1-[difluoro(trifluoromethoxy)methoxy]-1,2,2-trifluoroethylene

Endpoint summary

Description of key information

Skin Irritation / Corrosion

The potential of MOVE3 to be irritating to the skin was investigated in the in vitro skin irritation test [reconstructed human epidermis model EPISKIN model] conducted according to the method described in OECD guideline No. 439 and performed in compliance with good laboratory practices GLP.

MOVE 3 was found not-irritant and not-corrosive to the skin.

 

Eye Irritation / Corrosion

The potential of MOVE3 to be irritating to the eye was investigated in the in vitro eye irritation test [reconstructed human cornea-like epithelium EpiOCular model] conducted according to the method described in OECD guideline No. 492 and performed in compliance with good laboratory practices GLP.

MOVE 3 was found not-irritant and not-corrosive to the eye.

Respiratory Irritation

The potential of MOVE3 to be irritating to the respiratory tract was assessed basing on the results of the acute toxicity study and the combined repeated dose with reproduction/developmental toxicity screening study conducted by inhalation on rats.

Basing on the results MOVE 3 is not irritant to the respiratory tract following single exposure but it is considered potentially irritant for the respiratory system following prolonged repeated exposure.

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:

04-Jan-2016 to 11-Jan-2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

Deviations:

no

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:

MOVE 3 / Batch 150525V10 (2015)
- Name of test material (as cited in study report): MOVE3
- Substance type: Clear colourless liquid
- Physical state: Liquid
- Storage condition of test material: In refrigerator (2-8°C)

Test system:

human skin model

Source species:

human

Cell type:

other: keratinocytes. Prior to test the keratinocytes were cultured for 13 days, which results in a highly differentiated and stratified epidermis model comprising the main basal, supra basal, spinous and granular layers and a functional stratum corneum.

Cell source:

other: adult human donor

Justification for test system used:

In the interest of sound science and animal welfare, a sequential testing strategy is recommended to minimise the need of in vivo testing. The RhE method according to OECD 439 is one of the OECD validated skin irritation test, which is recommended in international guidelines (e.g. OECD and EC). The RhE-based test methods are able to identify Cat. 2 and No Cat. chemicals and can thus serve as stand-alone skin irritation methods for non-corrosives in countries where optional Cat. 3 is not implemented. In case RhE-based test methods result in Cat. 2, an in vitro skin corrosion test, if not performed upfront, is required to determine the final classification (Cat. 2 (irritant) or Cat. 1(A, B or C) (corrosive)).

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: Episkin Small model
- Tissue batch number(s): 16-EKIN-001
- Expiring date: 11 January 2016
- Experimental starting date: 04 January 2016
Experimental completion date: 11 January 2016

The test consists of topical application of MOVE3 on the skin tissue for 15 minutes. After exposure the skin tissue is thoroughly rinsed to remove the test item and transferred to fresh medium. After a 42 hour incubation period, determination of the cytotoxic (irritancy) effect is performed.
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.

On the day of receipt the tissues were transferred to 12-well plates and preincubated with prewarmed Maintenance Medium for 24 hours at 37°C (Figure 1). Maintenance medium and Assay medium were supplied by the Tissue supplier.

ENVIRONMENTAL CONDITIONS
All incubations, with the exception of the test item incubation of 15 minutes at room temperature, were carried out in a controlled environment, in which optimal conditions were a humid atmosphere of 80 - 100% (actual range 71 - 90%), containing 5.0 ± 0.5% CO2 in air in the dark at 37.0 ± 1.0°C (actual range 36.3 - 36.7°C). Temperature and humidity were continuously monitored throughout the experiment. The CO2 percentage was monitored once on each working day. Temporary deviations from the temperature, humidity and CO2 percentage may occur due to opening and closing of the incubator door. Based on laboratory historical data these deviations are considered not to affect the study integrity

TEST FOR INTERFERENCE OF THE TEST ITEM WITH MTT ENDPOINT:
1) TEST FOR COLOUR INTERFERENCE
MOVE3 was checked for possible colour interference before the study was started. Some non-coloured test items may change into coloured items in aqueous conditions and thus stain the skin tissues during the exposure. To assess the colour interference, approximately 10 μl of MOVE3 was added to 90 μl Milli-Q water. The mixture was mixed for approximately 15 minutes. A negative control, 10 μl Milli-Q water was tested concurrently. At the end of the shaking period a colour check was performed.
2) TEST FOR REDUCTION OF MTT
MOVE3 was checked for possible direct MTT reduction before the study was started. To assess the ability of the test item to reduce MTT, 25 μl of the test item was added to 2 ml MTT solution (0.3 mg/ml in PBS). The mixture was incubated for 3 hours at room temperature. A negative control, sterile Milli-Q water was tested concurrently.

