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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 481-150-8 | CAS number: 500011-86-9
- 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 chemico
- Remarks:
- direct peptide reactivity assay
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 14 February 2020 to 15 May 2020
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 020
- Report date:
- 2020
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 442C (In Chemico Skin Sensitisation: Direct Peptide Reactivity Assay (DPRA))
- Version / remarks:
- June 18, 2019
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of study:
- direct peptide reactivity assay (DPRA)
- Justification for non-LLNA method:
- This assay assesses the covalent binding potential of the test item to proteins.” The DPRA is proposed to address the molecular initiating event of the skin sensitisation AOP, namely protein reactivity, by quantifying the reactivity of test chemicals towards model synthetic peptides containing either Cysteine or Lysine. Cysteine and Lysine percent peptide depletion values are then used to categorise a substance in one of four classes of reactivity for supporting the discrimination between skin sensitisers and non-sensitisers.
Test material
- Reference substance name:
- -
- EC Number:
- 481-150-8
- EC Name:
- -
- Cas Number:
- 500011-86-9
- Molecular formula:
- C9H5BrClN3O2
- IUPAC Name:
- 3-bromo-1-(3-chloropyridin-2-yl)-1H-pyrazole-5-carboxylic acid
- Test material form:
- solid: particulate/powder
- Details on test material:
- Identification: 3-Bromo-1-(3-Chloro-2-Pyridinyl)-1H-Pyrazole-5-Carboxylic acid
CAS No.: 500011-86-9
Batch: 628-067-1117
Test item Content: 964.8 ± 1.5 g/kg
Production date: November 17, 2019
Expiry date: November 17, 2021
Supplier: ADAMA Makhteshim Ltd.
Appearance: White solid
Constituent 1
- Specific details on test material used for the study:
- Storage Condition (at JRF):
Storage Temperature: Room temperature (15 to 30 °C).
Storage Condition: Cool and dry conditions.
Storage Container: In original container as supplied by the Sponsor.
Storage Location: Test Item Control Office, JRF.
In chemico test system
- Details on the study design:
- DETAILS OF TEST SYSTEM
- Synthetic heptapeptides containing either Lysine (Ac-RFAAKAA-COOH) or Cysteine (Ac-RFAACAA-COOH) are used as the test system for the direct peptide reactivity assay.
- Synthetic heptapeptides used in this study were obtained from RS Synthesis, Louisville, KY 40270, USA. - Batch number of Cysteine and Lysine peptide was P181203-LC180433 and P170906-LC107617, respectively.
INSTRUMENTS AND EQUIPMENT
Micropipettes: Brand, Eppendorf AG.
Refrigerator: LG Electronics Inc..
Digital Balance: Ohaus (Capable of measuring 10 mg to 210 g).
Microbalance: Rodwag (Capable of measuring 1 mg to 5 g).
pH meter: Thermo Scientific.
Cyclomixer: Remi Electrotechnik.
Millipore Water
System: Millipore.
High Performance Liquid.
Chromatography (HPLC): Shimadzu.
Ultralow Temperature.
Freezer: SANYO.
SOLVENTS AND CHEMICALS
Milli Q Water: Prepared by using Millipore water purification system, (Elix-10 and Milli-Q gradient).
Acetonitrile: Finar (HPLC #292761121FS).
Trifluoro acetic acid: SDFCL (HPLC #F17A/0517/0905/53).
Sodium phosphate, monobasic dihydrat: Merk (# DL6D663605).
Sodium phosphate, dibasic heptahydrate: Sigma (BioReagent #SLBL6644V).
Ammonium acetat: Qualigens (HPLC #3293830219).
Isopropanol: Qualigens (#3126601218).
SOLVENT CONTROL: yes.
Isopropanol
CAS Number: 67-63-0.
Manufactured by: Qualigens.
Lot No: 3126601218.
Date of receipt: January 01, 2019.
Date of Expiry: December 2023.
Appearance: Clear Colourless Liquid.
Purity: 99.93%.
Storage: Room Temperature.
POSITIVE CONTROL USED: yes.
Cinnamaldehyde was used as positive control (PC) at a concentration of 100 mM in acetonitrile.
Name: Cinnamaldehyde
CAS Number: 104-55-2.
Density: 1.049 g/mL.
Manufactured by: Sigma-Aldrich.
Lot N°: MKBT8955V.
Date of receipt: February 12, 2016.
Date of Expiry: February 2020.
