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Registration Dossier
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EC number: 201-762-9 | CAS number: 87-66-1
- 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
Skin Irritation:
On the test item an in vitro toxicological study aimed to evaluate any potential cutaneous corrosion was carried out.
The percentage reduction of viability was used to predict the corrosive potential.
On the basis of the results, interpreted according to OECD 431:2016, the test items must be considered CORROSIVE to skin.
Eye irritation:
The study does not need to be conducted because the substance is classified as skin irritation and the available information indicates that it should be classified as eye irritation (Category 2)
Key value for chemical safety assessment
Skin irritation / corrosion
Link to relevant study records
- Endpoint:
- skin corrosion: in vitro / ex vivo
- Remarks:
- in vitro study
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- 04/04/2018 - 06/04/2018
- 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)
- Version / remarks:
- v. 2016
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 20170710
- Expiration date of the lot/batch: 1 year
- Purity test date: not specified
RADIOLABELLING INFORMATION (if applicable) not applicable
- Radiochemical purity: not applicable
- Specific activity: not applicable
- Locations of the label: not applicable
- Expiration date of radiochemical substance: not applicable
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room Temperature
- 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
- Treatment of test material prior to testing: no
- Preliminary purification step (if any): no
- Final dilution of a dissolved solid, stock liquid or gel: no
- Final preparation of a solid: MTT thiazolyl blue tetrazolium 5 mg/ml has been diluted with MTT diluent up to 1 mg/ml. The MTT solution was prepared at use, sheltered from light, and discarded at the end of the study
FORM AS APPLIED IN THE TEST (if different from that of starting material)
TYPE OF BIOCIDE/PESTICIDE FORMULATION (if applicable) not applicable
OTHER SPECIFICS:
Sample code ACE-2018-00019547 - Test system:
- artificial membrane barrier model
- Remarks:
- EpiDerm™ a 3D system of reconstructed epidermis of normal human keratinocytes
- Source species:
- human
- Cell type:
- non-transformed keratinocytes
- Details on animal used as source of test system:
- not relevant
- Justification for test system used:
- The Test Guideline makes use of reconstructed human epidermis (RhE) (obtained from human derived non-transformed epidermal keratinocytes) which closely mimics the histological, morphological, biochemical and physiological properties of the upper parts of the human skin, i.e. the epidermis and a commercially available RhE models was used.
- Vehicle:
- water
- Details on test system:
- RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Tissue batch number(s): MatTek Corporation
- Production date: no data
- Shipping date: no data
- Delivery date: no data
- Date of initiation of testing: no data
TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 3 minutes (at room temperature) and 60 minutes (at 37°C)
- Temperature of post-treatment incubation (if applicable): no data
REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: no data
- Observable damage in the tissue due to washing: no data
- Modifications to validated SOP: no data
MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 5 mg/ml
- Incubation time: 1 hour at 37°±1°C
- Spectrophotometer: no data
- Wavelength: no data
- Filter: no data
- Filter bandwidth: no data
- Linear OD range of spectrophotometer: no data
FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: % of cell viability = (ODti)/(ODnc)x100 and % of cell viability = (ODpc)/(ODnc)x100
- Barrier function: The model exhibits normal barrier functions (presence of a differentiated stratum corneum)
- Morphology: no data
- Contamination: no data
- Reproducibility: no data
NUMBER OF REPLICATE TISSUES:
CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- Fresh tissues / killed tissues no data
- Procedure used to prepare the killed tissues (if applicable): no data
- N. of replicates : no data
- Method of calculation used: The Average data for ODNC, ODPC and ODti , the Standard Deviation (SD) and coefficient of Variation (CV%) have been calculated
NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION:
PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be corrosive to skin if [complete, e.g. the viability after 3 minutes exposure is less than 50%, 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 if [complete, e.g. 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%.]
- Justification for the selection of the cut-off point(s) if different than recommended in TG 431 and 439: - Control samples:
- yes, concurrent negative control
- yes, concurrent positive control
- yes, concurrent MTT non-specific colour control
- Amount/concentration applied:
- TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 25 mg of test item have been used neat
- Concentration (if solution):
VEHICLE
- Amount(s) applied (volume or weight with unit): 25 ul of water have been added for wetting the test material
- Concentration (if solution):
- Lot/batch no. (if required): not specified
- Purity: not specified
NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 50 ul of H2O for injection have been used as Negative Control
- Concentration (if solution):
POSITIVE CONTROL
- Amount(s) applied (volume or weight): 50 ul of H2O for injection have been used as Negative Control
- Concentration (if solution): - Duration of treatment / exposure:
- The test item, the negative and positive control have been applied in duplicate on the tissues for 3 minutes (at room temperature) and 60 minutes (at 37°C, 5% CO2), to respect the exposure times the applications have been performed at intervals of 1 minute.
