Cyclopropanecarboxylic acid, 3-[(1Z)-2-chloro-3,3,3-trifluoro-1-propen-1-yl]-2,2-dimethyl-, (1R,3R)-rel-

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:

1986-11-14 to 1987-02-27

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

dissolved oxygen clearly low (38% of staturation); water temperature lower than recommended (only run 1); fish size higher than guideline recommendation; precipitation of the test material during the test.

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Test material proven to be stable in aqueous solution (Hill 1984), therefore solutions were only prepared at the beginning of the study (static bioassay)
- Samples were taken at 0, 45 (run 1) or 47 (run 2), and 96 hours; triplicate values were onl y reported for one concentration and time point: run 1, nominal 5.6 mg/L, 45 h; run 2, nominal 5.6 mg/L, 47 h. The triplicate results were practically identical (+/- 0.1 mg/L)
- Concentrations: nominal: 32, 18, 10, 5.6, 3.2 mg/L
- Sampling method and storage conditions before analysis: not reported

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Appropriate amount of test material dissolved in acetone (maximum 4 mL), added to dilution water in test vessel (40 L), stirred.
- Fresh test solution prepared only once, at beginning of exposure (static bioassay)
- Controls: freshwater and solvent (acetone, 100 microliter/L) controls
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone (2-propanone)
- Concentration of vehicle in test medium (final test solution(s) including solvent control): 100 microliter/L
- Evidence of undissolved material (e.g. precipitate, surface film, etc): After addition to water, the test material came back out of solution in all test vessels, particularly in the higher concentrations. Observations during the 96-hour exposure period indicated that the test material subsequently slowly re-dissolved; this could be proven analytically.

Test organisms (species):

Lepomis macrochirus

Details on test organisms:

TEST ORGANISM
- Common name: bluegill sunfish
- Age at study initiation: not reported; time on site 120 days (run 1) or 55 days (run 2)
- Length at study initiation (length definition, mean, range and SD): run 1: 31.3 mm (23 - 35 mm); run 2: 36.3 mm (30 - 40 mm)
- Weight at study initiation (mean and range, SD): run 1: 0.72 g (0.25 - 1.02 g); run 2: 0.95 g (0.58- 1.23 g)
- Method of breeding: Not reported
- Feeding during test: Not reported

ACCLIMATION
- Acclimation period: acclimatised to 22 +/- 1 °C for a period of 7 days
- Type and amount of food: Not reported
- Feeding frequency: Not reported
- Health during acclimation (any mortality observed): Not reported

QUARANTINE (wild caught)
- not applicable, fish obtained from breeding facility

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

None reported

Hardness:

Hardness: run 1: 56.0 mg/L as CaCO3, run 2: 54.0 mg/L as CaCO3,
Conductivity: run 1: 170 micro-Siemens/cm; run 2: 120 micro-Siemens/cm

Test temperature:

Run 1: 19.7 - 22.4 °C
Run 2: 21.4 - 23.0 °C

pH:

Run 1: 6.6 - 8.0
Run 2: 6.8 - 8.2

Dissolved oxygen:

Run 1: 3.0 - 9.0 mg/L (without aeration)
Run 2: 3.4 - 9.0 mg/L (gentle aeration)

Salinity:

Not determined (freshwater)

Nominal and measured concentrations:

Nominal: 3.2, 5.6, 10, 18, 32 mg/L
Analytical (mean of run 1 and run 2 at 0 h): 2.8, 2.9, 3.4, 2.9, 4.0 mg/L (after preparation of test solution)
Analytical (mean of run 1 and run 2 at 45/47 h): 3.2, 5.6, 9.9, 13.0, 17.0 mg/L (no change of test solution, static test)
Analytical (mean of run 1 and run 2 at 96 h): 3.7, 5.9, 10.0, 17.0, 24.5 mg/L (end of static test)

Details on test conditions:

TEST SYSTEM
- Test vessel: rectangular, open
- Material, size, headspace, fill volume: glass, 45x30x31 cm (capacity 42 L), nominal fill volume used: 40 L
- Aeration: run 1: none; run 2: gentle (not futher specified)
- Renewal rate of test solution (frequency): none (static)
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1 per run (1+2)
- No. of vessels per control (replicates): 1 per run (1+2)
- No. of vessels per vehicle control (replicates): 1 per run (1+2)
- Biomass loading rate: run 1: 0.18 g/L; run 2: 0.24 g/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Water from 20000 gallon reservoir, maintained at 22 +/- 1 °C
- No concentrations of dissolved substances (except hardness and conductivity) or water quality measurements reported

OTHER TEST CONDITIONS
- Adjustment of pH: none
- Photoperiod, light intensity: not reported

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- mortalities, symptoms of toxicity: at 24, 48, 72, and 96 hours of exposure

