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Description of key information

Skin irritation - Mean value of three test data gave a value of 78.3% mean issue viability in the EpiDerm skin test (OECD 439).

Accoridng to the test protocol, a value of >50% predicts Non-Irritant under GHS classification.

Human patch test data also indicated non-irritant at 10% in water over 72 hours.

Eye irritation - Under the conditions of the Matek EpiOcular test, the results indicate that the Colaliquid DC-5 100% Active test article, at 100%, has a non-irritating/minimally irritating classification. Non-Irritant under GHS classification.

The HET-CAM hen's egg test also confirmed non-irritant.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Deviations:
not specified
GLP compliance:
yes
Specific details on test material used for the study:
ColaLiquid DC-5, 100% active
Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
other: EpiDerm system - cultured
Vehicle:
unchanged (no vehicle)
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
30 microlitres of all articles.
Duration of treatment / exposure:
35 +/- 1 minute @ 37C and then placed in a steril hood until the 60 minute exposure period was reached.
Duration of post-treatment incubation (if applicable):
24 hours +/- 2 hours after washing.
Number of replicates:
Three
Irritation / corrosion parameter:
% tissue viability
Value:
78.3
Vehicle controls validity:
valid
Negative controls validity:
valid
Remarks:
Value is 2.287 and lies between the range of >=0.8 - <=2.8 as per OECD 439
Positive controls validity:
valid
Remarks:
Value is 7.4% which is <=20% as required by the method
Remarks on result:
no indication of irritation
Interpretation of results:
GHS criteria not met
Conclusions:
Mean value of three test data gave a value of 78.3% mean issue viability in the EpiDerm skin test (OECD 439).
Accoridng to the test protocol, a value of >50% predicts Non-Irritant under GHS classification
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
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: 48 hour patch test
Principles of method if other than guideline:
Fifty-three (53) subjects, male and female, ranging in age from 16 to 77 years, who qualified were selected for this evaluation. Fifty-two (52) subjects completed this study. The renaining subject discontinued her participation for personal reasons unrelated to the use of the test material
GLP compliance:
no
Remarks:
ICH Guideline E6 - Good Clinicla Practice
Specific details on test material used for the study:
Colamide DIPA - no specific details given
Species:
other: human
Details on test animals or test system and environmental conditions:
a. Male and female subjects, age 16 and over.
b. Absence of any visible skin disease which might be confused with a skin reaction from the test material.
c. Prohibition of use of topical or systemic steroids andlor antihistamines for at least seven days prior to study initiation.
d. Completion of a Medical History form and the understanding and signing of an Informed Consent form.
e. Considered reliable and capable of following directions.
Type of coverage:
semiocclusive
Preparation of test site:
other: area between scapula
Vehicle:
water
Amount / concentration applied:
The upper back between the scapulae served as the treatment area. Approximately 0.2 ml of the test material (10% in water) or an amount sufficient to cover the contact surface, was applied to the 1" x l" absorbent pad portion of a clem adhesive dressing*. When secured to the appropriate treatment site,this dressing formed a semi-occlusive patch.
Duration of treatment / exposure:
The test material remained in contact with the skin for a total of forty-eight hours. This site was then evaluated for gross changes. Absence of any visible skin change was assigned a zero value. The test site was reevaluated at seventv-two hours.
Number of animals:
53 humans
Irritation parameter:
primary dermal irritation index (PDII)
Basis:
mean
Time point:
72 h
Score:
0
Remarks on result:
no indication of irritation
Irritant / corrosive response data:
Evaluation Key: 0 = No visible skin reaction + : Barely perceptible or spotty erytherna
1 : Mild erythmna covering most of the test site
2 = Moderate erythema, possible presence of mild edema
J = Marked erytherna, possible edema
4 = Severe erythema, possible edema, vesiculationo bullae and/or
ulceration
Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of this study, test material, Colamide DIPA, did not indicate a potential for dermal irritation
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Referenceopen allclose all

