Reaction mass of N,N-dimethyldodecanamide and N,N-dimethyltetradecanamide

Endpoint summary

Description of key information

Irritation/corrosion

Skin: in vivo, rabbit, semiocclusive, 4h, OECD 404: Moderate to severe skin reactions, erythema 2.7 and oedema 2.8, full reversible within 21 days. (C10) (Cognis 1997, M.Hoyer)

Eye: in vivo, rabbit, OECD 405: Moderate to severe eye reactions, .(Cognis 1997, M.Hoyer), cornea opacity 1.0; iris lesion 0.0; redness of conjunctiva 2.6; oedema of conjunctiva 2.8, full reversible within 21 days. (C10) (Cognis 1997, M.Hoyer).

These data indicate that the test material was an irritant for skin and eyes.

It may be predicted from this that registered susbtance would share a similar toxicity profile and would also be predicted to be an irritant.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1997

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study according OECD guideline, GLP, well documented

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

GLP compliance:

yes

Species:

rabbit

Strain:

other: Mol:Russian

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Mollegaard Breeding and Research Centre A/S, Ejby, DK-4623
- Weight: 2.3-2.5 kg
- Housing: animal room 2, filtered Air, ppo cages
- Diet (e.g. ad libitum): ad libitum / Altromin 2123 , Altromin , Lage , Germany
- Water (e.g. ad libitum): ad libitum (acified, HCl)
- Acclimation period: at least one week


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20±3°C
- Humidity (%): 55±15%
- Air changes (per hr): 10times/h
- Photoperiod (hrs dark / hrs light):12h each

Type of coverage:

semiocclusive

Preparation of test site:

other: clipped

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5ml

Duration of treatment / exposure:

4h

Observation period:

1h, 24h, 48h, 72h, 7d, 14d, 21d

Number of animals:

3

Details on study design:

TEST SITE
- Clipped area: 10*10cm
- Area of exposure: 2.5cm*2.5cm
- % coverage: 100
- Type of wrap if used: 16-layer gaze fixed with 2.5cm adhesive Gothaplast tape

REMOVAL OF TEST SUBSTANCE
- Washing (if done): treated skin was cleaned with mild soap and lukewarm water
- Time after start of exposure: 4h

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: after 21 days rests of scales were observed

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

fully reversible within: after 21 days rests of scales were observed

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

fully reversible within: after 21 days rests of scales were observed

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: after 21 days rests of scales were observed

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

3.33

Max. score:

4

Reversibility:

fully reversible within: after 21 days rests of scales were observed

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

fully reversible within: after 21 days rests of scales were observed

Irritant / corrosive response data:

no evidence for corrosion reported

Interpretation of results:

Category 2 (irritant)

Remarks:

Migrated information Criteria used for interpretation of results: other: EU GHS EC 1272/2008

Conclusions:

irritating to skin

Executive summary:

The acute skin irritant effect of SAT 970 419 was investigated according to the method recommended in the OECD Guideline No. 404, "Acute Dermal Irritation/Corrosion", 1992, and EEC Guideline B.4 "Acute Toxicity (Skin Irritation)", 29.12.1992.

Three female albino rabbits were exposed to the test article at two skin sites on the back. After 4 hours of exposure the test article was removed and the skin was examined 1, 24, 48 and 72 hours as well as 7, 14 and 21 days after termination of exposure. Moderate to severe skin reactions were observed.

Under the experimental conditions described in this report, the mean score for erythema was 2.7 (2.00, 3.00, 3.00) and for oedema 2.8 (2.00, 3.33, 3,00). After 21 days rests of scales on both sides were observed in all animals.

Based on this result the substance has to be labeled as irritating to skin according to EU GHS EC 1272/2008.