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.8 (nominal)
- Justification for using less concentrations than requested by guideline: 5 concentrations used as recommended
- Range finding study: None (a preliminary test of solubility was performed)
- Results used for reporting: mean measured of 3 measurements at 0, 45/47, and 96 hours

Reference substance (positive control):

no

Duration:

24 h

Dose descriptor:

LC0

Effect conc.:

17 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Remarks:

mean of 3 measurements (0, 47, 96 h)

Basis for effect:

mortality (fish)

Remarks on result:

other: Run 2 at correct temperature: no mortality

Duration:

48 h

Dose descriptor:

LC10

Effect conc.:

11 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Remarks:

mean of 3 measurements (0, 47, 96 h)

Basis for effect:

mortality (fish)

Remarks on result:

other: Run 2 at correct temperature: no dose/mortality relation, one fish died at 11 mg/L analytical concentration

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

> 17 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Remarks:

mean of 3 measurements (0, 47, 96 h)

Basis for effect:

weight

Remarks on result:

other: Run 2 at correct temperature: no dose/mortality relation, one fish died at 11 mg/L analytical (48 h)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 17 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Remarks:

mean of 3 measurements (0, 47, 96 h)

Basis for effect:

mortality (fish)

Remarks on result:

other: Run 2 at correct temperature: no dose/mortality relation, one fish died at 11 mg/L analytical (48 h)

Duration:

24 h

Dose descriptor:

LC0

Effect conc.:

14 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Remarks:

mean of 3 measurements (0, 45, 96 h)

Basis for effect:

mortality (fish)

Remarks on result:

other: Run 1 at low temperature: no mortality

Duration:

48 h

Dose descriptor:

LC10

Effect conc.:

14 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Remarks:

mean of 3 measurements (0, 45, 96 h)

Basis for effect:

mortality (fish)

Remarks on result:

other: Run 1 at low temperature: One fish died at 14 mg/L analytical concentration

Duration:

72 h

Dose descriptor:

LC10

Effect conc.:

10 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Remarks:

mean of 3 measurements (0, 45, 96 h)

Basis for effect:

mortality (fish)

Remarks on result:

other: Run 1 at low temperature: One fish each died at 10 and 14 mg/L analytical concentration

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 14 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Remarks:

mean of 3 measurements (0, 45, 96 h)

Basis for effect:

mortality (fish)

Remarks on result:

other: Run 1 at low temperature: One fish each died at 10 mg/L (72 h) and 14 mg/L (48 h). Both concentrations analytical.

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

8 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Remarks:

mean of 3 measurements (0, 47, 96 h)

Basis for effect:

behaviour

Remarks on result:

other: Run 2 at proper temperature: no symptoms observed at any concentration; only effect seen: 1 mortality at 11 mg/L (48 h)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

7.5 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Remarks:

mean of 3 measurements (0, 45, 96 h)

Basis for effect:

behaviour

Remarks on result:

other: Run 1 at low temperature: no symptoms observed up to 10 mg/L analytical (one fish at 10 mg/L died without foregoing symptoms). At 14 mg/L, one fish died, 2-3 showed symptoms at various times.

Details on results:

- Temperature: Run 1 had had to be repeated because the temperature in the test vessels fell below 21°C. A second definitive test was run, but did not yield any significantly different results.
- Behavioural abnormalities: Run 1 (low temperature): 2-3 fish at top concentrations showed quiescence, loss of balancing, and surfacing; run 2 (correct temperature): no visible symptoms at any concentration
- Observations on body length and weight: none reported
- Other biological observations: none reported
- Mortality of control: none
- Other adverse effects control: none
- Abnormal responses: Run 2 (correct temperature): one mortality observed at 11 mg/L (analytical) at 48 hours, without any foregoing symptoms. No symptoms or mortalities at the highest dose.
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: Even though acetone was used as dispersing agent, some of the test material initially came out of solution and subsequently slowly re-dissolved during the 96 hour test period. Therefore all values reported (LC50, LC10 as well as NOEC) are based on mean analytical concentrations in the test solutions at 0, 45/47, and 96 hours.
- Effect concentrations exceeding solubility of substance in test medium: A preliminary test indicated a solubility limit < 32 mg/L. Analytically determined concentrations at the beginning of exposure (0 h) were consistently below the nominal values; retrieval rates (% of nominal) decreased monotonously from 86 % at 3.2 mg/L to only 13 % at 32 mg/L (nominal). At 45/47 h, the decrase was less pronounced: 100 % retrieval at 3.2 mg/L to 53 % at 32 mg/L. At the end of exposure (96 h), complete retrieval (94-116 %) was found in all concentrations but the highest, whereas at the nominal value 32/mg/L only 77 % the were found in solution.

Results with reference substance (positive control):

None

Reported statistics and error estimates:

Mean results (and percentages of nominal) are reported for test material concentrations in the test solutions (samples at 0, 45/47, and 96 hours).
Since only one mortality (10% of the exposed fish) was observed in run 2, at an intermediate concentration of 11 mg/L (65% of the maximum concentration), no mortality statistics was performed, and no confidence limits of the LC50 could be determined.

