(Z)-non-6-enal

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

December 20, 2019 - May 28, 2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2020

Report date:

2020

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

Due to the volatility of the test substance, the incubation period was performed under a safety hood. Additional test substance also had to be sent to the testing laboratory.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

(Z)-non-6-enal

EC Number:

218-900-9

EC Name:

(Z)-non-6-enal

Cas Number:

2277-19-2

Molecular formula:

C9H16O

IUPAC Name:

(Z)-non-6-enal

Test material form:

liquid

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Source strain:

other: 00267

Justification for test system used:

The test system is designed to mimic human skin, the target organ of interest.

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: Normal human epidermal keratinocytes
- Tissue batch number(s): 30858
- Date of initiation of testing: Jan. 9, 2020

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

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: 15 times with enough DPBS to fill the well, then submerged 3 times with 150 mL of DPBS
- Observable damage in the tissue due to washing: No
- Modifications to validated SOP:

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration:5 mg/mL in PBS
- Incubation time: 3 hour
- Wavelength: 570 nm
- Filter bandwidth: +/- 30 nm

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: 1.598
- Barrier function: 6.45 hr
- Contamination: Sterile

NUMBER OF REPLICATE TISSUES: 3

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- N. of replicates: 3

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 corrosive to skin if the viability is less than 50%.
- The test substance is considered to be non-corrosive to skin if the viability is greater than or equal to 50%.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 30 uL
- Concentration (if solution): 100%

VEHICLE
- Amount(s) applied (volume or weight with unit): None

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 30 uL
- Concentration (if solution): 100% DPBS

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

Duration of treatment / exposure:

25 minutes under a safety hood, then 35 minutes on a warming plate in a safety hood.

Duration of post-treatment incubation (if applicable):

24 hrs, then added to new wells containing 0.9 mL incubation medium and post-incubated for an additional 18 hrs.

Number of replicates:

3

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Average

Value:

3.7

Negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

	Relative Tissue Viability (%)	Average Viability (%)	Standard Deviation (%)
Negative Control 1	99.6	100	7.1
Negative Control 2	93.1
Negative Control 3	107.3
Positive Control 1	3.6	3.7	0.1
Positive Control 2	3.8
Positive Control 3	3.7
Test Item 1	3.6	3.7	0.3
Test Item 2	3.4
Test Item 3	4.0

 

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritant) based on GHS criteria

Conclusions:

The tissue viability was less than 50% at one hour of treatment. The test substance is therefore considered to be an irritant to skin.

Executive summary:

The potential for skin irritancy was studied in an OECD Guideline 439 in vitro human skin model test.  Model human epidermis was exposed to the test substance for one hour.  Due to volatility of the test substance, the incubation period was done under a safety hood.  Three replicates were performed.  The average percent viability for the test substance was 3.7%.  The test substance is therefore considered to be a category 2 skin irritant.