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EC number: 676-712-6 | CAS number: 68890-85-7
- 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
![](https://echa.europa.eu/o/diss-blank-theme/images/factsheets/A-REACH/factsheet/print_ecotoxicological-information.png)
Short-term toxicity to fish
Administrative data
Link to relevant study record(s)
- Endpoint:
- short-term toxicity to fish
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- From May 17, 2010 to June 21, 2010
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 203 (Fish, Acute Toxicity Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.1 (Acute Toxicity for Fish)
- Deviations:
- no
- GLP compliance:
- yes
- Specific details on test material used for the study:
- - Name of test material (as cited in study report): Photomer 4149 F
- Molecular formula: C21H32O9
- Molecular weight: 428.48 g/mol
- Physical state: Liquid, clear light yellowish
- Analytical purity: 100%
- Purity test date: 2010-03-19
- Lot/batch No.: S700040031
- Expiration date of the lot/batch: 2011-01-04
- Stability under test conditions: Not specified
- Storage condition of test material: 6 +/- 2 °C, protected from moisture and light - Analytical monitoring:
- yes
- Remarks:
- LC-MS/MS
- Details on sampling:
- - Concentrations: All test substance concentrations and the control were analytically verified at the beginning (new media, 0 h) and end (old media, 24 h) of the first sampling interval.
- Sampling method: For the second sampling interval (new media, 72h; old media, 96 h) only the test concentrations of nominal 0.625 – 2.50 mg/L and the control were analytically verified, due to 100 % mortality in the nominal concentrations of 5.00 and 10.0 mg/L.
- Sample storage conditions before analysis: All samples were stored at 6 ± 2 °C until start of analysis, if necessary. - Vehicle:
- no
- Details on test solutions:
- Preparation and application of test solution (especially for difficult test substances)
- Method: A stock solution of 10 mg/L was prepared and stirred with a magnetic stirrer for 2 h at 23 +/- 2 °C before further dilution was made.
- Eluate: Dilution water
- Differential loading: 0.564 – 1.06 – 2.14 – 3.85 – 8.82 mg/L
- Controls: 7 fish in dilution water (without test substance) were tested under the same test conditions as the test replicates. - Test organisms (species):
- Danio rerio (previous name: Brachydanio rerio)
- Details on test organisms:
- Test organism:
- Common name: Zebrafish
- Source: Niedersächsischer Landesbetrieb für Wasserwirtschaft, Küsten- und Naturschutz, Hildesheim, Germany
- Length at study initiation (length definition, mean, range and SD): Average body length 2.19 cm
- Weight at study initiation (mean and range, SD): Average body weight 0.176 g
- Method of breeding: Holding was performed at the test facility at 23 +/- 2 °C and diffuse light (0.1 - 10 µmol photons x m-2 x s-1, natural photoperiod). Tap water of local origin was used for holding and testing. The water was filterred on activated charcoal and aerated for at least 24 h to remove chlorine. Nominal water parameters: Total hardness: 10 - 250 mg CaCO3/L; pH-value: 6.0 - 8.5. The water is analysed biannula acc. to German tap water regulation. The water was changed at least once per week. the dissolved oxygen concentration was more than 80 % of the air saturation value.
- Feeding during test: No feeding during test
Acclimatization:
- Acclimation period: Zebrafish with al least 12 days of acclimatisation and mortality < 5% within these days before the study starts were used.
- Acclimation conditions (same as test or not): Same as test
- Type and amount of food: 4 % of the fish body weight per feeding day. The test fish were not fed 24 h before the test started. Food: Stoer perlets, SERA GmbH, 52518 Heidelberg, Germany
- Feeding frequency: 3 times per week
- Health during acclimation (any mortality observed): No mortality observed, no disease treatments were administered thwoughout holding and testing.
- Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 96 h
- Hardness:
- Total Hardness at day 0: 53 mg/L as CaCO3
- Test temperature:
- 23.3 - 24.5°C
- pH:
- 6.99 - 7.63
- Dissolved oxygen:
- 40 - 100%
- Salinity:
- Freshwater
- Conductivity:
- -
- Nominal and measured concentrations:
- Please refer to "Any other information on materials and methods"
- Details on test conditions:
- Test system:
- Test vessel: Glass aquaria
- Type (delete if not applicable): open, loosely covered by glass tops
- Material, size, headspace, fill volume: glass, 2 L test volume per vessel
- Aeration: Gentle aeration
- Renewal rate of test solution (frequency/flow rate): One application at test start
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: less than 1 g fish per L test solution
Test medium and water parameters:
- Source/preparation of dilution water: Tap water, local origin, pH: 6.0 - 8.5
- Total organic carbon: 10 - 250 mg CaCO3/L
- Chlorine: The dilution water was filtered as described above
- Culture medium different from test medium: No
- Intervals of water quality measurement: The water is analysed biannual
Other test conditions:
- Adjustment of pH: No
- Photoperiod: natural
- Light intensity: 0.1 - 10 µmol photons x m-2 x s-1
Effect parameters measured (with observation intervals if applicable):
Fish were considered dead if there was no visible movement (e.g. gill covers movement) and if touching of the caudal peduncle produced no reaction. Records were kept of visible abnormalities (e.g. loss of equilibrium, swimming behaviour, respiratory function, pigmentation, etc.). Observations were made after 24, 48, 72 and 96 h.
Test concentrations:
- Range finding study: Yes
Cumulative Mortality [%] in the Preliminary
(n = 5)
Nominal test substance concentration [mg/L] Test Duration [hours]
24 48 72 96
10 100 --- --- ---
1 0 0 0 0
Controls 0 0 0 0 - Reference substance (positive control):
- no
- Key result
- Duration:
- 96 h
- Dose descriptor:
- LC50
- Effect conc.:
- 1.95 mg/L
- Nominal / measured:
- meas. (arithm. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- mortality (fish)
- Remarks on result:
- other: Interval: 1.75-2.17 mg/L
- Key result
- Duration:
- 96 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 0.564 mg/L
- Nominal / measured:
- meas. (arithm. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- mortality (fish)
- Key result
- Duration:
- 96 h
- Dose descriptor:
- LC0
- Effect conc.:
- 0.564 mg/L
- Nominal / measured:
- meas. (arithm. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- mortality (fish)
- Key result
- Duration:
- 96 h
- Dose descriptor:
- LC100
- Effect conc.:
- 3.85 mg/L
- Nominal / measured:
- meas. (arithm. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- mortality (fish)
- Details on results:
- All effect levels are given based on mean measured test substance concentrations: At the test substance concentrations of 0.564 mg/L, no mortality was observed. Non lethal and lethal effects were observed in the test substance concentration of ≥ 1.06 mg/L. Therefore the NOEC was laid down as 0.564 mg/L and the LOEC was laid down as 1.06 mg/L.
- Results with reference substance (positive control):
- No reference substance tested
- Reported statistics and error estimates:
- LC50-values and 95 % confidence intervals after 72 and 96 hours exposure were calculated by sigmoidal dose-response regression. After 24 - 48 hours only 0 and 100 % mortality were found. No evaluation via regression analysis was carried out. Hence the LC50 is the mean measured value of the concentrations with 0 and 100 % mortality. The concentration causing 0 and 100 % mortality were chosen as lower and upper confidence limit. The concentrations causing 0 and 100 % mortality (LC0 and LC100) as well as the no effect level (NOEC) after 96 h were carried out using standard procedures.
- Sublethal observations / clinical signs:
Table 1. Observations in the test vessels
Mean measured test substance concentration [mg/L]
Effect
Test Duration [h]
2
24
48
72
96
8.82
(E)
-
7/7
---
---
---
(2.6)
7/7
-
---
---
---
3.85
(E)
-
-
7/7
---
---
(2.7)
-
6/7
---
---
---
(2.6)
-
4/7
---
---
---
(2.5)
-
1/7
---
---
---
(2.4)
-
6/7
---
---
---
(2.2)
-
1/7
---
---
---
(1)
7/7
-
---
---
---
2.14
(E)
-
-
-
1/7
3/6
(2.8)
-
-
-
6/7
3/3
(2.7)
-
-
-
6/7
3/3
(2.5)
-
-
-
-
3/3
(2.4)
-
-
-
6/7
-
(2.3)
-
-
1/7
-
-
(2.1)
-
-
-
-
1/3
(1)
7/7
7/7
6/7
-
-
1.06
(E)
-
-
-
-
1/7
(2.7)
-
-
-
-
6/7
(2.6)
-
-
-
-
6/7
(2.4)
-
-
-
-
6/7
(2.3)
-
-
-
1/7
-
(1)
7/7
7/7
7/7
6/7
-
0.564
(1)
7/7
7/7
7/7
7/7
7/7
Control
(1)
7/7
7/7
7/7
7/7
7/7
Table 2. Cumulative mortality [%] in the test vessels
Mean measured test substance concentration
[mg/L]
Test Duration [h]
2
24
48
72
96
8.82
0
100
100
100
100
3.85
0
0
100
100
100
2.14
0
0
0
14
57
1.06
0
0
0
0
14
0.564
0
0
0
0
0
Control
0
0
0
0
0
- Validity criteria fulfilled:
- yes
- Conclusions:
- Under the study conditions, the 96 h LC50 and NOEC (Danio rerio) were determined to be 1.95 and 0.564 mg/L (mean measured), respectively.
