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EC number: 202-223-0 | CAS number: 93-15-2
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Endpoint summary
Administrative data
Description of key information
4 -allylveratrole/Methyl Eugenol was tested for its possible skin irritation potential using a three dimestional Reconstructed Human Epidermis model, EpiSkin according to OECD 439 guideline. The tissue viability met the acceptance criterion. Mean OD570of negative control was 0,646587. The viability of culture treated by positive control 5% SDS was 6.9%. The positive control met the acceptance criterion: mean tissue viability less than 20%. Determined viability of culture treated by 4 -allylveratrole/Methyl Eugenol (123 %) fulfilled the criteria for non-irritancy.
4 -allylveratrole, was tested for its possible skin corrosion potential using a three dimensional Reconstructed Human Epidermis model, EpiSkin according to OECD 431 guideline. The absolute mean OD570of the negative control tissues was 0.55533, 0.67628 and 0.45058 for the 3 -minute, 60 -minute and 240 -minute exposures, respectively. The positive control has a mean cell viability of 8.95% after 240 -minute exposure. Thus, the results of the controls indicate that the test system functioned properly.
After 240 -minute exposure the test item showed tissue viability 177 %. Taken together, the test item 4 -allylveratrole is non-corrosive in the in vitro skin corrosion test.
In vitro eye irritation test was carried out using EpiOcularTM Human tissue model with an objective to evaluateEye Irritation potential of the test item 4 -allylveratrole. The study was conducted according OECD 492 guideline and fulfilled the acceptance criteria. As the test item-treated tissues showed viability of >60.0% relative to negative control-treated tissue viability, 4 -allylveratrole was concluded to be non-irritant.
Key value for chemical safety assessment
Skin irritation / corrosion
Link to relevant study records
- Endpoint:
- skin irritation: in vitro / ex vivo
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 27 October 2017-23 November 2017
- Reliability:
- 1 (reliable without restriction)
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
- GLP compliance:
- yes
- Test system:
- human skin model
- Cell type:
- non-transformed keratinocytes
- Vehicle:
- unchanged (no vehicle)
- Details on test system:
- RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
EpiSkin Small Model kit provided by SkinEthic
- Model used: EpiDerm™ (EPI-200-SIT)
- Tissue batch number(s): 17-EKIN-044
- Delivery date: 31 October , 2017
TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: incubator 37±1°C
- Temperature of post-treatment incubation (if applicable): incubator 37±1°C, 5±1% CO2 - Control samples:
- yes, concurrent negative control
- yes, concurrent positive control
- Amount/concentration applied:
- 10 ul
- Duration of treatment / exposure:
- 15 min
- Duration of post-treatment incubation (if applicable):
- 42 h
- Number of replicates:
- Three
- Details on study design:
- The test item was tested for possible skin irritation potentiial using RHE model, EpiSkin through topical application for 15 minutes. After 42 hour post-incubation perod, irritation potential of the test item was evaluated by assessing cytotoxic (irritancy) effect. Cytotoxicity is expressed as a reduction of mitochodrial dehydrogenase activity measured by formazan production from MTT at the end of the treatment.
- Irritation / corrosion parameter:
- % tissue viability
- Value:
- 122.59
- Vehicle controls validity:
- not applicable
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Interpretation of results:
- GHS criteria not met
- Conclusions:
- The test item, Methyl Eugenol/4-allylveratrole, is non-irritant in the in vitro skin irritation test using reconstructed human epidermis.
- Executive summary:
The test item 4 -allylveratrole/Methyl Eugenol was tested for its possible skin irritation potential using a three dimestional Reconstructed Human Epidermis model, EpiSkin. The magnitude of viability was quantified by using MTT test. Validity of the test method was ascertained by positive control 5% SDS. Three tissue replicates were used for each treatment (exposure time 15 minutes), including negative and positive control.
The tissue viability met the acceptance criterion. Mean OD570 of negative control was 0,646587. The viability of culture treated by positive control 5% SDS was 6.9%. The positive control met the acceptance criterion: mean tissue viability less than 20%. Determined viability of culture treated by 4 -allylveratrole/Methyl Eugenol (122,59%) fulfilled the criteria for non-irritancy. Therefore, the is considered to be non-irritant to the skin.
Criteria for interpretation
Classification UN GHS
Mean tissue viability is≤50%
Category 2 or Category 1
Mean tissue viability > 50%
Non-Irritant
- Endpoint:
- skin corrosion: in vitro / ex vivo
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 14 August 2017- 22 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)
- Test system:
- human skin model
- Source species:
- human
- Cell type:
- non-transformed keratinocytes
- Details on animal used as source of test system:
- EpiSkin
- Vehicle:
- unchanged (no vehicle)
- Details on test system:
- Source of the test system: SkinEthic Laboratories, France.
