Registration Dossier

Diss Factsheets

Administrative data

Description of key information

In an in vitro skin irritation study (OECD 439) test item was irritant to skin (GLP, Rel. K1)

In an in vitro eye irritation study (OECD 492) test item was considered non irritant to the eyes (GLP, Rel K1)

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:
01-16 March 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
GLP study conducted in compliance with OECD Guideline No. 439 without any deviation.
Qualifier:
according to guideline
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Version / remarks:
adopted 28 July 2015
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Version / remarks:
dated 23 July 2009
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes (incl. QA statement)
Remarks:
23 October 2015
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Robertet / 2563305
- Appearance: Yellow to orange liquid
- Manufacturing date: 07 September 2016
- Date received: 27 January 2017
- Expiration date of the lot/batch: 07 March 2018
- Purity test date: 03 April 2017

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature, darkness
Test system:
human skin model
Source species:
human
Cell type:
other: living human skin models and killed reconstructed human epidermis
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- The 0.50 cm2 reconstructed epidermises (Episkin SA, RHE/S/17 Batch No. 17 -RHE-031) were received on 14 March 2017. The same day, the insert (filter + epidermis) was gently removed from the agarose while avoiding leaving agarose on the polycarbonate filter. The inserts were placed in 6 wells culture plate which had been previously filled with 300 µL of growth medium (Episkin SA, batch No. 17 MPE 021) during 2 hours and 30 minutes. Then just before treatment, the inserts were placed in 24 wells culture plate which had been previously filled with 300 µL of maintenance medium (Episkin SA, batch No. 17 MA 017).
The two killed Reconstructed Human epidermis control tissue models, supplied by Episkin (Batch No. 17-RHE-023) frozen on 28 February 2017 were defrost to be used on 16 March 2017.

TREATMENT
- The test item was applied, as supplied, at the dose of 16 µL, to the epidermal surface of 3 living human skin models and 2 killed Reconstructed Human epidermis (for non-specific MTT reduction control) during 42 minutes at room temperature. To ensure a good contact with the epidermises, during all the treatment period, the test item was recovered with a nylon mesh provided by Episkin SA.

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: Test item was applied for 42 minutes at room temperature
- Temperature of post-treatment incubation: 41 hours and 15 minutes post-treatment incubation period in fresh medium at 37 °C, 5% CO2

REMOVAL OF TEST MATERIAL AND CONTROLS
42 minutes after the test item application, the nylon mesh was removed and the human epidermises were washed with 25 x 1 mL of DPBS. The rinsed tissues were checked for any coloration and noted to be slightly yellow instead of being whitish as for the coloration of the negative control tissues. They were incubated for a 41 hours and 15 minutes post-treatment incubation period in fresh medium at 37°C, 5% CO2. Then, the epidermises were put in contact with the MTT solution.

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- The cell viability was quantified by measurement of the cell succinate dehydrogenase activity. This enzyme is responsible for the MTT reduction into blue formazan crystal that is quantitatively measured by Optical Density (OD) after extraction from tissues. The measured OD are proportional to the number of living cells.
- The skin samples (included the 2 killed Reconstructed Human epidermis) were placed in 300 μL of a MTT solution at 1.0 mg/mL for 3 hours at 37 °C, 5% CO2. The precipitated blue formazan product
was then extracted using isopropanol during 1 hour and 57 minutes under gentle agitation in the dark, and the concentration of formazan was determined by measuring the OD at 570 nm, just after dilution of the extracts 1:2 in isopropanol.
- The OD was measured in triplicate of MTT extract. The measurement of OD at 570 nm of formazan extracts was performed in triplicate samples using the ELx800 absorbance microplate reader (controlled every year and calibrated if necessary) supplied by BioTek and the validated software Gen5 ELISA Vl.05.11 supplied by BioTek. The linearity range of optical density measured is validated for an optical density between 0 and 2.0.

VIABILITY:
Viability % = (mean OD test item / mean OD negative control) x100
As the test item was a direct MTT-reducer, true tissue viability was calculated as follows:
True Viability % = [(mean OD test item – mean OD non-specific MTT killed control)/ mean OD negative control] x 100
For each tissue, OD values and calculated percentage cell viability data for the test item, positive and negative controls, were reported in tabular form, including data from replicate repeat experiments as appropriate, mean and individual values.

