Registration Dossier
Registration Dossier
Data platform availability banner - registered substances factsheets
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.
The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.
Diss Factsheets
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 256-233-5 | CAS number: 45320-65-8
- 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
N,N’-[(methylimino)bis(trimethylene)]bis(oleamide) is not irritating to skin and the eye.
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:
- 2017-10-24 to 2017-11-27
- 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
- Test system:
- human skin model
- Source species:
- human
- Cell type:
- non-transformed keratinocytes
- Justification for test system used:
- Recommended by the OECD testing guideline 439
- Vehicle:
- unchanged (no vehicle)
- Details on test system:
- RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EPISKIN™ - 0.38 cm2
- Tissue batch number(s): 17-EKIN-047 (alive tissues) and 17-EKIN-028 (killed tissues)
- Production date:
- Shipping date:
- Delivery date: 21 November 2017
- Date of initiation of testing:
TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: room temperature
- Temperature of post-treatment incubation (if applicable): 37 °C
REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: each tissue was rinsed with approximately 25mL of sterile D-PBS
- Observable damage in the tissue due to washing:
MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 2 mL of MTT ready-to-use solution (0.3 mg/mL)
- Incubation time: 3 hours at 37 °C, 5% CO2
- Spectrophotometer: no further information
- Wavelength: 595 nm
- Linear OD range of spectrophotometer: yes
FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
no data available
NUMBER OF REPLICATE TISSUES: 3 for live tissues
CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- Fresh tissues / killed tissues : yes
- Procedure used to prepare the killed tissues (if applicable): stored at -20°C.
- N. of replicates : 2 for killed tissues
- Method of calculation used:
NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION:
PREDICTION MODEL / DECISION CRITERIA
Mean relative viability ≤ 50% UN GHS Category 2 or 1
Mean relative viability > 50% UN GHS No Category (for member states
that do not adopt optional category 3) - Control samples:
- yes, concurrent vehicle
- yes, concurrent positive control
- Amount/concentration applied:
- TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 20±2mg
NEGATIVE CONTROL
- Amount(s) applied (volume or weight): D-PBS 20 µL
POSITIVE CONTROL
- Amount(s) applied (volume or weight): 20 µL
- Concentration (if solution): 5% (w/v) SDS - Duration of treatment / exposure:
- 15±0.5 minutes
- Duration of post-treatment incubation (if applicable):
- 42 ± 1 hour
- Number of replicates:
- triplicates
- Irritation / corrosion parameter:
- % tissue viability
- Run / experiment:
- test item
- Value:
- 92
- 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:
- - OTHER EFFECTS:
- Direct-MTT reduction:
In a first step, the test item was assayed for the ability of reducing MTT per se. Purple precipitate was noted in the MTT solution at the end of the incubation period, indicating that the test item could direct interact with MTT.
- Colour interference with MTT:
In a second step, the test item was assayed for the ability of colouring water per se.
A colourless, opaque suspension was observed, indicating that the test item has no potential interfering ability.
DEMONSTRATION OF TECHNICAL PROFICIENCY:
Amounts of 20 mg or 20 μL of test item or positive control item were used for treatment during the demonstration of laboratory proficiency, in order to uniformly cover the epidermis surface while avoiding an infinite dose (as indicated in OECD 439, § 24).
Results obtained demonstrated that test method has been successfully established.
ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes
- Acceptance criteria met for variability between replicate measurements: yes - Interpretation of results:
- GHS criteria not met
- Conclusions:
- N,N'-[(methylimino)bis(trimethylene)] bis(oleamide) was not irritating in the in vitro skin irritation test.
- Executive summary:
The potential of the test item N,N’-[(methylimino)bis(trimethylene)]bis(oleamide) to be irritant to the skin was investigated according to OECD Guideline 439 (28 July 2015) through an in vitro skin irritation study, using a commercial reconstructed human epidermis (RhE) model named EPISKIN™.
The test item N,N’-[(methylimino)bis(trimethylene)]bis(oleamide) (100% a.i.) was tested for its ability to impair cell viability. The test item was applied as supplied by the sponsor in three tissue replicates at the treatment level of 20 ± 2 mg/epidermis unit, each one measuring 0.38cm² (treatment level: 53mg/cm²).