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
The read-across hypothesis proposed is that the organism is not exposed to common compounds but rather, as a result of structural similarity, that different compounds have similar toxicological and fate properties. In this case the ECHA Read-Across Assessment Framework (RAAF) Scenario 2 is used.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Source: N,N-Dimethyldecanamide [ EC 238-405-1; CAS 14433-76-2]
Target: Reaction Mass of N,N-Dimethyldodecanamide and N,N-Dimethyltetradecanamide [EC not assigned; CAS not assigned]

3. ANALOGUE APPROACH JUSTIFICATION
It may be concluded that both the registered substance (target) and the source molecules can be regarded as close structural analogues having both similar physical chemical and toxicological properties. It follows that where endpoints for the registered substance may not have experimental evidence, particularly those involving animal studies, that these can be addressed by read across grouping using an analogue approach. It is therefore proposed that, after careful assessment, experimental data from the source molecules may be used as surrogate evidence for read across to the registered substance. Please refer to attached document for information on the data available to support the read-across.

4. DATA MATRIX
Please refer to attached document

Reason / purpose for cross-reference:

read-across source

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: after 21 days rests of scales were observed

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

fully reversible within: after 21 days rests of scales were observed

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

fully reversible within: after 21 days rests of scales were observed

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: after 21 days rests of scales were observed

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

3.33

Max. score:

4

Reversibility:

fully reversible within: after 21 days rests of scales were observed

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

fully reversible within: after 21 days rests of scales were observed

Reference 3

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

03-12-2013 to 26-05-2014

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)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Name of test substance: N,N Dimethyldodecane-1-amide
Test-substance No.: 13/0555-1
Batch identification: 0009565072
CAS No.: 3007-53-2

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: EpiDermTM model

Source strain:

not specified

Details on animal used as source of test system:

EpiDermTM model consists of normal, human-derived epidermal keratinocytes

Justification for test system used:

Based on the results of ECVAM (European Center for Validation of Alternative Methods) funded validation studies, it was concluded by the ECVAM Scientific Advisory Committee that the EpiDerm™ human epidermis model is suitable to be used for distinguishing between corrosive and non-corrosive chemicals (ECVAM: ESAC statement on the application of the EpidermTM human skin model for skin corrosivity testing of 14-15 Mar 2000) as well as between irritant and non-irritant chemicals (ECVAM: ESAC statement on the scientific validity of in-vitro tests for skin irritation testing of 5 Nov 2008).

Vehicle:

unchanged (no vehicle)

Details on test system:

The EpiDermTM model consists of normal, human-derived epidermal keratinocytes which have been cultured to form a multi layered, highly differentiated model of the human epidermis. It consists of organized basal, spinous and granular layers, and a multi-layered stratum corneum containing intercellular lamellar lipid layers arranged in patterns analogous to those found in vivo. The EpiDermTM tissues (surface 0.6 cm²) are cultured on specially prepared cell culture inserts (MILLICELLs, 10 mm ) and commercially available as kits (EpiDerm™ 200), containing 24 tissues on shipping agarose.


Irritation test:

On the day of arrival in the laboratory, the tissues were transferred to sterile 6-well plates with 0.9 mL assay medium and preconditioned in the incubator at 37°C. After 1 hour the preincubation medium was replaced with fresh medium and preconditioning continued for 18 ± 3 hours. Three tissues were treated with the test substance, the PC and NC, respectively. Thirty microliter (30 μL) of the undiluted liquid test substance was applied using a pipette. A nylon mesh was placed carefully onto the tissue surface afterwards. Control tissues were concurrently applied with 30 μL of sterile PBS (negative control, NC) or with 30 μL of 5% SDS (positive control, PC). A nylon mesh was placed carefully onto the tissue surface afterwards.
The tissues were kept under the laminar flow hood at room temperature for 25 minutes overall and for 35 minutes in the incubator. The tissues were washed with sterile PBS to remove residual test material 1 hour after start of application. Rinsed tissues were blotted on sterile absorbent paper and transferred into new 6-well plates, pre-filled with 0.9 mL fresh medium. When all tissues were rinsed, the surface of each tissue was carefully dried with a sterile cotton swab. Subsequently, the tissues were incubated in the incubator at 37°C for 24 ± 2 hours. After 24 ± 2 hours the tissues were transferred into new 6-well plates pre-filled with 0.9 mL of fresh medium and placed into the incubator for additional 18 ± 2 hours post-incubation period.
After the post-incubation period, the assay medium was replaced by 0.3 mL MTT solution and the tissues were incubated in the incubator for 3 hours. After incubation, the tissues were washed with PBS to stop the MTT-incubation. The formazan that was metabolically produced by the tissues was extracted by incubation of the tissues in isopropanol. The optical density at a wavelength of 570 nm (OD570) of the extracts was determined spectrophotometrically. Blank values were established of 6 microtiter wells filled with isopropanol for each microtiter plate.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

Single topical application of 30 μL of the undiluted test substance to a reconstructed three dimensional human epidermis model (EpiDerm™).