Sublethal observations / clinical signs:

Table 1: Nominal and analytically determined concentrations of test solutions: Average of run 1 and 2

Nominal mg/L	0 h	% of Nominal		45/47 h	% of Nominal		96 h	% of Nominal		Mean	% of Nominal
32	4.0	12.5%	Mean	17.0	53.1%	Mean	24.5	76.6%	Mean	15.5	48.4%	Mean
18	2.9	16.1%	40.0%	13.0	72.2%	84.6%	17.0	94.4%	98.3%	10.5	58.3%	74.0%
10	3.4	33.5%	SD	9.9	98.5%	SD	10.0	99.5%	SD	7.8	77.5%	SD
5.6	2.9	51.8%	30.1%	5.6	99.1%	21.1%	5.9	105.4%	14.5%	4.8	85.7%	20.8%
3.2	2.8	85.9%		3.2	100.0%		3.7	115.6%		3.2	100.0%
Solvent c.	<0.32			<0.32			<0.32			<0.32
Water c.	<0.32			<0.32			<0.32			<0.32

Table 2: Mortality and Symptoms: Run 1

Nominal	Measured	Percent mortality and symptoms (see footnote) observed
conc. mg/L	conc. mg/L	24 h	48 h	72 h	96 h
32	14	0 (B)	10 (B)	10 (A)	10 (B)
18	10	0 (A)	0 (A)	10 (A)	10 (A)
10	7.5	0 (A)	0 (A)	0 (A)	0 (A)
5.6	4.7	0 (A)	0 (A)	0 (A)	0 (A)
3.2	3.2	0 (A)	0 (A)	0 (A)	0 (A)
Solvent c.	<0.32	0 (A)	0 (A)	0 (A)	0 (A)
Water c.	<0.32	0 (A)	0 (A)	0 (A)	0 (A)

A: no symptoms observed

B: symptoms in up to 30% of individuals: quiescence, loss of balance, surfacing

Table 3: Mortality and Symptoms: Run 2

Nominal	Measured	Percent mortality and symptoms (see footnote) observed
conc. mg/L	conc. mg/L	24 h	48 h	72 h	96 h
32	17	0 (A)	0 (A)	0 (A)	0 (A)
18	11	0 (A)	10 (A)	10 (A)	10 (A)
10	8	0 (A)	0 (A)	0 (A)	0 (A)
5.6	4.9	0 (A)	0 (A)	0 (A)	0 (A)
3.2	3.2	0 (A)	0 (A)	0 (A)	0 (A)
Solvent c.	<0.32	0 (A)	0 (A)	0 (A)	0 (A)
Water c.	<0.32	0 (A)	0 (A)	0 (A)	0 (A)

A: no symptoms observed

Validity criteria fulfilled:

yes

Conclusions:

The acute LC50 of the test substance to the bluegill sunfish (Lepomis macrochirus) was estimated to be > 17 mg/L at 24, 48, 72, and 96 hr of exposure (> 14 mg/L in a first run at lower temperature). Confidence limits could not be calculated. The study report is relevant and reliable with restrictions but not adequate for classification and labeling.

Executive summary:

The acute toxicity of the test material was assessed according to OECD 203 in the bluegill sunfish (Lepomis macrochirus), at nominal concentrations of 3.2, 5.6, 10, 18, and 32 mg/L, in a freshwater static test system. Appropriate amounts of the test substance were dissolved in acetone and mixed with 40 L of water to make up the test solutions, which remained unchanged until the end of exposure, and were analyzed by HPLC at 0, 45 or 47, and 96 hours. Some transient precipitation occurred, since the solubility limit of the test substance appears to be < 32 mg/L. Mean analytical concentrations at 0 hours were consistently below the nominal values (86% of nominal at 3.2 mg/L, only 13% at 32 mg/L). After 96 hours, recoveries ranged from 94-116 % of the nominal values in all but the highest concentrations; at 32 mg/L, only 77% were found in solution. All study results are based on mean analytical values.

The pH (6.6 - 8.2) and water hardness (54 - 56 mg/L) were within the recommendations of the guideline; a slightly low temperature (19.7 – 22.4 °C) in run1 was corrected to 21.4 – 23.0 °C in an independent run 2. The size of fish (means 31.3 and 36.3 mm) was slightly higher than recommended.

Dissolved oxygen was significantly low in run 1 without aeration (3 - 9 mg/L); gentle areation in run 2 did hardly improve the lower level (3.4 mg/L, 38% of saturation). Dose groups comprised 10 animals each; mortalities and signs of toxicity were recorded daily, at 24, 48, 72, and 96 hours of exposure.