The test chemical is applied topically to the three-dimensional RhE model. - Duration of post-treatment incubation (if applicable):
- At the end of the exposure time the product and the controls have been removed from the tissues and the tissues have been rinsed for 25 times with 1 ml of DPBS.
- Number of replicates:
- A 96-well plate has been prepared, transferring 200 μl in 3 replicates of positive and negative control and treated test item tissue, then the optical density (OD) at the microplate reader have been read. Isopropyl alcohol (IPA) - Desorb Solution has been used as blank
- Amount / concentration applied:
- not applicable
- Duration of treatment / exposure:
- not applicable
- Observation period:
- not applicable
- Number of animals:
- not applicable
- Details on study design:
- TEST SITE
- Area of exposure:the three-dimensional RhE model .
- % coverage: -
- Type of wrap if used: -
REMOVAL OF TEST SUBSTANCE
- Washing (if done):
Treatment
The test item, the negative and positive control have been applied in duplicate on the tissues for 3 minutes (at room temperature) and 60 minutes (at 37°C, 5% CO2), to respect the exposure times the applications have been performed at intervals of 1 minute.
At the end of the exposure time the product and the controls have been removed from the tissues and the tissues have been rinsed for 25 times with 1 ml of DPBS.
MTT test
After the treatment with the test item, the negative and positive control, the tissues have been treated for 3 hours at 37°C, 5% CO2 with MTT ready to use.
After incubation time, to each tissue 2 ml of isopropanol have been added an incubated for 2 hours at room temperature with gentle agitation for formazan extraction.
A 96-well plate has been prepared, transferring 200 μl in 3 replicates of positive and negative control and treated test item tissue, then the optical density (OD) at the microplate reader have been read. Isopropyl alcohol (IPA) - Desorb Solution has been used as blank.
- Time after start of exposure:
OBSERVATION TIME POINTS
(indicate if minutes, hours or days) : exposure time points t=3 and 60 minutes
SCORING SYSTEM:
- Method of calculation: Optical density measurements were performed. The absorbance has been measured on a microplate reader using a 570 nm wavelength using a GEN5 software (Biotek). The Average data for ODNC, ODPC and ODti , the Standard Deviation (SD) and
coefficient of Variation (CV%) have been calculated.
Moreover, for each replicate the true OD value of negative control, positive control and test item (ODNC, ODPC andODti) will be calculated against Average OD value of blank (ΔODblank). - Irritation / corrosion parameter:
- other: Optical density
- Remarks:
- The absorbance has been measured
- Run / experiment:
- Tissue 1 - Replicate 1
- Value:
- 0.24
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Irritation / corrosion parameter:
- other: Optical density (OD) at 570 nm
- Remarks:
- The absorbance has been measured
- Run / experiment:
- Tissue 1 - Replicate 2
- Value:
- 0.254
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Irritation / corrosion parameter:
- other: Optical density (OD) at 570 nm
- Remarks:
- The absorbance has been measured
- Run / experiment:
- Tissue 1 - Replicate 3
- Value:
- 0.255
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Irritation / corrosion parameter:
- other: Optical density (OD) at 570 nm
- Run / experiment:
- Tissue 2 - Replicate 1
- Value:
- 0.27
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Irritation / corrosion parameter:
- other: Optical density (OD) at 570 nm
- Run / experiment:
- Tissue 2 - Replicate 2
- Value:
- 0.292
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Irritation / corrosion parameter:
- other: Optical density (OD) at 570 nm
- Run / experiment:
- Tissue 2 - Replicate 3
- Value:
- 0.287
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Irritation / corrosion parameter:
- % tissue viability
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other: Not applicable
- Remarks:
- Viability not in range 20-100%
- Irritation / corrosion parameter:
- other: OD corrected by the non-specific reduction of MTT(NSMTT)
- Run / experiment:
- Tissue 1 - Replicate 1
- Value:
- 0.228
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- not examined
- Irritation / corrosion parameter:
- other: OD corrected by the non-specific reduction of MTT(NSMTT)
- Run / experiment:
- Tissue 1 - Replicate 2
- Value:
- 0.242
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- not examined