Before the Main Assay, a preliminary test was carried out to evaluate the compatibility of the test item with the test system. In a first step, the test item was assayed for the ability of reducing MTT per se. Purple precipitate was noted in the MTT solution at the end of the incubation period, indicating that the test item could direct interact with MTT. In a second step, the test item was assayed for the ability of colouring water per se. A colourless, opaque solution was observed, indicating that the test item has no potential interfering ability. Based on these results, an additional control for non specific MTT reduction (NSMTT) was added in the Main Assay, using two killed tissues and compared with negative control performed with alive tissues.
In the Main Assay, the negative control gave the expected baseline value (Optical Density values of the three replicates higher than 0.6) and variability [Standard Deviation (SD) of % viability lower or equal to 18], in agreement with the guideline indications. According to the method, the negative control mean value is considered the baseline value of the experiment and thus represents 100% of cell viability.
The positive control caused the expected cell death (5% of cell viability when compared to the negative control) and variability (SD of % viability equal to 0.8). Based on the stated criteria (mean viability ≤ 40% and SD of % viability ≤ 18), the assay was regarded as valid.
The NSMTT value was 1%, thus only the OD-blank background subtraction was performed. The test item did not induce cell death in any replicate, the mean cell viability after the blank subtraction was 92% when compared to the negative control. Intra-replicate variability was acceptable with a SD of % viability value equal to 1.7 (lower than 18, as stated in the Study Protocol).
Based on the results obtained, the test item N,N’-[(methylimino)bis(trimethylene)] bis(oleamide) is classified as non-irritant to the skin (UN GHS No Category).
Reference
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:
- 2018-09-03 to 2019-09-13
- 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)
- Details on test animals or tissues and environmental conditions:
- - Justification of the test method and considerations regarding applicability
The EpiOcular™ Eye Irritation Test (EIT) predicts the acute eye hazard potential of chemicals by measurement of its tissue damage caused by cytotoxic effects in the reconstructed human cornea-like tissue model.
Within a tired testing strategy, the EpiOcular™ EIT is used as areplacement of the in vivo Draize Eye Irritation Test.
It is utilized for the classification and labelling of ehernieals concerning their eye hazard potential.
The EpiOcular™ EIT can be used to identify ehernieals that do not require classification for eye irritation or serious eye damage according to the UN GHS classification system.
A limitation of this guideline is that it neither allows discrimination between eye irritation/reversible effects on the eye (Category 2) and serious eye damage/irreversible effects on the eye (Category 1), nor between eye irritants (optional Category 2A) and mild eye irritants (optional Category 2B). For these purposes, further testing with other suitable test methods is required.
- Description of the cell system used, incl. certificate of authenticity and the mycoplasma status of the cell live
The EpiOcular™ tissue consists of normal, human-derived keratinocytes which have been cultured to form a stratified squamous epithelium similar to that found in the human cornea.
It consists of highly organized basal cells. These cells are not transformed or transfected with genes to induce an extended life span.
The EpiOcular™ tissues are cultured in specially prepared cell culture inserts with a porous membrane through which nutrients can pass to the cells.
Certificate of analysis is available for tissues, confirming cell source, absence of biological contaminants and analysis of quality and functionality. - Vehicle:
- unchanged (no vehicle)
- Controls:
- yes, concurrent positive control
- yes, concurrent negative control
- Amount / concentration applied:
- Test substance:
50 mg
Positive and negative control: 50 µl for each - Duration of treatment / exposure:
- 6 hours
- Duration of post- treatment incubation (in vitro):
- 18 hours
- Number of animals or in vitro replicates:
- duplicate
- Details on study design:
- Details of the test procedure used
- RhCE tissue construct used, including batch number
EplOcular™Tissue, MatTek Lot: 27029
- Doses of test chemical and control substances used : 50 mg of test substance, each 50µL of negative and positive control
- Duration and temperature of exposure, post-exposure immersion and post-exposure incubation periods (where applicable)
After dosing the last tissue, all plates were transferred into the incubator for 6 hours at 37 ± 1 °C, 5 ± 1 % CO2 and ~ 95 % relative humidity.
