Registration Dossier

Administrative data

Description of key information

Skin Irritation

At the 1st minute, statistically significant differences were detected between control and cocamidopropyl betaine.

The SSWL values (g/sqmh) at the 1st and 25th minute for the test chemical were 44.7 and 12.1 respectively.

Based on these values, cocamidopropyl betaine can be considered to be irritating to human skin.

Eye Irritation

The average eye irritation index for the unwashed and washed eyes after 21 days of observation were 0.7 and 14.7 respectively.

The test chemical appears to be irritating to eyes. Washing of the eyes doesnot appear to be beneficial in reduction of the degree of irritation or the duration of irritation as seen in the unwashed eyes.

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
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
data is from peer reviewed journals
Qualifier:
according to
Guideline:
other: plastic occlusion stress test (POST) technique.
Principles of method if other than guideline:
Irritant effects and disturbance of water-holding capacity induced by Cocamidopropyl betaine were investigated using the plastic occlusion stress test (POST) technique.
GLP compliance:
not specified
Specific details on test material used for the study:
- Name of test material : 1-Propanaminium, 3-amino-N-(carboxymethyl)-N,N-dimethyl-, N-coco acyl derivs., hydroxides, inner salts- Common Name: Cocamidopropyl betaine (CAPB)- Molecular formula: C19H38N2O3- Molecular weight: 342.52 g/mol- Smiles notation: CCCCCCCCCCCC(=O)NCCCN{+}(C)(C)CC(=O)O{-}- Substance type: Organic - Physical state: Solid
Species:
other: humans
Strain:
not specified
Details on test animals and environmental conditions:
Details on test animalTEST ANIMALS- Source: Department of Dermatology, University of Pavia, Italy- Age: 32+/- 4 years
Type of coverage:
open
Preparation of test site:
not specified
Vehicle:
water
Controls:
yes, concurrent no treatment
Amount / concentration applied:
0.03 ml/sqcm of 7% cocamidopropyl betaine in distilled water
Duration of treatment / exposure:
Once daily for 3 days
Observation period:
24 hours after removal of POST
Number of animals:
8 healthy white men
Details on study design:
TEST SITE- Area of exposure: volar forearm- % coverage: 16 sq cm- Type of wrap if used: no data availableREMOVAL OF TEST SUBSTANCE- Washing (if done): not washedSCORING SYSTEM: skin surface water loss (SSWL) and transepidermal water loss (TEWL) were recorded using an evaporimeter. SSWL and TEWL readings were corrected for a skin temperature of 30 deg C.SSWL decay constants were calculated for each subject according to method described by Wagner
Irritation parameter:
other: SSWL
Basis:
mean
Time point:
other: 1 MINUTE
Score:
44.7
Reversibility:
not specified
Remarks on result:
positive indication of irritation
Irritation parameter:
other: SSWL
Basis:
mean
Time point:
other: 25 MINUTES
Score:
12.1
Reversibility:
not specified
Remarks on result:
positive indication of irritation
Irritant / corrosive response data:
At the 1st minute, statistically significant differences were detected between control and cocamidopropyl betaine

SSWL values(g/sqmh) corrected for skin temperature at the 1stand 25thminute(+/- standard error) and SSWL decay constants

Test chemical

1stminute

25thminute

Decay constant

Cocamidopropyl betaine

44.7±7.8*

12.0±2.0*

-0.046

Control

40.5±3.9*

8.7±1.*

-0.056

The differences between the surfactants are statistically significant (Friedman’s test p<0.01)

 

 

 

 

 

Interpretation of results:
Category 2 (irritant) based on GHS criteria
Conclusions:
At the 1st minute, statistically significant differences were detected between control and cocamidopropyl betaine.The SSWL values (g/sqmh) at the 1st and 25th minute for the test chemical were 44.7 and 12.1 respectively.Based on these values, cocamidopropyl betaine can be considered to be irritating to human skin.
Executive summary:

