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EC number: 222-394-5 | CAS number: 3458-72-8
- 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:
- 2019-06-27 to 2019-12-18 (provisional draft final report)
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 019
- Report date:
- 2019
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 438 (Isolated Chicken Eye 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:
- 2018
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU method B.48 (Isolated chicken eye test method for identifying occular corrosives and severe irritants)
- Version / remarks:
- 2017
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
Test material
- Reference substance name:
- Triammonium citrate
- EC Number:
- 222-394-5
- EC Name:
- Triammonium citrate
- Cas Number:
- 3458-72-8
- Molecular formula:
- C6H8O7.3H3N
- IUPAC Name:
- triammonium citrate
- Test material form:
- solid: crystalline
Constituent 1
Test animals / tissue source
- Species:
- chicken
- Strain:
- other: ROSS 308
- Details on test animals or tissues and environmental conditions:
- SOURCE OF COLLECTED EYES
- Source:
TARAVIS KFT. 9600 Sárvár, Rábasömjéni út 129. Hungary
- Storage, temperature and transport conditions of ocular tissue (e.g. transport time, transport media and temperature, and other conditions):
Head collection was performed by a slaughter house technician. Heads were removed immediately after sedation of the chickens (sedation was performed by electric current). The heads were transported to the testing facility. at the earliest convenience for use approximately within 2 hours from collection. The ambient temperature was optimal (19.3 ºC to 20.4 ºC) during the transport. All eyes used in the assay were from the same groups of eyes collected on one specific day.
After collection, the heads were inspected for appropriate quality and wrapped with paper moistened with saline, then placed in a plastic box that can be closed (4 5 heads/box).
- Time interval prior to initiating testing:
minimum 3h
- indication of any existing defects or lesions in ocular tissue samples:
No, after removing the head from the plastic box, it was put on soft paper. The eyelids were carefully cut away with scissors, avoiding damaging the cornea. One small drop of fluorescein solution 2 (w/v) % was applied onto the cornea surface for a few seconds and subsequently rinsed off with 20 mL isotonic saline. Then the fluorescein-treated cornea was examined with hand-held slit lamp or slit lamp microscope, with the eye in the head, to ensure that the cornea was not damaged. If the cornea was in good condition, the eyeball was carefully removed from the orbit.
- Indication of any antibiotics used: No
Test system
- Vehicle:
- unchanged (no vehicle)
- Controls:
- yes, concurrent positive control
- yes, concurrent negative control
- Amount / concentration applied:
- TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.03 g
- Duration of treatment / exposure:
- 10 sec
- Duration of post- treatment incubation (in vitro):
- 240 min
- Number of animals or in vitro replicates:
- 3 eyes for treatment, positive and negative control
- Details on study design:
- SELECTION AND PREPARATION OF ISOLATED EYES
After removing the head from the plastic box, it was put on soft paper. The eyelids were carefully cut away with scissors, avoiding damaging the cornea. One small drop of fluorescein solution 2 (w/v) % was applied onto the cornea surface for a few seconds and subsequently rinsed off with 20 mL isotonic saline. Then the fluorescein-treated cornea was examined with hand-held slit lamp or slit lamp microscope, with the eye in the head, to ensure that the cornea was not damaged. If the cornea was in good condition, the eyeball was carefully removed from the orbit.
The eyeball was carefully removed from the orbit by holding the nictitating membrane with surgical forceps, while cutting the eye muscles with bent scissors without cutting off the optical nerve too short. The procedure avoided pressure on the eye in order to prevent distortion of the cornea and subsequent corneal opacity. Once removed from the orbit, the eye was placed onto damp paper and the nictitating membrane was cut away with other connective tissue. The prepared eyes were kept on the wet papers in a closed box so that the appropriate humidity was maintained.
EQUILIBRATION AND BASELINE RECORDINGS
The appropriate numbers of eyes were selected and after being placed in the superfusion apparatus, the selected eyes were examined again with the slit lamp microscope to ensure that they were in good condition. The focus was adjusted to see clearly the isotonic saline which was flowing on the cornea surface. Eyes with a high baseline fluorescein staining (i.e., > 0.5) or corneal opacity score (i.e., > 0.5) were rejected. The cornea thickness was measured using the depth measuring device on the slit lamp microscope (Haag-Streit BQ 900) with the slit-width set at 9½, equalling 0.095 mm. Any eye with cornea thickness deviating more than 10 % from the mean value for all eyes, or eyes that showed any other signs of damage, were rejected and replaced. If the selected eyes were appropriate for the test, acclimatization started and was conducted for approximately 45 to 60 minutes. The temperature was verified to be in the range of 32 ± 1.5 °C in all chambers during the acclimatization and treatment periods.