In run 2, no symptoms and no mortalities were observed in any dose groups (up to 17 mg/L analytical), except one dead fish at 11 mg/L found at 48 hours (without any foregoing symptoms); hence a statistical calculation of the LC50 could not be performed. In run 1, clinical symptoms (quiescence, loss of balance, and surfacing) and 10% mortality each in the two highest dose groups were observed.

The authors estimate an acute LC50 of > 14 - 17 mg/L to Lepomis macrochirus at all four time points, 24, 48, 72, and 96 hours. No higher concentrations of the test material were applied.

From the report, it cannot be decided whether the significantly low oxygen saturation affected the results.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

disregarded due to major methodological deficiencies

Study period:

1983-10-01 to 1984-03-31

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

dissolved oxygen considerably reduced during exposure period (no aeration); water temperature and fish size higher than recommended; no concentrations > 10 mg/L tested in main study

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Fresh test solutions were prepared daily (semi-static bioassay)
- Two samples were taken from each batch of test water (four per vessel, water renewed daily at 24-hour intervals): at 0 hours (when the test water was freshly prepared), and at 24 hours (when the test water was changed)
- Concentration: nominal: 10 mg/L
- Sampling method and storage conditions before analysis: not reported

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Stock solution of test material (550 mg) in dilution water (20 L), by magnetic stirrer; further dilution to 55 L with water.
- Fresh test solutions prepared daily (semi-static bioassay)
- Controls: freshwater control
- Evidence of undissolved material (e.g. precipitate, surface film, etc): None. Analytically determined values at 0 and 24 hours almost identical with nominal concentration

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: carp
- Age at study initiation: 4-6 months (purchased one month after hatch, then kept for 2-4 months in holding tank on site; acclimatised for 3 weeks
- Length at study initiation (length definition, mean, range and SD): 56 +/- 1 mm
- Weight at study initiation (mean and range, SD): 4.4 +/- 0.3 g
- Method of breeding: Not reported
- Feeding during test: Not reported

ACCLIMATION
- Acclimation period: held in stock tanks for 2 - 4 months, treated with methylene blue / saline for 24 hours, then acclimatised for a period of 3 weeks
- Acclimation conditions: same as in test
- Type and amount of food: Not reported
- Feeding frequency: Not reported
- Health during acclimation (any mortality observed): Neither deaths nor abnormalities observed

QUARANTINE (wild caught)
- not applicable, fish bred in facility

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

None reported

Hardness:

Hardness: 48.8 mg/L as CaCO3
Alkalinity: 45.6 mg/L HCO3-
Conductivity: 121 micro-Siemens/cm

Test temperature:

24.1-25.0 °C

pH:

7.5 - 7.7 at 0 hours, 7.0 - 7.4 at 24 hours

Dissolved oxygen:

7.9 - 8.0 mg/L at 0 hours, 4.2 - 4.8 mg/L at 24 hours

Salinity:

Not determined (freshwater)

Nominal and measured concentrations:

Nominal: 10 ppm (equivalent to mg/L, assuming water density = 1 kg/L)
Analytical (0 h, after preparation of test solution): 9.3 - 10.3 ppm
Analytical (24 h, 24 hours later, before change of test solution): 9.7 - 10.8 ppm
Controls: all measurements below LOD (<0.5 ppm)

Details on test conditions:

TEST SYSTEM
- Test vessel: rectangular aquarium, open
- Material, size, headspace, fill volume: all glass, 60x30x36 cm (nominal 60 L), fill volume 55 L
- Aeration: dilution water aerated before introduction into test system, no aeration during test
- Renewal rate of test solution (frequency): every 24 hours (semi-static)
- No. of organisms per vessel: 10 (3 in preliminary range finding test)
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.8 g/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: tap water, dechlorinated with activated charcoal and aerated, then heated to 25°C prior to introduction into the test systems.
- Dissolved oxygen: 8.2 mg/L
- Total organic carbon: 3.2 mg/L
- Particulate matter: suspended solids <0.1 mg/L
- Metals (mg/L): As <0.01, Cd <0.005,Cr6+ <0.05, Cu <0.01, Fe 0.02, Pb <0.01, Hg <0.0005, Zn <0.01
- Pesticides: organic phosphate <0.1 mg/L
- Chlorine (residual): <0.1 mg/L
- Fluorine: <0.1 mg/L
- NH3 un-ionised: <0.03 mg/L
- Nitrate: 0.61 mg/L
- pH: 7.5
- Hardness (as CaCO3): 48.8 mg/L
- Alkalinity (as HCO3-): 45.6 mg/L
- Ca/Mg ratio: not reported
- Conductivity: 121 micro-Siemens/cm
- Intervals of water quality measurement: not reported

OTHER TEST CONDITIONS
- Adjustment of pH: none
- Photoperiod, light intensity: not reported
- Temperature: test tanks kept in a thermo-regulated room (25 +/- 1 °C)