Duration of treatment / exposure:

The irritation test was performed with three EpiDerm™ tissue samples, which were incubated with the test substance for 1 hour followed by a 42-hours post-incubation period.

Number of replicates:

3

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Irritation test

Value:

ca. 2.7 - ca. 3.5

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Interpretation of results:

Category 2 (irritant) based on GHS criteria

Conclusions:

Based on the observed results and applying the appropriate evaluation criteria it was concluded, that N,N Dimethyldodecane-1-amide shows a skin irritation potential in the EpiDerm™ skin corrosion/irritation test under the test conditions chosen.

Executive summary:

The potential of N,N Dimethyldodecane-1-amide to cause dermal irritation was assessed by a single topical application of 30 μL (irritation test) of the undiluted test substance to a reconstructed three dimensional human epidermis model (EpiDerm™). The irritation test was performed with three EpiDerm™ tissue samples, which were incubated with the test substance for 1 hour followed by a 42-hours post-incubation period.

Tissue destruction was determined by measuring the metabolic activity of the tissue after exposure/post-incubation using a colorimetric test. The reduction of mitochondrial dehydrogenase activity, measured by reduced formazan production after incubation with a tetrazolium salt (MTT) was chosen as endpoint. The formazan production of the test substance treated epidermal tissues is compared to that of negative control tissues. The quotient of the values indicates the relative tissue viability.

Irritation test:

The mean viability of the test-substance treated tissues determined after an exposure period of 1 hour with about 42 hours post-incubation was 3%.

Based on the observed results and applying the appropriate evaluation criteria it was concluded, that N,N Dimethyldodecane-1-amide shows a skin irritation potential in the EpiDerm™ skin corrosion/irritation test under the test conditions chosen.

Reference 4

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
The read-across hypothesis proposed is that the organism is not exposed to common compounds but rather, as a result of structural similarity, that different compounds have similar toxicological and fate properties. In this case the ECHA Read-Across Assessment Framework (RAAF) Scenario 2 is used.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Source: N,N-Dimethyldodecanamide [ EC 221-117-5; CAS 3007-53-2]
Target: Reaction Mass of N,N-Dimethyldodecanamide and N,N-Dimethyltetradecanamide [EC not assigned; CAS not assigned]

3. ANALOGUE APPROACH JUSTIFICATION
It may be concluded that both the registered substance (target) and the source molecules can be regarded as close structural analogues having both similar physical chemical and toxicological properties. It follows that where endpoints for the registered substance may not have experimental evidence, particularly those involving animal studies, that these can be addressed by read across grouping using an analogue approach. It is therefore proposed that, after careful assessment, experimental data from the source molecules may be used as surrogate evidence for read across to the registered substance. Please refer to attached document for information on the data available to support the read-across.

4. DATA MATRIX
Please refer to attached document

Reason / purpose for cross-reference:

read-across source

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Irritation test

Value:

ca. 2.7 - ca. 3.5

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

07-01-2014 to 26-05-2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

Version / remarks:

The BCOP test was conducted according to the following test guidelines:
- OECD Guideline for Testing of Chemicals No. 437, 26 July 2013 (“Bovine Corneal
Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye
Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye
Damage”)
- Commission Regulation (EU) No. 1152/2010 of 8 December 2010 amending for the
purpose of its adaptation to technical progress, Commission Regulation (EC) No
440/2008 of 30 May 2008 laying down test methods pursuant to Regulation (EC) No
1907/2006 of the European Parliament and of the Council on the Registration, Evaluation,
Authorisation and Restriction of Chemicals (REACH), Part B: Methods for the
determination of toxicity and other health effects: Bovine Corneal Opacity and
Permeability Test, Method for identifying ocular corrosives and severe irritants; Official
Journal of the European Union, No. L 324

