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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 258-887-7 | CAS number: 53956-04-0
- 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
Key value for chemical safety assessment
Skin sensitisation
Link to relevant study records
- Endpoint:
- skin sensitisation: in chemico
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 442C (In Chemico Skin Sensitisation: Direct Peptide Reactivity Assay (DPRA))
- Deviations:
- yes
- Remarks:
- The stock solution 100mM was prepared using the percentage of purity equal to 98.7% instead of 99.6%. The maximal concentration tested was therefore 100.9mM instead of 100mM, however, this difference does not affect the study result.
- GLP compliance:
- yes
- Type of study:
- direct peptide reactivity assay (DPRA)
- Justification for non-LLNA method:
- According to the column 2 of of the appendix VII of the Regulation EC 1907/2006, in vivo studies are not required if at least one or two in vitro study has been carried out and allow a classification for the substance.
- Details on the study design:
- The test item was prepared at 100 mM in water and the positive control was prepared at 100 mM in acetonitrile.
They were incubated in excess with the peptides at 1:10 and 1:50 ratio for cysteine and lysine peptides respectively.
Each sample was tested 3 times from 3 independent solutions.
*Preparation of the test item and positive control
Test item
According to the study plan, solubility of the test item in Acetonitrile was checked before the main test.
The test item being non soluble at 100 mM in acetonitrile, other suitable diluents have been tested given the order defined in the study plan. The chosen solvent was water.
The weight of test item was calculated according to the following formula:
"Weight (g) =" "0.01 × Vf × MW" /"Purity"
where MW = 840 g/mol and purity = 98.7%.
255 mg was pre-weighted in a glass vial in order to prepare, right for use, 3 ml of a limpid 100 mM solution.
Positive control
The volume of the positive control was calculated according to the following formula:
"Volume (ml) =" "0.01 × fV × MW" /"Purity × ¿"
MW= 132.16 g/mol
Purity = 99.1%
Density ¿ = 1.048
fV (final volume) = 3 ml
38.2 µl of the positive control were distributed in a 5 ml glass vial in order to prepare, right before use, 3 ml of a limpid 100 mM solution.
* Preparation of the peptide solution
Peptide solutions were prepared at 0.667 mM:
• 13.9 mg of cysteine peptide were pre-weighted then dissolved, right before the incubation, in 27.8 ml of phosphate buffer (100 mM; pH 7.5 ± 0.05).
• 14.5 mg of lysine peptide were pre-weighted then dissolved, right before the incubation, in 28 ml of ammonium acetate buffer (100 mM; pH 10.2 ± 0.05).
* Preparation of the sample for the test
Samples were dissolved immediately before use (100 mM positive control solution was kept for the 2 runs). Peptides were incubated with each sample (test item and positive control) at 1:10 and 1:50 ratio for cysteine and lysine respectively. All the replicates were prepared with the same peptide stock solutions.
The vials were capped and mixed carefully avoiding bubbling, then placed in the HPLC system sampler at 25°C ¿ 2.5°C. HPLC analysis started 24 hours ± 2 hours after addition of peptides.
Immediately upon addition of the test item solution to the peptide solution, just prior the beginning of the HPLC analysis and at the end of the analysis, samples vials were checked.
* Preparation of calibration curve
A calibration curve was generated for cysteine and lysine peptides. Peptides standards were prepared in 20% acetonitrile in buffer solution (phosphate buffer for cysteine peptide and ammonium acetate for lysine peptide).
Six standard solutions between 0.534 mM and 0.0167 mM (dilution factor 2) were prepared from the stock solution (0.667 mM) by serial dilution. The dilution buffer was also included as blank in the standard calibration curve.
* Preparation and manipulation of the HPLC system
The HPLC system was used according to the current working instruction IL MAT 24.
According to the guideline, the set-up parameters were adjusted to guarantee an appropriate elution and integration of the cysteine and lysine peptides.
Proficiency substances recommended in the OECD guideline were performed in these conditions. Results are shown at the end of the report.
The HPLC column was installed and equilibrated at 30°C with 50% of phase A and 50% of phase B, for at least 20 minutes before use. The gradient (cf. § 7.8) was performed at least one time before to use the column.
* HPLC analysis
A complete sequence was performed for each sample.
The column was re-equilibrated to initial conditions (90% Phase A and 10% of phase B) at least 4 minutes between each injection.
* Sequence of the analysis
The analysis was programmed according to the following principles:
• The reference controls B were placed at the beginning and at the end of the analysis (3 repetitions).