TREATMENT
The test was performed on a total of 3 tissues per test item together with negative and positive controls. An excess amount of test item was added into 12-well plates on top of the skin tissues. The test item was refilled continuously during the treatment for replacing the losses of test item due to volatility (the boiling point of test item is ca. 20°C). The refilling guaranteed the exposure of the tissues to the test item during the whole incubation period.
Three tissues were treated with 25 μl PBS (negative control) and 3 tissues with 25 μl 5% SDS (positive control) respectively. The positive control was re-spread after 7 minutes contact time.
After the exposure period of 15 ± 0.5 minutes at room temperature, the tissues were washed with phosphate buffered saline to remove residual test item.
After rinsing, the cell culture inserts were each dried carefully and moved to a new well on 2 ml pre-warmed maintenance medium until all tissues were dosed and rinsed. Subsequently the skin tissues were incubated for 42 hours at 37°C.

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

CELL VIABILITY MEASUREMENT
After incubation, cell culture inserts were dried carefully to remove excess medium and were transferred into a 12-wells plate prefilled with 2 ml MTT-solution (0.3 mg/ml in PBS). The tissues were incubated for 3 h at 37°C. After incubation the tissues were placed on blotting paper to dry the tissues. Total biopsy was made by using a biopsy punch. Epidermis was separated from the collagen matrix and both parts were placed in prelabeled microtubes and extracted with 500 μl isopropanol (Merck, Darmstadt, Germany). Tubes were stored refrigerated and protected from light for approximately 71 hours. The amount of extracted formazan was determined spectrophotometrically at 570 nm in duplicate with the TECAN Infinite® M200 Pro Plate Reader.
Cell viability was calculated for each tissue as a percentage of the mean of the negative control tissues. Skin irritation potential of the test item was classified according to remaining cell viability following exposure of the test item.

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 0.3 mg/ml in PBS
- Incubation time: 3 hours


NUMBER OF REPLICATE TISSUES: 3 tissues per test item / negative and positive controls.

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
not applicable

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION:
1

ACCEPTABILITY OF THE ASSAY
The in vitro skin irritation test is considered acceptable if it meets the following criteria:
a) The absolute mean OD570 (optical density at 570 nm) of the three tissues of the negative control should reasonably be within the laboratory historical control data range and the Standard Deviation value (SD) of the % viability should be ≤18.
b) The mean relative tissue viability of the positive control should be ≤50% relative to the negative control and the Standard Deviation value (SD) of the % viability should be ≤18.
c) The SD calculated from individual % tissue viabilities of the three identically treated replicates should be ≤18.

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- A test item is considered irritant in the skin irritation test if the relative mean tissue viability of three individual tissues after 15 minutes of exposure to the test item and 42 hours of post incubation is ≤ 50% of the mean viability of the negative controls.
- A test item is considered non-irritant in the in vitro skin irritation test if the relative mean tissue viability of three individual tissues after 15 minutes of exposure to the test item and 42 hours of post incubation is > 50% of the mean viability of the negative controls.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit):
- Concentration (if solution):

VEHICLE
- Amount(s) applied (volume or weight with unit):
- Concentration (if solution):
- Lot/batch no. (if required):
- Purity:

NEGATIVE CONTROL
- Amount(s) applied (volume or weight):
- Concentration (if solution):

POSITIVE CONTROL
- Amount(s) applied (volume or weight):
- Concentration (if solution):TEST MATERIAL
- Amount(s) applied (volume or weight with unit): excess amount of test item was added. The test item was refilled continuously during the treatment for replacing the losses of test item due to volatility (the boiling point of test item is ca. 20°C). The refilling guaranteed the exposure of the tissues to the test item during the whole incubation period.
- Concentration: Undiluted

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 25 μl Phosphate buffered saline

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 25 μl 5% (aq) Sodium dodecyl sulphate. The positive control was re-spread after 7 minutes contact time.