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- mortalities, symptoms of toxicity: observed at 24, 48, 72, and 96 hours of exposure
- observations: abnormal movements, other signs including abnormal myxeosis, haemorrhage, and a whitening of body surface colour

TEST CONCENTRATIONS
- Spacing factor for test concentrations: only one concentration tested
- Justification for using less concentrations than requested by guideline: Classification by the Japanese Agricultural Chemicals Control Law (fish toxicity 'A', ‘B' and 'C’) considers substances with a 48-h LC50 of >= 10 ppm as the lowest grade 'A'. The intention of the sponsor was to demonstrate that the 96-h LC50 of the test chemical was above 10 ppm, so this single dose level was chosen for the study, after discussion and agreement with the sponsor.
- Range finding study: 3 fish each, exposed to 6 and 12 ppm of the test material
- Results used to determine the conditions for the definitive study: No mortalities observed in preliminary study at either concentration, up to 96 hours of exposure

Reference substance (positive control):

no

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

> 10 other: ppm (equivalent to mg/L, with d=1 kg/L)

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

First test water, measured: 0 hours: 9.3 ppm (mg/L); 24 hours: 10.7 ppm (mg/L)

Basis for effect:

mortality (fish)

Remarks on result:

other: no mortality observed

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 10 other: ppm (equivalent to mg/L, with d=1 kg/L)

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

Second test water, measured: 0 hours: 10.1 ppm (mg/L); 24 hours: 10.8 ppm (mg/L)

Basis for effect:

mortality (fish)

Remarks on result:

other: no mortality observed

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

> 10 other: ppm (equivalent to mg/L, with d=1 kg/L)

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

Third test water, measured: 0 hours: 10.2 ppm (mg/L); 24 hours: 9.7 ppm (mg/L)

Basis for effect:

mortality (fish)

Remarks on result:

other: no mortality observed

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 10 other: ppm (equivalent to mg/L, with d=1 kg/L)

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

Fourth test water, measured: 0 hours: 10.3 ppm (mg/L); 24 hours: 10.3 ppm (mg/L)

Basis for effect:

mortality (fish)

Remarks on result:

other: no mortality observed

Duration:

24 h

Dose descriptor:

NOEC

Effect conc.:

10 other: ppm (equivalent to mg/L, with d=1 kg/L)

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

First test water, measured: 0 hours: 9.3 ppm (mg/L); 24 hours: 10.7 ppm (mg/L)

Basis for effect:

behaviour

Remarks on result:

other: no observable symptoms

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

10 other: ppm (equivalent to mg/L, with d=1 kg/L)

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

Fourth test water, measured: 0 hours: 10.3 ppm (mg/L); 24 hours: 10.3 ppm (mg/L)

Basis for effect:

behaviour

Remarks on result:

other: no observable symptoms

Details on results:

- Behavioural abnormalities: none observed
- Observations on body length and weight: none reported
- Other biological observations: none reported
- Mortality of control: none
- Other adverse effects control: none
- Abnormal responses: none
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: Measured concentrations immediately after renewal of the test water were almost identical with the nominal concentration; there were no significant changes in the concentrations after 24 hours.
- Effect concentrations exceeding solubility of substance in test medium: none

Results with reference substance (positive control):

None

Reported statistics and error estimates:

No statistics on analytical concentrations performed. Mean concentrations (test water 1-4) calculated by reviewer: at 0 hours: 10.0 +/- 0.5 ppm, at 24 hours: 10.4+/- 0.5 ppm
Since no mortality was observed, no statistics was performed, and no confidence limits of the LC50 were determined.

Sublethal observations / clinical signs:

Table 1: Test water: pH, dissolved oxygen, and temperature

Nominal	Test water	pH		Dissolved oxygen mg/L		Temperature °C
concentration	renewal	0 h	24 h	0 h	24 h	0 h	24 h
0 ppm	1	7.6	7.1	8.0	4.3	24.9	24.3
(control)	2	7.5	7.2	8.0	4.2	24.9	24.1
	3	7.5	7.4	7.9	4.8	25.0	24.4
	4	7.6	7.2	8.0	4.5	24.8	24.5
10 ppm	1	7.6	7.0	8.0	4.2	24.8	24.4
	2	7.5	7.2	7.9	4.4	25.0	24.2
	3	7.6	7.3	8.0	4.5	24.9	24.4
	4	7.7	7.1	7.9	4.7	24.8	24.5

Validity criteria fulfilled:

yes

Conclusions:

The acute LC50 of the test substance to the carp (Cyprinus carpio) was estimated to be > 10 mg/L at 24, 48, 72, and 96 hr of exposure. No higher concentrations were tested. Since no mortality was observed, confidence limits could not be calculated. The study report is relevant but not adequate and reliable for risk assessment, classification and labeling.