- Executive summary:
A study was conducted to determine the short-term toxicity of the test substance to fish according to OECD Guideline 203 and EU Method C.1, in compliance with GLP. In a semi-static test, Zebra fish (Danio rerio) were exposed for 96 h to 5 nominal concentrations of 0.625, 1.25, 2.50, 5.00 and 10.0 mg/L, corresponding to mean measured concentrations of 0.564, 1.06, 2.14, 3.85 and 8.82 mg/L. Seven test organisms were exposed at each dose level. Water quality parameters, pH, temperature and oxygen-saturation measured after 0, 2, 24, 48, 72 and 96 h were determined to be within the acceptable limits. The test substance concentrations were verified via LC-MS/MS at test start and end. The substance was clearly dissolved at all concentrations throughout exposure. Recovery rates of test substance in the new media were in the range of 82 - 106% and in the old media in the range of 69 – 85% of nominal values. All effect levels were given based on mean measured concentrations. Non-lethal and lethal effects were observed at a concentration of ca. 1.06 mg/L. Therefore, the NOEC was established as 0.564 mg/L and the LOEC was 1.06 mg/L. Further, based on a mortality rate at 96 h of 14 and 57% at 1.06 and 2.14 mg/L respectively, the 96 h LC50 (Danio rerio) was established at 1.95 mg/L (mean measured) (Sheerbaum, 2010).
- Endpoint:
- short-term toxicity to fish
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Study period:
- From May 17, 2010 to June 21, 2010
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- results derived from a valid (Q)SAR model and falling into its applicability domain, with adequate and reliable documentation / justification
- Justification for type of information:
- Refer to the section 13 for details on the read across justification. The short-term toxicity to fish study with the read across substance is considered sufficient to fulfil the information requirements as further explained in the provided endpoint summary.
- Reason / purpose for cross-reference:
- read-across: supporting information
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 203 (Fish, Acute Toxicity Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.1 (Acute Toxicity for Fish)
- Deviations:
- no
- GLP compliance:
- yes
- Analytical monitoring:
- yes
- Remarks:
- LC-MS/MS
- Details on sampling:
- - Concentrations: All test substance concentrations and the control were analytically verified at the beginning (new media, 0 h) and end (old media, 24 h) of the first sampling interval.
- Sampling method: For the second sampling interval (new media, 72h; old media, 96 h) only the test concentrations of nominal 0.625 – 2.50 mg/L and the control were analytically verified, due to 100 % mortality in the nominal concentrations of 5.00 and 10.0 mg/L.
- Sample storage conditions before analysis: All samples were stored at 6 ± 2 °C until start of analysis, if necessary. - Vehicle:
- no
- Details on test solutions:
- Preparation and application of test solution (especially for difficult test substances)
- Method: A stock solution of 10 mg/L was prepared and stirred with a magnetic stirrer for 2 h at 23 +/- 2 °C before further dilution was made.
- Eluate: Dilution water
- Differential loading: 0.564 – 1.06 – 2.14 – 3.85 – 8.82 mg/L
- Controls: 7 fish in dilution water (without test substance) were tested under the same test conditions as the test replicates. - Test organisms (species):
- Danio rerio (previous name: Brachydanio rerio)
- Details on test organisms:
- Test organism:
- Common name: Zebrafish
- Source: Niedersächsischer Landesbetrieb für Wasserwirtschaft, Küsten- und Naturschutz, Hildesheim, Germany
- Length at study initiation (length definition, mean, range and SD): Average body length 2.19 cm
- Weight at study initiation (mean and range, SD): Average body weight 0.176 g
- Method of breeding: Holding was performed at the test facility at 23 +/- 2 °C and diffuse light (0.1 - 10 µmol photons x m-2 x s-1, natural photoperiod). Tap water of local origin was used for holding and testing. The water was filterred on activated charcoal and aerated for at least 24 h to remove chlorine. Nominal water parameters: Total hardness: 10 - 250 mg CaCO3/L; pH-value: 6.0 - 8.5. The water is analysed biannula acc. to German tap water regulation. The water was changed at least once per week. the dissolved oxygen concentration was more than 80 % of the air saturation value.
