the application.

 

At removal of the dressing, very slight erythema was observed in the 2 males at the abraded and non-abraded skin sites, and a well-defined erythema (score 2) was observed in the female at both skin sites. No oedema was observed in the male rats, whereas the female presented a very slight oedema.

No cutaneous reactions were observed in any of the animals at 72 hours, at both the abraded and non-abraded skin sites.

 

Based on these results, Cocamidopropyl hydroxysultaine, as a 16% solids aqueous solution, was not considered a skin irritant.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
From 04 DEcember 2008 to 05 January 2009
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study well documented meeting accepted scientific principles, acceptable for assessment.
Reference:
Composition 0
Qualifier:
no guideline followed
Principles of method if other than guideline:
Hen's Egg Test - Utilizing the Chorioallantoic Membrane (HET-CAM). The test is a modification of that described by Kemper and Luepke (Kemper, F.M. & Luepke, N.P., (1986). The HET-CAM Test: An Alternative to the Draize Test. FD Chem. Toxic. 24, p. 495 - 496.).
The chick embryo has been used extensively in toxicology. The chorioallantoic membrane (CAM) of the chick embryo is a complete tissue with organoid elements from all germ cell layers. The chorionic epithelium is ectodermal and the allantoic epithelium is endodermal. The mesoderm located between these epithelia is a complete connective tissue including arteries, capillaries, veins and lymphatic vessels. The CAM responds to injury with a complete inflammatory reaction, comparable to that induced in the rabbit eye test. It is technically easy to study, and is without nerves to sense pain.
GLP compliance:
no
Test material information:
Composition 1
Species:
other: White Leghorn eggs
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Moyer's Chicks, Inc., in Quakertown, Pennsylvania, USA
- Age at study initiation: up to 7 days (stored at 13°C +/- 3°C)
- Weight at study initiation: not applicable
- Housing: For incubation the eggs were placed, on their sides, in a Kuhl, humidified incubator. The incubator is such that the eggs are automatically rotated once every hour. The eggs were incubated for 10 days. On day eight (8) the eggs were turned so that the acutely angled end faced down.
- Diet (e.g. ad libitum): not applicable
- Water (e.g. ad libitum): not applicable
- Acclimation period: not applicable

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 37° C (+/- 2° C)
- Humidity (%): not applicable
- Air changes (per hr): not applicable
- Photoperiod (hrs dark / hrs light): not applicable

IN-LIFE DATES: From: To: not applicable
Vehicle:
not specified
Controls:
other: reference articles: Johnson's Baby Shampoo (diluted at 50%) and Head & Shoulders Shampoo (diluted at 50%)
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.3 mL
- Concentration (if solution): 1.25%
Duration of treatment / exposure:
20 seconds
Observation period (in vivo):
All CAM's were observed immediately prior to test article administration and at 30 seconds, two and five minutes after exposure to the test article.
Number of animals or in vitro replicates:
4 CAMs were used for each conditions (test item, reference articles)
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): the test or reference article was rinsed from each CAM with 5 mL of physiological saline.
- Time after start of exposure: immediately after the end of the 20 seconds exposure period.

SCORING SYSTEM: The reactions of the CAM, the blood vessels, including the capillaries, and the albumin were examined and scored for irritant effects as detailed in the table 7.3.2/1. The numerical, time dependent scores were totaled for each CAM. Each reaction type can be recorded only once for each CAM, therefore the maximum score per CAM is 32. The mean score was determined for all CAM's similarly tested. Each article was then classified as indicated in the table 7.3.2/2.

TOOL USED TO ASSESS SCORE: no data
Irritation parameter:
other: Average CAM score
Basis:
other: CAM tested with Mackam LSB-50 (1.25%)
Time point:
other: overall 0.5; 2 and 5 min
Score:
12
Max. score:
32
Reversibility:
other: not applicable
Remarks on result:
other: see details in Table 7.3.2/3
Irritant / corrosive response data:
The test item was moderately irritating to the chorioallantoic membrane of Hen's Egg. See details in Table 7.3.2/3.
Other effects:
none

Table 7.3.2/3: Results of the HET-CAM test

Substance

CAM #

Scores

 

 

 

0.5 min

2 min

5 min

Total

Mackam LSB-50 (1.25%)

1

5  7

0

0

12

2

5  7

0

0

12

3

5  7

0

0

12

4

5  7

0

0

12

Average

12.00

Reference article Johnson's Baby shampoo (50%)

1

5  7

0

0

12

2

5  7

0

0

12

3

0

3  5

0

8

4

5  7

0

0

12

Average

11.00

Reference article Head & Shoulders shampoo (50%)

1

5  7

0

5  7

24

2

5  7

0

5  7

24

3

5  7

7

0

19

4

5  7

0

5

17

Average

21.00

 

Interpretation of results:
moderately irritating
Remarks:
Migrated information Criteria used for interpretation of results: not specified
Conclusions:
Under the test conditions, the test item Mackam LSB-50 diluted at 1.25% (vehicle unknown) induced moderate irritating effects on the chorioallantoic membrane of Hen's Egg.
Executive summary:

In an in vitro test on Chorioallantoic Membrane of Hen's Egg (HET-CAM test), 0.3 mL of Mackam LSB-50 diluted at 1.25% (vehicle unknown) was applied for 20 seconds on 4 Chorioallantoic Membranes from White Leghorn Hen’s egg. At the end of the exposure period, membranes were washed with physiological saline and observation of the membrane was done at 30 seconds, 2 and 5 minutes after the end of exposure. Different parameters such as Hyperemia, minimal hemorrhage (“feathering”), hemorrhage (obvious leakage) and coagulation and/or thrombosis were assessed and scored for each CAM at each time observation point. The mean score was determined for all CAM's similarly tested.