Irritant effects and disturbance of water‐holding capacity induced by Cocamidopropyl betaine were investigated using the plastic occlusion stress test (POST) technique. Doses for the test were determined on the basis of the irritant potential of the test chemical and its use in marketed products. 7% cocamidopropyl betaine in distilled water was applied to the open marked sites once daily for 3 days on the volar forearms of the 8 White male volunteers. The sites were allowed to dry without washing. On the 4thday, a 1inch diameter occlusive plastic device was applied for 24 hours on both the treated and an adjacent control site, in order to produce hydration for theplastic occlusion stress test (POST).

After removal of the devices, excess water was wiped with tissue papers and the SSWL was recorded continuously with an Evaporimeter. Skin temperature was recorded by a digital thermometer.

 

Statistical analysis was performed using the non parametric Freidman test and Fischer PLSD test.

At the 1stminute, statistically significant differences were detected between control and cocamidopropyl betaine. The SSWL values (g/sqmh) at the 1stand 25thminute for the test chemical were 44.7 and 12.1 respectively

Based on these values, cocamidopropyl betaine can be considered to be irritating to human skin.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (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:
secondary literature
Justification for type of information:
data is from safety assessment reports
Qualifier:
according to
Guideline:
other: FEDERAL HAZARDOUS SUBSTANCE ACT, 16 CFR 1500.42
Principles of method if other than guideline:
To determine the degree of irritation produced by Cocamidopropyl betaine when instilled into the eyes of albino rabbits
GLP compliance:
not specified
Specific details on test material used for the study:
- Name of test material : 1-Propanaminium, 3-amino-N-(carboxymethyl)-N,N-dimethyl-, N-coco acyl derivs., hydroxides, inner salts- Common Name: Cocamidopropyl betaine (CAPB)- Molecular formula: C19H38N2O3- Molecular weight: 342.52 g/mol- Smiles notation: CCCCCCCCCCCC(=O)NCCCN{+}(C)(C)CC(=O)O{-}- Substance type: Organic - Physical state: Solid
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
Details on test animalTEST ANIMALS- Source: Buckshire Corp, USA- Housing: individually housed in stainless elevated cages with wire mesh flooring- Diet (e.g. ad libitum): Wayne 15% rabbit ration, ad libitum- Water (e.g. ad libitum): tap water, ad libitum- Acclimation period: the rabbits were acclimated to the laboratory for at least 5 days prior to dosingENVIRONMENTAL CONDITIONS- Temperature (°C): 60-75 deg F- Humidity (%): 55+/-25%- Air changes (per hr): no data available- Photoperiod (hrs dark / hrs light): 112 hour light /dark cycleIN-LIFE DATES: From: To:
Vehicle:
water
Controls:
yes, concurrent no treatment
Amount / concentration applied:
0.1 ml of 50.7% w/v dilution in tap water
Duration of treatment / exposure:
single exposure
Observation period (in vivo):
1,2,3,4 and 7 days following instillation of the test chemical
Duration of post- treatment incubation (in vitro):
no data available
Number of animals or in vitro replicates:
9
Details on study design:
Details on study designREMOVAL OF TEST SUBSTANCE- Washing (if done): yes - Time after start of exposure: the treated eyes of 3 rabbits were washed after 30 seconds with tepid tap water( approx 1000ml/min) for 1 minuteSCORING SYSTEM: The treated eyes were examined at 1,2,3,4 and 7 days post instillation of the test chemical. The readings of the eyes were made in accordance with Draize technique for scoring of ocular lesions.
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
21 d
Score:
0.7
Max. score:
110
Reversibility:
not specified
Remarks on result:
positive indication of irritation
Remarks:
un washed eyes
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
21 d
Score:
18.7
Max. score:
110
Reversibility:
not specified
Remarks on result:
positive indication of irritation
Remarks:
washed eyes
Irritant / corrosive response data:
The test chemical appears to be irritating to eyes. Washing of the eyes doesnot appear to be beneficial in reduction of the degree of irritation or the duration of irritation as seen in the unwashed eyes.

