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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: - | CAS number: -
- 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
Skin sensitisation
Administrative data
- Endpoint:
- skin sensitisation: in vivo (LLNA)
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- from October 5 to October 26, 2016
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 016
- Report date:
- 2017
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.2600 (Skin Sensitisation)
- GLP compliance:
- yes (incl. QA statement)
- Type of study:
- mouse local lymph node assay (LLNA)
Test material
- Reference substance name:
- Reaction products of 3-Methylaniline with heptyl naphthalen-2-ol
- Cas Number:
- 1619917-05-3
- Molecular formula:
- Not applicable (UVCB Substance)
- IUPAC Name:
- Reaction products of 3-Methylaniline with heptyl naphthalen-2-ol
- Test material form:
- solid
Constituent 1
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- batch No.of test material: 78-231-15
- Expiration date of the batch: July 2017
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature, protected from light
In vivo test system
Test animals
- Species:
- mouse
- Strain:
- CBA/Ca
- Sex:
- female
- Details on test animals and environmental conditions:
- TEST ANIMALS
- Source: Envigo (formed partially from Harlan in September 2015), 5800 AN Venray, The Netherlands
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 8-9 weeks
Housing and Feeding Conditions
- Full barrier in an air-conditioned room
- Temperature: 22 ± 3 °C
- Relative humidity: 55 ± 10%
- Artificial light, sequence being 12 hours light, 12 hours dark
- Air change: at least 10 x / hour
- Free access to Altromin 1324 maintenance diet for rats and mice
- Free access to tap water, sulphur acidified to a pH value of approx. 2.8 (drinking water, municipal residue control, microbiological controls at regular intervals)
- The animals were kept in groups of 5 animals in IVC cages, type II L, polysulphone cages on Altromin saw fibre bedding
- Certificates of food, water and bedding are filed for two years at BSL Munich and afterwards archived at Eurofins Munich
- Adequate acclimatisation period (at least five days) under laboratory conditions
Study design: in vivo (LLNA)
- Vehicle:
- acetone/olive oil (4:1 v/v)
- Concentration:
- Based on the results observed in the prescreen test the following test item concentrations were selected for the main study:
3%, 6% and 12% (w/v)
The preparations were made immediately prior to each dosing. - No. of animals per dose:
- 15 / main test
5 / prescreen test - Details on study design:
- PRE-SCREEN TESTS:
The maximum technically applicable concentration of the test item in the vehicle was found to be 50% in AOO (Acetone, Sigma-Aldrich, lot no. SZBF3480V, expiry date: 08/2021; olive oil highly refined, Sigma-Aldrich, lot no. BCBQ4885V, expiry date: 17/10/2016).
In order to determine the highest tolerated and not excessively irritant test concentration a prescreen test was performed which was conducted under the same conditions as the main study, except there was no assessment of lymph node proliferation. The mice were observed daily for any clinical signs of systemic toxicity or local irritation at the application site. Body weights were recorded pre-test and prior to termination. Both ears were observed for erythema and scored.
Ear thickness measurements were performed on day 1 (pre-dose), day 3 (approximately 48 hours after the first dose) and day 6. Excessive local irritation was indicated by an erythema score ≥ 3 and/or ear swelling of ≥ 25%.
Two animals were treated by topical application with the test item on three consecutive days at a concentration of 50% (diluted with AOO) to the entire dorsal surface of each ear.
Two animals were treated by topical application with the test item on three consecutive days at a concentration of 12.5% (diluted with AOO) to the entire dorsal surface of each ear.
One further animal was treated with 100% AOO and served as negative control.
Immediately before the first application, approximately 48 hours after the first application and shortly before sacrificing the thickness of both ears of all animals was measured
During this period also all clinical signs were recorded.
Cageside observations included spontaneous activity, lethargy, recumbent position, convulsions, tremors, apnoea, asphyxia, vocalisation, diarrhoea, changes in the skin and fur, eyes and mucous membranes (salivation, discharge).
The animals treated with the test item at a concentration of 50% in AOO showed signs of systemic toxicity from day 2 to day 5 (moving the bedding) and signs of excessive irritation (increased ear thickness > 25%) at both application sites on day 3 and day 6.
The animals treated with the test item at a concentration of 12.5% in AOO showed slight signs of irritation at both application sites on day 6.
As in all animals both application sites were coloured red due to residual test item from day 2 onwards, skin irritation in terms of erythema could not be evaluated.
No signs of irritation were detected in the animal treated with the negative control.
