Fatty acids, C16-18, reaction products with diethanolamine

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation / corrosion, other

Remarks:

other: in vitro (Epiderm)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

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:

Fatty acids, C16-18, reaction products with diethanolamine

EC Number:

293-014-3

EC Name:

Fatty acids, C16-18, reaction products with diethanolamine

Cas Number:

91032-08-5

Molecular formula:

C22H45NO3 to C52H101NO5

IUPAC Name:

Fatty acids, C16-18, reaction products with diethanolamine

Test animals

Species:

other: EpiDerm reconstructed human epidermis model (MatTek Corp.)

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

Controls:

other: three tissues for negative control (30µL DPBS) and positive control (30µL 5% SDS)

Amount / concentration applied:

solids: 25 µL DPBS + 25 mg

Duration of treatment / exposure:

60 +/- 1 minutes

Observation period:

42 +/- 4 h

Number of animals:

three replicate tissues per dose group

Details on study design:

3 replicate tissues are dosed with the test item, the negative control (DPBS) and the positive control (5% SDS), respectively. The tissues are incubated for 35 +/-1 minutes in a humidified incubator (37+/- 1 °C, 5 +/- 1% CO2, 95% RH) and further 25 minutes at room temperature. Then the test item and the controls are rinsed off with DPBS and the tissues are post-incubated for 24 +/- 2 h and, after medium renewal, for further 18 +/- 2 h. Then the tissues are stained via MTT for 3 hours. Isopropanol extracts are measured photometrically at 550 nm.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

other: mean relative tissue viability

Basis:

other: mean tissue viability of the negative control tissues

Time point:

other: 60 min treatment 42 h post-incubation

Score:

> 50

Remarks on result:

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

Results 2

Irritation parameter:

other: mean relative tissue viability

Basis:

other: mean tissue viability of the negative control tissues

Time point:

other: 60 min treatment 42 h post-incubation

Score:

<= 50

Remarks on result:

other: irritant; EU CLP and UN GHS Category 2

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			TM
Tissue	1	2	3	1	2	3	1	2	3
OD550(blank-corrected)	1.320	2.141	1.843	0.123	0.117	0.117	1.961	2.564	2.449
	1.626	1.912	1.814	0.137	0.089	0.121	2.566	2.601	2.443
mean OD550of the replicates (blank-corrected)	1.473	2.026	1.828	0.130	0.103	0.119	2.263	2.582	2.446
total mean OD550of 3 replicate tissues (blank-corrected)	1.776			0.117			2.430
SD OD550		0.279			0.016			0.239
relative tissue viability [%]	82.9	114.1	103.0	7.3	5.8	6.7	127.5	145.4	137.7
SD tissue viability [%]^***	15.8			0.8			9.0
CV [% viability]		15.8			11.5			6.6
mean relative tissue viability [%]	100.0			6.6			136.9

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

^**      mean relative tissue viability of the three positive control tissues is < 20%

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

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

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

Executive summary:

In the in vitro skin irritation test using the EpiDerm human epidermis model 25 mg test item + 25 µL DPBS were applied topically for 60 minutes. After 42 h post-incubation cytotoxic effects were determined via MTT reduction assay.

The test item showed no irritant effects. The mean relative tissue viability (% negative control) was> 50% (137%) after 60 min treatment and 42 h post incubation.

This study is classified as acceptable:

Mean OD550 of the three negative control tissues is >=1 and <=2.5.

Mean relative tissue viability of the three positive control tissues is <= 20%.

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

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