Average ocular irritation scores

Condition of eyes

Duration of exposure

1 hour

1 day

2 day

3 day

4 day

7 day

14 days

21 day

Unwashed

16.0

18.8

18.3

14.5

12.2

5.0

2.3

0.7

Washed after 30 seconds

14.0

16.7

31.3

27.3

27.0

34.3

18.7

14.7

 

Fluorscein dye examination

Unwashed eyes

Observation period

Rabbit number

1

2

3

4

5

6

Pre dosing

-

-

-

-

-

-

1 day

+

+

+

+

+

+

2 day

+

+

+

+

+

+

3 day

+

+

+

+

+

+

4 day

+

+

+

+

+

+

7 day

+

+

+

+

-

+

14 day

+

-

-

-

-

 

21 day

+

 

 

 

 

 

 

Washed eyes

 

Observation period

Rabbit number

7

8

9

Pre dosing

-

-

-

1 day

+

+

+

2 day

+

+

+

3 day

-

-

+

4 day

+

+

+

7 day

+

+

+

14 day

+

+

+

21 day

 

-

+

 Where

 - = no corneal staining

+ = corneal staining

 

Interpretation of results:
Category 2 (irritating to eyes) based on GHS criteria
Conclusions:
The average eye irritation index for the unwashed and washed eyes after 21 days of observation were 0.7 and 14.7 respectively. The test chemical appears to be irritating to eyes. Washing of the eyes doesnot appear to be beneficial in reduction of the degree of irritation or the duration of irritation as seen in the unwashed eyes.
Executive summary:

A study was conducted to determine the degree of irritation produced by Cocamidopropyl betaine when instilled into the eyes of albino rabbits. 9 New Zealand White rabbits were used for the study.

0.1 ml of 50.7% w/v dilution in tap water was instilled into the eyes of 9 rabbits. The untreated eyes served as control. The eyes of 6 rabbits remained unwashed throughout the test, but eyes of the remaining 3 rabbitswere washed after 30 seconds with tepid tap water( approx 1000ml/min) for 1 minute.The treated eyes were examined at 1, 2, 3, 4 and 7 days following instillation of the test chemical. Any eyes exhibiting ocular irritation at 7 days were also examined at 14 days and again at 21 days if ocular irritation persisted at 14 days.

The treated eyes were examined at 1,2,3,4 and 7 days post instillation of the test chemical. The readings of the eyes were made in accordance with Draize technique for scoring of ocular lesions.

The average eye irritation index for the unwashed and washed eyes after 21 days of observation were 0.7 and 14.7 respectively.

The test chemical appears to be irritating to eyes. Washing of the eyes doesnot appear to be beneficial in reduction of the degree of irritation or the duration of irritation as seen in the unwashed eyes.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin Irritation:

In various studies,Cocamidopropyl betaine (CAPB)has been investigated for potential to cause dermal irritation to a greater or lesser extent. The studies are based on in vivo experiments in humans along with in vitro data for the target chemical. The results are summarized as follows:

Irritant effects and disturbance of water‐holding capacity induced by Cocamidopropyl betaine were investigated using the plastic occlusion stress test (POST) technique. Doses for the test were determined on the basis of the irritant potential of the test chemical and its use in marketed products. 7% cocamidopropyl betaine in distilled water was applied to the open marked sites once daily for 3 days on the volar forearms of the 8 White male volunteers. The sites were allowed to dry without washing. On the 4thday, a 1inch diameter occlusive plastic device was applied for 24 hours on both the treated and an adjacent control site, in order to produce hydration for theplastic occlusion stress test (POST).