Appearance: Yellow Liquid.
Purity: 99.1%.
Storage: Room Temperature.
Note: 38.10 µL of cinnamaldehyde was dissolved in 2961.90 µL of acetonitrile for the preparation of 100 mM solution (3 mL) for both Cysteine and Lysine peptides.
VEHICLE
3-Bromo-1-(3-Chloro-2-Pyridinyl)-1H-Pyrazole-5-Carboxylic acid was found to be soluble in isopropanol at 100 mM. Precipitation was not observed in isopropanol. Hence, isopropanol was selected as the vehicle for the experiment.
Results and discussion
- Positive control results:
- Cinnamaldehyde was used as the positive control in each assay with Cysteine and Lysine peptides. Value of the mean percent peptide depletion value of the positive control, viz., cinnamaldehyde, was 90% for Cysteine peptide and 66% for Lysine peptide. The relative Coefficient of Variability (RCV) for the positive control replicate was 1.79% for percent Cysteine depletion against the guideline limit of <14.9 % and 4.30% for percent Lysine depletion against the guideline limit of <11.6 %.
In vitro / in chemico
Results
- Key result
- Run / experiment:
- other: % Mean Depletion (Cysteine and Lysine)
- Parameter:
- other: DPRA Prediction
- Value:
- 3
- Positive controls validity:
- valid
- Remarks on result:
- no indication of skin sensitisation
- Remarks:
- DPRA Prediction: No or Minimal Reactivity (Negative)
- Other effects / acceptance of results:
- - Acceptance criteria met for positive control:: yes.
Any other information on results incl. tables
Reference Controls
The relative coefficient of variability (RCV) of peptide peak areas for the Reference Control A was 0.64 % for Cysteine peptide and 0.18 % for Lysine peptide. For Reference Control B relative coefficient of variability (RCV) of peptide peak areas was 0.62% for Cysteine peptide and 0.40 % for Lysine peptide Lysine, respectively.
Mean Reference Control Concentrations:
Peptide |
Mean Reference Control A (mM) |
Mean Reference Control B (mM) |
Mean Reference Control C (mM) |
Expected Concentration (mM) |
Cysteine |
0.50 |
0.50 |
0.50 |
0.45-0.55 |
Lysine |
0.50 |
0.50 |
0.51 |
0.45-0.55 |
Cysteine Peptide Stability in Acetonitrile
The stability of Cysteine peptide in acetonitrile was checked at 0, 24, and 48 h. The relative coefficient of variability (RCV) for the stability of Cysteine peptide in acetonitrile was 1.51% against the set standard of <15%. This showed that Cysteine peptide was stable in acetonitrile.
Data of Precipitation for Cysteine and Lysine Peptide:
Test Item Name |
Precipitation/Phase Separation (Yes/No) |
|
Cysteine |
Lysine |
|
3-Bromo-1-(3-Chloro-2-Pyridinyl)-1HPyrazole-5-Carboxylic acid-Replicate 1 |
No |
No |
3-Bromo-1-(3-Chloro-2-Pyridinyl)-1HPyrazole-5-Carboxylic acid-Replicate 2 |
No |
No |
3-Bromo-1-(3-Chloro-2-Pyridinyl)-1HPyrazole-5-Carboxylic acid-Replicate 3 |
No |
No |
3-Bromo-1-(3-Chloro-2-Pyridinyl)-1HPyrazole-5-Carboxylic acid- Co-elution control |
No |
No |
Standard Curve
A standard calibration curve was generated on each day of the HPLC analysis and the value of r2 obtained was 0.99997 for Cysteine and 0.99998 for Lysine containing peptides. This showed that system was suitable for assay.
Percent Peptide Depletion
% Mean Depletion of Peptides with Test Item:
Test Item Name |
Actual % Depletion (Cysteine) |
Actual % Depletion (Lysine) |
% Mean Depletion (Cysteine and Lysine) |
Maximum Standard Deviation (Cysteine) |
Maximum Standard Deviation(Lysine) |
DPRA Prediction |
3-Bromo-1-(3-Chloro-2-Pyridinyl)-1HPyrazole-5-Carboxylic acid |
3 |
3 |
3 |
0.07 |
0.62 |
No or Minimal Reactivity (Negative) |
Applicant's summary and conclusion
- Interpretation of results:
- GHS criteria not met
- Conclusions:
- From the results of this study, under the specified experimental conditions, 3-Bromo-1-(3-Chloro-2-Pyridinyl)-1HPyrazole-5-Carboxylic acid was identified as a Negative in the DPRA assay.