At the end of exposure time, the inserts were removed from the plates in one-minuteintervals using sterile forceps and rinsed immediately. The inserts were thoroughly rinsed with DPBS.
Then, the tissues were immediately transferred into 5 mL of assay medium in pre-Iabelled 12-well plate for 25 minutes post soak at room temperature.
For post-treatment incubation, the tissues were incubated for 18 hours at 37 ± 1 °C, 5 ± 1 % C02 and ~ 95 % relative humidity.
- Indication of controls used for direct MTT-reducers and/or colouring test chemicals (if applicable)
The test item was tested for the ability of direct MTT reduction.
The colour of the MTT turned blue/purple, therefore the test item is presumed to have reduced the MTT.
- Number of tissue replicates used per test chemical and controls (positive control, negative control, NSMTT, NSCliving and NSCkilled, if applicable)
duplicate tissues were used
- Wavelength and band pass (if applicable) used for quantifying MTT formazan, and linearity range of measuring device (e.g. spectrophotometer)
The wavelength used in the test is 570 nm.
Quarterly, a calibration of the microplate reader by using a calibration plate provided by the manufacturer is performed in order to ensure the linearity of photometric measurements.
The plate photometer has an indication range for optical density (OD) between 0.0 and 3.3 and a linear range of 0.1 - 2.5. Only measurements made within the linear range are used for the quantitative calculations.
- Description of the method used to quantify MTT formazan
Microtiter plate photometer, Anthos Reader 2010 Flexi, Anthos Microsysteme GmbH
- Reference to historical positive and negative control results demonstrating suitable run acceptance criteria
Values for negative control and for positive control were within the range of historical data of the test facility.
All validity criteria were met, therefore the experiment is considered valid.
- Demonstration of proficiency in performing the test method before routine use by testing of the proficiency chemicals
The validity of the EpiOcular™ test at test lab was demonstrated in a proficiency study.
For this purpose 15 proficiency ehernieals (indicated by the OECD 492 guideline) were tested. All of the 15 proficiency ehernieals were correctly categorized.
- Positive and negative control means and acceptance ranges based on historical data
% mean relative viability of positive control: < 50 % of negative control
- Acceptable variability between tissue replicates for positive and negative controls
< 20 % (test: 0.3% (negative control); 3.3% (positive control)
- Acceptable variability between tissue replicates for the test chemical
< 20 % (test: 1.1 % (test item) - Irritation parameter:
- other: % Tissue Viability Mean
- Run / experiment:
- Experiment 1
- Value:
- 98.3
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- no indication of irritation
- Other effects / acceptance of results:
- The test item did not show any colour development in isopropanol.
The result of this pre-test was that the colour of the MTT turned blue/purple, therefore the test item has shown its ability to reduce MTT.
Therefore, it was necessary to perform an additional test with freeze killed tissues that possess no metabolic activity but absorb and bind the test item like viable tissues.
Results showed that the MTT reduction by the test item did not influence the result of the main test.
All validity criteria were met. The criterion for optical density of the negative control was fulfilled: The OD value was 2.0 (> 0.8 and < 2.5).
The positive control induced a decrease in tissue viability as compared to the negative control to 28.0%.
The variation within the replicates of the controls and the test item was acceptable (< 20%).
For these reasons, the result of the test is considered valid. - Interpretation of results:
- GHS criteria not met
- Conclusions:
- Under the conditions of the test, N,N'-[(methylimino)bis(trimethylene)]bis(oleamide) is considered non-eye irritant in the EpiOcular™ Eye Irritation Test.
- Executive summary:
An in-vitro eye irritation study was conducted using the EpiOcular™ Reconstructed human Cornea-like Epithelium (RhCE) test method according to OECD guideline 492 with the test substance N,N'-[(methylimino)bis(trimethylene)]bis(oleamide).
Since in the pre-test, potential MTT reduction by the test item was observed, an additional test was performed to correct photometrical measurement values due to direct MTT reduction by the test item. The results of the additional test showed that the MTT reduction by the test item did not influence the result of the main test.