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Name of test substance: N,N Dimethyldodecane-1-amide
Test-substance No.: 13/0555-1
Batch identification: 0009565072
CAS No.: 3007-53-2

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

Isolated bovine cornea:

The test system (target tissue) is the isolated bovine cornea.
Bovine eyes are obtained as a by-product of freshly
slaughtered cattle (age of the animals: minimum 12 months,
maximum 60 months).

Supplier:

Schlachthof Bensheim, Am Schlachthof 7-9, 64625 Bensheim,
Germany

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

Single topical application of 750 μL of the undiluted test substance to the
epithelial surface of isolated bovine corneas.

Duration of treatment / exposure:

Three corneas were treated with the test substance for 10 minutes followed by a 2-hours
post-incubation period.

Duration of post- treatment incubation (in vitro):

2 hours

Number of animals or in vitro replicates:

3 corneas per dosage tested

Details on study design:

Corneas free of defects (opacity, scratches, pigmentation etc.) were dissected with a 2 to 3 mm rim of sclera. Isolated corneas were mounted in cornea holders that consists of anterior and posterior chambers. Both chambers were filled to excess with pre-warmed Eagles’s MEM (without phenol red) and then equilibrated in a vertical position at about 32 °C for at least 1 hour. After the equilibration period the medium in both chambers was replaced with fresh prewarmed medium and initial corneal opacity readings were taken for each cornea with an opacitometer. Any corneas that showed macroscopic tissue damage or an opacity value < 530 opacity units1 were discarded. Corneas with opacity values close to the median value of all corneas were selected as negative control (NC). The remaining corneas were then distributed into treatment and positive control groups. Each corneal holder was uniquely identified with a number on the chambers.
Application of the test substance and washing:
Each treatment group (test substance, NC and PC) consisted of 3 corneas. Before application the medium in the anterior chamber was removed using a syringe. 750 μL of the undiluted liquid test substance was applied into the anterior chamber using a pipette. Control tissues were concurrently applied into the anterior chamber with 750 μL of de-ionized water (negative control, NC) or with 750 μL of 100% dimethylformamide (positive control, PC) using a pipette.
The corneas were incubated in a horizontal position at about 32 °C for approximately 10 minutes (liquids and surfactants). The test substance, NC and PC were then removed from the anterior chamber using a syringe and the epithelium was washed at least 3 times with Eagle’s MEM (containing phenol red) and once with Eagle’s MEM (without phenol red). Both chambers were then refilled with fresh Eagle’s MEM (without phenol red).
The corneas were incubated for further 2 hours at about 32 °C. After the incubation period the medium was removed and both chambers were then refilled with fresh Eagle’s MEM.
Measurement of final corneal opacity:
Before measurement, each cornea was observed visually and observations were recorded. Final corneal opacity readings were taken for each cornea with an opacitometer.
Determination of permeability:
For determination of permeability the medium in the anterior chamber was replaced by 1 mL sodium fluorescein solution (4 mg/mL for liquid test substances and surfactants) and incubated for 90 ± 5 min in a horizontal position at about 32 °C. The amount of sodium fluorescein that permeated through the corneas into the posterior chamber was measured spectrophotometrically. Three aliquots per cornea were transferred to a 96-well microtiter plate and the optical density (OD490) was determined.
Histopathology:
After determination of opacity and permeability, the corneas will be fixed in 4% formaldehyde for at least 24h and transferred to the laboratory of General Pathology for further histotechnical processing and examination by light microscopy.

