Dodecyl nonan-1-oate

Endpoint summary

Description of key information

Skin irritation (similar to OECD 404, read across): not irritating

Eye irritation (OECD 492): not irritating

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:

11 - 21 Mar 1991

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

The exposure was performed under occlusive conditions, the analytical purity of the test substance was not specified.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

occlusive exposure

Principles of method if other than guideline:

The testing was performed equivalent to OECD guideline 404.

GLP compliance:

not specified

Species:

rabbit

Strain:

New Zealand White

Type of coverage:

occlusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Controls:

other: the untreated skin site of the same animal served as the control

Amount / concentration applied:

0.5 mL

Duration of treatment / exposure:

4 h

Observation period:

72 h
Reading time points: 1, 24, 48 and 72 h

Number of animals:

3 males

Details on study design:

TEST SITE
- Type of wrap if used: occlusive

REMOVAL OF TEST SUBSTANCE
- Washing (if done): the treated site was not washed after patch removal

SCORING SYSTEM: Draize scoring system

Irritation parameter:

erythema score

Basis:

animal #1

Remarks:

mean over

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Irritation parameter:

erythema score

Basis:

animal #2

Remarks:

mean over

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility not applicable

Irritation parameter:

erythema score

Basis:

animal #3

Remarks:

mean over

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility not applicable

Irritation parameter:

edema score

Basis:

animal #1

Remarks:

mean over

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h

Irritation parameter:

edema score

Basis:

animal #2

Remarks:

mean over

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility not applicable

Irritation parameter:

edema score

Basis:

animal #3

Remarks:

mean over

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility not applicable

Irritant / corrosive response data:

1 hour after exposure, all the animals had slight to well-defined erythema, while 2/3 had very slight to slight edema. These effects gradually reduced in severity. 24 hours after exposure ended, 1/3 animals had very slight erythema and slight edema. the edema persisted until the 48-hour reading time point.All skin irritation effects had cleared completely within 72 hours after the exposure ended.

Table 1: Erythema and edema scores for skin irritation of the individual animals at different reading time points:

Observation time	Rabbit no.
	1		2		3
	Erythema	Edema	Erythema	Edema	Erythema	Edema
1 h	2	2	1	0	1	1
24 h	1	2	0	0	0	0
48 h	0	1	0	0	0	0
72 h	0	0	0	0	0	0
Mean value 24 + 48 + 72 h	0.33	1.00	0	0	0	0
Mean value 24 + 48 + 72 h all animals	0.11	0.33

Interpretation of results:

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

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

09 May 1989 - 17 Jul 1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

(analytical purity of test substance not specified, occlusive exposition).

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

adopted in 1981

Deviations:

yes

Remarks:

occlusive exposition, analytical purity of test material not specified

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

adopted in 2015

Deviations:

yes

Remarks:

no rationale for in vivo testing

GLP compliance:

yes

Species:

rabbit

Strain:

other: Kleinrussen, Chbb:HM

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Thomae, Germany
- Age at study initiation: 8-12 months
- Weight at study initiation: approx. 2620 g
- Diet: ad libitum (Altromin-Haltungsdiat 2023)
- Water: ad libitum
- Acclimation period: at least 1 week


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-25
- Humidity (%): 45-70
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

occlusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Controls:

other: untreated site of the same animal served as control

Amount / concentration applied:

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

Duration of treatment / exposure:

4 h

Observation period:

Reading time points: 1, 24, 48, 72h and 7 days
7 days

Number of animals:

4

Details on study design:

TEST SITE
- Area of exposure: 6 cm²
- Type of wrap if used: inert plastic foil fixed with an adhesive patch (Leukosilk) and a special gauze bandage (Acrylastic)


REMOVAL OF TEST SUBSTANCE
- Washing (if done): No data


SCORING SYSTEM: Draize scoring system

Irritation parameter:

erythema score

Basis:

animal #1

Remarks:

mean over

Time point:

24/48/72 h

Score:

1.33

Max. score:

4

Reversibility:

fully reversible within: 7 d

Irritation parameter:

erythema score

Basis:

animal #2

Remarks:

mean over

Time point:

24/48/72 h

Score:

0.66

Max. score:

4

Reversibility:

fully reversible within: 7 d

Irritation parameter:

erythema score

Basis:

animal #3

Remarks:

mean over

Time point:

24/48/72 h

Score:

0.66

Max. score:

4

Reversibility:

fully reversible within: 7 d

Irritation parameter:

erythema score

Basis:

animal #4

Remarks:

mean over

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 7 d

Irritation parameter:

edema score

Basis:

animal #1

Remarks:

mean over

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Irritation parameter:

edema score

Basis:

animal #2

Remarks:

mean over

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Irritation parameter:

edema score

Basis:

animal #3

Remarks:

mean over

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Irritation parameter:

edema score

Basis:

animal #4

Remarks:

mean over

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Irritant / corrosive response data:

The test substance, when administered to rabbit skin for 24h, caused only slight irritations that were fully reversible within 7 days.

Erythema Score

Animal Number	1 h	24 h	48 h	72 h	7 d
102	1	1	1	2	0
128	1	1	0	1	0
129	1	1	0	1	0
130	1	0	0	1	0

 

Edema Score

Animal Number	1 h	24 h	48 h	72 h	7 d
102	1	1	0	0	0
128	0	0	0	0	0
129	2	0	0	0	0
130	0	0	0	0	0

Interpretation of results:

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

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

17 July - 30 Jan 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)

Version / remarks:

9 October 2017

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Hess. Ministerium für Umwelt, Klimaschutz, Landwirtschaft und Verbraucherschutz, Wiesbaden, Germany

Species:

human

Strain:

other: EpiOcularTM (OCL-200-EIT)

Details on test animals or tissues and environmental conditions:

- Justification of the test method and considerations regarding applicability
The EpiOcularTM Eye Irritation Test (EIT) predicts the acute ocular irritation potential of chemicals by measurement of its irreversible tissue damage caused by cytotoxic effects in the human cornea model. The EpiOcular ™ Eye Irritation Test (EIT) was validated by the European Union Reference laboratory for Alternatives to Animal Testing (EURL ECVAM) and cosmetics Europe between 2008 and 2013. It is utilized for the classification and labelling of chemicals concerning their eye irritation potential. The EpiOcularTM EIT is intended to differentiate substances that require “no classification" as eye irritant from those that require labelling as either GHS category 1 or 2 for serious eye damage respective eye irritation potential.

- Description of the cell system used, incl. certificate of authenticity and the mycoplasma status of the cell live
The EpiOcularTM tissue consists of normal, human-derived keratinocytes which have been cultured to form a stratified squamous epithelium similar to that found in the human cornea. It consists of highly organized basal cells, which progressively flatten out as the apical surface of the tissue is approached, analogous to the normal in vivo corneal epithelium. The EpiOcularTM tissues (surface 0.6 cm²) were cultured on specially prepared cell culture inserts (MILLICELL®, 10 mm Ø). Analysis for tissue functionality and for potential contaminants was passed.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 50 µL
- Concentration: undiluted

Duration of treatment / exposure:

30 min

Duration of post- treatment incubation (in vitro):

120 min

Number of animals or in vitro replicates:

2

Details on study design:

- Details of the test procedure used :
EpiOcularTM tissue model was used. After the assessment and exclusion of direct MTT reduction and colouring potential of the test substance, the well plates were prepared and the tissues were pre-incubated in warm medium under standard culture conditions (37 ± 1.5 °C, 5 ± 0.5 % CO2 and 95 % relative humidity). After overnight incubation, the tissues were pre-wetted with 20 μL DPBS buffer (Ca2+ Mg2+ free) and then incubated at standard culture conditions for 30 min. 50 μL of the controls and the neat test substance were then applied and incubated for 30 min at standard culture conditions. After exposure the tissue constructs were thoroughly rinsed and transferred to fresh medium for an immersion incubation at room temperature. Afterwards, the tissues were transferred to fresh medium and incubated for 120 min at standard culture conditions, prior to determination of the cytotoxic (irritancy) effect. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) at the end of the treatment.