The test item N,N'-[(methylimino)bis(trimethylene)]bis(oleamide) was applied to a three dimensional human cornea tissue model in duplicate for an exposure time of 6 hours.
After treatment, the test item was rinsed off the tissue; then, cell viability of the tissues was evaluated by addition of MTT, which can be reduced to formazan. The formazan production was evaluated by measuring the optical density (OD) of the resulting solution.
Demineralised water was used as negative control and methyl acetate was used as positive control.
After treatment with the negative control, the absorbance values were within the required acceptability criterion of mean OD > 0.8 and <2.5, the OD was 2.0.
The positive control showed clear eye irritating effects, the mean value of the relative tissue viability was 28.0% (< 50%).
The variation within tissue replicates of the controls and the test item was acceptable(< 20%).
After treatment with the test item, the mean value of relative tissue viability was 98.3%. This value is well above the threshold for eye irritation potential (≤ 60%).
Test items that induce values above the threshold are considered non-eye irritant.
Under the conditions of the test, N,N'[(methylimino)bis(trimethylene)]bis(oleamide) is considered non-eye irritant in the EpiOcular™ Eye Irritation Test.
Reference
Absorbance Values Negative Control, Positive Control and Test Item (OD at 570 nm)
|
Measurement |
Negative Control |
Positive Control |
Test substance |
Tissue 1 |
1 |
1.989 |
0.545 |
2.005 |
|
2 |
2.044 |
0.572 |
1.935
|
Tissue 2 |
1 |
2.025 |
0.623 |
1.987 |
|
2 |
1.997 |
0.624 |
1.998 |
Absorbance Values of Negative Control and Test Item with Freeze-killed Tissues (OD at 570 nm)
|
Measurement |
Negative Control |
Test substance |
Tissue 1 |
1 |
0.078 |
0.066 |
2 |
0.076 |
0.063 |
|
Tissue 2 |
1 |
- |
0.073 |
2 |
- |
0.073 |
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (not irritating)
Additional information
Skin:
The potential of the test item N,N’-[(methylimino)bis(trimethylene)]bis(oleamide) to be irritant to the skin was investigated according to OECD Guideline 439 (28 July 2015) through an in vitro skin irritation study, using a commercial reconstructed human epidermis (RhE) model named EPISKIN™.
The test item did not induce cell death in any replicate, the mean cell viability after the blank subtraction was 92% when compared to the negative control.
The validity criteria were fulfilled, the assay was regarded as valid.
Based on the results obtained, the test item N,N’-[(methylimino)bis(trimethylene)] bis(oleamide) is classified as non-irritant to the skin (UN GHS No Category).
Eye:
Data from an OECD 437 guideline study withN,N’-[(methylimino)bis(trimethylene)]bis(oleamide) are available. Under the conditions of this study, the test item N,N'-[(methylimino)bis(trimethylene)]bis(oleamide) showed effects on the cornea of the bovine eye.
The calculated IVIS (in vitro irritancy score) is 15.70.
According to OECD Guideline no. 437 (Jul. 2013), a substance with an IVIS > 3 and ≤ 55 induces effects on the cornea, that cannot be classified in an UN GHS Category.
Thus, no prediction can be made based on study results.
Therefore an in-vitro eye irritation study was conducted using the EpiOcular™ Reconstructed human Cornea-like Epithelium (RhCE) test method according to OECD guideline 492.
After treatment with the test item, the mean value of relative tissue viability was 98.3%. This value is well above the threshold for eye irritation potential (≤ 60%). Test items that induce values above the threshold are considered non-eye irritant.
The validity criteria were fulfilled, the assay was regarded as valid.
Under the conditions of the test, N,N'[(methylimino)bis(trimethylene)]bis(oleamide) is considered non-eye irritant in the EpiOcular™ Eye Irritation Test.
Justification for classification or non-classification
N,N’-[(methylimino)bis(trimethylene)]bis(oleamide) does not need to be classified for skin or eye irritation according to the criteria of CLP, EU GHS (Regulation (EC) No 1272/2008), based on data from a study according to OECD Guideline 439 for skin irritation and data from a study according to OECD Guideline 492 for eye irritation.
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.