NUMBER OF REPLICATE TISSUES:
3 living human skin models and 2 killed Reconstructed Human epidermis (for non-specific MTT reduction control)

PREDICTION MODEL / DECISION CRITERIA
- The OD values obtained for each test sample were used to calculate a percentage of viability relative to the negative control, which was arbitrarily set at 100%. The cut-off values for the prediction of irritation associated with the RHE models were as follows:
The test item is considered as non-irritant to skin: if the mean percent viability after 42 minutes exposure and 42 hours of post-treatment incubation is > 50%.
The test item is considered to be irritant to skin: if the mean percent tissue viability after 42 minutes exposure and 42 hours of post-treatment incubation is ≤ 50% and the result of a skin corrosion test is “non-corrosive”.
The test item is considered to be irritant or corrosive to skin: if the mean percent tissue viability after 42 minutes exposure and 42 hours of post-treatment incubation is ≤ 50% and in absence of information on a skin corrosion test.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 16 μL
- Concentration (if solution): Undiluted
Duration of treatment / exposure:
42 minutes at room temperature
Duration of post-treatment incubation (if applicable):
41 hours and 15 minutes post-incubation period at 37 °C, 5% CO2
Number of replicates:
3 living human skin models and 2 killed Reconstructed Human epidermis (for non-specific MTT reduction control)
Irritation / corrosion parameter:
% tissue viability
Remarks:
mean
Run / experiment:
Main test (42 minutes)
Value:
-1.2
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Remarks:
mean corrected percent viability of the treated tissues was -1.2% (considered as 0%)
Other effects / acceptance of results:
VIABILITY
- The mean corrected percent viability of the treated tissues was -1.2% (considered as 0%), versus 0.9% in the positive control (5% Sodium Dodecyl Sulfate).

ACCEPTANCE OF RESULTS:
- The mean percent tissue viabilities obtained with the negative control and positive controls are within the range of historical data and therefore validate the experiment.

Table 7.3.1/1: Assessment of the skin irritation - individual and average values of OD after 42 minutes exposure

 

Items

Skin

OD

Mean OD/disc#

Mean OD/product

Viability%

Mean viability%

SD

Conclusion

Negative control

1

1.195

1.272

1.090

116.7

100.0

16.4

-

1.294

1.327

2

1.070

1.084

99.4

1.095

1.088

3

0.936

0.915

83.9

0.933

0.878

Positive control

4

0.008

0.008

0.010

0.7

0.9

0.2

Irritant

0.009

0.008

5

0.010

0.010

0.9

0.010

0.011

6

0.012

0.012

1.1

0.012

0.013

Test item

7

0.018

0.019

0.018

1.7

1.7

0.1

-

0.020

0.020

8

0.019

0.018

1.7

0.019

0.018

9

0.017

0.017

1.6

0.017

0.017

Test item

Killed tissues

10

0.031

0.031

0.031

2.8

2.8

0.0

0.032

0.032

11

0.031

0.031

2.8

0.030

0.032

Test item

corrected

-

-1.2

-

Corrosive or irritant

 

≠: mean of 3 values (triplicate of the same extract)

OD: optical density

 

Acceptability criteria:

• SD≤18%

• Negative control: OD value of the 3 replicates in the range 0.8 and ≤ 3.0.

The optical density was measured after a 1:2 dilution of the formazan extracts in isopropanol: the acceptability criteria should be in the range 0.4 and ≤1.5 for the negative control.

Interpretation of results:
other: Category 2 “Irritant” or Category 1 “Corrosive” based on GHS criteria
Conclusions:
Under these experimental conditions and in accordance with the Regulation EC No.1272/2008 and in absence of information on a skin corrosion, the test item has to be classified in Category 2 “Irritant” or in Category 1 “Corrosive”.
Executive summary:

An in vitro skin irritation study was performed according to OECD Guideline 439 and in compliance with GLP to evaluate the possible irritating effects of the test item Tagete essential oil Oil after topical application on in vitro human reconstructed epidermis (SkinEthic RHE®model).

 

The test item was applied as supplied, at the dose of 16 µL, to 3 living and 2 killed Reconstructed Human epidermis (SkinEthic RHE® model) during 42 minutes, followed by a rinse with 25 mL of DPBS and a 41 hours and 15 minutes post-incubation period at 37 °C, 5% CO2. Cell viability was then measured by enzymatic conversion of the vital dye MTT into a blue formazan salt that was quantitatively measured after extraction from tissues.

 

The mean percent tissue viabilities obtained with the negative control and positive controls are within the range of historical data and therefore validate the experiment.

 

The mean corrected percent viability of the treated tissues was -1.2% (considered as 0%), versus 0.9% in the positive control (5% Sodium Dodecyl Sulfate).

