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EC number: 948-778-9 | 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
Acute Toxicity: dermal
Administrative data
- Endpoint:
- acute toxicity: dermal
- Data waiving:
- study scientifically not necessary / other information available
- Justification for data waiving:
- the study does not need to be conducted because the substance does not meet the criteria for classification as acute toxicity or STOT SE by the oral route and no systemic effects have been observed in in vivo studies with dermal exposure (e.g. skin irritation, skin sensitisation)
Cross-referenceopen allclose all
- Reason / purpose for cross-reference:
- data waiving: supporting information
Reference
- Endpoint:
- skin irritation: in vitro / ex vivo
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2019
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
- 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: 23ADB/50
- Expiration date of the lot/batch: 21/11/2023
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room Temperature - Test system:
- human skin model
- Source species:
- human
- Cell type:
- non-transformed keratinocytes
- Cell source:
- foreskin from multiple donors
- Justification for test system used:
- Method in compliance with current regulatory requirements
- Vehicle:
- unchanged (no vehicle)
- Details on test system:
- RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDermTM from MatTek Corporation (3D system of reconstructed epidermis of normal human keratinocytes)
- Tissue batch number(s): keratinocyte strain: 00267
- Production date: 16-01-2019
- Shipping date: n/a
- Delivery date: n/a
- Date of initiation of testing: 16-01-2019
TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37±1°C
- Temperature of post-treatment incubation (if applicable): 37±1°C
REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: At the end of the exposure time the product and the controls were removed and the tissues rinsed for several times with DPBS (up to 25 times if necessary).
- Observable damage in the tissue due to washing: n/a
- Modifications to validated SOP: n/a
MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: MTT thiazolyl blue tetrazolium 5 mg/ml (MTT-100-CON) has been diluted with MTT diluent (MTT-100-DIL) up to 1 mg/ml.
- Incubation time: 3h
- Spectrophotometer: The absorbance is measured at 570 nm by microplate reader using a GEN5 software (Biotek).
NUMBER OF REPLICATE TISSUES: 3
NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 3
PREDICTION MODEL / DECISION CRITERIA
-The test chemical is identified as requiring classification and labelling according to UN GHS (Category 2 or Category 1) if the mean percent tissue viability after exposure and post-treatment incubation is less than or equal (≤) to 50%.
In case the test chemical is found to be non-corrosive (e.g., based on TG 430, 431 or 435), and shows tissue viability after exposure and post-treatment incubation is less than or equal (≤) to 50%, the test chemical is considered to be irritant to skin in accordance with UN GHS Category 2.
Depending on the regulatory framework in member countries, the test chemical may be considered as non-irritant to skin in accordance with UN GHS No Category if the tissue viability after exposure and post-treatment incubation is more than (>) 50%.
According to EU classification, the irritancy potential of test substances is predicted to distinguish between H315 skin irritating (category 2) and not classified test substances (EU CLP).
In this study the irritancy potential of test substances is predicted by mean tissue viability of tissues exposed to the test substance.
The test substance is considered to be irritant to skin (H315), if the mean relative viability after 60 minutes exposure and 42 hours post incubation is less or equal to 50% of the negative control. - Control samples:
- yes, concurrent negative control
- yes, concurrent positive control
- Amount/concentration applied:
- TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 30 µl
- Concentration (if solution): n/a applied neat
NEGATIVE CONTROL
- Amount(s) applied (volume or weight): (H2O) 30 µl
- Concentration (if solution): n/a
POSITIVE CONTROL
- Amount(s) applied (volume or weight): 30 µl
- Concentration (if solution):SDS 5% in DPBS - Duration of treatment / exposure:
- 25 min at room temperature
residual incubation time to 60 minutes has been done at 37±1°C, 5% CO2. - Duration of post-treatment incubation (if applicable):
- At the end of the exposure time the product and the controls were removed and the tissues rinsed for several times with DPBS (up to 25 times if necessary).
Each tissue was then transferred in 6-well plate with 1 ml of Assay Medium and incubated for 24±2 hours at 37±1°C, 5% CO2, then the tissues will be transfer in a renewed Assay Medium for 18±2 h at 37±1°C, 5% CO2 before the MTT test. - Number of replicates:
- 3
- Irritation / corrosion parameter:
- % tissue viability
- Run / experiment:
- Mean value
- Value:
- 73.3
- Vehicle controls validity:
- not applicable
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- no indication of irritation
- Other effects / acceptance of results:
ACCEPTABILITY CRITERIA
Negative control: the mean OD570nm value of the negative control tissues should be ≥0.8 and ≤2.8.
