Hexadecyl dihydrogen phosphate

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

March 6, 2019 to March 8, 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Version / remarks:

The MatTek Corporation In Vitro EpiDermTM Skin Irritation Test (EPI-200-SIT) (OECD 439) Protocol: In Vitro EpiDermTM Skin Irritation Test (EPI-200-SIT)
Protocol Date: 11/7/17

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Hexadecyl dihydrogen phosphate

EC Number:

222-581-1

EC Name:

Hexadecyl dihydrogen phosphate

Cas Number:

3539-43-3

Molecular formula:

C16H35O4P

IUPAC Name:

hexadecyl dihydrogen phosphate

Test material form:

solid: particulate/powder

Details on test material:

Product commercial name is ColaFax CPE

Specific details on test material used for the study:

1-Hexadecanol, 1-(dihydrogen phosphate), (ColaFax CPE), 100% Active; Lot Number: 64252F18

In vitro test system

Test system:

human skin model

Remarks:

The reconstructed human epidermal model EpiDerm consists of normal, human-derived epidermal keratinocytes which have been cultured to form a multilayered, highly differentiated model of the human epidermis.

Source species:

human

Cell type:

non-transformed keratinocytes

Justification for test system used:

EpiDerm, 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-yl]-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 tetrazolium salt. The amount of MTT reduced by a culture is therefore proportional to the number of viable cells

Vehicle:

unchanged (no vehicle)

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

After the appropriate tissue preparation, milligrams of the test article, at 100%, 30 microliters of the positive control article and 30 microliters of the negative control article were each added to each of three Millicells containing the EpiDerm samples

Duration of treatment / exposure:

After dosing the last tissue, the six (6) well plates containing the dosed EpiDerm samples were incubated at 37°C, five (5)% carbon dioxide and > 95% humidity for 35 + 1 minutes.
After the 35 + 1 minute exposure period, the six well plates were removed from the incubator and placed in the sterile hood until the 60 minute exposure period was reached for the first dosed tissue.

Duration of post-treatment incubation (if applicable):

The EpiDerm samples were then returned to the incubator for 24 + 2 hours

Number of replicates:

Six

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Mean of three runs (6 plates)

Value:

78.8

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

As per MatTek and OECD 439, the assay meets the negative control criteria if the mean OD-570 for the negative control tissues is > 0.8 and < 2.8. The value is 1.473 and meets the acceptance criteria.
As per MatTek, the assay meets the positive control criteria if the mean viability of the positive control tissues, expressed as the percent of the negative control tissues is < 20%. The value is 3.4 and meets the acceptance criteria.
As per MatTek and OECD 439, the assay meets the standard deviation criteria if the standard deviation calculated from individual percent tissue viabilities of the 3 identically treated replicates is < 18%. The values are 14.11, 5.90 and 0.21 and they meet the acceptance criteria.

Any other information on results incl. tables

According to the EU and the Globally Harmonized System of Classification and Labeling of Chemicals, an irritant is predicted if the mean relative tissue viability of three individual tissues exposed to the test substance is reduced to below 50% of the mean viability of the negative controls.

In vitro result                                                 In vivo prediction

Mean tissue viability < 50%                            Irritant, (R38 or GHS category 2)

Mean tissue viability > 50%                                   Non-irritant

Test Article Mean tissue viability = 78.8%              Non-irritant

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of this test, the test article, 1-Hexadecanol, 1-(dihydrogen phosphate), (ColaFax CPE), 100% Active; Lot Number: 64252F18, at 100%, with a mean viability of 78.8%, is not considered an irritant.