• The reference controls C were placed at the beginning of each repetition.
• The standards of the calibration curve and the reference controls A were placed in order to be analyzed progressively throughout the sequence.
Lysine and cysteine analysis were conducted on separate day and test item was freshly prepared for both assays on each day. The analysis was timed to assure that the injection of the first sample starts 22 to 26 hours after the test item was mixed with the peptide solution. The HPLC analysis time was less than 30 hours - Key result
- Parameter:
- other: Depletion in lysine peptide (%)
- Value:
- 0
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks:
- (55.31%)
- Key result
- Parameter:
- other: depletion in cysteine peptide (%)
- Value:
- 5.66
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks:
- (73.06%)
- Interpretation of results:
- other: Based on a part of integrated approach, these results support a non-classification.
- Conclusions:
- The sensitivity of the test item is determined by calculating the mean percentage of depletion of Lysine and Cysteine.
The test item, GLYCYRRHIZATE MONOAMMONIACAL, shows mean depletion of 0% for Lysine and 5.66% for Cysteine, i.e. an overall average of 2.83% reflecting no or minimal reactivity and therefore a negative prediction of DPRA.
Based on OECD 442C, GLP study, results are considered scientifically valid to be used as part of an integrated approach to support a non classification.
Please refer to endpoint summary for conclusion. - Endpoint:
- skin sensitisation: in vitro
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 442D (In Vitro Skin Sensitisation: ARE-Nrf2 Luciferase Test Method)
- Deviations:
- yes
- Remarks:
- The stock solution dilution was prepared 4X in treatment medium-4% DMSO instead of 100X. This has no impact on the study result because the test item was tested in the expected final condition (i.e 2000 µM as maximal concentration).
- GLP compliance:
- yes (incl. QA statement)
- Type of study:
- activation of keratinocytes
- Justification for non-LLNA method:
- According to the column 2 of of the appendix VII of the Regulation EC 1907/2006, in vivo studies are not required if at least one or two in vitro study has been carried out and allow a classification for the substance.
- Specific details on test material used for the study:
- Storage conditions: Darkness + room temperature + hygroscopic
Color : White
Physical state at 20°C : Solid
Purity : 98.7%
Homogeneity : Yes
Molecular weight : 840 g/mol
pH : 4.2-4.8 (solution at 1% in ethanol 50%) - Details on the study design:
- Cells suspension are adjusted to a density of 8.104 cells/ml in seeding medium.
125 µl of the cell suspension at 8.104 cells/ml (i.e. 104 cells per well) were distributed in three white plates for the induction measurement and two transparent plates to assess the cytotoxicity. The seeded plates were incubated 24 hours ± 1 hour at 37°C, 5% CO2.
The second day, in the 5 seeded plates, the medium was aspirated and replaced with 150 µl of treatment medium. Then the 4 X plate was replicated 5 times: 50 µl from the 4 X plate were placed in each of the three white plates and in the two transparent plates. The plates (1 X) were covered with an adhesive plastic foil to prevent evaporation and incubated for 48 hours ± 1 hour (37°C, 5% CO2).
After 48 hours, the luciferase activity was measured with an integration time of 2 seconds.
Viability was assessed measuring absorbance at 540 nm. - Positive control results:
- Positive control : cinnamaldehyde
Mean Imax : 3.94 - Key result
- Parameter:
- other: Mean Imax
- Value:
- 1.28
- Vehicle controls validity:
- valid
- Negative controls validity:
- other: negative control is also vehicle control
- Positive controls validity:
- valid
- Interpretation of results:
- other: Based on a part of integrated approach, these results support a non-classification.
- Conclusions:
- Based on OECD 442D, GLP study, results are considered scientifically valid to be used as part of an integrated approach to support a non classification.
Please refer to endpoint summary for conclusion.
Referenceopen allclose all
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (not sensitising)
Justification for classification or non-classification
Skin sensitisation
According to the column 2 of the appendix VII of the Regulation EC 1907/2006, in vivo studies are not required if at least one or two in vitro study has been carried out and allow a classification for the substance. Therefore, one in chemico study DPRA assay and one in vitro study Keratinosens assay address have been conducted according to OECD 442 C and 442 D, respectively. Studies were carried out on GLYCYRRHIZATE MONOAMMONIACAL and both of them provided negative results.
Based on a part of integrated approach, these results support a non-classification, so, it can be concluded GLYCYRRHIZATE MONOAMMONIACAL is not a skin sensitiser.
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.