Irritation parameter:

cornea opacity score

Run / experiment:

Opacity score of the test substance

Value:

ca. 11.1 - ca. 14.9

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Run / experiment:

Opacity score of the negative control

Value:

ca. 2.4 - ca. 6.6

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Run / experiment:

Opacity score of the positive control

Value:

ca. 105.4 - ca. 113.5

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Irritation parameter:

in vitro irritation score

Run / experiment:

In Vitro Irritancy score (IVIS) test substance

Value:

ca. 5.6 - ca. 10.2

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Irritation parameter:

in vitro irritation score

Run / experiment:

In Vitro Irritancy score (IVIS) negative control

Value:

ca. 0 - ca. 2.1

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Irritation parameter:

in vitro irritation score

Run / experiment:

In Vitro Irritancy score (IVIS) positive control

Value:

ca. 100.2 - ca. 100.8

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the observed results and applying the appropriate evaluation criteria it
was concluded, that N,N Dimethyldodecane-1-amide does not causes ocular corrosion
or severe irritation in the Bovine Corneal Opacity and Permeability Test (BCOP Test) under
the test conditions chosen.

Executive summary:

The potential of N,N Dimethyldodecane-1-amide to cause serious damage to the eyes was assessed by a single topical application of 750 μL of the undiluted test substance to the epithelial surface of isolated bovine corneas. Three corneas were treated with the test substance for 10 minutes followed by a 2-hours post-incubation period. In addition to the test substance a negative control (NC; de-ionized water) and a positive control (PC; 100% dimethylformamide) were applied to three corneas, each. Corneal opacity was measured quantitatively as the amount of light transmitted through the cornea. Permeability was measured quantitatively as the amount of sodium fluorescein dye that passes across the full thickness of the cornea. Both measurements were used to calculate an In Vitro Irritancy Score of the test substance. In addition H&E-stained cross sections were evaluated for the irritation potential of the test substance.

The results were:

Test substance	Mean Opacity Value	Mean Permeability Value	MeanIn Vitro Irritancy Score
N,N Dimethyldodecane-1-amide	8.0	0.809	20.2
Negative control	1.8	0.00	1.8
Positive control	102.3	2.125	135.2

Based on the observed results and applying the appropriate  evaluation criteria it was concluded, that N,N Dimethyldodecane-1-amide does not causes ocular corrosion or severe irritation in the Bovine Corneal Opacity and Permeability Test (BCOP Test) under the test conditions chosen.

Reference 2

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
The read-across hypothesis proposed is that the organism is not exposed to common compounds but rather, as a result of structural similarity, that different compounds have similar toxicological and fate properties. In this case the ECHA Read-Across Assessment Framework (RAAF) Scenario 2 is used.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Source: N,N-Dimethyldodecanamide [ EC 221-117-5; CAS 3007-53-2]
Target: Reaction Mass of N,N-Dimethyldodecanamide and N,N-Dimethyltetradecanamide [EC not assigned; CAS not assigned]

3. ANALOGUE APPROACH JUSTIFICATION
It may be concluded that both the registered substance (target) and the source molecules can be regarded as close structural analogues having both similar physical chemical and toxicological properties. It follows that where endpoints for the registered substance may not have experimental evidence, particularly those involving animal studies, that these can be addressed by read across grouping using an analogue approach. It is therefore proposed that, after careful assessment, experimental data from the source molecules may be used as surrogate evidence for read across to the registered substance. Please refer to attached document for information on the data available to support the read-across.

4. DATA MATRIX
Please refer to attached document

Reason / purpose for cross-reference:

read-across source

Irritation parameter:

cornea opacity score

Run / experiment:

Opacity score of the test substance

Value:

ca. 11.1 - ca. 14.9

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Run / experiment:

Opacity score of the negative control

Value:

ca. 2.4 - ca. 6.6

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Run / experiment:

Opacity score of the positive control

Value:

ca. 105.4 - ca. 113.5

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Irritation parameter:

in vitro irritation score

Run / experiment:

In vitro irritancy score of test item

Value:

ca. 5.6 - ca. 10.2

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Irritation parameter:

in vitro irritation score

Run / experiment:

In vitro irritancy score of negative control

Value:

ca. 0 - ca. 2.1

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Irritation parameter:

in vitro irritation score

Run / experiment:

In vitro irritancy score of the positive control

Value:

ca. 100.2 - ca. 100.8

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Reference 3

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

27-11-2013 to 26-05-2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Qualifier:

no guideline followed

Principles of method if other than guideline:

There are no official national or international guidelines for the EpiOcularTM test yet; however,
the study was performed according to the methods described in the following publications:
- MatTek Corporation, Ashland, MA 01721, USA: EpiOcularTM human cell construct:
Procedure details, Version 3.1a of February 10, 2010.
- Harbell J.W. et al. (2009): COLIPA Program on Optimization of Existing In Vitro Eye
Irritation Assays for Entry into Formal Validation: Technology Transfer and Intra/Inter
Laboratory Evaluation of EpiOcular Assay for Chemicals. Poster # 378, Society of
Toxicology, March 2009.
In addition the study follows the testing strategy for determination of eye irritation/corrosion
as given in the following OECD guideline:
- OECD Guideline for Testing of Chemicals No. 405, October 2, 2012 (“Acute Eye
Irritation/Corrosion”)

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Name of test substance: N,N Dimethyldodecane-1-amide
Test-substance No.: 13/0555-1
Batch identification: 0009565072
CAS No.: 3007-53-2

Species:

human

Strain:

not specified

Details on test animals or tissues and environmental conditions:

The EpiOcularTM model (OCL-200) is a three-dimensional non-keratinized tissue construct
composed of normal human derived epidermal keratinozytes used to model the human
corneal epithelium.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

other: MTT reduction control (KC): De-ionized water, sterile or test substance

Amount / concentration applied:

Undiluted test material

Number of animals or in vitro replicates:

2 tissue samples per sample tested were used.

Irritation parameter:

other: mean % viability of test substance treated tissues

Value:

ca. 28.6 - ca. 30.5

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Interpretation of results:

study cannot be used for classification

Conclusions:

Based on the observed results and applying the appropriate evaluation criteria cited was concluded, that N,N Dimethyldodecane-1-amide shows an eye irritation potential in the EpiOcular™ eye irritation test under the test conditions chosen.

Executive summary:

The potential of N,N Dimethyldodecane-1-amide to cause ocular irritation was assessed by a single topical application of 50μL of the undiluted test substance to a reconstructed three dimensional human cornea model (EpiOcular™). Two EpiOcular™ tissue samples were incubated with the test substance for 30 minutes followed by a 2-hours post-incubation period.

Tissue destruction was determined by measuring the metabolic activity of the tissue after exposure/post-incubation using a colorimetric test. The reduction of mitochondrial dehydrogenase activity, measured by reduced formazan production after incubation with a tetrazolium salt (MTT) was chosen as endpoint. The formazan production of the test substance treated epidermal tissues is compared to that of negative control tissues. The quotient of the values indicates the relative tissue viability.

The EpiOcular™ eye irritation test showed the following results:

The test substance is able to reduce MTT directly. However, this ability of direct MTT reduction did not impair the study result as demonstrated by the concurrently performed exposure of control tissues inactivated by freezing. The mean viability of the test-substance treated tissues was 30%.

Based on the observed results and applying the evaluation criteria cited in chapter 3.8 it was concluded, that N,N Dimethyldodecane-1-amide shows an eye irritation potential in the EpiOcular™ eye irritation test under the test conditions chosen. The test method does not yet allow for the evaluation of serious eye damage. The result does not exclude a serious eye irritation potential of the test substance.

Reference 4

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

supporting study

Justification for type of information:

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
The read-across hypothesis proposed is that the organism is not exposed to common compounds but rather, as a result of structural similarity, that different compounds have similar toxicological and fate properties. In this case the ECHA Read-Across Assessment Framework (RAAF) Scenario 2 is used.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Source: N,N-Dimethyldodecanamide [ EC 221-117-5; CAS 3007-53-2]
Target: Reaction Mass of N,N-Dimethyldodecanamide and N,N-Dimethyltetradecanamide [EC not assigned; CAS not assigned]

3. ANALOGUE APPROACH JUSTIFICATION
It may be concluded that both the registered substance (target) and the source molecules can be regarded as close structural analogues having both similar physical chemical and toxicological properties. It follows that where endpoints for the registered substance may not have experimental evidence, particularly those involving animal studies, that these can be addressed by read across grouping using an analogue approach. It is therefore proposed that, after careful assessment, experimental data from the source molecules may be used as surrogate evidence for read across to the registered substance. Please refer to attached document for information on the data available to support the read-across.