- Executive summary:
This study was conducted to evaluate the skin sensitisation potential of 3-Bromo-1-(3-Chloro-2-Pyridinyl)-1H-Pyrazole-5-Carboxylic acid, using synthetic heptapeptides. The method followed was as per OECD Test Guideline No. 442C.
3-Bromo-1-(3-Chloro-2-Pyridinyl)-1H-Pyrazole-5-Carboxylic acid was found to be soluble in isopropanol at 100 mM. Therefore, isopropanol was selected as the vehicle for this study.
Synthetic heptapeptides containing either Lysine (Ac-RFAAKAA-COOH) or Cysteine (Ac-RFAACAACOOH), were used as the test system in this assay. Cysteine and Lysine containing peptides were incubated with a positive control and the test item for 24 ± 2 hours at 22.5-30 ºC (in the dark), separately. Relative peptide concentration was measured, using the high-performance liquid chromatography (HPLC) with gradient elution and UV detection at 220 nm. Cysteine and Lysine peptide percent depletion values were calculated and used in a prediction model which allows assigning the test item to one of four reactivity classes used to support the discrimination between sensitisers and non-sensitisers (OECD, 2019).
HPLC system suitability was determined by the standard calibration curve and the value of r2 obtained was 0.99997 for cysteine and 0.99998 for lysine peptides, against the set standard of r2 > 0.99, as per the guideline.
The mean peptide concentration of Reference Control A, B and C were mentioned below:
Peptide
Mean Reference Control A (mM)
Mean Reference Control B (mM)
Mean Reference Control C (mM)
Expected Concentration
(mM)
Cysteine
0.50
0.50
0.50
0.45-0.55
Lysine
0.50
0.50
0.51
0.45-0.55
The relative coefficient of variability (RCV) of peptide peak areas for the Reference Control A was 0.64 % for Cysteine peptide and 0.18 % for Lysine peptide. For Reference Control B relative coefficient of variability (RCV) of peptide peak areas was 0.62% for Cysteine peptide and 0.40 % for Lysine peptide Lysine, respectively. The Relative Coefficient of Variability (RCV) for the Reference Control C was 0.11% for cysteine peptide and 2.33% for lysine peptide.
The percent peptide depletion obtained for Cysteine and Lysine is as tabulated below:
Sample Name and Date
Percent Peptide Depletion (Cysteine)
Percent Peptide Depletion (Lysine)
%
Depletion
SD
RCV
Expected
Depletion
%
Depletion
SD
RCV
Expected
Depletion
Cinnamaldehyde
(Positive control)
90
3312.01
1.79
60.8-100
66
24472.88
4.30
40.2-69
3-Bromo-1-(3-Chloro-2-Pyridinyl)-1HPyrazole-5-Carboxylic acid
3
1172.48
0.07
-
3
10062.57
0.62
-
Percentage peptide depletion values of 3-Bromo-1-(3-Chloro-2-Pyridinyl)-1H-Pyrazole-5-Carboxylic acid for Cysteine and Lysine were 3% and 3%, respectively. The mean percent depletion for 3-Bromo-1-(3-Chloro-2-Pyridinyl)-1H-Pyrazole-5-Carboxylic acid was 3%.
The relative coefficient of variability (RCV) for the stability of Cysteine peptide in acetonitrile was 1.51% against the set standard of <15% as per the guideline, indicating that Cysteine is stable in acetonitrile. The relative coefficient of variability (RCV) for the stability of Lysine peptide in acetonitrile was 0.40% against
the set standard of <15% as per the guideline, indicating that Lysine peptide was stable in acetonitrile over the analysis time.
From the results of this study, under the specified experimental conditions, 3-Bromo-1-(3-Chloro-2-Pyridinyl)-1H-Pyrazole-5-Carboxylic acid is predicted by DPRA to be a non-sensitiser (Negative).
From the results of the study “In Chemico Skin Sensitisation: Direct Peptide Reactivity Assay (DPRA) of 3-Bromo-1-(3-Chloro-2-Pyridinyl)-1H-Pyrazole-5-Carboxylic acid using Synthetic Heptapeptides”, under the specified experimental conditions, 3-Bromo-1-(3-Chloro-2-Pyridinyl)-1H-Pyrazole-5-Carboxylic acid was identified as a Negative in the DPRA assay.
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.