Registration Dossier

Administrative data

Description of key information

Skin irritation/corrosion: Not corrosive (OECD 431/GLP)

Skin irritation/corrosion: Not irritating (OECD 439/GLP)

Serious eye damage/eye irritation: Not irritating (OECD 437/GLP)

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
30th October 2017 - 27th November 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: O’Laughlin (Nantong) Fine Chemicals Co., Ltd.; NTA375
- Expiration date of the lot/batch: Sep 25, 2020
- Purity: CAS No. 67634-00-8: 79.45 %; CAS No.: 67634-01-9 20.28 %

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Store in cool place. Keep container tightly closed in a dry and well-ventilated place.
Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
The reconstructed human epidermal model EpiDerm™ (EPI-200, MatTek, Bratislava, SR) consists of normal human-derived epidermal keratinocytes, which have been cultured to form a multilayered highly differentiated model of the human epidermis. The EpiDerm™ tissues (surface 0.63 cm²) are cultured on specially prepared cell culture inserts and shipped as kits, containing 24 tissues on shipping agarose together with the necessary amount of culture media. The certificate of analysis from the EpiDerm™ model (Lot No. 25851, kit B) is presented in Annex 1.

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 3 mins at room temperature and 60 minutes 37±1°C

REMOVAL OF TEST MATERIAL AND CONTROLS
- Number of washing steps: After exposition, tissues were thoroughly rinsed and blotted to remove the test item/controls.
- Observable damage in the tissue due to washing: None
- Modifications to validated SOP:None

DYE BINDING METHOD
- Dye used in the dye-binding assay: MTT
- Spectrophotometer: Libra S22
- Wavelength:OD570

NUMBER OF REPLICATE TISSUES: In each time interval three tissues were used per test item, three tissues for the positive control (PC) and three tissues for negative control (NC).

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE

Direct MTT reduction - functional check in tubes
Functional check for this possibility was performed as follows: 50 µL of the test item was added to 1 mL MTT medium (red) and it was incubated in the incubator (37±1 °C, 5±1 % CO2, humidified) for 1 hour. At the end of
the exposure time, the presence and intensity of the staining (if any) was observed.

Concurrent MTT non-specific colour control
For this purpose 50 µL of the test item were added to 1.0 mL of water and it was kept at culture conditions (37±1 °C, 5±1 % CO2, humidified) for 1 hour. Another 50 µL of the test item were added to 2 mL of isopropyl
alcohol and it was left at 2 hours at room temperature.

PREDICTION MODEL / DECISION CRITERIA
Corrosivity potential of the test item is predicted from the relative mean tissue viabilities obtained after 3 minutes and 60 minutes of treatment compared to the negative control tissues concurrently treated with H2O.

According to the OECD TG 431 as well as to the EU Method B.40, the test item is considered to be corrosive to skin:
i) if the viability after 3-minute exposure is less than 50 %, or
ii) if the viability after 3-minute exposure is greater than or equal to 50 % and the viability after 1-hour exposure is less than 15 %.
The test item is considered to be non-corrosive to skin:
i) if the viability after 3 minute exposure is greater than or equal to 50 % and the viability after 1-hour exposure is greater than or equal to 15 %.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
yes, concurrent MTT non-specific colour control
other: Direct MTT reduction
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 50 µL

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 50 μL

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 50 μL
- Concentration (if solution): 8N KOH
Duration of treatment / exposure:
3 mins at room temperature
60 minutes at 37±1°C
Number of replicates:
3
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
AAG (3 mins)
Value:
95.2
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
AAG (60 mins)
Value:
104.4
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Positive Control (3 mins)
Value:
7.5
Vehicle controls validity:
not applicable
Negative controls validity:
not applicable
Positive controls validity:
not applicable
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Positive Control (60 mins)
Value:
4.6
Vehicle controls validity:
not examined
Negative controls validity:
not applicable
Positive controls validity:
not applicable
Other effects / acceptance of results:
- OTHER EFFECTS:
- Visible damage on test system: None
- Direct-MTT reduction: The test item did not change colour of MTT medium from red to blue (see Figure 1). The test item does not reduce MTT directly.
- Colour interference with MTT: The colour of water/isopropyl alkohol did not change, so colour of the test item did not interfere with evaluation (see Figures 2 a, b).