After removal of the devices, excess water was wiped with tissue papers and the SSWL was recorded continuously with an Evaporimeter. Skin temperature was recorded by a digital thermometer. Statistical analysis was performed using the non parametric Freidman test and Fischer PLSD test.

At the 1stminute, statistically significant differences were detected between control and cocamidopropyl betaine. The SSWL values (g/sqmh) at the 1stand 25thminute for the test chemical were 44.7 and 12.1 respectively

Based on these values, cocamidopropyl betaine can be considered to be irritating to human skin.

This is supported by the results of a Human patch test performed to determine the irritation potential of the test chemical. The study was conducted at the Dermatology Clinical Testing Facility, University of California, USA. The participants in this initial clinical study were 66 healthy subjects, male and female, 18–65 years of age. Cocamidopropyl betaine1% weight/vol in distilled water) and distilled water alone (as a control) were tested in closed patch sites for 24 hours. No positive control was included in the study. Sites were graded for erythema and edema using a simplified 4-point grading scale.

The Sum of scores for 22 observations was 4 after 24 hours. The mean value for erythema/edema was 0.17

Based on the scores, Cocamidopropyl betaine can be considered to be irritating to skin.

These results are supported by the results of another Human clinical closed patch test performed to assess the dermal irritation potential of Cocamidopropyl betaine

10% concentration of the test material was tested in a 24 hour closed patch test on human volunteers. The untreated skin served as control. The dermal reactions were scored  according to the standard Draize scoring system (Draize, 1945).

The clinical score for dermal irritation potential of Cocamidopropyl Betaine in humans was 1.08.

Based on the scores, Cocamidopropyl betaine can be considered to be irritating to skin.

These results are further supported by the study performed according to FEDERAL HAZARDOUS SUBSTANCE ACT, CFR 16, SECTION 1500.14 to determine the degree of irritation produced by Cocamidopropyl betaine when applied to the skin of albino rabbits. 6 New Zealand white rabbits were used for the study. Prior to application, the backs of 6 rabbits were clipped free of hair over a wide area. Two sites on each rabbit were used. One site was left intact and the other was abraded sufficiently deep to penetrate the stratum corneum but not enter the derma to produce bleeding.

0.5 ml of 50.7% w/v dilution of the test chemical in tap water was applied to one intact and one abraded site on each rabbit. Gauze patches were placed over the treated sites and wrapped with a suitable occlusive dressing. The wrapping was removed after 24 hours of exposure, and the treated sites where then wiped free of the excess test chemical. The treated sites were observed, scored for erythema and edema according to Draize method.

Eschar formation was observed in 3 of 6 rabbits. The test chemical was considered to be corrosive to the skin of rabbits within the definition of the F.H.S.A, CFR 16, 1500.14. This conclusion was not based on the primary irritation score of 4.54.

These results are also supported by a Human patch test performed to determine the irritation potential of the test chemical.

40 microliters of 24% CAPB was applied on 5 different sites on both volar forearms of 24 healthy female volunteers, following a rotating pattern of application. After 30 min, patches were removed and the skin was rinsed with tap water. Test sites were covered again with an empty chamber until the 1stmeasurement.Instrumental evaluations were performed before the beginning of the experiment, and 24 h and 72 h after patch test removal.

The Evaporimeter EP1 (Servomed, Sweden) was used to measure transepidermal water loss, and the Chroma Meter CR200 (Minolta, Japan) to evaluate erythema employing the parameter a*. Transepidermal water loss[TEWL] measurements were performed according to the guidelines of the Standardization Group of the European Society.

TheTransepidermal water loss (TEWL)values at 24 h and 72 hours were 1.39±1.64, 1.82±1.87 respectively.

Cocamidopropyl betaine [CAPB] induced a significant increase as compared to baseline, at 24 h and 72 h for TEWL and at 72 h for a* values.

Hence, Cocamidopropyl betaine [CAPB] can be considered to be irritating to skin.