Positive control: the mean viability value of positive control tissues, expressed as % against negative control tissues should be ≤20%.
Standard deviation (SD): the SD calculated from individual % tissue viabilities of the 3 treated replicates should be ≤18%.- Interpretation of results:
- GHS criteria not met
- Conclusions:
- On the basis of the results, interpreted according to OECD 439 the test item “TRISODIUM (2S)-2,6-BIS(3-CARBOXYLATOPROPANAMIDO)HEXANOATE” must be considered NOT IRRITANT for the skin.
- Executive summary:
On the test item “TRISODIUM (2S)-2,6-BIS(3-CARBOXYLATOPROPANAMIDO)HEXANOATE” an in
vitro toxicological study aimed to evaluate any potential cutaneous irritation was carried out.
The following test was performed:
-In vitro skin irritation test on Reconstructed Human Epidermis according to OECD N. 439:2015.
To perform the in vitro skin irritation test, three-dimensional Reconstructed Human Epidermis (RHE) tissues, consisting of normal human keratinocytes cultured for 17-days on an inert 0.63 cm2polycarbonate filter at the air-liquid interface, were used.
The test item was topically applied on three tissues replicates for 60 minutes. Exposure was followed by rinsing with phosphate buffer saline (DPBS) and dried, then tissues were transferred to fresh medium and incubated for 42 additional hours, 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 in viability is used to predict the irritation potential.
On the basis of the results, interpreted according to OECD 439:2015 the test item “TRISODIUM (2S)-2,6-BIS(3-CARBOXYLATOPROPANAMIDO)HEXANOATE” must be considered NOT IRRITANT for the skin.
RESULTS
Optical density (OD Value) at 570 nm.
|
Tissue 1 |
Tissue 2 |
Tissue 3 |
||||||
Replicates |
1 |
2 |
3 |
1 |
2 |
3 |
1 |
2 |
3 |
Negative control |
2,283 |
2,307 |
2,283 |
2,386 |
2,339 |
2,346 |
2,480 |
2,460 |
2,471 |
Positive control |
0,111 |
0,110 |
0,111 |
0,127 |
0,127 |
0,128 |
0,119 |
0,116 |
0,117 |
Sample |
1,588 |
1,599 |
1,572 |
1,867 |
1,835 |
1,845 |
1,840 |
1,803 |
1,803 |
ASSAY VALIDITY CRITERIA |
Value |
Acceptability |
Result |
|
Negative control |
Mean OD value |
2.33 |
≥0.8 and≤2.8 |
Complies |
Positive control |
Mean Viability % |
3.28 |
≤ 20 |
Complies |
SD |
0.347 |
≤ 18 |
Complies |
|
Sample |
SD |
6.139 |
Complies |
SAMPLE |
% VIABILITY |
TRISODIUM (2S)-2,6-BIS(3- |
73.30 |
- Reason / purpose for cross-reference:
- data waiving: supporting information
Reference
- Endpoint:
- acute toxicity: oral
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2019
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.1100 (Acute Oral Toxicity)
- GLP compliance:
- yes (incl. QA statement)
- Test type:
- fixed dose procedure
- Limit test:
- yes
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 23ADB/50
- Expiration date of the lot/batch: 21/11/2023
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature
TREATMENT OF TEST MATERIAL PRIOR TO TESTING
In order to get the test item in a solution or suspension, which is applicable to the animals, several vehicles (sterile water, corn oil) were evaluated and preparation and solubility tests were performed. The test material formed an emulsion in corn oil which could not be applied. The preparation with sterile water yielded an applicable solution and considered to be adequate.