 

Under these experimental conditions and in accordance with the Regulation EC No.1272/2008 and in absence of information on a skin corrosion, the test item has to be classified in Category 2 “Irritant” or in Category 1 “Corrosive”.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (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:
01 - 09 March 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
GLP study performed according to OECD Guideline 492 without any deviation.
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:
adopted 28 July 2015
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes (incl. QA statement)
Remarks:
27 April 2017
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Robertet / 2563305
- Appearance: Yellow to orange liquid
- Manufacturing date: 07 September 2016
- Date received: 27 January 2017
- Expiration date of the lot/batch: 07 March 2018
- Purity test date: 21 December 2016

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature, darkness
Species:
other: Reconstructed human Cornea-like Epithelium
Details on test animals or tissues and environmental conditions:
- Description of the cell system used:
Reconstructed human Cornea-like Epithelium (RhCE) tissue constructs: The 0.60 cm² Reconstructed human Cornea-like Epithelia (EpiOcularTM OCL-212-ver2.0, supplied by MatTek Corporation, batch No. 23770) were received on 08 March 2017.
The killed Reconstructed human Cornea-like Epithelia (EpiOcularTM OCL-212-ver2.0, supplied by MatTek Corporation, batch No. 23706) were defrost on 03 March 2017, re-freeze on 03 March 2017 and defrost to be used on 08 March 2017.
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 50 µL
- Concentration (if solution): Undiluted
Duration of treatment / exposure:
30 minutes at 37°C, 5% CO2, 95% humidity (standard culture conditions).
Duration of post- treatment incubation (in vitro):
After 30 minutes of exposure, extensive rinsing with DPBS at room temperature, a 12 minutes post-exposure immersion period at room temperature and a 2 hours and 02 minutes post-exposure incubation at standard culture conditions.
Number of animals or in vitro replicates:
Test item, negative and positive controls were applied on duplicate tissues (2 living RhCE tissue replicates and 2 killed DPBS pre-treated Reconstructed human Cornea-like Epithelia tissue replicates).
Details on study design:
- RhCE tissue construct used, including batch number: Reconstructed human Cornea-like Epithelium (RhCE) tissue constructs: The 0.60 cm² Reconstructed human Cornea-like Epithelia (EpiOcularTM OCL-212-ver2.0, supplied by MatTek Corporation, batch No. 23770) were received on 08 March 2017. The killed Reconstructed human Cornea-like Epithelia (EpiOcularTM OCL-212-ver2.0, supplied by MatTek Corporation, batch No. 23706) were defrost on 03 March 2017, re-freeze on 03 March 2017 and defrost to be used on 08 March 2017.
The same day, the 12 tissues in their 24-well shipping container were equilibrated at room temperature for 15 minutes. Then the inserts (filter + epithelium) were gently removed from the agarose while avoiding leaving agarose on the polycarbonate filter. They were placed in 6 wells culture plate which had been previously filled with 1 mL of 37°C pre-warmed assay medium (MatTek Corporation, batch No. 030617MGKA) and incubated during 19 hours and 55 minutes at standard culture conditions.

- Evaluation of direct interaction with MTT: The direct interaction of MTT with the test item was checked by adding 50 µL of the test item to 1 mL of solution of MTT at 1 mg/mL. A purple solution was observed after 3 hours of incubation between 36.4 °C and 37.8 °C, 5% CO2. Therefore, the test item was identified as a direct MTT reducer and two killed control tissue models were added to the study which underwent the entire testing procedure to generate a non-specific MTT reduction control.

- Doses of test chemical and control substances used: 50 µL of test substance, negative or positive controls used.

- Pre-treatment: After the overnight incubation, the tissues were pre-wetted with 20 μL of Ca2+Mg2+Free-DPBS. The tissues were incubated at standard culture conditions for 30 minutes.

Treatment of tissues
- Duration and temperature of exposure, post-exposure immersion and post-exposure incubation periods:
- The test item was applied, as supplied, at the dose of 50 µL, to the entire surface of 2 living RhCE tissue replicates and 2 killed DPBS pre-treated Reconstructed human Cornea-like Epithelia tissue replicates during 30 minutes at standard culture conditions. In the same experimental conditions, the positive and negative controls were carried out.
- After the treatment, the test item and control substances were carefully washed from the RhCE tissues by extensive rinsing with Ca2+Mg2+Free-DPBS. The rinsed tissues were checked for any coloration and noted to be of comparable colour with the negative control treated tissues (whitish). This rinsing step was followed by a 12-minute post-exposure immersion period at room temperature in 5 mL of fresh medium to remove any test item absorbed into the tissue. The RhCE constructs were then incubated for a 2 hours and 02 minutes post-exposure incubation at standard culture conditions in 1 mL of fresh medium at 37°C, 5% CO2.