The above in vivo results are further supported by the results of an in vitro study whereEpiDerm Epi-100 was used to predict the dermal irritation potential of Cocamidopropyl betaine. EpiDermTM Epi-100, a human epidermal model, was obtained from MatTek Corporation (Ashland, MA, USA) and maintained according to the manufacturers' instructions. Normal human-derived epidermal keratinocytes (NHEK) are cultured to form this multilayered differentiated model of the epidermis.

Skin equivalents were exposed in triplicate to 100 microliter topically applied test material, for 1 hr.. Following exposure, the test articles were removed by repeated rinsing with phosphate buffered saline (PBS, Gibco-BRL), and the skin equivalents placed in fresh medium. Medium was collected 24 hr post treatment and stored at 20 deg C.

Viability was determined 24 hr post treatment using the novel tetrazolium compound (3-(4,5- dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenenyl)- 2-(4-sulfophenyl)-2H-tetrazolium, inner salt), MTS (Promega Corporation, Madison, USA), MTS (Promega), or alamarBlueTM (Alamar Biosciences,USA), using manufacturers' protocols, and a UVMax Plate Reader (Molecular Devices, USA) or a Cytofluor II Fluorescent Plate Reader (PerSeptive Biosystems, USA), respectively.

Viability was used as a first screen to identify the moderate/severe irritants. Materials causing greater than 20% cytotoxicity were classified as moderate to severe irritants. The mean tissue viability of Cocamidopropyl betaine (expressed as % untreated) was 92. Since the mean viability was greater than 20%, cocamidopropyl betaine can be considered to be irritating to skin.

The results of in vivo and in vitro studies are in mutual agreement with each other indicating a very strong possibility of Cocamidopropyl betaine [CAPB] being irritating to eyes. Hence, Cocamidopropyl betaine [CAPB] was considered to be irritating to skin.Comparing the above annotations with the criteria of CLP regulation,Cocamidopropyl betaine [CAPB]can be classified under the category “Category 2”.

 

 

Eye Irritation:

In various studies,Cocamidopropyl betaine (CAPB)has been investigated for potential to cause ocular irritation to a greater or lesser extent. The studies are based on in vivo experiments in rabbits along with in vitro data for the target chemical. The results are summarized as follows:

A study was conducted to determine the degree of irritation produced by Cocamidopropyl betaine when instilled into the eyes of albino rabbits. 9 New Zealand White rabbits were used for the study.

0.1 ml of 50.7% w/v dilution in tap water was instilled into the eyes of 9 rabbits. The untreated eyes served as control. The eyes of 6 rabbits remained unwashed throughout the test, but eyes of the remaining 3 rabbits were washed after 30 seconds with tepid tap water( approx 1000ml/min) for 1 minute. The treated eyes were examined at 1, 2, 3, 4 and 7 days following instillation of the test chemical. Any eyes exhibiting ocular irritation at 7 days were also examined at 14 days and again at 21 days if ocular irritation persisted at 14 days.

The treated eyes were examined at 1,2,3,4 and 7 days post instillation of the test chemical. The readings of the eyes were made in accordance with Draize technique for scoring of ocular lesions.

The average eye irritation index for the unwashed and washed eyes after 21 days of observation were 0.7 and 14.7 respectively.

The test chemical appears to be irritating to eyes. Washing of the eyes does not appear to be beneficial in reduction of the degree of irritation or the duration of irritation as seen in the unwashed eyes.

This result is supported by a study performed according to Draize method to assess the irritation potential of cocamidopropyl betaine. The study was. Undiluted test chemical was instilled in the eyes of 4 rabbits and observed for signs of irritation till 7 days. The reactions observed were scored according to Draize method. Mean scores are calculated for each animal from gradings at 24, 48, and 72 h after instillation of the test chemical and these “severity scores” are then used to determine the classification of the test chemical.