- Species:
- rat
- Strain:
- Wistar
- Sex:
- female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Charles River, 97633 Sulzfeld, Germany
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 8 – 9 weeks
- Weight at study initiation: animal no. 1: 173 g; animal no. 2: 184 g; animal no. 3: 153 g; animal no. 4: 176 g; animal no. 5: 170 g
- Fasting period before study: n/a
- Housing: Full barrier in an air-conditioned room
- Diet: ad libitum
- Water : ad libitum
- Acclimation period: Adequate acclimatisation period (at least five days) under laboratory conditions
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 3 °C
- Humidity (%): 55 10%
- Air changes (per hr): 10 x / hour
- Photoperiod (hrs dark / hrs light): Artificial light, sequence being 12 hours light, 12 hours dark - Route of administration:
- oral: gavage
- Vehicle:
- water
- Remarks:
- Aqua ad injectionem (sterile water, DELTAMEDICA, lot no. 807634, expiry date: 06/2021). This vehicle was chosen due to its non-toxic characteristics.
- Details on oral exposure:
- Animal No. /
Sex Dose(mg/kg bw) Body Weight on Day of Administration (g) Volume of Test Item Preparation in Vehicle Administered per Animal (mL)
1 / Female 2000 173 1.7
2 / Female 2000 184 1.8
3 / Female 2000 153 1.5
4 / Female 2000 176 1.8
5 / Female 2000 170 1.7 - Doses:
- 2000 mg/kg bw
- No. of animals per sex per dose:
- 5 animals, Fixed dose
- Control animals:
- no
- Details on study design:
- - Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The animals were weighed on day 1 (prior to the administration) and on days 8 and 15. A careful clinical examination was made several times on the day of dosing (at least once during the first 30 minutes and with special attention given during the first 4 hours post-dose). Thereafter, the animals were observed for clinical signs once daily until the end of the observation period.
- Necropsy of survivors performed: yes
- Other examinations performed: Cageside observations included changes in the skin and fur, eyes and mucous membranes. Also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour pattern were examined. Particular attention was directed to observations of tremor, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
All animals were subjected to gross necropsy and examined macroscopically for gross pathological changes. In the absence of gross pathological changes no tissues were preserved for a possible histopathological evaluation. - Preliminary study:
- Sighting Study
The starting dose for the sighting study was 2000 mg/kg body weight.
No compound-related mortality was recorded for the first animal dosed. - Sex:
- female
- Dose descriptor:
- LD50
- Effect level:
- > 5 000 mg/kg bw
- Based on:
- test mat.
- Mortality:
- none
- Clinical signs:
- other: none
- Gross pathology:
- No specific gross pathological changes were recorded for any animal.
- Interpretation of results:
- GHS criteria not met
- Conclusions:
- Under the conditions of the present study, a single oral application of the test item Trisodium (2S)-2,6-bis(3-carboxylatopropanamido) hexanoate_Disuccinyl lysine sodium salt to rats at a dose of 2000 mg/kg body weight was not associated with signs of toxicity or mortality.
The median lethal dose of Trisodium (2S)-2,6-bis(3-carboxylatopropanamido) hexanoate_Disuccinyl lysine sodium salt after a single oral administration to female rats, observed over a period of 14 days is:
LD50 cut-off (rat): ˃ 5000 mg/kg bw - Executive summary:
Five females WISTAR Crl: WI(Han) rats, were treated with the test item by oral gavage administration at a dosage of 2000 mg/kg body weight. The test item was dissolved in the vehicle aqua ad injectionem (sterile water) at a concentration of 0.2 g/mL and administered at a dose volume of 10 mL/kg.
All animals used in the study after their entrance at BSL were allowed to acclimatise to the laboratory conditions for at least 5 days. The animals were observed on delivery, on inclusion in the study and before administration for mortality/morbidity and other clinical signs. All animals were examined for clinical signs several times on the day of dosing and once daily until the end of the observation period. Their body weights were recorded on day 1 (prior to the administration) and on days 8 and 15. All animals were necropsied and examined macroscopically.
Table 1: Results
Animal No. / Sex
Dose
(mg/kg bw)
Number of Animals
Number of
Intercurrent DeathsSighting Study
1 / Female
2000
1
0
Main Study
2 / Female
2000
1
0
3 / Female
1
0
4 / Female
1
0
5 / Female
1
0
All animals survived until the end of the study without showing any signs of toxicity.Throughout the 14-day observation period, the body weight gain of the test animals was within the normal range of variation for this strain. At necropsy, no treatment-related macroscopic findings were observed in any animal of any step.
LD50cut-off (rat): ˃5000 mg/kg bw
Data source
Materials and methods
Results and discussion
Applicant's summary and conclusion
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.
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