Reaction product of Fatty acids, C18-unstaturated, dimer with 4,7,10-trioxa-1,13-tridecane-diamine

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2015

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Remarks:

No deviations ocurred that adversely impacted the integrity of the study.

GLP compliance:

no

Remarks:

This study was conducted prior to the intention to register the substance for REACH. The study was well-documented and is scientifically valid.

Test material

Test material information

Constituent 1

Reference substance name:

Reaction product of Fatty acids, C18-unstaturated, dimer with 4,7,10-trioxa-1,13-tridecane-diamine

EC Number:

701-270-9

Cas Number:

68911-25-1

Molecular formula:

N.A. UVCB substance

IUPAC Name:

Reaction product of Fatty acids, C18-unstaturated, dimer with 4,7,10-trioxa-1,13-tridecane-diamine

Test material form:

liquid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 3M Company, Lot 20125
- Expiration date of the lot/batch: No data
- Purity test date: 21 January, 2015

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: No data

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: None, dosed neat

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

other: Human Keratinocytes cultivated in vitro to resemble human epidermis.

Cell source:

other: Proprietary EpiDerm reconstructed human epidermis tissues were purchased from MatTek Corporation

Details on animal used as source of test system:

SOURCE ANIMAL
Proprietary EpiDerm reconstructed human epidermis tissues were purchased from MatTek Corporation

Justification for test system used:

EpiDerm is an appropriate test system per OECD 439.

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm Skin Irritation Test (EPI-200-SIT)
- Tissue batch number(s): No data
- Production date: No data
- Shipping date: No data
- Delivery date: 03 February, 2015
- Date of initiation of testing: 02 February, 2015

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37 C
- Temperature of post-treatment incubation (if applicable): 37 C

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: The tissue inserts were filled and emptied 20 times with fresh DPBS from a wash bottle.
- Observable damage in the tissue due to washing: No
- Modifications to validated SOP: None

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: No data
- Incubation time: 3 hours
- Spectrophotometer: No data
- Wavelength: 570 nm
- Filter: No data
- Filter bandwidth: No data
- Linear OD range of spectrophotometer: No data

NUMBER OF REPLICATE TISSUES: 3

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- Killed tissues
- Procedure used to prepare the killed tissues (if applicable): Freezing
- N. of replicates : 2
- Method of calculation used: % Viability of negative test system control

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 3

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be an irritant if the mean tissue viability (as a percentage of the negative control) was less than or equal to 50%.

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 30 uL
- Concentration (if solution): Dosed undilluted.

VEHICLE : None

NEGATIVE CONTROL : DPBS
- Amount(s) applied (volume or weight): 30 uL
- Concentration (if solution): No data

POSITIVE CONTROL : SDS
- Amount(s) applied (volume or weight): 30 uL
- Concentration (if solution): 5%

Duration of treatment / exposure:

60 minutes

Duration of post-treatment incubation (if applicable):

42 hours

Number of replicates:

3

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

1

Value:

8

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Results 2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

2

Value:

5

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Results 3

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3

Value:

19

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Other effects / acceptance of results:

- OTHER EFFECTS:
- Visible damage on test system: No
- Direct-MTT reduction: No
- Colour interference with MTT: No

DEMONSTRATION OF TECHNICAL PROFICIENCY: Yes

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes
- Acceptance criteria met for positive control: Yes
- Acceptance criteria met for variability between replicate measurements: Yes

Any other information on results incl. tables

Mean % tissue viablity for the test article was 11%.

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritant) based on GHS criteria

Conclusions:

Based on the results of the study (11% mean tissue viability following 60 minute exposure), MTDID 18990 is a skin irritant.

Executive summary:

The dermal irritation potential of MTDID 18990 was evaluated in the EpiDerm reconstructed human epidermal tissue model. The study was conducted according to OECD 439. EpiDerm tissues (n=3) were exposed to 30 uL of test article (neat), DPBS (negative control) or SDS (postive control) for 60 minutes at 37C. Following exposure, the tissues were washed 20 times by filling the plate wells with a gentle stream of DPBS and emptying. The plate wells were then filled with fresh media and returned to an incubator at 37C for 24 hours. At 24 hours, the media was replaced with fresh media and incubated for another 18 hours. Following incubation the tissues were patted dry with a tissue and transferred to plates containing 300 uL of MTT solution per well. The tissues were incuabted with MTT for 3 hours. Following incubation they were washed and the wells were filled with MTT extraction soltuion and extracted for 2 hours at room temperature. The OD at 570 nm was measured for each solution and normalized to the mean of the negative control group to calculate % tissue viability. MTDID 18990 -exposed tissues had % viability values of 8, 5, and 19% for a mean tissue viability of 11%. Based on the results of the study (11% mean tissue viability following 60 minute exposure), MTDID 18990 is a skin irritant.