All animals showed the expected weight development, which includes a weight loss of up to 2 g throughout the duration of the prescreen test
MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
The animals were randomly selected using the validated departmental computerised system E WorkBook (version 9.4.0, ID Business Solutions Ltd.).
Identification was ensured by cage number and individual marking (tail).
- Criteria used to consider a positive response:
The proliferative response of lymph node cells was expressed as the number of radioactive disintegrations per minute per lymph node (DPM/NODE) and as the ratio of 3H-methyl thymidine - incorporation into lymph node cells of test group animals relative to that recorded for control group animals (STIMULATION INDEX). Before DPM/NODE values were determined, background values were subtracted.
EC3 values, calculated concentrations which induce stimulation indices of three, are determined by linear interpolation, EC3=c+[(3-d)/(b-d)]x(a c), between two points of the stimulation indices axis, one above (a,b) and one below (c,d) the stimulation index of three [10], [11]. If all measured points are above or below the stimulation index of three, no EC3 value can be stated.
In certain situations where the dose response does not incorporate a data point lying below the SI value of three, provided the data are of good quality (relatively close to an SI of three and evidence of a dose response), an EC3 value may be estimated by using the two doses closest to the SI value of three. The EC3 value is estimated by log-linear interpolation between these two points on a plane where the x-axis represents the dose level and the y-axis represents the SI. The point with the higher SI is denoted (a,b) and the point with the lower SI is denoted (c,d). The formula for the EC3 estimate is as follows: EC3=2^{(log2(c)+(3-d)/(b-d)*[(log2(a)-log2(c)]}, by log-transforming the doses, EC3 estimates will never fall below zero [11], [12].
A substance is regarded as a 'sensitiser' in the LLNA if at least one concentration of the test item results in a 3-fold or greater increase in 3H-methyl thymidine - incorporation into lymph node cells of the test group animals, relative to that recorded for the lymph nodes of control group animals (Stimulation Index equal to or greater than 3.0).
On the basis of the test results, the test substance may be classified into one of the following categories in conformity with the criteria given in Commission Regulation (EU) No 286/2011 [8] as well as in GHS - Globally Harmonised System of Classification and Labelling of Chemicals, sixth revised edition, 2015 [10]:
Skin sensitiser
Category 1:
A substance is classified as a skin sensitiser
a) if there is evidence in humans that the substance can lead to sensitisation by skin contact in a substantial number of persons, or
b) if there are positive results from an appropriate animal test.
WARNING, exclamation mark. May cause an allergic skin reaction.
Sub-category 1A:
Substances showing a high frequency of occurrence in humans and/or a high potency in animals can be presumed to have the potential to produce significant sensitisation in humans. Severity of reaction may also be considered.
EC3 value ≤ 2%
WARNING, exclamation mark. May cause an allergic skin reaction.
Sub-category 1B:
Substances showing a low to moderate frequency of occurrence in humans and/or a low to moderate potency in animals can be presumed to have the potential to produce sensitisation in humans. Severity of reaction may also be considered.
EC3 value > 2%
WARNING, exclamation mark. May cause an allergic skin reaction.
TREATMENT PREPARATION AND ADMINISTRATION:
Topical Application
Each mouse was treated by topical application of 25 µL of the selected solution to the entire dorsal surface of each ear.
Topical applications were performed once daily over three consecutive days. The first treatment day is defined as study day 1.
Administration of 3H-Methyl Thymidine
Five days after the first topical application all mice were dosed with 20 µCi 3H-methyl thymidine by intravenous injection (tail vein) of 250µL of 3H-methyl thymidine, diluted with PBS to a working concentration of 80µCi/mL.
Preparation of Cell Suspension
Approximately 5 hours after the injection of 3H-methyl thymidine all mice were sacrificed by cervical dislocation. The draining auricular lymph nodes were excised, individually pooled for each animal (2 lymph nodes per animal) and collected in phosphate buffered saline (PBS). A single cell suspension of pooled lymph node cells was prepared by gentle mechanical disaggregation through polyamide gauze (200 mesh size). After washing the gauze with PBS the cell suspension was pelleted in a centrifuge. The supernatant was discarded and the pellets were resuspended with PBS. This washing procedure was repeated.
After the final wash each pellet was resuspended in approx. 1 mL 5% TCA at approx. 4° C for approximately 18 hours for precipitation of macromolecules. Each precipitate was once washed again, resuspended in 1 mL 5% TCA and 7 mL scintillation fluid was added. Then this solution was transferred into scintillation vials and stored at room temperature overnight.
