2,2,2-trifluoroethanol

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from 1985.03.25 to 1985.03.29

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

Study not performed according to international standard nor under GLP but following a methodology comparable to an OECD 203 guideline. The report is not very well documented but all the relevant information is reported.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Principles of method if other than guideline:

This test on 2, 2, 2 trifluoroethanol is part of a serie of similar tests on fish performed between 1978 and 1985 by Geiger. The majority of tests were conducted at the U.S. EPA Environmenatl Research Laboratory-Duluth with the reminder completed at the University of Wisconsin-Superior campus.

GLP compliance:

no

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 0 (control), 62.9, 96.8, 149, 229, 352
- Sampling method: All test exposure chambers were sampled at approximatively mid-depth at 0, 24, 48, 72 and 96h.
- Sample storage conditions before analysis: all samples were analysed immediately or adequately preserved for later analysis.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: diluted stock of 1900 mg/l (stirred)
- Eluate: not specified
- Differential loading: not specified
- Controls: yes
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): none
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): not applicable
- Evidence of undissolved material (e.g. precipitate, surface film, etc): none

Test organisms (species):

Pimephales promelas

Details on test organisms:

TEST ORGANISM
- Common name: Fathead Minnow
- Strain: not specified
- Source:US EPA Environmental research laboratory Duluth and the University of Wisconsin superior campus
- Age at study initiation (mean and range, SD): 31-32 days
- Length at study initiation (length definition, mean, range and SD): 17.7 mm (2.029)
- Weight at study initiation (mean and range, SD): 0.076 g (0.0284)
- Method of breeding: modified mini-diluters
- Feeding during test: no

ACCLIMATION
- Acclimation period: yes
- Acclimation conditions (same as test or not): yes
- Type and amount of food: frozen adult brine shrimp (Artemia sp.)
- Feeding frequency: not specified
- Health during acclimation (any mortality observed): only groups of fish having healthy appearance and no history of unusual thermal exposure of abnormally high mortality rate were used for toxicant exposure.

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

None

Hardness:

46.3 (1.19) mg/l CaCO3

Test temperature:

24.7 (0.69) °C

pH:

7.9 (0.02)

Dissolved oxygen:

7.5 (0.21) mg/l
Daily measured in all tanks.

Salinity:

Not applicable

Nominal and measured concentrations:

Date Control A B C D E
Nominal (mg/l)0 62.9 96.8 149 229 352
Day 0 <1.0 67.2 97.1 128 197 308
Day 1 <1.0 76.3 111 141 204 294
Day 2 <1.0 67.9 97.8 130 202 307
Day 3 <1.0 66.0 100 130 202 301
Day 4 <1.0 67.4 98.9 138 203 304
Average 1.0 69.0 101 133 202 303
Cor Average 1.02 70.1 107 136 205 308

Details on test conditions:

TEST SYSTEM
- Test vessel: modified mini-diluters
- Material, size, headspace, fill volume: 2 litres
- Aeration: not specified
- Type of flow-through (e.g. peristaltic or proportional diluter): not specified
- Renewal rate of test solution (frequency/flow rate): 18 volume additions/day
- No. of organisms per vessel: 20
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): not applicable
- Biomass loading rate: 0.76 g/l

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Filtered Lake superior water or Dechlorinated water from City of Superior, WI.
- Total organic carbon: Not specified
- Particulate matter: < 1 mg/l
- Pesticides: Not specified
- Chlorine: 1.2- 4.1 mg/l (chloride)
- Alkalinity: Not specified
- Ca/mg ratio: 12.9-14.7 mg/l
- Conductivity: 78-138 µmhos/cm
- Culture medium different from test medium: no
- Intervals of water quality measurement: Not specified

OTHER TEST CONDITIONS
- Adjustment of pH: yes
- Photoperiod: 16 hr light/ 8 hr dark
- Light intensity: no data, light controlled photoperiod

