Reaction mass of hexadecyl dihydrogen phosphate and dihexadecyl hydrogen 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:

Jan - Feb 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: EU Method B.46 (Skin Irritation)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany

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 animals

Species:

other: EpiSkin reconstructed human epidermis model (SkinEthic Laboratories)

Details on test animals or test system and environmental conditions:

organotypic reconstructed three-dimensional model of the human epidermis

Test system

Type of coverage:

other: direct application

Amount / concentration applied:

10 mg + 5 µL aqua dest.
The amount of aqua dest. was increased to 50 µL to allow for evenly spreading of the test item to match size of the tissue.

Duration of treatment / exposure:

15 minutes

Observation period:

42 +/- 1 h

Details on study design:

3 replicate tissues are dosed with the test item, the negative control (PBS) and the positive control (5% SDS), respectively. After 15 minutes treatment period at room temperature the test item and the controls are rinsed off with PBS and the tissues are post-incubated for 42 +/- 1 h. Then the tissues are stained via MTT for 3 hours. Isopropanol extracts are measured photometrically at 550 nm.

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

other: other: mean relative tissue viability

Value:

> 50

Remarks on result:

other:

Remarks:

Basis: other: mean tissue viability of the negative control tissues. Time point: 15 min treatment 42 h post-incubation. Remarks: non-irritant; EU CLP and UN GHS: No Category. (migrated information)

Results 2

Irritation / corrosion parameter:

other: other: mean relative tissue viability

Value:

<= 50

Remarks on result:

other:

Remarks:

Basis: other: mean tissue viability of the negative control tissues. Time point: 15 min treatment 42 h post-incubation. Remarks: irritant; EU CLP and UN GHS Category 2. (migrated information)

In vivo

Irritant / corrosive response data:

If mean tissue viability is > 50% relative to the mean negative control, the test item is classified as non-irritant (EU CLP and UN GHS: No Category).
If mean tissue viability is <= 50% relative to the mean negative control, the test item is classified as irritant (EU CLP and UN GHS: Category 2).

Other effects:

The test item showed no direct MTT reducing capability and no colouring potential.

Any other information on results incl. tables

 

Name	NC			PC			TI
Tissue	1	2	3	1	2	3	1	2	3
absolute OD550	0.913	0.905	0.854	0.061	0.058	0.061	1.076	1.059	1.121
	0.929	0.892	0.908	0.060	0.061	0.064	1.077	1.070	1.151
blank-corrected OD550	0.868	0.860	0.809	0.016	0.013	0.015	1.031	1.014	1.076
	0.884	0.847	0.863	0.014	0.015	0.018	1.032	1.025	1.106
mean OD550of the duplicates (blank-corrected)	0.876	0.853	0.836	0.015	0.014	0.017	1.031	1.019	1.091
total mean OD550of 3 replicate tissues (blank-corrected)	0.855^*			0.015			1.047
SD OD550	0.03			0.00			0.04
relative tissue viabilities [%]	102.5	99.8	97.7	1.8	1.6	2.0	120.6	119.2	127.6
mean tissue viability [%]	100.0			1.8^**			122.5
SD tissue viability [%]^***	2.4			0.2			4.5
CV [% viability]	2.4			9.1			3.7

*       Corrected mean OD₅₅₀of the negative control corresponds to 100% absolute tissue viability.

^**      Mean relative tissue viability of the three positive control tissues is <= 40%

^***     Standard deviation (SD) obtained from the three concurrently tested tissues is < 18%

Applicant's summary and conclusion

Interpretation of results:

other: non-irritant; EU CLP and UN GHS: No Category

Remarks:

Criteria used for interpretation of results: other: OECD 439

Conclusions:

The test item is classified as "non-irritant" (No Category).

Executive summary:

In the in vitro skin irritation test using the EpiSkin human epidermis model 10 mg test item + 50 µl A. dest. were applied topically for 15 minutes. After 42 h post-incubation cytotoxic effects were determined via MTT reduction assay.

The mean relative tissue viability (% negative control) was > 50%.

The test item is therefore classified as "non-irritant" (EU CLP and UN GHS: No Category).

This study is classified as acceptable:

OD550 of the blank is < 0.1 (0.045).

Mean OD550 of the three negative control tissues is >= 0.6 and <= 1.5 (0.855).

Mean relative tissue viability of the three positive contol tissues is <= 40% (1.8%).

Standard deviation (SD) of relative tissue viability obtained from each three concurrently tested tissues is < 18% (0.2% - 4.5%).

This study satisfies the requirement for Test Guideline OECD 439 for in vitro skin irritation.