Executive summary:

The acute toxicity of the test material was assessed according to OECD 203 in the carp (Cyprinus carpio), at one nominal concentration of 10 ppm (equivalent to 10 mg/L, assuming water density 1 kg/L)), in a freshwater semi-static test system. The test substance (550 mg) was mechanically mixed with 20 L water, and further diluted to make up 55 L of test solution, which was changed (prepared freshly) every 24 hours and analyzed by HPLC before and after exposure (0 and 24 hours). Analytical concentrations were 10.0 and 10.4 ppm (both +/- 0.5 ppm), respectively. Dissolved oxygen decreased considerably, from 7.9 – 8.0 mg/L at 0 h to 4.2 – 4.8 mg/L at 24 hours. The pH (7.0 – 7.7) and water hardness (48.8 mg/L) were within the recommendations of the guideline; the temperature (24.1 – 25.0 °C) was slightly higher than recommended. Due to a delayed start, the size of fish (5.6 cm) exceeded the recommendation of OECD 203 (3 +/- 1 cm), but were in the range (for carp: 5 cm) suggested by a Japanese MAFF Standard Method. The dose and control groups comprised 10 animals each; mortalities and signs of toxicity were recorded daily.

No mortalities were observed, so that a statistical calculation of the LC50 could not be performed. No clinical symptoms were detected in any of the fish.

The author estimates an acute LC50 of >10 ppm (mg/L) to Cyprinus carpio at all four time points, 24, 48, 72, and 96 hours. No higher concentrations of the test material were investigated, although neither the limit of solubility nor the condition for an OECD limit test were reached. Rationale: The intention of the sponsor was to establish the fish toxicity classification 'A' for the test material, according to the Japanese Agricultural Chemicals Control Law (48-h LC50 of >= 10 ppm), so this single dose level was deemed sufficient for the study.

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1984-03-12 to 1984-05-11

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

water temperature and fish size lower than recommended, short acclimation period; shortened exposure periods, 87.5 and 91.5 h, precipitation of the test material during the test.

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Fresh test solutions were prepared daily (semi-static bioassay)
- Two samples were taken from each test vessel, i.e. four times during the study: "ON" (when the test solution was freshly prepared), "OFF" (after 24 hours, when the test solution was changed)
- Concentrations: nominal: 18, 10, 7.5, 5.6, 3.2 mg/L
- Sampling method and storage conditions before analysis: not reported

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Stock solution of test material (100 mg/g) in acetone, further diluted as appropriate; necessary amounts added to test vessel and mixed with water to 10 L. For the highest test concentration, 180 mg test material were dissolved in 1 g of acetone and added to the test vessel.
- Fresh test solutions prepared daily (semi-static bioassay)
- Controls: freshwater and solvent controls (acetone, 100 mg/L)
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone (2-propanone)
- Concentration of vehicle in test medium (final test solution(s) including solvent control): 100 mg/L
- Evidence of undissolved material (e.g. precipitate, surface film, etc): White precipitate observed in the higher concentrations tested, gradually redissolved during the test. Analytical determinations: "ON" values (freshly prepared test solutions) vary from 32.8 % to 87.5 % of nominal, showing a clear concentration-dependent increase of losses, whereas "OFF" values (test solutions before changing after 24 hours) amount to 93.9 % of the nominal concentration (range 87.8 - 97.3 %, no concentration dependence)

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: rainbow trout
- Age at study initiation: 71 days (hatched on 1984-01-01)
- Length at study initiation (length definition, mean, range and SD): 36.95 mm (31 - 40 mm, no SD reported)
- Weight at study initiation (mean and range, SD): 0.63 g (0.33 - 0.83 g, no SD reported)
- Method of breeding: Not reported
- Feeding during test: Not reported

ACCLIMATION
- Acclimation period: held in stock tanks for 2 months, then acclimatised to the test temperature for a period of 5 days
- Acclimation conditions: same as in test for 5 days
- Type and amount of food: Not reported
- Feeding frequency: Not reported
- Health during acclimation (any mortality observed): Not reported

QUARANTINE (wild caught)
- not applicable, fish bred in facility

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

91.5 h

Remarks on exposure duration:

top concentration (18 mg/L) and controls; lower doses were only exposed for 87.5 hours

Post exposure observation period:

None reported

Hardness:

Hardness: 65.2 mg/L (ppm) as CaCO3, range 60.6 - 68.3 mg/L
Alkalinity: 28.4 mg/L CaCO3 (one measurement)
Conductivity: 120 micro-siemens/cm

Test temperature:

11.4 - 13.0 °C

pH:

7.4 - 8.0

Dissolved oxygen:

10.0 - 11.2 mg/L

Salinity:

Not determined (freshwater)

Nominal and measured concentrations:

Nominal: 3.2, 5.6, 7.5, 10, 18 mg/L
Analytical ("ON"): 2.8, 4.0, 4.8, 5.8, 5.9 mg/L (after preparation of test solution)
Analytical ("OFF"): 3.1, 5.3, 7.3, 9.3, 15.8 mg/L (24 hours later, before change of test solution)

