ColaLiquids DC5, DL, DM (Commercial names)Cocamide DIPA (INCI name)

Eye irritation

Administrative data

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

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

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

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of N,N-Bis(2-hydroxypropyl)(C6-C18 even numbered and C18 unsatd)amides and bis(2-hydroxypropyl)ammonium myristate

Molecular formula:

No molecular formula. UVCB reaction mass.

IUPAC Name:

Reaction mass of N,N-Bis(2-hydroxypropyl)(C6-C18 even numbered and C18 unsatd)amides and bis(2-hydroxypropyl)ammonium myristate

Test material form:

liquid

Test animals / tissue source

Species:

human

Test system

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

Results and discussion

In vitro

Results

Results 1

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

Results 2

Irritation parameter:

other: Draize equivalent

Value:

4.5

Vehicle controls validity:

not specified

Negative controls validity:

valid

Positive controls validity:

not applicable

Any other information on results incl. tables

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.

Applicant's summary and conclusion

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.