Two reference articles were also tested: Johnson's Baby Shampoo and Head & Shoulders Shampoo, both diluted at 50%.

The mean score for the CAM exposed to the test item Mackam LSB-50 diluted at 1.25% was 12.00 (max achievable score of 32.00). The mean score obtained for the CAM treated with the reference article Jonhson's baby shampoo at 50% was 11.00 (considered as moderate irritant) whereas the one obtained for the second reference article Head & Shoulders shampoo at 50% was 21.00 (considered as severe irritant).

Under the test conditions, the test item Mackam LSB-50 diluted at 1.25% induced moderate irritating effects on the chorioallantoic membrane of Hen's Egg. It can be assumed that the undiluted test item Mackam LSB-50 (containing the registered substance at ca. 42%) would induce more severe irritating effects. Therefore the classification as severe irritating to eyes (Eye Dam. 1, H318) of the registered substance, pure lauramidopropylhydroxysultaine, is warranted.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Only one RIPT test in human and one in vitro HET-CAM assay are available on the registered substance. In the absence of studies conducted according to OECD guidelines, a read-across was therefore performed with the cocamidopropyl hydroxysultaine which contains alkylamidopropylhydroxysultaine with carbon chain from C8 to C18. The main component of Cocamidopropylhydroxysultaine C8-18 is lauramidopropylhydroxysultaine. Justification for the read-across is documented in a separate document attached in Iuclid Section 13.

Skin irritation:

Cocamidopropyl hydroxysultaine was tested in vivo in a Klimisch score 2 study (1995), used as a key study. In this study, the test substance was applied to rabbits at 41.5% under semi-occlusive conditions for 4 hours. Very slight erythema was observed, which was fully reversible within 72 hours.

In another study (Klimisch score 2, 1989), used as supportive, Cocamidopropyl hydroxysultaine had been applied to rabbits at 16% solids under occlusive conditions for 24 hours, either to intact or abraded skin. Whatever conditions, very slight erythema and/or edema were observed, which were fully reversible within 72 hours.

This correlates with the results of Repeated Insult Patch Tests in healthy human volunteers (see section 7.10.3. Direct observations), where the repeated dermal application of a 2.5% aqueous solution of C8 -C18 amidopropylhydroxysultaine to 44 healthy human volunteers under occlusive conditions and the repeated dermal application of a 12% aqueous solution of Lauramidopropylhydroxysultaine to 54 healthy volunteers did not result in any skin reaction indicative of irritation.

Eye irritation:

Cocamidopropyl hydroxysultaine was tested in vivo in a Klimisch score 2 study (1995), used as a key study. In this study, the test substance was instillated at 41.5% in the eyes of rabbits observed for up to 21 days. Corneal and conjunctival injuries persisted for up to day 14 inclusive in some animals. Conjunctival chemosis was still observed in one of the rabbits 21 days after instillation.

In two other studies (Klimisch score 2, 1988 or 1989), used as supportive, Cocamidopropyl hydroxysultaine had been instillated in rabbit eyes at 10% or 16% solids, the animals being observed for up to 7 days. Although the period of observation was not extended up to 21 days, corneal opacity and conjunctival redness, chemosis or discharge were still observed in some animals 7 days after instillation. The severity of the ocular findings was concentration-related.

An in vitro test on chorioallantoic membrane of Hen's egg was available on the registered substance itself and used as supporting study. In this test, a solution at 1.25% of the commercial product containing the registered substance induced moderate irritating effects of the membrane, confirming therefore that the pure registered substance is considered as severe eye irritant.


Justification for selection of skin irritation / corrosion endpoint:
Recent GLP and OECD test guideline-compliant study (Klimisch score 2)

Justification for selection of eye irritation endpoint:
Recent GLP and OECD test guideline-compliant study (Klimisch score 2)

Effects on eye irritation: highly irritating

Justification for classification or non-classification

Skin irritation:

Based on the data available on Cocoamidoproplhydroxysultaine and Lauramidopropylhydroxysultaine, the registered substance does not need to be classified for skin irritation.

Eye irritation:

Based on the data available on Cocoamidoproplhydroxysultaine and Lauramidopropylhydroxysultaine, the registered substance

needs to be considered a severe eye irritant and classified accordingly as Xi; 41 (according to Dir. 67/548/EEC criteria) or Eye damage 1, H318 (according to Reg. 1272/2008 (CLP) criteria).