- Feeding during test: No feeding during test
Acclimatization:
- Acclimation period: Zebrafish with al least 12 days of acclimatisation and mortality < 5% within these days before the study starts were used.
- Acclimation conditions (same as test or not): Same as test
- Type and amount of food: 4 % of the fish body weight per feeding day. The test fish were not fed 24 h before the test started. Food: Stoer perlets, SERA GmbH, 52518 Heidelberg, Germany
- Feeding frequency: 3 times per week
- Health during acclimation (any mortality observed): No mortality observed, no disease treatments were administered thwoughout holding and testing.
- Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 96 h
- Hardness:
- Total Hardness at day 0: 53 mg/L as CaCO3
- Test temperature:
- 23.3 - 24.5°C
- pH:
- 6.99 - 7.63
- Dissolved oxygen:
- 40 - 100%
- Salinity:
- Freshwater
- Conductivity:
- -
- Nominal and measured concentrations:
- Please refer to "Any other information on materials and methods"
- Details on test conditions:
- Test system:
- Test vessel: Glass aquaria
- Type (delete if not applicable): open, loosely covered by glass tops
- Material, size, headspace, fill volume: glass, 2 L test volume per vessel
- Aeration: Gentle aeration
- Renewal rate of test solution (frequency/flow rate): One application at test start
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: less than 1 g fish per L test solution
Test medium and water parameters:
- Source/preparation of dilution water: Tap water, local origin, pH: 6.0 - 8.5
- Total organic carbon: 10 - 250 mg CaCO3/L
- Chlorine: The dilution water was filtered as described above
- Culture medium different from test medium: No
- Intervals of water quality measurement: The water is analysed biannual
Other test conditions:
- Adjustment of pH: No
- Photoperiod: natural
- Light intensity: 0.1 - 10 µmol photons x m-2 x s-1
Effect parameters measured (with observation intervals if applicable):
Fish were considered dead if there was no visible movement (e.g. gill covers movement) and if touching of the caudal peduncle produced no reaction. Records were kept of visible abnormalities (e.g. loss of equilibrium, swimming behaviour, respiratory function, pigmentation, etc.). Observations were made after 24, 48, 72 and 96 h.
Test concentrations:
- Range finding study: Yes
Cumulative Mortality [%] in the Preliminary
(n = 5)
Nominal test substance concentration [mg/L] Test Duration [hours]
24 48 72 96
10 100 --- --- ---
1 0 0 0 0
Controls 0 0 0 0 - Reference substance (positive control):
- no
- Key result
- Duration:
- 96 h
- Dose descriptor:
- LC50
- Effect conc.:
- 1.95 mg/L
- Nominal / measured:
- meas. (arithm. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- mortality (fish)
- Remarks on result:
- other: Interval: 1.75-2.17 mg/L
- Key result
- Duration:
- 96 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 0.564 mg/L
- Nominal / measured:
- meas. (arithm. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- mortality (fish)
- Key result
- Duration:
- 96 h
- Dose descriptor:
- LC0
- Effect conc.:
- 0.564 mg/L
- Nominal / measured:
- meas. (arithm. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- mortality (fish)
- Details on results:
- All effect levels are given based on mean measured test substance concentrations: At the test substance concentrations of 0.564 mg/L, no mortality was observed. Non lethal and lethal effects were observed in the test substance concentration of ≥ 1.06 mg/L. Therefore the NOEC was laid down as 0.564 mg/L and the LOEC was laid down as 1.06 mg/L.
- Results with reference substance (positive control):
- No reference substance tested
- Reported statistics and error estimates:
- LC50-values and 95 % confidence intervals after 72 and 96 hours exposure were calculated by sigmoidal dose-response regression. After 24 - 48 hours only 0 and 100 % mortality were found. No evaluation via regression analysis was carried out. Hence the LC50 is the mean measured value of the concentrations with 0 and 100 % mortality. The concentration causing 0 and 100 % mortality were chosen as lower and upper confidence limit. The concentrations causing 0 and 100 % mortality (LC0 and LC100) as well as the no effect level (NOEC) after 96 h were carried out using standard procedures.