Soon after reciept, the epidermis units were transferred to 12-well plates containing 2ml of pre-warmed medium and incubated in CO2 incubator for 43 hours. - Control samples:
- yes, concurrent negative control
- yes, concurrent positive control
- Amount/concentration applied:
- 50 ul
- Duration of treatment / exposure:
- 3 min, 60 min and 240 min
- Number of replicates:
- 2
- Irritation / corrosion parameter:
- % tissue viability
- Run / experiment:
- 3-minutes exposure
- Value:
- 173.15
- Vehicle controls validity:
- not applicable
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Irritation / corrosion parameter:
- % tissue viability
- Run / experiment:
- 60-min exposure
- Value:
- 129.94
- Vehicle controls validity:
- not applicable
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Irritation / corrosion parameter:
- % tissue viability
- Run / experiment:
- 240-min exposure
- Value:
- 176.58
- Vehicle controls validity:
- not applicable
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks:
- 8.95
- Interpretation of results:
- GHS criteria not met
- Conclusions:
- Since the tissue viability is > 35% after the 240 minute exposure, the test item 4-allyveratrole is non-corrosive under the experimental conditions described.
- Executive summary:
The test item, 4 -allylveratrole, was tested for its possible skin corrosion potential using a three dimesnsional Reconstructed Human Epidermis model, EpiSkin, through topical application. The absolute mean OD570 of the negative control tissues was 0.55533, 0.67628 and 0.45058 for the 3 -minute, 60 -minute and 240 -minute exposures, respectively. The positive control has a mean cell viability of 8.95% after 240 -minute exposure. Thus, the results of the controls indicate that the test system functioned properly.
After 240 -minute exposure the test item showed tissue viability 177 %. Taken together, the test item 4 -allylveratrole is non-corrosive in the in vitro skin corrosion test.
Referenceopen allclose all
Table 1. Skin irritation potential of 4 -allylveratrole after 15 min exposure in reconstructed human epidermis (RHe)
Sample | OD Mean | SD of OD | Viability mean % | In vivo prediction |
Negative control | 0,646587 | 0,006166 | 100 | NI |
Positive control | 0,03837 | 0,001103 | 5,93 | I |
Test item | 0,79262 | 0,001477 | 122,59 | NI |
OD= optical density
S=standard deviation
NI=non-irritant
I=irritant
OD values of individual epidermis units.
Table 1. 3 -min exposure
Absorption (OD570) | ||
R1 | R2 | |
Negative control | 0,5604 |
0,55025 |
Test item |
0,95955 |
0,9635 |
Table 2. 60 -minutes exposure
Absorption (OD570) | ||
R1 | R2 | |
Negative control | 0,66795 | 0,6846 |
Test item | 0,771 | 0,9862 |
Table 3. 240 -minutes exposure
Absorption (OD570) | ||
R1 | R2 | |
Negative control | 0,4484 | 0,4575 |
Test item | 0,79745 | 0,79385 |
Positive control | 0,04195 | 0,03865 |
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (not irritating)
Eye irritation
Link to relevant study records
- Endpoint:
- eye irritation: in vitro / ex vivo
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- Study initiation date: 18 Septermber 2017. Study completion date: 30 November 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)
- GLP compliance:
- yes (incl. QA statement)
- Species:
- human
- Details on test animals or tissues and environmental conditions:
- EpiOcular TM Tissue from MatTek, lot number 27005. Keraticocyte strain 4F1188.
Tissues were incubated at 37±1oC, 5% CO2 and humidified atmosphere. - Vehicle:
- unchanged (no vehicle)
- Controls:
- yes, concurrent positive control
- yes, concurrent negative control
- Amount / concentration applied:
- 50 ul
- Duration of treatment / exposure:
- 30 minutes
- Duration of post- treatment incubation (in vitro):
- 120+-15 minutes
- Number of animals or in vitro replicates:
- Two tissues, two aliquots
- Details on study design:
- Assessment of Direct MTT reduction by the Test Item
A quantity of 50 μL of liquid test item was added to 1 mL of MTT solution (1.0 mg / mL) in a 6 well plate and incubated in dark at standard culture conditions for 3 hours. Concurrent negative control (50 μL sterile deionized
water) was tested along with test item. Post incubation, test item did not convert the MTT solution to blue or purple color, indicates test item is not reducing the MTT solution.
Assessment of Colored or Staining Materials
Colored test item or test items which become colored after application to the tissues may interfere with the quantitative MTT measurement. Therefore, the test items are checked for its colorant properties.
In this context, chemicals which absorb light and appear red, yellow, green, and blue dark purple and black are considered as intrinsic colorants.
a. Blue, dark purple and black test items: As the test item is not colored,test was not performed.
b. Other intrinsically colored test items (e.g. red, yellow, green colorants): A volume of 50 μL test item was added to 2 mL of isopropanol, incubated in 6-well plates, and placed on an orbital plate shaker for 2 hours and 50 minutes at room temperature. Two 200 μL aliquots of isopropanol solutions (mixed with test item) and of pure isopropanol was transferred to a 96-well plate and the absorbance was measured with a plate reader and tested for their ability to absorb significantly light at 570 nm (wavelength used for the MTT determination). 200 μL aliquots of the mixture was used for measurement and measured the OD at 570 nm.