- RhCE tissue construct used, including batch number :
Keratinocyte strain: 4F1188, Lot Number: 27002

- Doses of test chemical and control substances used :
50 μL of undiluted test substance, 50 μL of demineralised water (negative control), 50 μL of methyl acetate (positive control)

- Duration and temperature of exposure, post-exposure immersion and post-exposure incubation periods:
30 min at 37.0 ± 1.5 °C, 12 min immersion incubation at room temperature, 120 min at 37 ± 1.5 °C post-exposure incubation

- Number of tissue replicates used per test chemical and controls (positive and negative control) : 2

- Wavelength used for quantifying MTT formazan : 570 nm

- Description of the method used to quantify MTT formazan : The amount of extracted formazan was determined spectrophotometrically at 570 nm in duplicate with a plate reader (Versamax® Molecular Devices, Ismaning, Germany). Then the mean OD of the blank isopropanol (OD Blk) was calculated, followed by the subtraction of mean OD Blk of each value of the same experiment (corrected values).
The mean OD of the two replicates for each tissue the mean OD of the two relating tissues for controls and test substance were then calculated.

To calculate the relative absorbance, the following equation was used: viability (%) = (OD corrected of test substance or positive control/ OD corrected of mean negative control)*100

- Description of evaluation criteria used including the justification for the selection of the cut-off point for the prediction model :

The in vitro eye irritation test is considered valid if it meets the following criteria:
a) mean OD570 of the negative control: > 0.8 and < 2.5
b) mean relative tissue viability of the positive control: < 50% relative to the negative control.
c) variation within replicates: <20%

If the test item-treated tissue viability is > 60% relative to the negative control treated tissue viability, the test item is labeled non-irritant (no Category according to UN GHS).
If the test item-treated tissue viability is ≤ 60% relative to negative control treated tissue viability, the test item is labeled irritant (Category 2 or Category 1 according to UN GHS; no differentiation between the categories possible).

- Reference to historical positive and negative control results demonstrating suitable run acceptance criteria : negative contrtol range: 1.27 - 2.16; positive control range: 6.9 - 43.4%

- Complete supporting information for the specific RhCE tissue construct used :
Tissue viability: 1.269 ± 0.039 (accepted range: 1.1 - 3.0)
Barrier function: ET-50: 24.36 min (accepted range: 12.2 - 37.5)

Irritation parameter:

other: mean viability (%)

Run / experiment:

mean value of test substance

Value:

95

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Irritation parameter:

other: mean viability (%)

Run / experiment:

mean value of positive control

Value:

36.5

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:

DEMONSTRATION OF TECHNICAL PROFICIENCY: yes, but not in this report

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes, mean OD = 1.691 (> 0.8 and < 2.5)
- Acceptance criteria met for positive control: yes, % viability = 36.5% (< 50%)
- Range of historical values if different from the ones specified in the test guideline: 0.6 - 2.1% (< 20%)

Table 1: Results of MTT assay

	Tissue No.	OD570	Mean (OD570)	Mean (OD570) - blank	Mean tissue viability (% of negative control)
Negative control	1	1.674	1.714	1.676	99.1	100
		1.753
	2	1.735	1.744	1.706	100.9
		1.753
Positive control	1	0.626	0.660	0.622	36.8	36.5
		0.693
	2	0.647	0.649	0.611	36.2
		0.652
Test substance	1	1.614	1.626	1.588	93.9	95.0
		1.638
	2	1.676	1.662	1.624	96.1
		1.649