- Indication of controls used for direct MTT-reducers and/or colouring test chemicals (if applicable):
Spectral analysis of the test item in isopropanol: The spectral properties at 570 nm of test item in isopropanol were checked by adding 50 µL of the test item to 2 mL of isopropanol (same conditions as in the main test). A yellow solution was obtained after 2 hours of incubation at ambient temperature with gentle shaking. The mean of the corrected OD (blank subtracted) was 0.024 which is less than 0.08 (value corresponding to approximately twice the OD (Optical Density) of the extracting solvent). Therefore the test item was considered not to interfere with the MTT assay and there was no need to add non-specific coloration controls to the study.

- Cell viability measurements: Following the exposure to the test item, viability measurements are performed immediately after the post-exposure incubation period of the rinsed tissues in fresh medium. This period allows both for recovery from weak cytotoxic effects and for appearance of clear cytotoxic effects.
The RhCE tissue viability was measured by enzymatic conversion of the vital dye MTT by the viable cells of the tissue into a blue MTT formazan salt that is quantitatively measured after extraction from tissues.

- Optical Density measurements: The RhCE constructs were placed in 300 µL of a MTT solution at 1.0 mg/mL for 3 hours and 05 minutes at standard culture conditions. The precipitated blue formazan product was then extracted from the tissues by placing each insert in 2 mL of isopropanol during 17 hours and 28 minutes at 6±3°C in the dark. The concentration of formazan was measured by determining the OD at 570 nm, just after dilution of the extractions in isopropanol (1:2). The OD at 570 nm was measured in triplicate samples of formazan extracts. The measured OD are proportional to the number of living cells. The measurement of OD was performed using the ELx800 absorbance microplate reader (controlled every year and calibrated if necessary) supplied by BioTek and the validated software Gen5 ELISA V1.05.11 supplied by BioTek.

- Description of evaluation criteria used including the justification for the selection of the cut-off point for the prediction model: The OD values obtained with the replicate tissue extracts for each test item were used to calculate the mean percent tissue viability normalized to the negative control, which was set to 100%. The percentage tissue viability cut-off value distinguishing classified from non-classified test items is 60%. Results are interpreted as follows:
The test item is identified as not requiring classification and labelling according to UN GHS No Category: if the mean percent tissue viability after exposure and post-exposure incubation is > 60%. In this case, no further testing in other test methods is required.
The test item is identified as potentially requiring classification and labelling according to UN GHS (Category 2 or Category 1): if the mean percent tissue viability after exposure and post-exposure incubation is ≤ 60%.
When the final mean percent tissue viability is ≤ 60%, further testing with other test methods will be required because the RhCE test method shows a certain number of false positive results and cannot resolve between UN GHS Categories 1 and 2.

- Reference to historical positive and negative control results demonstrating suitable run acceptance criteria: The mean percent tissue viabilities obtained with the positive control and negative controls are within the range of historical data and therefore validate the experiment.
Irritation parameter:
other: % mean viability of the tissues
Run / experiment:
Main test - 30 minutes
Value:
69.55
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Other effects / acceptance of results:
MEAN PERCENT TISSUE VIABILITIES
- The mean corrected percent tissue viability of the RhCE replicates treated with the test item was 69.55 %, versus 39.16% in the positive control (Methyl acetate).

ACCEPTANCE OF RESULTS:
- The mean percent tissue viabilities obtained with the positive control and negative controls are within the range of historical data and therefore validate the experiment.

Table 7.3.2/1: Assessment of the eye irritation potential individual and average values of OD after 30 minutes exposure

 

Items

Tissues

OD

Mean OD/disc#

Mean OD / product

Viability%

Mean viability%

Difference of viability%

Conclusion

Negative control

1

0.947

0.912

0.959

95.10

100.00

9.80

-

0.902

0.887

2

1.162

1.006

104.90

0.966

0.891

Positive control

3

0.318

0.366

0.376

38.16

39.16

1.98

UN GHS category 2 or 1

0.372

0.408

4

0.382

0.385

40.15

0.394

0.379

Test item

7

0.749

0.733

0.726

76.434

75.70

1.46

-

0.728

0.722

8

0.710

0.719

74.974

0.720

0.727

Test item

killed control

11

0.060

0.059

0.059

6.152

6.15

0.00

0.059

0.057

12

0.060

0.059

6.152

0.059

0.058

Test item

-

69.55

-

No category

#: mean of 3 values (triplicate of the same extract)

OD: optical density

Interpretation of results:
GHS criteria not met
Conclusions:
Under the experimental conditions adopted and in accordance with the Regulation EC No. 1272/2008, the test item does not require classification for eye irritation or serious eye damage. It corresponds to the UN GHS No Category.
Executive summary:

An in vitro eye irritation test, reconstructed human cornea-like epithelium tissues (EpiOcularTM tissue model) was performed according to the OECD Guideline 492 and in compliance with GLP to evaluate the eye hazard potential of the test item Tagete Essential Oil.