After subtraction of the OD for isopropanol, the OD of the test item solution is not > 0.08 (approximately 5% of the mean viability of the negative control), hence test item is considered as non-interactive with the
MTT measurement and an additional test on colorant controls (CC) was not performed.
c. Non coloured test item: As the test item is non-colored after contact with water or isopropanol, it was considered as non-interactive with the MTT. Measurement and an additional test on colorant controls (CC) were not
performed.
Preparation of EpiOcular Tissues for Treatment
On the day of tissue receipt (19 September 2017) the tissues in its 24-well plate shipping container, were equilibrated to room temperature for about 15 minutes. EpiOcular™ assay medium (1 mL/well) was added to 6 well plates and warmed to approximately 37ºC in a CO2 incubator. Each 12 well shipping containers were removed from the plastic bag under sterile conditions and surface disinfected by wiping with 70% ethanol. Each tissue in the 12 well plates was inspected for air bubbles between the insert and the agarose gel. The tissues were removed carefully from the 12 well plate using sterile forceps, agarose adhering to insert was removed gently by blotting on to sterile filter paper and placed in the 6 well plate containing 1 mL of EpiOcular™ assay solution for 1 hour. After 1 hour, assay medium was replaced with fresh assay medium and incubated further for 16 hours at standard culture conditions - Irritation parameter:
- in vitro irritation score
- Run / experiment:
- The mean tissue viability (%)
- Value:
- 144.08
- Vehicle controls validity:
- not applicable
- Negative controls validity:
- valid
- Remarks:
- 100.0
- Positive controls validity:
- valid
- Remarks:
- 30.75
- Interpretation of results:
- GHS criteria not met
- Conclusions:
- The test item showed viability of >60% relative to negative control-treated viability. Thus, the test item, 4-allylveratrole, was concluded to be non-irritant in the EpiOcularTM test.
- Executive summary:
A guideline compliant eye irritation study (OECD 492) –in vitro eye irritation in Reconstructed Human Cornea-like Epithelium (RhCE)) was conducted. Aim of the study was to examine the acute eye irritation potential of the test item by measurement of its cytotoxic effect on the EpiOcular™ cornea epithelial model. The EpiOcular tissue construct is a nonkeratinized epithelium prepared from normal human keratinocytes (MatTek).
In this assay, the test item was applied to the surface of the cornea epithelial construct for 30 minutes. After exposure period, each tissue was extensively rinsed, and the tissue was allowed to express the resulting damage. Two construct tissues were used for the treatment with test item and for negative and positive controls. The MTT assay was performed to determine tissue viability. The tissues were transferred in the 24-wells plate containing MTT medium and incubated in a humidified atmosphere for 3 hours. After incubation the blue formazan salt formed was extracted in isopropanol and the optical density of extracted formazan was determined using a spectrophotometer at 540 nm. Validity of the test method was ascertained by positive control (methyl acetate) and negative control (deionized sterile water). The tissue viability met the acceptance criterion as the mean OD of negative controlwas in between > 0.8 and < 2.5. The positive control met the acceptance criterion as the viability of culture treated by methyl acetate was 30.8% (less than 60%).The viability of culture treated by 4-allylveratrole was 144.1%. Therefore, 4-allylveratrole is considered to be non-irritant to the eye.
Reference
Table 1. OD values of individual epiocular tissues.
Tissue 1 | Tissue 2 | |||
Sample | R1 | R2 | R1 | R2 |
Negative control | 1.3252 | 1.2966 | 0.9017 | 0.8696 |
Positive control | 0.3901 | 1.6199 | 1.5355 | 1.5177 |
Test item | 1.5845 | 1.6199 | 1.5355 | 1.5177 |
Table 2. The percentage viablity of the epiocular tissues.
Viability Tissue 1 |
Viability Tissue 2 | ||||||||
Sample | Ali 1 | Ali 2 | Mean 1 | Ali 1 | Ali 2 | Mean | Mean 1&2 | Diff 1 & 2 | Classification |
NC | 121,46 | 118,75 | 120,11 | 81,41 | 78,38 | 79,89 | 100,00 | 40,21 | NI |
PC | 33,03 | 31,24 | 32,13 | 28,95 | 29,80 | 29,37 | 30,75 | 2,76 | I |
Test item | 145,98 | 149,33 | 147,65 | 141,35 | 139,66 | 140,50 | 144,08 | 7,15 | NI |
NC: Negative control (deionized water), PC: Positive control (methyl acetate), I: Irritant, NI:Non-irritant, Ali: Aliquot, Diff: difference of viability between two tissues.
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (not irritating)
Additional information
Justification for classification or non-classification
The results of the key studies (OECD439 & 431) do not indicate the substance to be classified for skin irritant according to CLP Regulation 1272/2008.
The result of the key study (OECD492) does not indicate the substance to be classified for eye irritant according to CLP Regulation 1272/2008.
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