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Qualifier:
according to guideline
Guideline:
other: MatTek Corporation Epiocular ia vitro toxicity testing systern.
Principles of method if other than guideline:
"MatTek's patented Epiocular comeal Model consists of normal, human-derived epidermal keratinocytes which have been cultured to form a stratified, squarnous epithelium similm to that found in the cornsa. The epidermal cells, which are cultured on specially prepared cell culture inserts using serum free medium, differentiate to form a multilayered structure which closely parallelsthecornealepithelium..."This system"...providesapredictive,morphologically relevant in vitro means to assess ocular irritancy."
EpiOcular, when used with the recommended cell metabolism assay, can quickly provide toxicological profiles. The procedure utilizes a water-soluble, yellow, tetrazolium salt (MTT {3-[4,5-dimethylthiazol-2-yll-2,5-diphenyl-tetrazolium bromide])" which is reduced by succinate dehydrogenase in the mitochondria of viable cells to a purple, insoluble formazan derivative.Substances which damage this mitochondrial enzyme inhibit the reduction of the tehazolium salt.The amount of MTT reduced by a culture is therefore proportional to the number of viable cells
GLP compliance:
yes
Species:
human
Vehicle:
water
Controls:
yes, concurrent negative control
Amount / concentration applied:
The Sponsor requested the test article's irritation potential at l00%. The test article, at 100%, is water soluble and has a specific Savity greater than 0.95 g/ml. Therefore, as per MatTek's protocolo the test article was diluted to 20% in distilled watsr. After the appropriate tissue preparation, 100 microliters of the test article and the negative control (distilled water) were added to the Millicells containing the Epi0cular samples.
Duration of treatment / exposure:
Total time of 4 hours
Number of animals or in vitro replicates:
Six
Details on study design:
The six (6) well plates containing the dosed EpiOcular samples were then incubated at 37C, five (5)% carbon dioxide and > 90% humidity. After the appropriate exposure period, each insert was individually removed from its plate and rinsed with phosphate buffered saline (PBS) to remove any residual material. Each was then rinsed a second and third time
Irritation parameter:
other: % viability (ET-50) in minutes
Value:
122.1
Vehicle controls validity:
not specified
Negative controls validity:
valid
Positive controls validity:
not applicable
Irritation parameter:
other: Draize equivalent
Value:
4.5
Vehicle controls validity:
not specified
Negative controls validity:
valid
Positive controls validity:
not applicable

A Molecular Devices SpectraMax M5 Microplate Reader was used to determine the absorbance of each extract at 570nm. With the absorbance of the negative control (distilled water) defined as 100%, the percent absorbencies of the test article were determined. The

percentages listed below directly correlate with the cell metabolism in the EpiOcular samples.

(20% - 4 hrs.) - % viability = 39, % inhibition = 61

(20% - 1 hr.) - % viability = 61, % inhibition = 39

(20% - 20 mins.) - % viability = 75, % inhibition = 25

When possible, using a semilog scalg the percent viabilities for the article were plotted on the linear y axis versus the dosing time on the log x axis. By interpolation, the time at which the percent viability would be 50% was determined (ET-50).

As a general guideline (provided by MatTek) the following equation can be used to estimate the rabbit Draize eye score:

Draize : 4.7 4 + 101.7/(ET-50)^0.5

Based on the literature (Kay, J.H. and Calan&a J.C., "lnterpretation of eye irritation tests,"J Soc. Cosmetic Chem., 13, 281-289 (1962)), the ocular initancy estimated potential has been categorized by MatTek into the following groups, based on the Draize score:

Under the conditions of this test, the Colaliquid DC-5 100% Active test article, at 20%, elicited in vitro results which indicate that its ET-50 is 122.1 minutes. Therefore, at 100%, the test article's estimated Draize ocular iritation score is approximately 4.5 with a "minimally irritating" irritancy classification.

Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of this test, the results indicate that the Colaliquid DC-5 100% Active test article, at 100%, has a non-irritating/minimally irritating classification.
Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2004
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
equivalent or similar to guideline
Guideline:
EU method B.48 (Isolated chicken eye test method for identifying occular corrosives and severe irritants)
Deviations:
not specified
GLP compliance:
yes
Remarks:
Performed under Good Laboratory Practice principles (including govenunent regulations to the extent applicable) and in accordance with standard operating procedures and applicable standard protocols. Quality assessed.
Specific details on test material used for the study:
Colamide DIPA 10% active
Species:
other: hen's egg
Strain:
other: white leghorn
Details on test animals or tissues and environmental conditions:
Not applicable -no live animals used.
Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
10% active solution
Duration of treatment / exposure:
The test or.reference article, at a dosage of three-tenths of one milliliter (0.3 ml) of a liquid or three-tenths of one gram (0.3 g) of a solid was then administered to each of four (4) CAM's. Twenty seconds later, the test or control article was rinsed from each CAM with five (5) milliliters of physiological saline. All CAM's •were observed immediately prior to test article administration and at 30 seconds. two (2) and five (5) minutes after exposure to the test article. The reactions of the CAM, the blood vessels, including the capillaries, and the albumin were examined and scored for irritant effects.
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.
Two reference materials were used as a comparison to the ColaMoist 200 (5% active aqueous sample).
These were (1) Johnson's baby shampoo and (2) Prell shampoo (no data on pH of this material). Both reference materials are commercially available shampoos that have a similar % active ingredient level to the ColaMoist 200 (5% active) material
Details on study design:
Fresh, fertile, White Leghorn eggs were obtained from Avian Services in Frenchtown, New Jersey. They were stored at this facility for up to seven (7) days, at 13° C, before being incubated. For incubation the eggs were placed, on their sides, in a Kuhl incubator. The incubator is such that the eggs are automatically rotated once every hour. The temperature was controlled at 99° F (± I 0 with a relative humidity of 60 - 70% for the ten (I0) days of incubation. On day eight (8) the eggs were turned so that the acutely angled end faced down.

On day ten ( I 0) each egg was removed from the incubator and placed in a Plexiglas work enclosure. This enclosure had been preheated and humidified so that its environment approached that of the incubator. A cut was made in the Iaeger end of each egg, where the air sack is located. A Dremel11 Moto-Flex Tool (model 232-5) equipped with a Dremel-,; Cut-Off Wheel (No. 409) was used to make each cut. Forceps were then used to remove the shell down to the shell­
membranc junction. The inner egg membrane was then hydrated with a warm, phyiological saline solution. The saline was removed after a two (2) to five (5) minute exposure. Utilizing pointed forceps, the inner egg membrane was then carefully removed to reveal the CAM.

The test or.reference article, at a dosage of three-tenths of one milliliter (0.3 ml) of a liquid or three-tenths of one gram (0.3 g) of a solid was then administered to each of four (4) CAM's. Twenty seconds later, the test or control article was rinsed from each CAM with five (5) milliliters of physiological saline. All CAM's •were observed immediately prior to test article administration and at 30 seconds. two (2) and five (5) minutes after exposure to the test article. The reactions of the CAM, the blood vessels, including the capillaries, and the albumin were examined and scored for irritant effects.
Irritation parameter:
other: Mean CAM score
Run / experiment:
Colamide DIPA (10% active) test sample
Value:
2
Vehicle controls validity:
not applicable
Negative controls validity:
not applicable
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Irritation parameter:
other: Mean CAM score
Run / experiment:
Johnson's baby Shampoo - Reference 1
Value:
11
Vehicle controls validity:
not applicable
Negative controls validity:
not applicable
Positive controls validity:
valid
Irritation parameter:
other: Mean CAM score
Run / experiment:
Prell shampoo - Reference 2
Value:
24.25
Vehicle controls validity:
not applicable
Negative controls validity:
not applicable
Positive controls validity:
valid
Other effects / acceptance of results:
Each article was classified using the following scoring system
Irritation Potential - Practically None - Score 0 - 4.9
- Slight - Score 5.0 - 9.9
- Moderate - Score 10.0 - 14.9
- Severe - Score 15 - 32

Previous studies have shown that the CAM of the hen's egg is more sensitive to lquid irritants than is the rabbit eye. Therefore, 50% dilutions of the liquid test and reference articles were used.

Historical in-vivo testing on Johnson's reference product has shown this material to be 'moderatley irritating' eliciting scores approaching 10 at 24 hours when dosed at 100% and tested using the draize ocular irritation method.

The Prell reference product under the same in-vivo Draize testing has given scores approaching 30 at 24 hours when dosed at 100%.

Interpretation of results:
GHS criteria not met
Conclusions:
The scores from the CAM test measured against known reference materials whose in-vivo eye irritation results are known show that Colamide DIPA (10% active solution) has little or no irritant effect by the Hen's Egg in-vitro method.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Additional information

Justification for classification or non-classification