 OD = optical density

Interpretation of results:

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

Conclusions:

CLP: not classified

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Read across justification

There are no available data on the skin irritation effects of Lauryl nonanoate (CAS 17671-26-0). The assessment was therefore based on studies conducted with analogue substances as part of a read across approach, which is in accordance with Regulation (EC) No. 1907/2006, Annex XI, 1.5. For each specific endpoint the source substance(s) structurally closest to the target substance is/are chosen for read-across, with due regard to the requirements of adequacy and reliability of the available data. Structural similarities and similarities in properties and/or activities of the source and target substance are the basis of read-across. A detailed justification for the analogue read-across approach is provided in the technical dossier (see IUCLID Section 13).

Skin irritation 

CAS 95912-86-0

A skin irritation study similar to OECD guideline 404 (key study, 1991) was performed with Fatty acids, C8-10, C12-18-alkyl esters (CAS 95912-86-0). 0.5 mL of the undiluted test substance was applied to the shaved skin of 3 male rabbits for 4 hours of exposure under occlusive conditions. The local skin effects were scored 1, 24, 48, and 72 h after exposure ended. At the 24-hour reading time point 1/3 rabbits showed erythema (score 1), which was fully reversible within 48 hours. Oedema was seen in 1/3 rabbits with score 2 at the 24-hour reading and score 1 at the 48 hour reading, which was fully reversible within 72 hours.

The mean erythema scores (over 24, 48 and 72 hours) were 0.33, 0.0 and 0.0 while the mean oedema scores (over 24, 48 and 72 hours) were 1.0, 0.0 and 0.0. The test substance is not considered to be irritating to the skin.

CAS 135800-37-2

A supporting skin irritation study similar to OECD guideline 404 (supporting study, 1990) was performed with Fatty acids, C8-16, 2- ethylhexyl esters (CAS 135800-37-2). The test substance was applied to the shaved skin of 4 rabbits under occlusive conditions for 4 hours of exposure. The local skin effects were scored 1, 24, 48 and 72 h, and 7 days after exposure ended. At the 24-hour reading time point 3/4 rabbits showed slight erythema (score 1), which persisted as slight erythema for 48 hours in 1/4 animals. At the 72-hour reading time point, slight to well-defined erythema was observed in 4/4 rabbits. The erythema had cleared completely within 7 days after the exposure ended. Oedema was observed in 1/4 animals 1 and 24 h after patch removal; the effect had cleared completely within 48 hours. The mean erythema scores (over 24, 48 and 72 hours) were 0.75, 0.25, 1.25, while the mean edema scores (over 24, 48 and 72 hours) were 0.25, 0, 0 and 0. The test substance is not considered to be irritating to the skin.

Eye irritation

CAS 17671-26-0

The eye irritation properties of Lauryl nonanoate were tested in a study according to OECD 492 and in compliance with GLP (key study, 2018). The EpiOcular™ cornea epithelial model was exposed to 50 μL of the neat/ undiluted test substance for 30 minutes and was incubated for another 2 hours after rinsing and removal of the test substance and 12 minutes of post-soak immersion.After treatment with the negative control the absorbance values were well within the required acceptability criterion of OD > 0.8 and < 2.5 thus showing the quality of the tissues. Treatment with the positive control induced a decrease below 50% compared with the negative control value in the relative absorbance thus ensuring the validity of the test system.The viability test (MTT) with the test substance revealed a mean tissue viability of 95% compared with the value of the negative control. Based on the obtained results, Lauryl nonanoate (CAS 17671-26-0) is not eye irritating.

 

Overall conclusion for skin and eye irritation

The available data on suitable source substances conclude that the source substances are not skin irritating. Therefore, the target substance Lauryl nonanoate is also not expected to be a skin irritant. No eye irritating potential was found for the target substance and is thus considered to be not an eye irritant.