Details on test conditions:

TEST SYSTEM
- Test vessel: rectangular, open
- Material, size, headspace, fill volume: glass, 39x28x28 cm (nominal 28 L), fill volume 10 L
- Aeration: continuous, compressed air (approx. 1 bar = 15 psi) passing through Pasteur pipettes
- Renewal rate of test solution (frequency): every 24 hours (semi-static)
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1
- Biomass loading rate: 0.63 g/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Water supply maintained at 12 +/- 2 °C
- No concentrations of dissolved substances (except hardness and alkalinity) or water quality measurements reported

OTHER TEST CONDITIONS
- Adjustment of pH: none
- Photoperiod, light intensity: not reported

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- mortalities, symptoms of toxicity
- observation in controls and highest dose at 2.5, 19.5, 26.5, 43.5, 50.5, 67.5, 74.5 and 91.5 hours of exposure
- observation in intermediate doses at 15.5, 22.5, 39.5, 46.5, 63.5, 70.5 and 87.5 hours of exposure

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.3 - 1.8
- Justification for using less concentrations than requested by guideline: 5 concentrations used as recommended
- Range finding study: None
- Results used to determine the conditions for the definitive study: mean measured of 4 measurements of "OFF" concentration (after each 24-h exposure)

Reference substance (positive control):

no

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

> 15.8 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Remarks:

measured "OFF" values in test solutions after each 24 hour exposure period

Basis for effect:

mortality (fish)

Remarks on result:

other: no dose/mortality relation could be established

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 15.8 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Remarks:

measured "OFF" values in test solutions after each 24 hour exposure period

Basis for effect:

mortality (fish)

Remarks on result:

other: no dose/mortality relation could be established

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

> 15.8 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Remarks:

measured "OFF" values in test solutions after each 24 hour exposure period

Basis for effect:

mortality (fish)

Remarks on result:

other: no dose/mortality relation could be established

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 15.8 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Remarks:

measured "OFF" values in test solutions after each 24 hour exposure period

Basis for effect:

mortality (fish)

Remarks on result:

other: no dose/mortality relation could be established

Duration:

24 h

Dose descriptor:

NOEC

Effect conc.:

15.8 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Remarks:

measured "OFF" values in test solutions after each 24 hour exposure period

Basis for effect:

behaviour

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

9.3 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Remarks:

measured "OFF" values in test solutions after each 24 hour exposure period

Basis for effect:

behaviour

Remarks on result:

other: One fish of the 7.3 mg/L group (analytical "OFF" concentration) died at 70.5 hours. Since the animal showed no foregoing symptoms, and there was no dose-mortality relation, it is unclear whether this death is related to the treatment.

Details on results:

- Behavioural abnormalities: rapid respiration at the highest concentration only (15.8 mg/L analytical "OFF" concentration), onset at 43.5 hours (5/10 fish), mitigation towards end of study (3/10)
- Observations on body length and weight: none reported
- Other biological observations: none reported
- Mortality of control: none
- Other adverse effects control: none
- Abnormal responses: one mortality observed at 7.3 mg/L (analytical) at 70.5 hours, without any foregoing symptoms. It is unclear whether this is treatment-related.
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: A white precipitate was observed
in the higher concentrations tested. Analytical measurements showed that this precipitate gradually redissolved during the test. Therefore all values reported (LC50 as well as NOEC) are based on mean analytical concentrations in test solutions after 24 hours of exposure ("OFF" concentrations)
- Effect concentrations exceeding solubility of substance in test medium: at the end of each 24 hour exposure period, mean analytical concentrations ranged from 87.8 - 97.3% of the nominal concentration, which indicates that the re-dissolution of the test substance was almost complete

Results with reference substance (positive control):

None

Reported statistics and error estimates:

Mean results (and percentages of nominal) are reported for test material concentrations in the four test solutions, immediately after preparation (samples "ON") and after 24 hours, before the solutions were changed (samples "OFF").
Since only one mortality (10% of the exposed fish) was observed, at an intermediate concentration of 7.3 mg/L (46% of the maximum concentration), no mortality statistics was performed, and no confidence limits of the LC50 could be determined.

Sublethal observations / clinical signs:

Table 1: Nominal and analytically determined concentrations of test solutions: Summary

Nominal mg/L	"ON" mg/L	% of Nominal		"OFF" mg/L	% of Nominal
18	5.9	32.8%	Mean	15.8	87.8%	Mean
10	5.8	58.0%	62.7%	9.3	93.0%	93.9%
7.5	4.8	64.0%	SD	7.3	97.3%	SD
5.6	4	71.4%	20.1%	5.3	94.6%	3.9%
3.2	2.8	87.5%		3.1	96.9%

Table 2: Nominal and analytically determined concentrations of test solutions: Detail