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The assay meets the acceptance criterion - OD570 of the NC tissues was 1.790 (3 min) and 1.747 (60 min) which is ≥ 0.8 and ≤ 2.8.
- Acceptance criteria met for positive control: Viability of tissues treated with 8N KOH after 60 minutes of treatment was 4.6 % which is < 15 %.
- Acceptance criteria met for variability between replicate measurements: CV values in all triplets of tissues were ≤ 0.3.
Interpretation of results:
GHS criteria not met
Conclusions:
In the in vitro skin corrosion test using the reconstructed human epidermal model EpiDerm, Allyl Amyl Glycolate was not corrosive.
Executive summary:

In an in vitro skin corrosion assay using a human epidermal model EpiDerm (17 -698), reconstructed human epidermis tissue was exposed to 50 µL mg of Allyl Amyl Glycolate for 3 mins at room temperature and 60 minutes at 37±1°C. Water was used for the negative control and 8N KOH was used for the positive control. After removal of the test substance, tissues were incubated with MTT for 3 hours incubation. The OD570 of isopropyl alcohol extracts was measured on a spectrophotometer. Relative cell viability was calculated for each tissue as % of the mean viability of the negative control tissues.

The colour of the test substance did not interfere with the endpoint. The test substance is not directly MTT reducing. The average viability of Allyl Amyl Glycolate-treated tissues was 95.2 % of the negative control average value after 3 minutes of treatment and 104.4 % after 1 hour of treatment. The average viability of 8N KOH-treated tissues was 7.5 % of the negative control average value after 3 minutes of treatment and 4.6 % after 1 hour of treatment According to these results, Allyl Amyl Glycolate is not corrosive.

This in vitro skin corrosion assay in the human epidermal model EpiDerm is acceptable and satisfies the guideline requirement for an OECD 431 study.

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
20th November 2017 to 19th Decemer 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: O’Laughlin (Nantong) Fine Chemicals Co., Ltd.; NTA375
- Expiration date of the lot/batch: Sep 25, 2020
- Purity: CAS No. 67634-00-8: 79.45 %; CAS No.: 67634-01-9 20.28 %

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Store in cool place. Keep container tightly closed in a dry and well-ventilated place.
Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
The reconstructed human epidermal model EpiDerm™ (EPI-200, MatTek, Bratislava, SR) consists of normal human-derived epidermal keratinocytes, which have been cultured to form a multilayered highly differentiated model of the human epidermis. The EpiDerm™ tissues (surface 0.63 cm²) are cultured on specially prepared cell culture inserts and shipped as kits, containing 24 tissues on shipping agarose together with the necessary amount of culture media. The certificate of analysis from the EpiDerm™ model (Lot No. 25860 kit A) is presented in Annex 1.

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 60±1 minutes (25 minutes at room temperature and the remaining 35 minutes at culture conditions).
- Temperature of post-treatment incubation (if applicable): 23 hours and 46 minutes

REMOVAL OF TEST MATERIAL AND CONTROLS
- Number of washing steps Tissues were then thoroughly rinsed with PBS
- Observable damage in the tissue due to washing: None
- Modifications to validated SOP:None

DYE BINDING METHOD
- Dye used in the dye-binding assay: MTT
- Spectrophotometer: Libra S22
- Wavelength:OD570
- Filter: None
- Filter bandwidth: 2-3 nm

NUMBER OF REPLICATE TISSUES: In each time interval three tissues were used per test item, three tissues for the positive control (PC) and three tissues for negative control (NC).

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE

The test was performed as a part of Study No. 467/17/4AC: Allyl Amyl Glycolate - In vitro Skin Corrosion Test (EpiDermTM Model); VUOS CETA Report No.: 17-698. The test item does not reduce MTT directly.

The test was performed as a part of Study No. 467/17/4AC: Allyl Amyl Glycolate - In vitro Skin Corrosion Test (EpiDermTM Model); VUOS CETA Report No.: 17-698. The test item does not interfere with the endpoint.

PREDICTION MODEL / DECISION CRITERIA
The cut-off values for the prediction of irritation are given below; these values are stated in OECD Test Guideline No. 439, par. 36:
- In case the test chemical is found to be non-corrosive (e.g., based on TG 430, 431 or 435), and shows tissue viability after exposure and post-treatment incubation is less than or equal (≤) to 50 %, the test chemical is considered to be irritant to skin in accordance with UN GHS * Category 2.
- The test chemical may be considered as non-irritant to skin in accordance with UN GHS No Category if the tissue viability after exposure and post-treatment incubation is more than (>) 50 %.
A single testing run composed of three replicate tissues should be sufficient for a test chemical when the classification is unequivocal. However, in cases of borderline results, such as non-concordant replicate measurements and/or mean percent viability equal to 50 ± 5%, a second run should be considered, as well as a third one in case of discordant results between the first two runs.




Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 30 μL

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 30 μL

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 30 μL
- Concentration (if solution):5%
Duration of treatment / exposure:
60±1 minutes (25 minutes at room temperature and the remaining 35 minutes at 37±1°C).
Duration of post-treatment incubation (if applicable):
42 hours
Number of replicates:
3
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
AAG
Value:
104.8
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Postive Control
Value:
2.4
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Other effects / acceptance of results:
- OTHER EFFECTS:
- Visible damage on test system: None
- Direct-MTT reduction: The test item does not reduce MTT directly based on the test conducted for study No. 467/17/4AC.

- Colour interference with MTT: The test item does not interfere with the endpoint based on the test conducted for study No. 467/17/4AC.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The mean OD570 of the NC tissue was 1.893 which meets the acceptance criteria of ≥ 0.8 and ≤ 2.8. The 95% confidence interval of historical negative control is 1.676-2.117 (44 entries).

- Acceptance criteria met for positive control:
The mean viability of the PC tissues expressed as % of the negative control tissues was 2.4 % which meets the acceptance criterion of ≤ 20 %. The 95% confidence interval of historical positive control is 0.039-0.138 (44 entries).

- Acceptance criteria met for variability between replicate measurements:
The SD calculated from individual % tissue viabilities of the 3 identically treated replicates was 0.2 % for the positive control, 3.0 % for negative control and 4.5 % for the test item what is < 18 % in all cases.




Interpretation of results:
GHS criteria not met
Conclusions:
In the in vitro skin irritation test using the reconstructed human epidermal model EpiDerm, Allyl Amyl Glycolate was not irritating.
Executive summary:

In an in vitro skin irritation assay in a human epidermal model EpiDerm (17-776), reconstructed human epidermis tissue was exposed to 30 μL of Allyl Amyl Glycolate for 60±1 minutes (25 minutes at room temperature and 35 minutes at 37±1°C). PBS was used for the negative control and 5% SDS was used for the positive control. After removal of the test substance, tissues were post-incubated for approximately 42 hours. After removal of the test substance, tissues were incubated with MTT for 3 hours incubation. The OD570 of isopropyl alcohol extracts was measured on a spectrophotometer. Relative cell viability was calculated for each tissue as % of the mean viability of the negative control tissues.

The colour of the test substance did not interfere with the endpoint. The test substance is not directly MTT reducing. The average viability of tissues treated by the positive control, 5% SDS, was 2.4 % of the negative control average value. The average viability of tissues treated by the test item, Allyl Amyl Glycolate, was 104.8 % of the negative control average value i.e. viability was > 50 %. According to these results, the test substance is not irritating.

This in vitro skin irritation study in the human epidermal model EpiDerm is acceptable and satisfies the guideline requirement for an OECD 439 study.

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:
21st November 2017 - 21st December 2017
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)
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: O’Laughlin (Nantong) Fine Chemicals Co., Ltd.; NTA375
- Expiration date of the lot/batch: Sep 25, 2020
- Purity: CAS No. 67634-00-8: 79.45 %; CAS No.: 67634-01-9 20.28 %

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Store in cool place. Keep container tightly closed in a dry and well-ventilated place.

Species:
cattle
Details on test animals or tissues and environmental conditions:
SOURCE OF COLLECTED EYES
- Source: Breeding service CHOVSERVIS a.s., division TORO® Hlavečník, Hradec Králové, Czech Republic
Eyes were collected by slaughterhouse employees. The eyes were enucleated as soon as possible after death. No detergent was used. Only healthy animals (12 to 30 months old) considered suitable for entry into the human food chain were used as a source of corneas for use in the BCOP test. The risk of contamination was minimized (e.g., by keeping the container containing the eyes on ice, by adding antibiotics to the HBSS used to store the eyes during transport (e.g., penicillin at 100 IU/mL and streptomycin at 100 μg/mL).
The time interval between collection of the eyes and use of corneas in the BCOP was minimized (typically collected and used on the same day). The results were based on the selection criteria for the eyes, as well as the positive and negative control responses. All eyes used in the assay were from the same group of eyes collected on a specific day.
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): 750 µL
Duration of treatment / exposure:
10 minutes
Number of animals or in vitro replicates:
3
Details on study design:
SELECTION AND PREPARATION OF CORNEAS:
The eyes, once they arrived at the laboratory, were carefully examined for defects including scratched, and neovascularisation. Only corneas from eyes free of such defects were used.
The isolated corneas, after achieve normal metabolic activity (inductive incubation at 32 ± 1°C for one hour), were examined again. The corneas that show macroscopic tissue damage (e.g., scratches, pigmentation, neovascularization) or a baseline opacity >7 opacity units were discarded.