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Death was the major test endpoint. The number of dead fish was noted every 24 hr after the beginning of a test, at which time they were also removed. For tests completed prior to 1983, only general behavioral changes were noted when they occurred. Therefore, an EC50 value was not calculable based upon this data. In tests completed after 1983, observations of fish behavior and toxic signs were made at 2-8, 24, 48, 72 and 96 hr. The changes in behavior and body morphology are briefly outlined in the remarks section for nearly every chemical tested. Unique behavior was also recorded using a color video camera and 0.5 inch video tape recorder.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.5
- Justification for using less concentrations than requested by guideline: no data
- Range finding study: not specified
- Test concentrations: 0, 62.9, 96.8, 149, 229, 352 mg/l
- Results used to determine the conditions for the definitive study: no data

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

119 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL=105-135

Key result

Duration:

96 h

Dose descriptor:

EC50

Effect conc.:

119 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

behaviour

Remarks on result:

other: 95% CL=105-135

Details on results:

Affected fish lost schooling behavior and swam near the tank surface. They were hyperactive and overreactive to external stimuli, had increased respiration, were darkly colored and lost equilibrium prior to death. Cumulative mortalities and toxic effects are reported in details inTable 1 and 2 of "Any information on results incl. tables" field.

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

No data

Sublethal observations / clinical signs:

Table 2: Cumulated mortalities of fish exposed to 2.2.2 trifluoroethanol

Nominal concentration (mg/l)	0	62.9	96.8	149	229	352
Initial	20	20	20	20	20	20
24h			2	5	16	19
48h			7	7	18	19
96h		1	10	10	19	20

Table 3: Cumulated toxic effects observed on fish exposed to 2.2.2 trifluoroethanol

Nominal concentration (mg/l)	0	62.9	96.8	149	229	352
Initial	20	20	20	20	20	20
24h			2	7	16	19
48h			8	7	18	19
96h		1	10	10	19	20

Validity criteria fulfilled:

yes

Conclusions:

The LC50 and EC50 (96h) were 119 mg/L based on mortality and behaviour, respectively. Affected fish were hyperactive and overreactive to external stimuli, had increased respiration, were darkly colored and lost equilibrium prior to death.

Executive summary:

The acute toxicity of 2,2,2 trifluoroethanol to the Fathead Minnow (Pimephales promelas) was dermined under flow-through conditions according to a methodology comparable to the OECD 203 Guideline, without GLP statement. This test is part of a series of similar tests on fish performed between 1978 and 1985 by Geiger. The majority of these tests were conducted at the U.S. EPA Environmenatl Research Laboratory-Duluth with the remainder completed at the University of Wisconsin-Superior campus. The nominal concentrations of 2,2,2 trifluoroethanol were 62.9, 96.8, 149, 229 and 352 mg/L with one control. The concentrations were analytically verified and corresponded to at least 80 per cent of the nominal concentration throughout the test. The treatment group and the control group had one replicate, each containing 20 fish and the test duration was 96h.

During the exposure, the temperature was 24.7 ± 0.69 °C, the pH was 7.9 ± 0.02 and the dissolved oxygen concentration was 7.5 ± 0.21 mg/l on average. Daily values of these parameters were not reported however the using of a high flow through system (18 vol/day) and the low biomass load (0.76g/l) give some evidence that its had been fairly constant during the test.

The number of dead fish was noted every 24 hr after the beginning of the test and they were also removed. Observations of fish behavior and toxic signs were made at 2-8, 24, 48, 72 and 96 hr. Affected fish were hyperactive and overreactive to external stimuli, had increased respiration, were darkly colored and lost equilibrium prior to death.

The LC50 and EC50 (96h) for mortality and behaviour were determined at 119 mg/L.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

Not specified

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Study performed according to OECD guidelines and without GLP statement. All validity criteria were fulfilled except for tested concentration because its not measured throughout the test. However, no detail on analytical purity of test substance were specified in this report.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

, no measurement of test substance were performed and no detail on analytical purity of test substance were specified in this report