 

The test item was applied, as supplied, at the dose of 50 µL, to 2 living and 2 killed DPBS pre-treated RhCE (EpiOcularTM tissue model) during 30 minutes at 37 °C, 5% CO2, 95% humidity (standard culture conditions). The exposure period was followed by extensive rinsing with DPBS at room temperature, a 12 minutes post-exposure immersion period at room temperature and a 2 hours and 02 minutes post-exposure incubation at standard culture conditions. The tissue viability was measured by performing an MTT assay.

 

The mean corrected percent tissue viability of the RhCE replicates treated with the test item was 69.55 %, versus 39.16% in the positive control (Methyl acetate). The mean percent tissue viabilities obtained with the positive control and negative controls are within the range of historical data and therefore validate the experiment.

 

In conclusion, under the experimental conditions adopted and in accordance with the Regulation EC No. 1272/2008, the test item does not require classification for eye irritation or serious eye damage. It corresponds to the UN GHS No Category.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation:

An in vitro skin irritation study was performed according to OECD Guideline 439 and in compliance with GLP to evaluate the possible irritating effects of the test item Tagete essential oil Oil after topical application onin vitrohuman reconstructed epidermis (SkinEthic RHE®model).

 

The test item was applied as supplied, at the dose of 16 µL, to 3 living and 2 killed Reconstructed Human epidermis (SkinEthic RHE®model) during 42 minutes, followed by a rinse with 25 mL of DPBS and a 41 hours and 15 minutes post-incubation period at 37 °C, 5% CO2. Cell viability was then measured by enzymatic conversion of the vital dye MTT into a blue formazan salt that was quantitatively measured after extraction from tissues.

 

The mean percent tissue viabilities obtained with the negative control and positive controls are within the range of historical data and therefore validate the experiment.

 

The mean corrected percent viability of the treated tissues was -1.2% (considered as 0%), versus 0.9% in the positive control (5% Sodium Dodecyl Sulfate).

 

Under these experimental conditions and in accordance with the Regulation EC No.1272/2008 and in absence of information on a skin corrosion, the test item has to be classified in Category 2 “Irritant” or in Category 1 “Corrosive”.

Eye irritation:

An in vitro eye irritation test,reconstructed human cornea-like epithelium tissues (EpiOcularTMtissue model)was performed according to the OECD Guideline 492 and in compliance with GLPto evaluate the eye hazard potential of the test item Tagete Essential Oil.

 

The test item was applied, as supplied, at the dose of 50 µL, to 2 living and 2 killed DPBS pre-treated RhCE (EpiOcularTMtissue model) during 30 minutes at 37 °C, 5% CO2, 95% humidity (standard culture conditions). The exposure period was followed by extensive rinsing with DPBS at room temperature, a 12 minutes post-exposure immersion period at room temperature and a 2 hours and 02 minutes post-exposure incubation at standard culture conditions. The tissue viability was measured by performing an MTT assay.

 

The mean corrected percent tissue viability of the RhCE replicates treated with the test item was 69.55 %, versus 39.16% in the positive control (Methyl acetate). The mean percent tissue viabilities obtained with the positive control and negative controls are within the range of historical data and therefore validate the experiment.

 

In conclusion, under the experimental conditions adopted and in accordance with the Regulation EC No. 1272/2008, the test item does not require classification for eye irritation or serious eye damage. It corresponds to the UN GHS No Category.

Justification for classification or non-classification

Self-classification:

Based on the available information (OECD 492 test), the substance is not classified for eye irritation/corrosion according to the Regulation (EC) No. 1272/2008.

Based on the available information (OECD 439 test), the substance has to be classified as Category 2 "irritant" or category 1 "corrosive" to the skin. However the substance is not classified as eye irritant, that can theorically be considered as a worst case for skin irritation/corrosion. In addition, none of the constituents are currently known to have skin corrosive properties. Furthermore, the three major constituents (z beta ocimene, dihydrotagetone and e/z tagetenone) are not classified for skin irriation in notified classification and labelling. As a consequence, the test item is very unlikely to be corrosive to the skin and can be classified as irritant to the skin (H315) according to CLP regulation 1272/2008 (CE).

No data was available regarding respiratory irritation.