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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 228-762-1 | CAS number: 6358-09-4
- 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 irritation / corrosion
Administrative data
- Endpoint:
- skin corrosion: in vitro / ex vivo
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- From 24 April 2017 to 8 June 2017
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 017
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
- Version / remarks:
- Deviation 1
A third correction group was required to be added to the study design due to the potential interference with the MTT reduction assay by the test item. Following an inconclusive result during the pre-test for the direct reduction of MTT, a further correction group (in addition to the water-killed tissues for correcting direct MTT reduction and viable tissues for correcting color interference) was necessary in order to avoid a double correction.
Assessment of Color Interference in non-viable tissues:
For each exposure period, the additional group was composed of two test item treated freeze killed tissues that underwent the entire testing procedure with the exception that the MTT incubation period was replaced by incubation with assay medium (therefore without MTT). For each exposure period, two freeze killed tissues were treated with sterile water for negative control purposes. The optical density measurements from these tissues were then assessed for possible quantitative correction of the results.
Deviation 2
An assessment of the test item’s capability to directly reduce MTT was inconclusive due to the intrinsic color of the test item and therefore, an additional procedure using freeze-killed tissues was performed. However, the results obtained showed that negligible interference due to direct reduction of MTT occurred.
Deviation 3
The solution containing the test item was a yellow to green color, therefore additional color correction tissues were incorporated into the testing procedure. However, the results obtained showed that no color interference occurred. It was therefore considered unnecessary to use the results of the color correction tissues for quantitative correction of results or for reporting purposes. As the results of the color correction tissues were not used it was therefore unnecessary to use the results of the killed color correction tissues, as no double correction for color interference would have occurred. - Deviations:
- yes
- Remarks:
- See remarks above
- GLP compliance:
- yes (incl. QA statement)
Test material
- Reference substance name:
- 2-amino-6-chloro-4-nitrophenol
- EC Number:
- 228-762-1
- EC Name:
- 2-amino-6-chloro-4-nitrophenol
- Cas Number:
- 6358-09-4
- Molecular formula:
- C6H5ClN2O3
- IUPAC Name:
- 2-amino-6-chloro-4-nitrophenol
- Test material form:
- solid: particulate/powder
Constituent 1
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Batch 7215607388
- Expiration date of the lot/batch: 04 August 2017
- Purity test date: 15.12.2016
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature in the dark
- Stability under test conditions: not specified
- Solubility and stability of the test substance in the solvent/vehicle: not specified
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: not specified
TREATMENT OF TEST MATERIAL PRIOR TO TESTING
The test item was used as supplied
FORM AS APPLIED IN THE TEST (if different from that of starting material)
The test item was used as supplied
In vitro test system
- Test system:
- artificial membrane barrier model
- Source species:
- human
- Cell type:
- other: Reconstructed Human Epithelium
- Justification for test system used:
- This model incorporates several features, which make it advantageous in the study of potential dermal corrosivity. The target cells are epithelial, derived from human skin, and formed into a stratified, cornified epithelium. Test items are applied to the culture surface, at the air interface, so that undiluted and/or end use dilutions can be tested directly.
- Vehicle:
- unchanged (no vehicle)
- Details on test system:
- RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDermTM
- Tissue batch number(s): 25817
- Production date: not specified
- Shipping date: not specifed
- Delivery date: 06 June 2017
- Date of initiation of testing: 24 April 2017
TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37°C
- Temperature of post-treatment incubation (if applicable): 37°C
REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: Rinsing was achieved by filling and emptying each tissue under a constant soft stream of DPBS to gently remove any residual test item. Excess DPBS was removed by blotting the bottom of the tissue insert with tissue paper. Rinsing was performed at the end of the 3 minutes and at the 60 minutes exposure periods.
- Observable damage in the tissue due to washing: not specified
- Modifications to validated SOP: not specified
MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1mg/mL
- Incubation time: 3 hours
- Spectrophotometer: Labtech LT-4500 microplate reader
- Wavelength: 570 nm
- Filter: not specified
- Filter bandwidth:not specified
- Linear OD range of spectrophotometer: not specified
FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
Not specified
NUMBER OF REPLICATE TISSUES: duplicates were used
CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- Killed tissues
- Procedure used to prepare the killed tissues (if applicable): Freeze-killed tissues were prepared by placing untreated EPIDERMTM tissues in an empty 12 well plate and storing in a freezer (-14 to -30 °C) for a minimum of 24 hours. Before use each tissue was thawed by placing in 0.9 mL of assay medium for approximately 1 hour at room temperature.
- N. of replicates : Two freeze-killed tissues were used
- Method of calculation used: relative viability (%) = (Mean OD570 of test item / Mean OD570 of negative control) x 100
NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 2 Independant test sequencies were performed : Pre test and Main test
PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be corrosive to skin if the viability after 3 minutes exposure is less than 50% and or if the viability after 3 minutes exposure is greater than or equal to 50 % and the viability after 1 hour exposure is less than 15%.
