Nonanamide, N,N-dimethyl-

Endpoint summary

Description of key information

The registration substance, Genagen PA/N,N-Dimethylnonanamide  was evaluated for the skin irritation property in an in-vitro test system using reconstructed human epidermal model according to the Guideline OECD 439. A clear reduction of tissue viability was obtained and thus the registration substance was found to be irritating to skin.

The eye irritation property of the registration substance Genagen PA/ N, N-Dimethylnonanamide was evaluated based on the eye irritation data on the structurally related substances N,N-Dimethylalkyl(C8/C10)amide and N,N-Dimethylalkyl(C10)amide. These read-across source substances were investigated according to the Guideline OECD 405.

N,N-Dimethylalkyl(C8/C10)amide induced clear adverse effects in eye of a rabbit that were not reversible within 7 days.

N,N-Dimethylalkyl(C10)amide induced clear adverse effects in eyes of three rabbits that were reversible within 21 days.

Genagen PA/ N, N-Dimethylnonanamide is to be classified as eye irritant.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

22 December 2016 to 09 March 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

OECD Guidelines for the testing of chemicals No. 439, “In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method”, adopted on 28 July 2015.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and batch No.of test material:Clariant Produkte Deutschland GmbH and 112-2016-11 D
- Expiration date of the batch:May 2018
- Purity test date: 98.3%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Cool and dry (+2 to +8°C)


FORM AS APPLIED IN THE TEST (if different from that of starting material): Liquid

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: Human Epidermal Model {EpiDerm™ (EPI-200-SIT)}

Justification for test system used:

As recommended in OECD Guideline No. 439, Reconstructed Human Epidermal Model EpiDerm™ (EPI-200-SIT) has been selected as test system for in vitro skin irritation.

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE:
- Model used: The Reconstructed Human Epidermal Model - EpiDerm™ (EPI-200-SIT)
- Tissue Lot number(s): 23387
- Production date: 21 Dec-2016
- Delivery date:23 December 2016
- Date of initiation of testing: 23 December 2016

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37±1°C
- Temperature of post-treatment incubation (if applicable): 37±1°C

REMOVAL OF TEST MATERIAL AND CONTROLS
washing steps: After 60 minutes of test item exposure, the tissues were rinsed with sterile DPBS by filling and emptying the tissue insert for 15 times to remove any residual test item.
- Observable damage in the tissue due to washing: No
- Modifications to validated SOP: No

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL
- Incubation time: 3 hours 5 minutes
- Spectrophotometer: Spectromax M5e
- Wavelength: 570 nm


FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability:MTT QC Assay-4 hours, 1.354±0.216
- Barrier function:5.19 hours (Pass)
- Contamination: Sterile (Pass)


NUMBER OF REPLICATE TISSUES: Three

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
The assay met the acceptance criterion as the mean OD570 of the NC tissues is 1.762 which in the range of ≥ 0.8 and ≤ 2.8.
The assay met the acceptance criterion as the mean viability of PC tissues is 2.7% which were ≤ 20% of the negative control tissues and the SD of the three tissues replicates is 0.42 (below the 18%).
The assay met the acceptance criterion as the standard deviation (SD) calculated from individual % tissue viabilities of the 3 identically treated replicates is < 18% i.e., in the range of 0.13 to 17.36
The test item is considered as “irritant” to skin in accordance with UN GHS Category 2, as the tissue viability after exposure and post-treatment incubation is ≤ 50%.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume with unit): 30 µL test item (Genagen PA / N, N-Dimethylnonanamide).

NEGATIVE CONTROL
- Amount(s) applied (volume ): 30 µL each of DPBS (negative control: NC),

POSITIVE CONTROL
- Amount(s) applied (volume ):30 µL of 5% aq. SDS solution (positive control: PC)

Duration of treatment / exposure:

60 minutes

Duration of post-treatment incubation (if applicable):

23 hours and 30 minutes

Number of replicates:

Three

Details on test animals or test system and environmental conditions:

Not Applicable

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

Not Applicable

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Experiment

Value:

< 50

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:

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

Interpretation of results:

Category 2 (irritant) based on GHS criteria

Conclusions:

The registration substance, Genagen PA/N,N-Dimethylnonanamide was evaluated for the skin irritation property in an in-vitro test system using reconstructed human epidermal model according to the Guideline OECD 439. A clear reduction of tissue viability was obtained and thus the registration substance was found to be irritating to skin.

Executive summary:

The registration substance, Genagen PA/N,N-Dimethylnonanamide  was evaluated for the skin irritation property in an in-vitro test system using reconstructed human epidermal model according to the Guideline OECD 439. A clear reduction of tissue viability was obtained and thus the registration substance was found to be irritating to skin.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

GLP compliance:

yes

Species:

rabbit

Strain:

other: russian

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

0.1 ml

Observation period (in vivo):

up to 21 days

Number of animals or in vitro replicates:

3

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 21 days

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

fully reversible within: 21 days

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

2.6

Max. score:

3

Reversibility:

fully reversible within: 21 days

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

2.8

Max. score:

4

Reversibility:

fully reversible within: 21 days

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

The eye irritation property of N,N-Dimethylalkyl(C10)amide was investigated according to the Guideline OECD 405. Clear adverse effect on the eyes was observed that was reversible within 21 days.

Executive summary:

The eye irritation property of N,N-Dimethylalkyl(C10)amide was investigated according to the Guideline OECD 405.

0.1ml of N,N-Dimethylalkyl(C10)amide was applied to the one eye of each three rabbits and observed for up to 21 days.

The calculated scores (average of 24h, 48h and 72h) are: cornea opacity 1.0, iritis 0.0, conjunctivia redness 2.6 and chemosis 2.8. Effects observed were reversible within 21 days.

N,N-Dimethylalkyl(C10)amide is to be classified as eye irritant.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Justification for classification or non-classification

The registration substance, Genagen PA/N,N-Dimethylnonanamide  was evaluated for the skin irritation property in an in-vitro test system using reconstructed human epidermal model according to the Guideline OECD 439. A clear reduction of tissue viability was obtained and thus the registration substance was found to be irritating to skin.

The eye irritation property of the registration substance Genagen PA/ N, N-Dimethylnonanamide was evaluated based on the eye irritation data on the structurally related substances N,N-Dimethylalkyl(C8/C10)amide and N,N-Dimethylalkyl(C10)amide. These read-across source substances were investigated according to the Guideline OECD 405.

N,N-Dimethylalkyl(C8/C10)amide induced clear adverse effects in eye of a rabbit that were not reversible within 7 days.

N,N-Dimethylalkyl(C10)amide induced clear adverse effects in eyes of three rabbits that were reversible within 21 days.

Genagen PA/ N, N-Dimethylnonanamide is to be classified as eye irritant.