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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 205-181-1 | CAS number: 135-16-0
- 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
Eye irritation
Administrative data
- Endpoint:
- eye irritation: in vitro / ex vivo
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2017-08-03 to 2018-01-03
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 018
- Report date:
- 2018
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)
- Version / remarks:
- adopted 09 October 2017
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
Test material
- Reference substance name:
- N-[4-[[(2-amino-1,4,5,6,7,8-hexahydro-4-oxo-6-pteridinyl)methyl]amino]benzoyl]-L-glutamic acid
- EC Number:
- 205-181-1
- EC Name:
- N-[4-[[(2-amino-1,4,5,6,7,8-hexahydro-4-oxo-6-pteridinyl)methyl]amino]benzoyl]-L-glutamic acid
- Cas Number:
- 135-16-0
- Molecular formula:
- C19H23N7O6
- IUPAC Name:
- (2S)-2-[(4-{[(2-amino-4-oxo-1,4,5,6,7,8-hexahydropteridin-6-yl)methyl]amino}phenyl)formamido]pentanedioic acid
- Test material form:
- solid
Constituent 1
Test animals / tissue source
- Species:
- cattle
- Details on test animals or tissues and environmental conditions:
- SOURCE OF COLLECTED EYES
- Number of animals: not applicable
- Storage, temperature and transport conditions of ocular tissue (e.g. transport time, transport media and temperature, and other conditions): transported in HBSS + Pen/Strep on ice
- Time interval prior to initiating testing: 1h at 32 ± 1°C
Test system
- Vehicle:
- physiological saline
- Controls:
- yes, concurrent vehicle
- yes, concurrent positive control
- Amount / concentration applied:
- TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.75 mL
- Concentration (if solution): 20% solution in physiological saline
VEHICLE
- Concentration (if solution): 0.9% NaCl solution
- Lot/batch no. (if required): 17296405 - Duration of treatment / exposure:
- 4h ± 5 min
- Observation period (in vivo):
- 90 min
- Number of animals or in vitro replicates:
- 3 corneae per treatment
- Details on study design:
- SELECTION AND PREPARATION OF CORNEAS
Immediately after arrival of the eyes, cornea preparation was initiated. The eyes were carefully examined for defects and any defective eyes were discarded. The tissue surrounding the eyeball was carefully pulled away and the cornea was excised leaving a 2 to 3 mm rim of sclera. The isolated corneas were stored in a petri dish containing HBSS. Before the corneas were mounted in corneal holders (Duratec GmbH) with the endothelial side against the O-ring of the posterior chamber, they had been visually examined for defects and any defective cornea had been discarded.
NUMBER OF REPLICATES
3 corneae for the test item
3 corneae as negative controls treated with physiological saline 0.9% NaCl
3 corneae as positive controls treated with imidazole 20% in physiological saline 0.9% NaCl
APPLICATION DOSE AND EXPOSURE TIME
750 µl of the test item preparation or the control substance was introduced into the anterior chamber
TREATMENT METHOD: closed chamber
POST-INCUBATION PERIOD: yes, 90 min
REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: at least three times with MEM containing phenol red and once rinsed with RPMI without phenol red.
- POST-EXPOSURE INCUBATION: After the washing steps the anterior chamber was filled with RPMI and an illuminance measurement was performed.
METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: The corneal opacity was determined by an opacimeter (BASF-OP3.0, Duratec GmbH). Three glass filter were calibrated for the measurement. F2, readout 540-560 lux, F3, readout 300-310 lux and F4 readout 95-105 lux.
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of UV/VIS spectrophotometry (Jenway 6405 UV/VIS; OD490)
- Others: each cornea was observed visually and pertinent observations were recorded.
SCORING SYSTEM: In Vitro Irritancy Score (IVIS) . The following formula was used to determine the in vitro irritation score (IVIS):
IVIS = mean opacity value + (15 x mean permeability OD490 value)
DECISION CRITERIA: The BCOP assay is considered to be valid if the in vitro irritation score obtained with the positive control falls within the two standard deviations of the current historical mean.
The negative control responses should result in opacity and permeability values that are less than the established upper limits for background bovine corneas treated with the respective negative control.
Results and discussion
In vitro
Results
- Irritation parameter:
- in vitro irritation score
- Remarks:
- 20% solution in physiological saline
- Value:
- 1.05
- Vehicle controls validity:
- valid
- Remarks:
- IVIS = 0.35
- Negative controls validity:
- not examined
- Positive controls validity:
- valid
- Remarks:
- IVIS = 102.44
- Remarks on result:
- no indication of irritation
- Other effects / acceptance of results:
- ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes, the BCOP assay is considered to be valid if the in vitro irritation score obtained with the positive control falls within the two standard deviations of the current historical mean.
- Acceptance criteria met for positive control: yes, the negative control responses should result in opacity and permeability values that are less than the established upper limits for background bovine corneas treated with the respective negative control.
Any other information on results incl. tables
Table 2: Opacity
Cornea No. |
Test Item |
Initial Opacity |
Final Opacity |
Change of Opacity Value |
Corrected Opacity Value |
1 |
Negative control |
1.26 |
1.29 |
0.04 |
|
2 |
1.40 |
1.43 |
0.04 |
||
3 |
1.26 |
1.72 |
0.47 |
||
MV |
1.30 |
1.48 |
0.18 |
||
4 |
Positive control |
2.20 |
72.73 |
70.53 |
70.35 |
5 |
2.76 |
81.45 |
78.68 |
78.50 |
|
6 |
2.57 |
94.26 |
91.69 |
91.51 |
|
MV |
2.51 |
82.81 |
80.30 |
80.12 |
|
7 |
Test item |
0.70 |
1.72 |
1.03 |
0.85 |
8 |
1.87 |
2.96 |
1.09 |
0.91. |
|
9 |
1.01 |
2.46 |
1.45 |
1.27 |
|
MV |
1.19 |
2.38 |
1.19 |
1.01 |
Table 3: Permeability
Cornea No. |
Test Item |
OD490 |
Corrected OD490 Value |
1 |
Negative control |
0.010 |
|
2 |
0.013 |
||
3 |
0.012 |
||
MV |
0.012 |
||
4 |
Positive control |
1.296 |
1.284 |
5 |
1.363 |
1.351 |
|
6 |
1.840 |
1.828 |
|
MV |
1.500 |
1.488 |
|
7 |
Test item |
0.013 |
0.001 |
8 |
0.024 |
0.012 |
|
9 |
0.006 |
-0.006 |
|
MV |
0.014 |
0.003 |
Table 4: In Vitro Irritation Score
Cornea No. |
Test Item |
Corrected Opacity |
Corrected OD490 Value |
IVIS |
1 |
Negative control |
0.04 |
0.01 |
0.35 |
2 |
0.04 |
0.013 |
||
3 |
0.47 |
0.012 |
||
MV |
0.18 |
0.012 |
||
4 |
Positive control |
70.35 |
1.284 |
102.44 |
5 |
78.50 |
1.351 |
||
6 |
91.51 |
1.828 |
||
MV |
80.12 |
1.488 |
||
7 |
Test item |
0.85 |
0.001 |
1.05 |
8 |
0.91 |
0.012 |
||
9 |
1.27 |
-0.006 |
||
MV |
1.01 |
0.003 |
Applicant's summary and conclusion
- Interpretation of results:
- GHS criteria not met
- Conclusions:
- The eye irritancy potential of Tetrahydrofolic acid (THFA) was investigated in the bovine corneal opacity and permeability assay. According to the evaluation criteria the test item Tetrahydrofolic acid (THFA) is classified into UN GHS No Category.
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.