Duration of treatment / exposure:

15 minutes

Duration of post-treatment incubation (if applicable):

42 hours

Number of replicates:

3

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): An excessive amount of the liquid test item was applied undiluted directly on top of the tissue. The test item was applied three times to completely cover the skin tissue during the whole incubation period. In order to reduce the volatilisation during the treatment, the test item was kept in the refrigerator (2-8°C) until the treatment.

NEGATIVE CONTOL:
- Amount(s) applied (volume or weight with unit): 25 µl Phosphate buffered saline

POSITIVE CONTROL
- Amount(s) applied (volume or weight with unit): 25 µl
- Concentration (if solution): 5% (aq) Sodium dodecyl sulphate

Duration of treatment / exposure:

Exposure:15 minutes
Post incubation period: 42 hours

Details on study design:

TEST SITE
- Area of exposure: human epidermis model
- % coverage: 0.38 cm2

REMOVAL OF TEST SUBSTANCE
- Washing (if done): phosphate buffered saline
- Time after start of exposure: 15 minutes

POST INCUBATION PERIOD
- 42 hours

SCORING SYSTEM:
- After a 42 hour incubation period, determination of the cytotoxic (irritancy) effect was performed. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT at the end of the treatment. Cell viability was calculated for each tissue as a percentage of the mean of the negative control tissues.

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Mean of 3 runs (see details below)

Value:

96

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

The positive control had a mean cell viability of 7% after 15 ± 0.5 minutes exposure.
The absolute mean OD570(optical density at 570 nm) of the negative control tissues was within the laboratory historical control data range.
The standard deviation value of the percentage viability of three tissues treated identically was less than 8%.
The test system functioned properly and the test is valid.

Absorption Result
	Replicate 1 OD570	Replicate 2 OD570	Replicate 3 OD570	Mean OD570	SD
Negative control	0,832	0,758	0,721	0,770	0,057
MOVE3	0,744	0,772	0,710	0,742	0,031
Positive control	0,064	0,063	0,039	0,055	0,014
Tissue viability
	Replicate 1	Replicate 2	Replicate 3	Mean viability	SD
MOVE3	96,582	100,216	92,168	96,322	4,031
Positive control	8,308	8,178	5,063	7,183	1,837
Negative control	108,005	98,399	93,596	100,000	7,337

Interpretation of results:

GHS criteria not met

Conclusions:

The in vitro skin irritation test was conducted according to OECD 439 guideline and GLP principles.
It is concluded that this test is valid and that MOVE3 is non-irritant in the in vitro skin irritation test.

Executive summary:

This study assessed the ability of MOVE3 to induce skin irritation on a human three dimensional epidermal model (EPISKIN Small Model (EPISKIN-SMTM)). The possible skin irritation potential of MOVE3 was tested through topical application for 15 minutes.

The study procedures described in this report were based on the most recent OECD and EC guidelines.

MOVE3 was applied undiluted directly on top of the skin tissue for 15 ± 0.5 minutes.

MOVE3 is a volatile liquid, therefore the test item was refilled continuously during the treatment for replacing the losses of test item due to volatility. The refilling guaranteed the exposure of the tissues to the test item during the whole incubation period.

After a 42 hour post-incubation period, determination of the cytotoxic (irritancy) effect was performed. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT at the end of the treatment.

Skin irritation is expressed as the remaining cell viability after exposure to the test item. The relative mean tissue viability obtained after 15 ± 0.5 minutes treatment with MOVE3 compared to the negative control tissues was 96%. Since the mean relative tissue viability for MOVE3 was above 50% after 15 ± 0.5 minutes treatment MOVE3 is considered to be non-irritant.

The positive control had a mean cell viability of 7% after 15 ± 0.5 minutes exposure. The absolute mean OD570(optical density at 570 nm) of the negative control tissues was within the laboratory historical control data range. The standard deviation value of the percentage viability of three tissues treated identically was less than 8%, indicating that the test system functioned properly.

It is concluded that this test is valid and that MOVE3 is non-irritant in thein vitro skin irritation test.

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:

March 2016 - May 2016

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:

28 July 2015

Deviations:

yes

Remarks:

Tissues treated with test item were incubated at room temperature instead of at standard culture conditions (due to the volatility of the substance).

GLP compliance:

yes

Specific details on test material used for the study:

The test item has a boiling point of ca. 20°C and it is volatile.