4. DATA MATRIX
Please refer to attached document

Reason / purpose for cross-reference:

read-across source

Irritation parameter:

other: mean % viability of test substance treated tissues

Value:

ca. 28.6 - ca. 30.5

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Reference 5

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1997

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted according to OECD guideline, GLP, well documented

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

other: Mol:Russian

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Mollegaard Breeding and Research Centre A/S
- Weight: 2.4-2.6kg
- Housing: animal room 2, filtered Air, ppo cages
- Diet (e.g. ad libitum): ad libitum / Altromin 2123 , Altromin , Lage , Germany
- Water (e.g. ad libitum): ad libitum (acified, HCl)
- Acclimation period: at least one week


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20±3°C
- Humidity (%): 55±15%
- Air changes (per hr): 10times/h
- Photoperiod (hrs dark / hrs light):12h each

Vehicle:

unchanged (no vehicle)

Controls:

other: right eye served as control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1ml

Observation period (in vivo):

1h, 24h, 48h, 72h, 7d, 14d, 21d

Number of animals or in vitro replicates:

3

Irritation parameter:

cornea opacity score

Remarks:

corne opacity

Basis:

mean

Remarks:

three animals

Time point:

24/48/72 h

Score:

<= 1

Max. score:

4

Reversibility:

fully reversible within: 21 days in 2 of 3 animals

Remarks on result:

other: When Fluorescein has been applied only the highest score for degree of corneal opacity from three readings are included

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

other: normal reaction observed

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

other: normal reaction observed

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

other: normal reaction observed

Irritation parameter:

conjunctivae score

Remarks:

redness of conjunctiva

Basis:

mean

Remarks:

three animals

Time point:

24/48/72 h

Score:

<= 2.6

Max. score:

3

Reversibility:

fully reversible within: 21 days in 2 of 3 animals

Irritation parameter:

chemosis score

Remarks:

oedema of conjunctiva (chemosis)

Basis:

mean

Remarks:

three animals

Time point:

24/48/72 h

Score:

<= 2.8

Max. score:

4

Reversibility:

fully reversible within: 21 days in 2 of 3 animals

Interpretation of results:

other: Category 2

Remarks:

Criteria used for interpretation of results: other: EU GHS EC 1272/2008

Conclusions:

Eye irritant potential

Executive summary:

The eye irritant effect of SAT 970 419 (N,N-Dimethyldecan-1-amide) was investigated according to the method recommended in the OECD Guideline No. 405, "Acute Eye Irritation/Corrosion", Feb. 1987, and EEC Guideline B.5 "Acute Toxicity (Eye Irritation)", 29.12.1992.

Three female albino rabbits were exposed to 0.1 ml of the test article in the left eye. The eyes were examined and the changes were graded according to a numerical scale 1, 24, 48 and 72 hours as well as 7, 14 and in two animals 21 days after dosing.

Moderate to severe signs of eye irritation were observed among the rabbits.

The following mean values, based on the results from the 24, 48 and 72 hour readings, were calculated:

Cornea opacity 1.0

Iris lesion 0.0

Redness of conjunctiva 2.6

Oedema of conjunctiva 2.8

After 14 days all animals showed hairless areas around the eye. Treatment related adverse eye reactions were fully reverse within 14 (1 animal) and 21 days.

Based on the result of this study the test substance is classified as eye irritant

Reference 6

Endpoint:

eye irritation: in vivo

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
The read-across hypothesis proposed is that the organism is not exposed to common compounds but rather, as a result of structural similarity, that different compounds have similar toxicological and fate properties. In this case the ECHA Read-Across Assessment Framework (RAAF) Scenario 2 is used.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Source: N,N-Dimethyldecanamide [ EC 238-405-1; CAS 14433-76-2]
Target: Reaction Mass of N,N-Dimethyldodecanamide and N,N-Dimethyltetradecanamide [EC not assigned; CAS not assigned]

3. ANALOGUE APPROACH JUSTIFICATION
It may be concluded that both the registered substance (target) and the source molecules can be regarded as close structural analogues having both similar physical chemical and toxicological properties. It follows that where endpoints for the registered substance may not have experimental evidence, particularly those involving animal studies, that these can be addressed by read across grouping using an analogue approach. It is therefore proposed that, after careful assessment, experimental data from the source molecules may be used as surrogate evidence for read across to the registered substance. Please refer to attached document for information on the data available to support the read-across.