- Irritation / corrosion parameter:
- other: OD corrected by the non-specific reduction of MTT(NSMTT)
- Run / experiment:
- Tissue 1 - Replicate 3
- Value:
- 0.24
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- not examined
- Irritation / corrosion parameter:
- other: OD corrected by the non-specific reduction of MTT(NSMTT)
- Run / experiment:
- Tissue 2 - Replicate 1
- Value:
- 0.28
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- not examined
- Irritation / corrosion parameter:
- other: OD corrected by the non-specific reduction of MTT(NSMTT)
- Run / experiment:
- Tissue 2 - Replicate 3
- Value:
- 0.298
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- not examined
- Irritation / corrosion parameter:
- % tissue viability
- Run / experiment:
- 3 minutes of treatment
- Value:
- 2.49
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- not examined
- Irritation / corrosion parameter:
- other: Optical density (OD) at 570 nm (60 minutes of treatment)
- Run / experiment:
- Tissue 1 - Replicate 1
- Value:
- 0.339
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Irritation / corrosion parameter:
- other: Optical density (OD) at 570 nm (60 minutes of treatment)
- Run / experiment:
- Tissue 1 - Replicate 2
- Value:
- 0.361
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Irritation / corrosion parameter:
- other: Optical density (OD) at 570 nm (60 minutes of treatment)
- Run / experiment:
- Tissue 1 - Replicate 3
- Value:
- 0.366
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Irritation / corrosion parameter:
- other: Optical density (OD) at 570 nm (60 minutes of treatment)
- Run / experiment:
- Tissue 2 - Replicate 1
- Value:
- 0.331
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Irritation / corrosion parameter:
- other: Optical density (OD) at 570 nm (60 minutes of treatment)
- Run / experiment:
- Tissue 2 - Replicate 2
- Value:
- 0.347
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Irritation / corrosion parameter:
- other: Optical density (OD) at 570 nm (60 minutes of treatment)
- Run / experiment:
- Tissue 2 - Replicate 3
- Value:
- 0.342
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Irritation / corrosion parameter:
- % tissue viability
- Run / experiment:
- 3 minutes treatment
- Vehicle controls validity:
- not examined
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other: Not applicable
- Remarks:
- Viability not in range 20-100%
- Irritation / corrosion parameter:
- other: OD corrected by the non-specific reduction of MTT(NSMTT)
- Remarks:
- 60 minutes treatment
- Run / experiment:
- Tissue 1 - Replicate 1
- Value:
- 0.364
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- not examined
- Irritation / corrosion parameter:
- other: OD corrected by the non-specific reduction of MTT(NSMTT)
- Run / experiment:
- Tissue 1 - Replicate 2
- Value:
- 0.393
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- not examined
- Irritation / corrosion parameter:
- other: OD corrected by the non-specific reduction of MTT(NSMTT)
- Run / experiment:
- Tissue 1 - Replicate 3
- Value:
- 0.392
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- not examined
- Irritation / corrosion parameter:
- other: OD corrected by the non-specific reduction of MTT(NSMTT)
- Run / experiment:
- Tissue 2 - Replicate 2
- Value:
- 0.361
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- not examined
- Irritation / corrosion parameter:
- other: OD corrected by the non-specific reduction of MTT(NSMTT)
- Run / experiment:
- Tissue 2 - Replicate 2
- Value:
- 0.386
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- not examined
- Irritation / corrosion parameter:
- other: OD corrected by the non-specific reduction of MTT(NSMTT)
- Run / experiment:
- Tissue 2 - Replicate 3
- Value:
- 0.377
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- not examined
- Irritation / corrosion parameter:
- % tissue viability
- Run / experiment:
- 60 minutes of treatment
- Value:
- 1.95
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- not examined
- Irritation / corrosion parameter:
- % tissue viability
- Run / experiment:
- 3 minutes treatment
- Value:
- 2.49
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- not examined
- Other effects / acceptance of results:
- - OTHER EFFECTS:
- Visible damage on test system: no data
- Direct-MTT reduction: no data
- Colour interference with MTT: the test item has been interfered with MTT it was necessary to evaluate the OD corrected by the non-specific reduction of MTT(NSMTT) using killed tissues.