- Source and lot/batch No.of test material: 150525V10
- Purity/composition correction factor: No correction factor for purity/composition applied.
- Test item storage: In refrigerator (2-8°C)
- Stability under test conditions: chemically stable.

Species:

human

Details on test animals or tissues and environmental conditions:

The EpiOcular tissue construct is a nonkeratinized epithelium prepared from normal human keratinocytes. It models the cornea epithelium with progressively stratified, but not cornified cells. A cell suspension is seeded into the insert in specialized medium. After an initial period of submerged culture, the medium is removed from the top of the tissue so that the epithelial surface is in direct contact with the air. This allows the test material to be directly applied to the epithelial surface in a fashion similar to how the corneal epithelium would be exposed in vivo.

Vehicle:

unchanged (no vehicle)

Remarks:

all the tissues were pre-wetted with 20 μL of Ca2+Mg2+-Free-DPBS

Controls:

yes

Amount / concentration applied:

TEST MATERIAL
Since the test item had a boiling point of ca. 20°C and was volatile, an excess amount of test item was applied to the tissue. During the 30 min-exposure period it was regularly checked if the skin was completely covered and if needed additional substance was added to guarantee that during the whole exposure period the tissue was covered with test item. During the exposure approximately 450 μl of test item was used per skin tissue. During the exposure the temperature of the room was approximately 20°C.

NEGATIVE CONTROL
- Amount applied: 50 µl of MilliQ water per tissue

POSITIVE CONTROL
Amount(s) applied (volume or weight with unit): 50 µl Methyl Acetate per tissue

Duration of treatment / exposure:

30 ± 2 minutes

Number of animals or in vitro replicates:

Test item: 2 tissues at room temperature
Negative control (MilliQ water): 2 tissues treated at room temperature and 2 tissues treated at standard culture conditions
Positive control (Methyl Acetate): 2 tissues treated at room temperature and 2 tissues treated at standard culture conditions

Details on study design:

TEST SYSTEM
- EpiOcular™ (OCL-200-EIT MatTek Corporation, Lot: 23426 kit B)
The EpiOcular tissue construct is a non-keratinized epithelium (0.6 cm2) prepared from normal human keratinocytes.

Interference of the test item with the MTT endpoint
- Test item was checked for possible colour intereference and direct MTT reduction before the study started

- All incubations, with the exception the exposure and post-exposure immersion at room temperature, were carried out in a controlled environment set to maintain:
- Humidity: 80 - 100%
- CO2: 5.0 ± 0.5%
- Temperature: 37.0 ± 1.0°C.

- Before the assay was started all the tissues were pre-wetted with 20 μL of Ca2+Mg2+-Free-DPBS and incubated at standard culture conditions for 30 ± 2 minutes.
- Exposure: 30 ± 2 minutes at room temperature (2 negative and 2 positive control tissues were exposed at standard culture conditions).
- Removal test item: rinsing with Ca2+Mg2+-free D-PBS (brought to room temperature)
- Post-exposure immersion: After rinsing the cell culture inserts were each dried carefully and immediately transferred to and immersed in 5 ml Assay Medium for a 12 ± 2 minute immersion incubation at room temperature.
- Post-exposure incubation: After the Post-exposure immersion period cell culture inserts were each dried carefully and transferred to a 6-well plate containing 1.0 ml of warm Assay Medium and were incubated for 120 ± 15 minutes at 37°C.

CELL VIABILITY MEASUREMENT
After incubation, cell culture inserts were dried carefully to remove excess medium. The cell culture inserts were transferred into a 24-wells plate prefilled with 0.3 ml MTT-medium (1.0 mg/ml). The tissues were incubated for 180 ± 10 minutes at 37°C. After incubation with MTT-medium the tissues were placed on blotting paper to dry the tissues. Formazan was extracted with 2 ml isopropanol refrigerated for 18 ± 2 hours in the dark. The amount of extracted formazan was determined spectrophotometrically at 570 nm in duplicate with the TECAN Infinite® M200 Pro Plate Reader. The amount of extracted formazan was determined spectrophotometrically at 570 nm in duplicate with the TECAN Infinite® M200 Pro Plate Reader.