QUALITY CHECK OF THE ISOLATED CORNEAS:
From 24 eyes the 5 eyes were eliminated after inductive incubation, because the baseline opacity values were >7. Nine corneas were used for the study (the corneas No. 1, 2, 3, 7, 8, 9, 13, 14 and 15), 4 eyes were superfluous and 6 corneas were used for testing of other substances.

NUMBER OF REPLICATES: 3

NEGATIVE CONTROL USED: 0.9% NaCl

POSITIVE CONTROL USED: 100% DMFA

APPLICATION DOSE AND EXPOSURE TIME: As a liquid, the test item was tested undiluted for 10 minutes.

TREATMENT METHOD: Closed-chamber method was used, because the test substance was applicable by micropipette. The test substance (750 µL of application form) to cover the epithelial side of the cornea is introduced into the anterior chamber through the dosing holes on the top surface of the chamber, and the holes were subsequently sealed with the chamber plugs during the exposure.

POST-INCUBATION PERIOD: Yes, two hours at 32 ± 1 ºC

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: After the exposure period, the test item was removed from the anterior chamber with EMEM (containing phenol red - the effectiveness of rinsing acidic or alkaline materials). The corneas were given a final rinse with EMEM (without phenol red). The EMEM (without phenol red) was used as a final rinse to ensure removal of the phenol red from the anterior chamber prior to the opacity measurement. The anterior chamber was then refilled with fresh EMEM without phenol red. The corneas were incubated for an additional two hours
at 32 ± 1 ºC with EMEM. At the end of the post-exposure incubation period, the opacity and permeability of each cornea were recorded.


METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: Corneal opacity was measured quantitatively with the aid of an opacitometer (Opacitometer, MC2 - Le spécialiste du laboratoire – France) resulting in opacity values measured on a continuous scale

- Corneal permeability: 1 mL sodium fluorescein solution (5 mg/mL) was added to the anterior chamber of the corneal holder, which interfaced with the epithelial side of the cornea, while the posterior chamber, which interfaced with the endothelial side of the cornea, is filled with fresh EMEM. The holder was incubated in horizontal position for 1.5 hours at 32 ± 1 ºC. The amount of sodium fluorescein that crosses into the posterior chamber was quantitatively measured with the aid of UV/VIS spectrophotometry (Spectrophotometer GENESYSTM 10 UV/VIS Scanning). The values of absorbance measured at 490 nm were recorded as optical density (OD490) values. This term was used because the measuring is performed with visible light spectrophotometer using a standard 1 cm path length

SCORING SYSTEM: IVIS = mean opacity value + (15 x mean permeability OD490 value)

DECISION CRITERIA: A test is considered acceptable if the positive control gives IVIS that falls within one standard deviations of the current historical mean, which is to be updated at least every three months, or each time an acceptable test is conducted in laboratories where tests are conducted infrequently. The negative or solvent/vehicle control responses should result in opacity and permeability values that are less than the established upper limits for background opacity and permeability values for bovine corneas treated with the respective negative or solvent/vehicle control.

The IVIS cut-off value for identifying the test item as including serious eye damage (UN GHS Category 1) and the test item not requiring classification for eye irritation or serious damage (UN GHS No Category) is given in Table 1 below.
Irritation parameter:
cornea opacity score
Run / experiment:
AAG
Value:
1.58
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Remarks:
58.12
Irritation parameter:
fluorescein retention score
Run / experiment:
AAG
Value:
0.022
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Remarks:
0.171
Irritation parameter:
in vitro irritation score
Run / experiment:
AAG
Value:
1.91
Vehicle controls validity:
not examined
Negative controls validity:
valid
Remarks:
0.85
Positive controls validity:
valid
Remarks:
60.68
Other effects / acceptance of results:
OTHER EFFECTS:
- Visible damage on test system: Appearance of corneas was observed before and after application of the test item, negative and positive control (see table 2 and 3). No macroscopic damage was observed on corneas before application. Corneal opacity was observed on the corneas treated by positive control. The corneas treated by negative control and the test item were without macroscopic damage.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The value of opacity for negative control (0.9% NaCl) obtained during the study was 0.73 and value of permeability was 0.008. The values obtained during this study not exceeded upper limits (2.86 and 0.0446, respectively), so the study is considered acceptable.