4. DATA MATRIX
Please refer to attached document

Reason / purpose for cross-reference:

read-across source

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

three animals

Time point:

24/48/72 h

Score:

<= 1

Max. score:

4

Reversibility:

fully reversible within: 21 days in 2 of 3 animals

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

other: normal reaction observed

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

other: normal reaction observed

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

other: normal reaction observed

Irritation parameter:

conjunctivae score

Basis:

mean

Remarks:

three animals

Time point:

24/48/72 h

Score:

<= 2.6

Max. score:

3

Reversibility:

fully reversible within: 21 days in 2 out of 3 animals

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

three animals

Time point:

24/48/72 h

Score:

<= 2.8

Max. score:

4

Reversibility:

fully reversible within: 21 days in 2 out of 3 animals

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Valid experimental analogue data are available for the assessment of skin and eye irritation. These data are read across to the registered substance.

Skin:

The acute skin irritant effect of N,N-Dimethyldecan-1-amide was investigated according to OECD TG 404 and EEC Guideline B.4 (Cognis 1997, M.Hoyer). Three female albino rabbits were exposed to the test article at two skin sites on the back. After 4hours of exposure the test article was removed and the skin was examined 1, 24, 48 and 72 hours as well as 7, 14 and 21 days after termination of exposure.Moderate to severe skin reactions were observed. Under the experimental conditions described in this report, the mean score for erythema was 2.7 (2.00, 3.00, 3.00) and for oedema 2.8 (2.00, 3.33, 3,00). After 21 days rests of scales on both sides were observed in all animals.

This study is supported by in vitro corrosivity and irritation tests performed using N,N-dimethyldodecan-1-amide according to OECD TG 431 and OECD TG 439. These studies concluded that the test item was not corrosive to skin, but was a skin irritant.

Eye:

The acute eye irritant effect of N,N-Dimethyldecan-1-amide was investigated according to the method recommended in the OECD TG 405 and EEC Guideline B.5 (Cognis 1997, M. Hoyer).Three female albino rabbits were exposed to 0.1 ml of the test article in the left eye. The eyes were examined and the changes were graded according to a numerical scale 1, 24, 48 and 72 hours as well as 7, 14 and in two animals 21 days after dosing.Moderate to severe signs of eye irritation were observed among the rabbits. The following mean values, based on the results from the 24, 48 and 72 hour readings, were calculated: Cornea opacity 1.0; Iris lesion 0.0; Redness of conjunctiva 2.6; Oedema of conjunctiva 2.8.After 14 days all animals showed hairless areas around the eye. Treatment related adverse eye reactions were fully reverse within 14 (1 animal) and 21 days.

This study is supported by in vitro eye irritation tests performed using N,N-dimethyldodecan-1-amide according to OECD TG 437 and the EpiOcular test (no guideline available at the time of the test). Taking both these studies together, it was concluded that the test item should be classified as an eye irritant.

Justification for classification or non-classification

Skin:

Clear skin irritating effects were observed in an in vivo skin irritation study performed using the C10 analogue: The scores obtained from the study led to a classification as an irritant to skin (cat. 2) due to criteria of GHS (Regulation (EU) 1272/2008). Supported by in vitro testing using the C12 analogue.

Eye:

Clear eye irritating effect were observed in an in vivo eye irritation study: The scores obtained in this study led to a classification as an irritating to eye (cat. 2) due to criteria of GHS (Regulation (EU) 1272/2008). Supported by in vitro testing using the C12 analogue.

It may be predicted from this that the registered substance would share a similar toxicity profile and would also be predicted to be an irritant.