Acceptability of the assay
The in vitro eye irritation test is considered acceptable if it meets the following criteria:
a) The absolute mean OD570 of the two tissues of the negative control should reasonably be > 0.8 and < 2.5 and the Standard Deviation value (SD) of the % viability should be ≤20%.
b) The mean relative tissue viability of the positive control should be <50% relative to the negative control and the Standard Deviation value (SD) of the % viability should be ≤20%.
c) The SD calculated from individual % tissue viabilities of the two identically treated replicates should be <20%.

Data evaluation and statistical procedures
A test item is considered to be positive in the in vitro eye irritation test if:
The relative mean tissue viability of two individual tissues after exposure to the test item is ≤ 60% of the mean viability of the negative controls, requiring classification for eye irritation or serious eye damage (UN GHS Category 1 or 2).
A test item is considered to be negative in the in vitro eye irritation test if:
The relative mean tissue viability of two individual tissues after exposure to the test item is > 60% of the mean viability of the negative controls, requiring no classification for eye irritation or serious eye damage (UN GHS No Category).

Irritation parameter:

mean percent tissue viability 

Run / experiment:

Single run

Value:

99

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: None

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes.
The absolute mean OD570 of the negative control tissues treated at room temperature and at standard culture conditions was 1.865 and 1.773, respectively (within 0.8 and 2.5). The standard deviation value of the percentage viability of two tissues treated identically was less than 4%, indicating that the test system functioned properly.
- Acceptance criteria met for positive control: Yes.
The positive control tissues treated at standard culture condition had a viability of 41% (below 50%). The positive control tissues treated at room temperature had a viability of 51%, this result is considered acceptable considering the applied deviation from the standard procedure which requires that exposure of tissues is performed at 37 ± 1 °C. Moreover the mean viability is just above the acceptance criterion of 50% and still below 60% (positive in case of test item treatment), therefore it is considered that test system functioned properly.

Interpretation of results:

GHS criteria not met

Remarks:

according to EC 1272/2008

Conclusions:

Based on the evaluation of the eye hazard potential with MOVE3 using the EpiOcular cornea epithelial model performed according to OECD guideline and GLP principles, it is concluded that MOVE3 is not irritant for the eye.

Executive summary:

An evaluation of the eye hazard potential with MOVE3 using the EpiOcular™ cornea epithelial model was performed with MOVE3 according to OECD guideline 492 and GLP principles. Since MOVE3 has a boiling point of approximately 21°C an excess amount of MOVE3 was applied undiluted directly on top of the tissue at room temperature and during the 30 min-treatment period additional substance was added in order to guarantee that the tissue was covered with MOVE3 during the entire exposure period. Negative and positive controls were incubated at room temperature and at standard culture conditions with the aim of confirming the appropriateness of the test system. The positive control had a mean relative cell viability of 51% after 30±2 minutes exposure at room temperature and 41% after 30±2 minutes exposure at standard culture conditions (The mean relative cell viability of the positive control treated at room temperature is considered acceptable considering the applied deviation from the standard procedure which requires that the exposure of tissues is performed at 37 ± 1 °C and that the value is just above the acceptance criterion of 50% and still below 60%). The absolute mean OD570 of the negative control tissues was within 0.8 and 2.5 and the standard deviation value of the percentage viability of two tissues treated identically was less than 4%, indicating that the test conditions were adequate and the test system functioned properly. The relative mean tissue viability after treated with MOVE3 was 99%. Since the mean relative tissue viability for MOVE3 was above 50% after 30±2 minutes treatment MOVE3 is considered to be non-irritant and no classification is required for eye irritation or serious eye damage according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2011) (including all amendments) and the Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures (including all amendments).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Additional information

Skin Irritation / Corrosion

The potential of MOVE 3 to be irritating to the skin was investigated in a study conducted according to OECD guideline 439, Reconstructed Human Epidermis Test Method and in compliance with good laboratory practices (GLP).

The in vitro method described in the OECD guideline 439 allows to discriminates skin irritants (Cat. 2) from chemicals not classified for skin irritation (No Cat.) according to the EU GHS classification system.

Irritant chemicals are identified by their ability to decrease tissue viability below 50% of the negative control. According to OECD No. 439 method, a result indicating skin irritation (Cat. 2) does not allow excluding corrosion (Cat. 1) therefore in case the test showed a tissue viability below 50% of the negative control for the test chemical, an in vitro skin corrosion test would be required to determine the final classification (Cat. 2 (irritant) or Cat. 1(A, B or C) (corrosive)).This was not the case for the tissues treated with MOVE3 which showed a mean tissue viability = 96% compared to the negative control, therefore no further test is necessary and it can be concluded that the substance does not require classification and labelling according to EU GHS Criteria.