DEMONSTRATION OF TECHNICAL PROFICIENCY: no data
ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The negative control data meet the acceptance criteria if the mean OD value of the 3 tissues at 570 nm is ≥0.8 and ≤2.8.
- Acceptance criteria met for positive control: The positive control data meet the criteria if the mean viability, expressed as % of the NC, is <15%.
The Coefficient of variation value will be considered valid if it is < 30% according to the performance standard when the % viability is in range between 20-100%; if %viability is not in the range 20-100%, CV% will be not calculated and considered “Not Applicable”.
- Acceptance criteria met for variability between replicate measurements: The Coefficient of variation value will be considered valid if it is < 30% according to the performance standard when the % viability is in range between 20-100%; if %viability is not in the range 20-100%, CV% will be not calculated and considered “Not Applicable”.
ACCEPTABILITY CRITERIA for the blank: the blank data meet the acceptance criteria if every replicate is <0.1.
- Range of historical values if different from the ones specified in the test guideline: no data - Interpretation of results:
- Category 1 (corrosive) based on GHS criteria
- Conclusions:
- On the basis of the results, interpreted according to OECD 431, the test items must be considered CORROSIVE to skin.
- Executive summary:
On the test item an in vitro toxicological study aimed to evaluate any potential cutaneous corrosion was carried out.
The following test was performed:
- In vitro skin corrosion test on Reconstituted Human Epithelium according to OECD N. 431:2016.
To perform the in vitro skin corrosion test, three-dimensional Reconstituted Human Epithelium (RHE) tissues, consisting of normal human keratinocytes cultured for 17-days on an inert 0.6 cm2, polycarbonate filter at the air-liquid interface, were used.
The test item was topically applied on two tissues replicates for 3 minutes (at room temperature) and 60 minutes (at 37°C, 5% CO2), exposure was followed by rinsing with phosphate buffer saline (DPBS) and dried, then tissues were transferred to MTT for 3 hours.
The aim of this assay was to assess quantitatively the effects of the tested product on cell survival through the MTT assay.
Cell viability determination is based on cellular dehydrogenase activity, measured by MTT reduction and conversion into blue formazan salt that is quantified after extraction from tissues. The percentage reduction of viability is used to predict the corrosive potential.
On the basis of the results, interpreted according to OECD 431:2016, the test items must be considered CORROSIVE to skin.
Reference
Optical density measurements:The absorbance has been measured on a microplate reader using a 570 nm wavelength using a GEN5 software (Biotek). The Average data for ODNC, ODPC and ODti , the Standard Deviation (SD) and coefficient of Variation (CV%) have been calculated.
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed (irritating)
Eye irritation
Link to relevant study records
- Endpoint:
- eye irritation: in vitro / ex vivo
- Data waiving:
- study scientifically not necessary / other information available
- Justification for data waiving:
- the study does not need to be conducted because the substance is classified as skin irritation and the available information indicates that it should be classified as eye irritation (Category 2)
Reference
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed (irritating)
Respiratory irritation
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
Justification for classification or non-classification
Data in vitro ( OECD guideline 431 and 439) show that the substance should be classified as corrosive.
Nevertheless, an in vivo acute dermal toxicity study showed no irritation of the skin, contrasting with the results obtained in OECD 431 and 439 studies.This last discrepancy between the results of different test methods might be explained considering the possibility of a false positive result in the MTT tests performed according to OECD 431-439. These in vitro tests are based on the transformation of MTT in a coloured derivate and on its UV-absorbance. The test might have limitations for 2 main reasons: it is performed in an in vitro “simplified” system and it is very sensitive to unspecific development of coloured compounds, possibly interfering with the wavelength used for MTT-formazan. In the case of Pyrogallol, the study report indicates that MTT solution in contact with the test substance became dark blue, thus it has been necessary to evaluate the optical density due to a non-specific reduction of the MTT and this might have corrupted the test results.
Moreover, the expert system Toxtree did not highlight any alert related to skin irritation/corrosion for this molecule, further supporting the findings in vivo.
For the purpose of registration, in order to keep a conservative approach and considering a weight of evidence of in vitro and in vivo tests, the classification as eye and skin irritant will be applied.
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.