Fatty acids, C8-10-(even numberd), Esters with pentaerythritol and adipic acid

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

12 - 16 Nov 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:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: ECVAM international validation study on in vitro tests for acute skin irritation: Report on the validity of the EPISKIN and EpiDerm assays and on the Skin Integrity Function Test (Altern Lab Anim. 2007 Dec.; 35 (6): 559-601)

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Protocol for: IN VITRO EpiDerm™ SKIN IRRITATION TEST (EPI-200-SIT), Rev. 3/23/2009, MatTek Corporation, Ashland, USA

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Landesamt für Umwelt, Wasserwirtschaft und Gewerbeaufsicht, Mainz, Germany

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, C8-10, mixed esters with adipic acid and trimethylolpropane

EC Number:

306-085-3

EC Name:

Fatty acids, C8-10, mixed esters with adipic acid and trimethylolpropane

Cas Number:

95912-89-3

IUPAC Name:

95912-89-3

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: EpidermTM, reconstructed three-dimensional human epidermis model (EPI-200)

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: MatTek, In Vitro Life Science Laboratories, Bratislava, Slovakia
- Tissue batch number(s): 16854
- Delivery date: 13 Nov 2012

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 35 min at 37 °C, 25 min at room temperature
- Temperature of post-treatment incubation: 37°C

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: The tissues were rinsed in one-minute intervals.

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1mg/mL
- Incubation time: 3 h
- Spectrophotometer: Photometer MRX
- Wavelength: 570 nm

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Contamination: No biological contamination ofHIV-1 virus, Hepatitis B virus, Hepatitis C virus or bacteria, yeast and fungi were detected.

PREDICTION MODEL / DECISION CRITERIA
- The test substance is considered to be irritant to skin if the viability after 1 h exposure is less than 50%.
- The test substance is considered to be non-irritant to skin if the viability after 1 h exposure is greater than 50%.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied: 30 µL

NEGATIVE CONTROL
- Amount(s) applied: 30 µL

POSITIVE CONTROL
- Amount(s) applied: 30 µL
- Concentration: 5% solution in deionised water

Duration of treatment / exposure:

35 min at 37 °C, 25 min at room temperature

Duration of post-treatment incubation (if applicable):

42 h

Number of replicates:

Triplicate tissues; from each tissue, 2 absorbance measurements after MTT incubation were performed

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Mean of 3 tissues

Value:

99.4

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The negative control OD (1.329, 1.057 and 1.049) was in the range of ≥ 1.0 and ≤ 2.5 for all three tissues. As the negative control result fell within the range defined in the acceptance criteria, the result is considered to be valid.
- Acceptance criteria met for positive control: Exposure to the positive control induced a decrease in the relative absorbance as compared to the negative control to 14.7% and thus, lower than threshold of 20%.
- Acceptance criteria met for variability between replicate measurements: The standard deviations between the three tissue replicates of the treatment group, the negative control and the positive control were 4.4%, 13.9% and 0.0%, respectively, which is ≤ 18%.

Any other information on results incl. tables

Table 2. Results of MTT assay after 1 h exposure

	Negative control			Test item			Positive control
Tissue sample	1	2	3	1	2	3	1	2	3
OD570	1.466 1.277	1.113 1.087	1.096 1.088	1.298 1.179	1.164 1.161	1.140 1.146	0.212 0.209	0.210 0.212	0.211 0.210
OD570(mean)	1.329	1.057	1.049	1.196	1.120	1.100	0.168	0.168	0.168
OD570(mean values of replicates±%RSD)	1.145 ± 13.9			1.139 ± 4.4			0.168 ± 0.0
Viability (%)	100			99.4			14.7

OD = optical density; RSD = relative standard deviation

Applicant's summary and conclusion

Interpretation of results:

other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No. 1272/2008

Conclusions:

Under the conditions of the RHE test method the test substance did not show irritant properties.
CLP: not classified