 

 

Eye Irritation / Corrosion

The potential of MOVE 3 to be irritating to the eye was investigated in an in vitro study conducted according to OECD Guideline 492, Reconstructed Human Cornea-like Epithelium (RhCE) Test Method and in compliance with good laboratory practices (GLP).

The method described in the OECD guideline 492 allows toIdentify Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage according to EU GHS and UN GHS classification system.

According to OECD Guideline 492 method, chemicals not requiring classification and labelling according to UN GHS are identified as those having a tissue viability higher than (>) 60% compard to the one of the negative control.

The method is not intended to differentiate between UN GHS Cat. 1 (serious eye damage) and UN GHS Cat. 2 (eye irritation), so in case the viability of the tissues treated with the test item was below than (<) 60% further evaluation would be needed. This was not the case for the tissues treated with MOVE3 which showed a mean tissue viability = 99% compared to the negative control, therefore no further test is necessary and it can be concluded that MOVE 3 does not require classification and labelling according to EU GHS Criteria.

 

 

Respiratory Irritation

MOVE 3 was tested by inhalation on rats in one acute study and in one combined repeated dose with reproduction/developmental toxicity screening test. No human exposure data by inhalation are available.

 In the acute toxicity study, no clinical signs of respiratory tract irritation and no macroscopic finding on the respiratory system were observed in rat during and/or following exposure to the vapours of MOVE 3 at the limit concentration of 20.65 mg/L.

In the combined repeated dose test and reproduction/developmental toxicity screening test study no clinical signs of respiratory irritation were observed during and following exposure and no gross pathological findings on the respiratory tract were observed at the macroscopy examination however, at the histopathological examination performed at the end of the exposure degeneration of the olfactory epithelium in the dorsal meatus of several levels in the nose (levels 3, 4, 5, and 6) was observed in in all males and females of the highest concentration (12 mg/L) group. The degeneration was minimal (12 males, 11 females) to mild (1 female) and it was characterized by disarrangement of the nuclei, increased vacuolation and decreased size of the apical part of the olfactory epithelial cells. The effects appeared not to be present in the mid and low concentration groups.

According to the described results MOVE 3 appears to be able to excert cytotoxic effect on the upper repiratory tract of the rat following repeated exposure. The relevance of the observed effect to human is unkown as the olfactory epithelium in rats is much more developed than in humans and it can be exposed to a larger air flow, potentially resulting in a more severe effects.

Basing on the results and considering that adverse effect on the olfactory ephitelium cannot be excluded in human, MOVE 3 is considered potentially irritant for the respiratory system following prolonged repeated exposure.

Justification for classification or non-classification

Skin Irritation / Corrosion

The in vitro study described in the OECD guideline 439 allows to discriminates skin irritants (Cat. 2) from chemicals not classified for skin irritation (No Cat.) in member countries or regions that do not adopt the optional UN GHS Category 3 (mild irritants), like Europe.

Irritant chemicals are identified by their ability to decrease tissue viability below 50% of the negative control. According to the results, MOVE 3 was found not-irritant to the skin (mean tissue viability = 96% compared to the negative control) therefore not requiring classification and labelling according toRegulation (EC) No. 1272/2008(EU GHS).

 

 

Serious Eye Damage / Eye Irritation

The in vitro study described in the OECD guideline 492 allows to Identify Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage according to EU GHS and UN GHS classification system.

According to OECD Guideline 492 method, chemicals not requiring classification and labelling according to UN GHS are identified as those having a tissue viability higher than (>) 60% compard to the one of the negative control. According to the results, MOVE 3 was found not requiring Classification and Labelling for Eye Irritation or Serious Eye Damage (mean tissue viability = 99% compared to the negative control) according toRegulation (EC) No. 1272/2008(EU GHS).

 

Respiratory Irritation

MOVE 3 is considered potentially irritant for the respiratory system following prolonged repeated exposure, nevertheless no signs of respiratory irritation were observed after single exposure by inhalation in the acute toxicity study conducted at the limit concentration, therefore MOVE 3 does not fulfill the classification criteria for respiratory tract irritation - STOT SE 3 according to theRegulation (EC) No. 1272/2008(EU GHS) criteria.