GLP compliance:

no

Analytical monitoring:

no

Details on sampling:

not applicable

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: 10.0 g of test substance was mixed and dissolved in pure water. This solution was made up to 100 mL with pure water to prepare a test solution of 100 000 mg/L. For each concentration (300, 170, 95, 55 and 30 mg/L), the corresponding amount of stock solution of test substance was added to dilution water an made up to 3.0 L to give each test solution.
- Eluate: no data
- Differential loading: about 1.7
- Controls: For the control concentration, 3.0 L of dilution water was used.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): not applicable
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): not applicable
- Evidence of undissolved material (e.g. precipitate, surface film, etc): no data

Test organisms (species):

Oryzias latipes

Details on test organisms:

TEST ORGANISM
- Common name: Japanese killifish, medaka, etc.
- Strain: Oryzias latipes
- Source: Sankyo Suisan CO. Ltd (1-1 Ichigayatamachi, Shinjuku, Tokyo)
- Age at study initiation (mean and range, SD): no data
- Length at study initiation (length definition, mean, range and SD): Approx. 2.0 cm
- Weight at study initiation (mean and range, SD): Approx. 0.2 g
- Method of breeding: no data
- Feeding during test: no
- Food type: not applicable
- Amount: not applicable
- Frequency: not applicable
- Other: batch number = K98-H-0108

ACCLIMATION
- Acclimation period: Animals were kept and acclimatised under the same conditions as the test conditions (water quality, temperature, etc) for at least one week.
- Acclimation conditions (same as test or not): same as test
- Type and amount of food: Commercial Tetramin fish food (made by Tetra Werke)
- Feeding frequency: Food was given once per day (excluding national holidays) up to 24 hours before the test.
- Health during acclimation (any mortality observed): In the 7 days before the test, acclimatised fish had no apparent malformations and had a survival rate of >= 95%.

QUARANTINE (wild caught)
- Duration: not applicable
- Health/mortality: not applicable

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Remarks on exposure duration:

none

Post exposure observation period:

No

Hardness:

No data

Test temperature:

Test temperature was performed using Water temperature meter Yokogawa Electric Corporation 2455 02.
Temperature of test water was measured at start of exposure period and before and after renewal of water at each 24 hour period. This temperature varied between 23.1 and 24.2°C. See table 1 in "Any other information on materials and methods incl. tables" field.

pH:

pH was measured using pH meter Toa Denpa Co. Ltd. HM-40V.
pH of test water was measured at start of exposure period and before and after renewal of water at each 24 hour period. This pH varied between 6.9 and 8.4. See table 2 in "Any other information on materials and methods incl. tables" field.

Dissolved oxygen:

Dissolved oxygen was measured using dissolved oxygen meter Denki Kagaku Keiki Co Ltd DOL-10.
Dissolved oxygen concentration of test water was measured at start of exposure period and before and after renewal of water at each 24 hour period. This dissolved oxygen concentration varied between 5.0 and 8.4 mg O2/L. See table 3 in "Any other information on materials and methods incl. tables" field.

Salinity:

Not applicable

Nominal and measured concentrations:

Nominal concentrations: 0 (control), 30, 55, 95, 170 and 300 mg test substance/L.
No measured concentrations were performed.

Details on test conditions:

TEST SYSTEM
- Test vessel: glass beaker
- Type (delete if not applicable): no data
- Material, size, headspace, fill volume: 3.0 L
- Aeration: none
- Type of flow-through (e.g. peristaltic or proportional diluter): no data
- Renewal rate of test solution (frequency/flow rate): whole of test solution renewed every 24 hours after start of exposure
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): no replicate
- No. of vessels per control (replicates): no replicate
- No. of vessels per vehicle control (replicates): not applicable
- Biomass loading rate: no data

TEST MEDIUM / WATER PARAMETERS
Dilution water : Yokohama city tap water dechlorinated, etc. by activated charcoal treatment and heated to 24°C

OTHER TEST CONDITIONS
no data

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortalities (animals were considered to be dead if they showed no response after being lightly touched on the caudal peduncle with the tip of strainless steel tweezers) were counted at 24, 48, 72 and 96 hours after start of test.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: about 1.7
- Justification for using less concentrations than requested by guideline: no data
- Range finding study: test concentrations were chosen following a preliminary test.
- Test concentrations: 0 (control), 30, 55, 95, 170 and 300 mg/L
- Results used to determine the conditions for the definitive study: no data on the preliminary test.