The corneal opacity persisted till day 21 in all the 4 rabbits; also the conjunctival redness observed in some rabbits was fully reversible by 7 to 14 days. Based on these observations, cocamidopropyl betaine was considered to highly irritating to rabbit eyes, was classified under the category “Category 1”.

These results are also supported by another study performed according to Draize method to assess the irritation potential of cocamidopropyl betaine. Undiluted test chemical was instilled in the eyes of rabbits and observed for signs of irritation till 24 hours. The Maximum average score after 24 hours of observation was 18.0, which according to Draize classification criteria was mild irritating

Hence, Cocamidopropyl betaine was considered to be irritating to rabbit eyes.

The above in vivo results are further supported by the results of an in vitro study where EpiOcularTM(OCL-200) model system was used to predict the eye irritation potential of Cocamidopropyl betaine.

EpiOcularTMtissue was purchased from MatTek Corporation as tissue inserts (MillicellsTM). Each batch of EpiOcularTMtissue (OCL-200) was tested with the negative control, ultra-pure (18 Mohm) water, and the positive control,0.3% Triton X-100.

100 microliters of 50% test chemical solution was applied to the tissue surface in duplicates. Following sample application, tissues were incubated for specific time intervals at 37 deg C in 5% CO2/ 95% air. Tissues treated with Triton X-100 (positive control) were incubated for 20 min and 60 min. In the initial phase of the evaluation, tissues treated with samples classified in vivo as ``minimal'' irritants were incubated for 60, 120 or 240 min; samples classified as ``mild'' irritants were incubated for 5, 20 or 60 min; while samples classified as ``moderate±severe'' irritants were incubated for 2, 10 or 30 min. In later stages of this evaluation, tissues for all samples were incubated for 3 min, 30 min and 60 min. Deionized water (negative control) was incubated at the longest time period (i.e. 60 or 240 min). Following incubation with a raw ingredient, tissues were removed from the six-well plates, rinsed thoroughly with DPBS, and placed into 12-well plates containing 5 ml assay medium per well. Tissues remained in medium for at least 10, but not more than 20 min, rinsed with DPBS and transferred to 24-well plates containing 0.3 ml of diluted MTT solution/well. These MTT plates were further incubated for 3 hr at 378C in 5% CO2. After incubation, the tissues were again rinsed with DPBS. MTT dye was extracted from the tissues by adding 2.0 ml extraction medium to each well at room temperature, or storing the plates overnight, in 2.0 ml extraction medium, at 48C in sealed plastic bags. In either case, plates were then gently rocked (IKA- Shuttler MTS 4 plate rocker) at room temperature for 2 hr to extract the dye.

Following extraction, 200 microliter extraction solution for each sample was transferred to a 96-well plate. The plate was immediately read at 540 nm in a 7520 Microplate Reader (Cambridge Technology, Inc., USA). Optical densities (OD) were converted to % viability using the following formula:

% viability =100*[OD{sample}/ (OD{negative control})]

The time for 50% viability (ET50) was determined by linear interpolation between two points, one which exceeded 50% viability, the other less than 50% viability.

The ET50 value for Cocamidopropyl betaine when tested at 25% in the EpiOcularTM(OCL-200) assay was <3. According to the classification based on the ET50 values, Cocamidopropyl betaine can be considered to be severely irritating to eyes.

The results of in vivo and in vitro studies are in mutual agreement with each other indicating a very strong possibility of Cocamidopropyl betaine [CAPB] being irritating to eyes. Hence, Cocamidopropyl betaine [CAPB] was considered to be irritating to eyes.Comparing the above annotations with the criteria of CLP regulation,Cocamidopropyl betaine [CAPB]can be classified under the category “Category 2”. 

Justification for classification or non-classification

Available data for Cocamidopropyl betaine (CAPB) suggests that it is likely to cause moderate to severe irritation to eyes and skin.

Cocamidopropyl betaine (CAPB) can be classified under the category “Category 2” as per CLP regulation.