Determination of Incorporated 3H -Methyl Thymidine
The 3H-methyl thymidine – incorporation was measured in a ß-counter and expressed as the number of disintegrations per minute (DPM). Similarly, background 3H-methyl thymidine levels were also measured (5% TCA). Determination of radioactivity was performed individually for each animal. - Positive control substance(s):
- other: phenylene-diamine in AOO
Results and discussion
- Positive control results:
- The positive-control substance exceeded the stimulation index of 3 confirming the reliability of the test system .
In vivo (LLNA)
Resultsopen allclose all
- Parameter:
- SI
- Value:
- 5
- Test group / Remarks:
- at a concentration of 3%
- Parameter:
- SI
- Value:
- 10.3
- Test group / Remarks:
- at concentration of 6%
- Parameter:
- SI
- Value:
- 16.4
- Test group / Remarks:
- at concentration of 12%
- Cellular proliferation data / Observations:
- Each of the three tested concentrations of the test item exceeded the stimulation index of 3.
DETAILS ON STIMULATION INDEX CALCULATION
Ratio of 3H-methyl thymidine - incorporation into lymph node cells of test group animals relative to that recorded for control group animals
EC3 CALCULATION
EC3 values, calculated concentrations which induce stimulation indices of three, are determined by linear interpolation, EC3=c+[(3-d)/(b-d)]x(a c), between two points of the stimulation indices axis, one above (a,b) and one below (c,d) the stimulation index of three [10], [11]. If all measured points are above or below the stimulation index of three, no EC3 value can be stated.
In certain situations where the dose response does not incorporate a data point lying below the SI value of three, provided the data are of good quality (relatively close to an SI of three and evidence of a dose response), an EC3 value may be estimated by using the two doses closest to the SI value of three. The EC3 value is estimated by log-linear interpolation between these two points on a plane where the x-axis represents the dose level and the y-axis represents the SI. The point with the higher SI is denoted (a,b) and the point with the lower SI is denoted (c,d). The formula for the EC3 estimate is as follows: EC3=2^{(log2(c)+(3-d)/(b-d)*[(log2(a)-log2(c)]}, by log-transforming the doses, EC3 estimates will never fall below zero
CLINICAL OBSERVATIONS:
All animals survived throughout the test period without showing any clinical signs.
BODY WEIGHTS
All animals showed the expected weight development, which includes a weight loss of up to 2 g throughout the study.
Applicant's summary and conclusion
- Interpretation of results:
- Category 1B (indication of skin sensitising potential) based on GHS criteria
- Conclusions:
- There was no mortality and there were no significant clinical observations or effects on body weights.
Each of the three tested concentrations exceeded the stimulation index of 3.
The stimulation index at a concentration of 3% was 5.0
The stimulation index at a concentration of 6% was 10.3
The stimulation index at a concentration of 12% was 16.4
The EC3 value (estimated by linear interpolation) was calculated to be at a test item concentration of 2.3%.
Consequently, according to OECD 429 solutions or preparations containing more than 2.3% RED 2596 are expected to have a stimulation index of >3 and are therefore considered to be dermal sensitisers.
According to Commission Regulation (EU) No 286/2011 [7] as well as GHS (Globally Harmonized Classification System) [10] the test item RED 2596 has obligatory labelling requirement for skin sensitisation and is classified into Category 1B. - Executive summary:
Based on the results of the prescreen test the test item was assessed for sensitising properties at concentrations of 3%, 6% and 12% (w/v), each diluted with AOO 4:1 (v/v), 4 parts acetone and 1 part olive oil.
Species/strain: Mice, CBA/CaOlaHsd
Number of animals: 15 / main test; 5 / prescreen test
Vehicle: AOO (4:1 (v/v) acetone/olive oil)
There was no mortality and there were no significant clinical observations or effects on body weights.
Each of the three tested concentrations exceeded the stimulation index of 3.
The stimulation index at a concentration of 3% was 5.0
The stimulation index at a concentration of 6% was 10.3
The stimulation index at a concentration of 12% was 16.4
The EC3 value (estimated by linear interpolation) was calculated to be at a test item concentration of 2.3%.
Consequently, according to OECD 429 solutions or preparations containing more than 2.3% RED 2596 are expected to have a stimulation index of >3 and are therefore considered to be dermal sensitisers.
According to Commission Regulation (EU) No 286/2011 [7] as well as GHS (Globally Harmonized Classification System) [10] the test item RED 2596 has obligatory labelling requirement for skin sensitisation and is classified into Category 1B.
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.