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% confidence interval = 93 to 160 mg/L

Key result

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

300 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

55 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

See table 4 in "Any other information on results incl. tables" field.

- Behavioural abnormalities: no data
- Observations on body length and weight: no data
- Other biological observations: no data
- Mortality of control: no mortalities were observed in the control group during the test
- Other adverse effects control: no data
- Abnormal responses: no data
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: no data
- Effect concentrations exceeding solubility of substance in test medium: no data

Results with reference substance (positive control):

not applicable

Reported statistics and error estimates:

96h- Median lethal concentration (LC50) were determined by using Probit statistical method.

Sublethal observations / clinical signs:

Table 4 : Cumulative mortalities

Concentration of test substance (mg/L)	Number of mortalities (mortality rate)
	24 hrs	48 hrs	72 hrs	96 hrs
0 (control)	0 (0)	0 (0)	0 (0)	0 (0)
30	0 (0)	0 (0)	0 (0)	0 (0)
55	0 (0)	0 (0)	0 (0)	0 (0)
95	0 (0)	0 (0)	1 (10)	4 (40)
170	0 (0)	0 (0)	3 (30)	7 (70)
300	0 (0)	0 (0)	4 (40)	10 (100)

Number of test animals: 10 per concentration

Validity criteria fulfilled:

yes

Remarks:

No measurement of test substance was performed, but TFE is expected to be stable. Mortality in the control < 10%. O2 concentration >= 60% of the air saturation value and constant conditions was maintained, throughout the test

Conclusions:

Acute toxicity test of Trifluoroethanol (TFEA) to Japanese killifish, according the OECD Guideline 203, at 5 nominal test concentrations and one control concentration in dilution water demonstrated that LC50 (96h) was 120 mg/L (93 - 160 mg/L); the LC100(96h) and LC0(96h) were 300 and 55 mg/L, respectively.

Executive summary:

In a 96-h acute toxicity study under semi-static conditions following the OECD Guideline 203, Japanese killifish (Oryzias latipes, n=10) were exposed to nominal concentrations at 300, 170, 95, 55 and 30 mg of trifluoroethanol and with one control. No analytical verifications were performed during the test. During the test, the test water had a temperature of 23.1 to 24.2°C, pH of 6.9 to 8.4 and dissolved oxygen concentration of 5.0 to 8.4 mg/L. No mortalities were observed in the control group during the test but mortalities appeared at a test concentration of 95 mg/L after 72 hours. The results demonstrated that LC50(96h) was 120 mg/L (93 - 160 mg/L); the LC100(96h) and NOEC(96h) were 300 and 55 mg/L, respectively. All validity criteria were fulfilled (i.e. mortality in fish control < 10%, constant conditions were maintained throughout the test, dissolved oxygen concentration was >= 60% of the air saturation value throughout the test).

Description of key information

Two studies are available to cover this endpoint. The key study on the Fathead Minnow (Pimephales promelas) was realized in flow-through conditions according to a methodology comparable to the OECD 203 Guideline without GLP statement. The nominal concentrations of 2,2,2 -Trifluoroethanol were 62.9, 96.8, 149, 229 and 352 mg/L with one control. Twenty fish per vessel were exposed at these concentrations during 96 hours. The concentrations were analytically verified and corresponded to at least 80 per cent of the nominal concentrations throughout the test. No mortality occurred in the control during the test and the dissolved oxygen concentration was greater than 60% of the air saturation value throughout the test. Hence, the test was considered to be valid. The LC50 and EC50 (96h) for mortality and behaviour were determined at 119 mg/L.

Another supportive test was performed during 96h under semi-static conditions on the Japanese killifish (Oryzias latipes).The test followed the OECD Guideline 203 but no information on the substance purity is reported and no analytical verification was performed during the test. The results demonstrated that the LC50(96h) was 120 mg/L (93 - 160 mg/L). The LC100(96h) and NOEC (96h) were 300 and 55 mg/L, respectively. These results are consistent with the results of the key study.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

119 mg/L