NUMBER OF REPLICATES
3
NEGATIVE CONTROL USED
physiological saline (NaCl 0.9%)
POSITIVE CONTROL USED
Imidazole (0.03g)
APPLICATION DOSE AND EXPOSURE TIME
0.03 g of the test item for 10 sec
OBSERVATION PERIOD
240 min
REMOVAL OF TEST SUBSTANCE
- Volume and washing procedure after exposure period:
The time of application was monitored, then after an exposure period of 10 seconds from the end of the application the cornea surface was rinsed thoroughly with 20 mL saline solution at ambient temperature, while taking care not to damage the cornea but attempting to remove all the residual test item if possible. The eye in the holder was then returned to its chamber. The time while the eye was out of the chamber was limited to the minimum.
- Indicate any deviation from test procedure in the Guideline
: No differences reported
METHODS FOR MEASURED ENDPOINTS:
The control and test eyes were evaluated pre-treatment and at approximately 30, 75, 120, 180 and 240 minutes after the post-treatment rinse. Minor variations within ± 5 minutes were considered acceptable. The cornea thickness and cornea opacity were measured at all time points. Fluorescein retention was determined at baseline (t=0) and 30 minutes after the post-treatment rinse.
- Corneal opacity:
Corneal opacity was measured according to the following formulae:
ΔCO at time t = CO at time t – CO at t=0
Mean CO(at time t) = FEΔCO (at time t)+ SEΔCO(at time t) + TEΔCO(at time t)/3
with
CO = Cornea opacity
ΔCO = Difference between cornea opacity and cornea opacity reference value
FEΔCO = Difference between first eye cornea opacity and first eye cornea opacity reference value
SEΔCO = Difference between second eye cornea opacity and second eye cornea opacity reference value
TEΔCO = Difference between third eye cornea opacity and third eye cornea opacity reference value
Mean CO = The mean corneal opacity value
at time t = Observation time at 30, 75, 120, 180 or 240 minutes after the post-treatment rinse
at t=0 = Reference value
- Damage to epithelium based on fluorescein retention:
Fluorescein retention was calculated according to the following formulae:
ΔFR at time t = FR at time t – FR at t=0
Mean FR = FEΔFR (at time t) + SEΔFR(at time t) + TEΔFR at time t)/3
with:
FR = Fluorescein retention
ΔFR = Difference between fluorescein retention and fluorescein retention reference value
FEΔFR = Difference between first eye fluorescein retention and first eye fluorescein retention reference value
SEΔFR = Difference between second eye fluorescein retention and second eye fluorescein retention reference value
TEΔFR = Difference between third eye fluorescein retention and third eye fluorescein retention reference value
Mean FR = The mean fluorescein retention value
at time t = Observation time at 30 minutes after the post-treatment rinse
at t=0 = Reference value
- Swelling: measured with optical pachymeter on a slit-lamp microscope; slit-width setting:
Cornea swelling was calculated according to the following formulae
CS at time t = CT at time t – CT at t=0 X100/CT at t=0
Mean CS at time t = FECS(at time t)+ SECS (at time t) + TECS (at time t)/3
with
CS = Cornea swelling
CT = Cornea thickness
FECS = First eye cornea swelling
SECS = Second eye cornea swelling
TECS = Third eye cornea swelling
Mean CS = The mean percentage of corneal swelling
at time t = Observation time at 30, 75, 120, 180 or 240 minutes after the post-treatment rinse
at t=0 = Reference value
- Others (e.g, histopathology):
After consultation with the Sponsor no histopathology evaluation was performed. Corneas are discarded 2 months after the final report.
DECISION CRITERIA: Decision criteria from OECD Guideline 438 was used.
Results and discussion
In vitro
Resultsopen allclose all
- Irritation parameter:
- percent corneal swelling
- Remarks:
- Mean maximum corneal swelling at up to 75 min
- Run / experiment:
- Mean of three treated eyes
- Value:
- 13
- Vehicle controls validity:
- not examined
- Negative controls validity:
- valid
- Remarks:
- 3% Mean maximum corneal swelling at up to 75 min
- Positive controls validity:
- valid
- Remarks:
- 30% Mean maximum corneal swelling at up to 75 min
- Irritation parameter:
- percent corneal swelling
- Remarks:
- Mean maximum corneal swelling at up to 240 min
- Run / experiment:
- Mean of three treated eyes
- Value:
- 13
- Vehicle controls validity:
- not examined
- Negative controls validity:
- valid
- Remarks:
- 3% Mean maximum corneal swelling at up to 240 min
- Positive controls validity:
- valid
- Remarks:
- 37% Mean maximum corneal swelling at up to 240 min
- Irritation parameter:
- cornea opacity score
- Remarks:
- Mean maximum corneal opacity
- Run / experiment:
- Mean of three treated eyes
- Value:
- 1.7
- Vehicle controls validity:
- not examined
- Negative controls validity:
- valid
- Remarks:
- 0.5 Mean maximum corneal opacity
- Positive controls validity:
- valid
- Remarks:
- 4.0 Mean maximum corneal opacity
- Irritation parameter:
- fluorescein retention score
- Remarks:
- Mean fluorescein retention
- Run / experiment:
- Mean of three treated eyes
- Value:
- 1.3
- Vehicle controls validity:
- not examined
- Negative controls validity:
- valid
- Remarks:
- 0.0 Mean fluorescein retention
- Positive controls validity:
- valid
- Remarks:
- 2.8 Mean fluorescein retention
- Other effects / acceptance of results:
- OTHER EFFECTS:
- Visible damage on test system:
Not reported
DEMONSTRATION OF TECHNICAL PROFICIENCY:
Prior to routine use of the method the test facility demonstrated the technical proficiency in a separate study using the ten Proficiency Chemicals according to OECD Test Guideline No. 438.
ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control:
Yes
- Acceptance criteria met for positive control: Yes
According to OECD guideline 438 A test is considered acceptable if the concurrent negative or vehicle/solvent control and the concurrent positive control are identified as GHS Non-Classified and GHS Category 1, respectively.
- Range of historical values if different from the ones specified in the test guideline: please refer to any other information on results incl. tables for the historical data range
Any other information on results incl. tables
Historical control data of Positive control
IMIDAZOLE HISTORICAL CONTROLDose level: 30 mg / eye |
||||||||||||||
n=318 |
Relative observation time (min)→ |
Corneal thickness |
Corneal opacity score |
Fluorescein retention |
||||||||||
30 |
75 |
120 |
180 |
240 |
|
30 |
75 |
120 |
180 |
240 |
|
ΔFR |
||
Maximum swelling (%): |
34 |
45 |
49 |
54 |
55 |
Max. OS: |
4.0 |
4.0 |
4.0 |
4.0 |
4.0 |
Max. FR: |
3.0 |
|
Minimum swelling (%): |
3 |
9 |
12 |
14 |
15 |
Min. OS: |
2.8 |
3.3 |
3.5 |
3.5 |
3.5 |
Min. FR: |
2.7 |
|
Average: |
21 |
28 |
32 |
35 |
37 |
Average: |
3.6 |
3.9 |
3.9 |
4.0 |
4.0 |
Avarage: |
3.0 |
Historical control data of Negative control
NaCl (9 g/L saline) SOLUTION HISTORICAL CONTROL Dose level: 30 µL / eye |
|
|||||||||||||||
|
n=210 |
Relative observation time (min)→ |
Corneal thickness |
Corneal opacity score |
Fluorescein retention |
|||||||||||
|
30 |
75 |
120 |
180 |
240 |
|
30 |
75 |
120 |
180 |
240 |
|
ΔFR |
|||
|
Maximum swelling (%): |
3 |
5 |
5 |
5 |
5 |
Max. OS: |
0.5 |
0.5 |
0.5 |
0.5 |
0.5 |
Max. FR: |
0.5 |
||
|
Minimum swelling (%): |
0 |
0 |
0 |
0 |
0 |
Min. OS: |
0 |
0 |
0 |
0 |
0 |
Min. FR: |
0.0 |
||
|
Average: |
0.3 |
0.4 |
0.5 |
0.5 |
0.5 |
Average: |
0.0 |
0.1 |
0.1 |
0.1 |
0.1 |
Average: |
0.0 |
Remark:
n= number of eyes
ΔFR= Difference between fluorescein retention and fluorescein retention reference value
OS= Opacity score
FR= Fluorescein retention
Applicant's summary and conclusion
- Interpretation of results:
- study cannot be used for classification
- Conclusions:
- In this ICET, Triammonium Citrate did not cause ocular corrosion or severe irritation in the enucleated chicken eyes. The overall ICE classes were once II (based on the fluorescein retention of 1.3) and twice III (based on the corneal swelling of 13% within 75 minutes and corneal opacity score of 1.7).
Positive and negative controls showed the expected results. The experiment was considered to be valid.
According to the guideline OECD 438, Triammonium Citrate overall in vitro classification is neither UN GHS Classification Category I (an ocular corrosive or severe eye irritant) nor No Category. Thus, according to the guideline OECD 438, test item has been categorized as “No prediction can be made”. - Executive summary:
In this ICET, Triammonium Citrate did not cause ocular corrosion or severe irritation in the enucleated chicken eyes. The overall ICE classes were once II (based on the fluorescein retention of 1.3) and twice III (based on the corneal swelling of 13% within 75 minutes and corneal opacity score of 1.7).
Positive and negative controls showed the expected results. The experiment was considered to be valid.
According to the guideline OECD 438, Triammonium Citrate overall in vitro classification is neither UN GHS Classification Category I (an ocular corrosive or severe eye irritant) nor No Category. Thus, according to the guideline OECD 438, test item has been categorized as “No prediction can be made”.
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