- Sublethal observations / clinical signs:
Table 1. Observations in the test vessels
Mean measured test substance concentration [mg/L]
Effect
Test Duration [h]
2
24
48
72
96
8.82
(E)
-
7/7
---
---
---
(2.6)
7/7
-
---
---
---
3.85
(E)
-
-
7/7
---
---
(2.7)
-
6/7
---
---
---
(2.6)
-
4/7
---
---
---
(2.5)
-
1/7
---
---
---
(2.4)
-
6/7
---
---
---
(2.2)
-
1/7
---
---
---
(1)
7/7
-
---
---
---
2.14
(E)
-
-
-
1/7
3/6
(2.8)
-
-
-
6/7
3/3
(2.7)
-
-
-
6/7
3/3
(2.5)
-
-
-
-
3/3
(2.4)
-
-
-
6/7
-
(2.3)
-
-
1/7
-
-
(2.1)
-
-
-
-
1/3
(1)
7/7
7/7
6/7
-
-
1.06
(E)
-
-
-
-
1/7
(2.7)
-
-
-
-
6/7
(2.6)
-
-
-
-
6/7
(2.4)
-
-
-
-
6/7
(2.3)
-
-
-
1/7
-
(1)
7/7
7/7
7/7
6/7
-
0.564
(1)
7/7
7/7
7/7
7/7
7/7
Control
(1)
7/7
7/7
7/7
7/7
7/7
Table 2. Cumulative mortality [%] in the test vessels
Mean measured test substance concentration
[mg/L]
Test Duration [h]
2
24
48
72
96
8.82
0
100
100
100
100
3.85
0
0
100
100
100
2.14
0
0
0
14
57
1.06
0
0
0
0
14
0.564
0
0
0
0
0
Control
0
0
0
0
0
- Validity criteria fulfilled:
- yes
- Conclusions:
- Based on the results of the read across study, the 96 h LC50 and NOEC (Danio rerio) were determined to be 1.95 and 0.564 mg/L (mean measured), respectively.
- Executive summary:
A study was conducted to determine the short-term toxicity of the read across substance, TMP(EO)TA, to fish according to OECD Guideline 203 and EU Method C.1, in compliance with GLP. In a semi-static test, Zebra fish (Danio rerio) were exposed for 96 h to 5 nominal concentrations of 0.625, 1.25, 2.50, 5.00 and 10.0 mg/L, corresponding to mean measured concentrations of 0.564, 1.06, 2.14, 3.85 and 8.82 mg/L. Seven test organisms were exposed at each dose level. Water quality parameters, pH, temperature and oxygen-saturation measured after 0, 2, 24, 48, 72 and 96 h were determined to be within the acceptable limits. The test substance concentrations were verified via LC-MS/MS at test start and end. The substance was clearly dissolved at all concentrations throughout exposure. Recovery rates of test substance in the new media were in the range of 82 - 106% and in the old media in the range of 69 – 85% of nominal values. All effect levels were given based on mean measured concentrations. Non-lethal and lethal effects were observed at a concentration of ca. 1.06 mg/L. Therefore, the NOEC was established as 0.564 mg/L and the LOEC was 1.06 mg/L. Further, based on a mortality rate at 96 h of 14 and 57% at 1.06 and 2.14 mg/L respectively, the 96 h LC50 (Danio rerio) was established at 1.95 mg/L (mean measured) (Sheerbaum, 2010).
Referenceopen allclose all
Description of key information
Key value for chemical safety assessment
Fresh water fish
Fresh water fish
- Effect concentration:
- 1.95 mg/L
Additional information
A study was conducted to determine the short-term toxicity of the read across substance, TMP(EO)TA, to fish according to OECD Guideline 203 and EU Method C.1, in compliance with GLP. In a semi-static test, Zebra fish (Danio rerio) were exposed for 96 h to 5 nominal concentrations of 0.625, 1.25, 2.50, 5.00 and 10.0 mg/L, corresponding to mean measured concentrations of 0.564, 1.06, 2.14, 3.85 and 8.82 mg/L. Seven test organisms were exposed at each dose level. Water quality parameters, pH, temperature and oxygen-saturation measured after 0, 2, 24, 48, 72 and 96 h were determined to be within the acceptable limits. The test substance concentrations were verified via LC-MS/MS at test start and end. The substance was clearly dissolved at all concentrations throughout exposure. Recovery rates of test substance in the new media were in the range of 82 - 106% and in the old media in the range of 69 – 85% of nominal values. All effect levels were given based on mean measured concentrations. Non-lethal and lethal effects were observed at a concentration of ca. 1.06 mg/L. Therefore, the NOEC was established as 0.564 mg/L and the LOEC was 1.06 mg/L. Further, based on a mortality rate at 96 h of 14 and 57% at 1.06 and 2.14 mg/L respectively, the 96 h LC50 (Danio rerio) was established at 1.95 mg/L (mean measured) (Sheerbaum, 2010).
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