- The test substance is considered to be non-corrosive to skin the viability after 3 minutes exposure is greater than or equal to 50% and the viability after 1 hour exposure is greater than or equal to 15%. - Control samples:
- yes, concurrent negative control
- yes, concurrent positive control
- Amount/concentration applied:
- TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 25 mg
- Concentration (if solution): pure
NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 50 µL
- Concentration (if solution): pure
POSITIVE CONTROL
- Amount(s) applied (volume or weight): 50 µL
- Concentration (if solution): 8.0N - Duration of treatment / exposure:
- 3 and 60 minutes
- Duration of post-treatment incubation (if applicable):
- 3 hours (MTT incubation)
- Number of replicates:
- Duplicates were used
Results and discussion
In vitro
Resultsopen allclose all
- Irritation / corrosion parameter:
- % tissue viability
- Run / experiment:
- Main Test - Negative Control at 3 and 60 minutes of exposure
- Value:
- 100
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Irritation / corrosion parameter:
- % tissue viability
- Run / experiment:
- Main test - Positive Control at 3 and 60 minutes of exposure
- Value:
- 3.9
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Irritation / corrosion parameter:
- % tissue viability
- Run / experiment:
- Main test - Test Item 3 minutes of exposure
- Value:
- 105.8
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Irritation / corrosion parameter:
- % tissue viability
- Run / experiment:
- Main test - Test Item 3 minutes of exposure
- Value:
- 97.9
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Other effects / acceptance of results:
- - OTHER EFFECTS:
- Visible damage on test system: not specifed
- Direct-MTT reduction: An assessment of the test item’s capability to directly reduce MTT was inconclusive due to the intrinsic color of the test item and therefore, an additional procedure using freeze-killed tissues was performed. However, the results obtained showed that negligible interference due to direct reduction of MTT occurred.
- Colour interference with MTT: The solution containing the test item was a yellow to green color, therefore additional color correction tissues were incorporated into the testing procedure. However, the results obtained showed that no color interference occurred. It was therefore considered unnecessary to use the results of the color correction tissues for quantitative correction of results or for reporting purposes.
DEMONSTRATION OF TECHNICAL PROFICIENCY: The positive control ( 8.0N Potassium Hydrxide) induced positive response to the test system
ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The mean OD570 for the negative control treated tissues was 1.556 for the 3 Minute exposure period and 1.507 for the 60 Minute exposure period. The negative control acceptance criteria were therefore satisfied.
- Acceptance criteria met for positive control:The relative mean tissue viability for the positive control treated tissues was 3.9% relative to the negative control following the 60 Minute exposure period. The positive control acceptance criterion was therefore satisfied.
- Acceptance criteria met for variability between replicate measurements:In the range 20 to 100% viability the Coefficient of Variation between the two tissue replicates of each treatment group did not exceed 30%. The acceptance criterion was therefore satisfied.
Any other information on results incl. tables
Table1 :Mean OD570Values and Viabilities for the Negative Control Item, Positive Control Item and Test Item
Tissue |
Exposure Period |
MeanOD570of individual tissues |
Mean OD570of duplicate tissues |
Standard Deviation |
Coefficient ofVariation |
Relative Mean Viability (%) |
Negative Control |
3 Minutes |
1.511 |
1.556 |
0.063 |
4.0 |
100* |
1.600 |
||||||
60 Minutes |
1.488 |
1.507 |
0.026 |
1.7 |
||
1.525 |
||||||
Positive Control |
3 Minutes |
0.062 |
0.061 |
0.002 |
na |
3.9 |
0.059 |
||||||
60 Minutes |
0.066 |
0.060 |
0.009 |
na |
3.9 |
|
0.053 |
||||||
Test Item |
3 Minutes |
1.636 |
1.646 |
0.014 |
0.9 |
105.8 |
1.656 |
||||||
60 Minutes |
1.380 |
1.475 |
0.134 |
9.1 |
97.9 |
|
1.570 |
OD = Optical density
*= The mean percentage viability of the negative control tissue is set at 100%
na = Not applicable
Applicant's summary and conclusion
- Interpretation of results:
- other: not classified as corrosive substance according CLP criteria.
- Conclusions:
- Under the experimental condition of this study, the registered substance 4-amino-6-chloro-4-nitrophenol did not led to corrosion when applied on EpiDermTM Reconstructed Human Epithelium. Hence, the B099 was not classified as corrosive substance according CLP criteria.
- Executive summary:
The purpose of this GLP compliant study is to evaluate the corrosivity potential of the test item using the EpiDerm™ Human Skin Model after treatment periods of 3 and 60 minutes according the OECD 431 guideline method.
Duplicate tissues were treated with the test item for exposure periods of 3 and 60 minutes. Negative and positive control groups were treated for each exposure period. The test item was found to have the potential to cause color interference and therefore additional tissues were incorporated for color correction purposes. At the end of the exposure period the test item was rinsed from each tissue before each tissue was taken for MTT loading. After MTT loading each tissue was placed in 2 mL Isopropanol for MTT extraction.
The relative viability after the 3 and 60 minutes exposure periods were respectively 105.8% and 97.9%. The positive and negative control values were withinthe acceptance criterias.
Under the experimental condition of this study, the registered substance 4-amino-6-chloro-4-nitrophenol did not led to corrosion when applied on EpiDermTM Reconstructed Human Epithelium. Hence, the B099 was not classified as corrosive substance according CLP criteria.
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.