- Acceptance criteria met for positive control: The value of IVIS for positive control (100% DMFA) obtained during the study was 60.68. This value is within the acceptance limit (one standard deviations of the current historical mean; 73.89±14.95), so the study is considered acceptable.
Interpretation of results:
GHS criteria not met
Conclusions:
In the Bovine Corneal Opacity and Permeability (BCOP) assay, the IVIS for AAG was 1.91. As the IVIS is ≤ 3, the classification of the test substance according to UN GHS criteria for eye irritation or serious eye damage is: No category.
Executive summary:

In the Bovine Corneal Opacity and Permeability (BCOP) assay (17-759), isolated bovine corneas were exposed to 100%  Allyl Amyl Glycolate  for 10  minutes using the closed chamber method. 0.9% sodium chloride was used for the negative control and 100% Dimethylformamide was used for the positive control. The corneas were rinsed twice and then incubated for an additional two hours at 32 ± 1 ºC in medium. The opacity and permeability (via sodium fluorescein dye) of each cornea were recorded.

There was no macroscopic damage to any corneas treated with the test substance There was no macroscopic damage to any corneas treated with the test substance (Table 3). The mean opacity value for the test substance was 1.58. The mean permeability OD490 for the test substance was 0.022. The IVIS for the test substance was 1.91. The positive control gave the appropriate response. The IVIS for AAG is ≤ 3 therefore the classification of test substance according to UN GHS criteria for eye irritation or serious eye damage is: No category.

This Bovine Corneal Opacity and Permeability (BCOP) is acceptable and satisfies the guideline requirement for an OECD 437 study.

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irriation/corrosion:

There is one in vitro skin corrosion test and one in vitro skin irritation test available.

In an in vitro skin corrosion assay using a human epidermal model EpiDerm (OECD 431/GLP), reconstructed human epidermis tissue was exposed to 50 µL mg of Allyl Amyl Glycolate for 3 mins at room temperature and 60 minutes at 37±1°C. Water was used for the negative control and 8N KOH was used for the positive control. After removal of the test substance, tissues were incubated with MTT for 3 hours incubation. The OD570 of isopropyl alcohol extracts was measured on a spectrophotometer. Relative cell viability was calculated for each tissue as % of the mean viability of the negative control tissues. The colour of the test substance did not interfere with the endpoint. The test substance is not directly MTT reducing. The average viability of Allyl Amyl Glycolate-treated tissues was 95.2 % of the negative control average value after 3 minutes of treatment and 104.4 % after 1 hour of treatment. The average viability of 8N KOH-treated tissues was 7.5 % of the negative control average value after 3 minutes of treatment and 4.6 % after 1 hour of treatment According to these results, Allyl Amyl Glycolate is not corrosive.

In an in vitro skin irritation assay in a human epidermal model EpiDerm (OECD 439/GLP), reconstructed human epidermis tissue was exposed to 30 μL of Allyl Amyl Glycolate for 60±1 minutes (25 minutes at room temperature and 35 minutes at 37±1°C). PBS was used for the negative control and 5% SDS was used for the positive control. After removal of the test substance, tissues were post-incubated for approximately 42 hours. After removal of the test substance, tissues were incubated with MTT for 3 hours incubation. The OD570 of isopropyl alcohol extracts was measured on a spectrophotometer. Relative cell viability was calculated for each tissue as % of the mean viability of the negative control tissues. The colour of the test substance did not interfere with the endpoint. The test substance is not directly MTT reducing. The average viability of tissues treated by the positive control, 5% SDS, was 2.4 % of the negative control average value. The average viability of tissues treated by the test item, Allyl Amyl Glycolate, was 104.8 % of the negative control average value i.e. viability was > 50 %. According to these results, the test substance is not irritating.

Serious eye damage/eye irritation:

There is one in vitro eye irritation test available.

In the Bovine Corneal Opacity and Permeability (BCOP) assay (OECD 437/GLP), isolated bovine corneas were exposed to 100%  Allyl Amyl Glycolate  for 10  minutes using the closed chamber method. 0.9% sodium chloride was used for the negative control and 100% Dimethylformamide was used for the positive control. The corneas were rinsed twice and then incubated for an additional two hours at 32 ± 1 ºC in medium. The opacity and permeability (via sodium fluorescein dye) of each cornea were recorded.

There was no macroscopic damage to any corneas treated with the test substance There was no macroscopic damage to any corneas treated with the test substance. The mean opacity value for the test substance was 1.58. The mean permeability OD490 for the test substance was 0.022. The IVIS for the test substance was 1.91. The positive control gave the appropriate response. The IVIS for AAG is ≤ 3 therefore the classification of test substance according to UN GHS criteria for eye irritation or serious eye damage is: No category.

The results of these tests are suitable for use in the human health hazard assessment.

Justification for classification or non-classification

Based on the available information in the dossier, the substance Allyl Amyl Glycolate (EC No. 916-328-0) does not need to classified  for skin irritation/corrosion and does not need to classified for serious eye damage/eye irritation when considering the criteria outlined in Annex I of 1272/2008/EC.