Nominal mg/L	Date of sample	Exp. time h	"ON" analyt. mg/L	Mean	% of nominal	Date of sample	Exp. time h	"OFF" analyt. mg/L	Mean, Std. Dev.	% of nominal
Fresh-	12.03.1984	0	<0.2	Mean		13.03.1984	24	<0.2	Mean
water	13.03.1984	25	<0.2	<0.2	-	14.03.1984	46	<0.2	<0.2	-
	14.03.1984	49	<0.2			15.03.1984	70	<0.2
	15.03.1984	72	<0.2			16.03.1984	94	<0.2
Acetone	12.03.1984	0	<0.2	Mean		13.03.1984	24	<0.2	Mean
100	13.03.1984	25	<0.2	<0.2	-	14.03.1984	46	<0.2	<0.2	-
mg/L	14.03.1984	49	<0.2			15.03.1984	70	<0.2
	15.03.1984	72	<0.2			16.03.1984	94	<0.2
3.2	12.03.1984	0	3.1	Mean		13.03.1984	20	3.1	Mean
	13.03.1984	21	2.5	2.8	87.5	14.03.1984	42	3.2	3.1	96.9
	14.03.1984	45	2.8	SD		15.03.1984	66	3.1	SD
	15.03.1984	68	2.7	0.3		16.03.1984	90	3.1	0.1
5.6	12.03.1984	0	4.2	Mean		13.03.1984	20	5.4	Mean
	13.03.1984	21	3.7	4.0	71.4	14.03.1984	42	5.2	5.3	94.6
	14.03.1984	45	4.1	SD		15.03.1984	66	5.3	SD
	15.03.1984	68	4.0	0.2		16.03.1984	90	5.4	0.1
7.5	12.03.1984	0	5.4	Mean		13.03.1984	20	7.4	Mean
	13.03.1984	21	4.7	4.8	64.0	14.03.1984	42	6.8	7.3	97.3
	14.03.1984	45	4.0	SD		15.03.1984	66	8.1	SD
	15.03.1984	68	5.1	0.6		16.03.1984	90	6.7	0.6
10	12.03.1984	0	6.5	Mean		13.03.1984	20	9.4	Mean
	13.03.1984	21	5.4	5.8	58.0	14.03.1984	42	9.2	9.3	93.0
	14.03.1984	45	5.7	SD		15.03.1984	66	9.4	SD
	15.03.1984	68	5.5	0.5		16.03.1984	90	9.0	0.2
18	12.03.1984	0	6.9	Mean		13.03.1984	24	15.9	Mean
	13.03.1984	25	4.7	5.9		14.03.1984	46	15.7	15.8	87.8
	14.03.1984	49	5.8	SD		15.03.1984	70	16.0	SD
	15.03.1984	72	6.3	0.9	32.8	16.03.1984	94	15.4	0.3

Table 3: Mortality and Symptoms:

12.03.84	Exposure time (h)**	2.5	2.5	-**	-**	-**	-**	2.5
16:00	Nominal conc (mg/L)	FW*	Solv*	3.2	5.6	7.5	10.0	18.0
	Dead in vessel	0	0	0	0	0	0	0
	Normal behaviour	10	10					10
	Respiration rapid
13.03.84	Exposure time (h)**	19.5	19.5	15.5	15.5	15.5	15.5	19.5
09:00	Nominal conc (mg/L)	FW*	Solv*	3.2	5.6	7.5	10.0	18.0
	Dead in vessel	0	0	0	0	0	0	0
	Normal behaviour	10	10	10	10	10	10	10
	Respiration rapid
13.03.84	Exposure time (h)**	26.5	26.5	22.5	22.5	22.5	22.5	26.5
16:00	Nominal conc (mg/L)	FW*	Solv*	3.2	5.6	7.5	10.0	18.0
	Dead in vessel
	Normal behaviour	10	10	10	10	10	10	10
	Respiration rapid
14.03.84	Exposure time (h)**	43.5	43.5	39.5	39.5	39.5	39.5	43.5
09:00	Nominal conc (mg/L)	FW*	Solv*	3.2	5.6	7.5	10.0	18.0
	Dead in vessel
	Normal behaviour	10	10	10	10	10	10	5
	Respiration rapid							5
14.03.84	Exposure time (h)**	50.5	50.5	46.5	46.5	46.5	46.5	50.5
16:00	Nominal conc (mg/L)	FW*	Solv*	3.2	5.6	7.5	10.0	18.0
	Dead in vessel
	Normal behaviour	10	10	10	10	10	10	4
	Respiration rapid							6
15.03.84	Exposure time (h)**	67.5	67.5	63.5	63.5	63.5	63.5	67.5
09:00	Nominal conc (mg/L)	FW*	Solv*	3.2	5.6	7.5	10.0	18.0
	Dead in vessel
	Normal behaviour	10	10	10	10	10	10	7
	Respiration rapid							3
15.03.84	Exposure time (h)**	74.5	74.5	70.5	70.5	70.5	70.5	74.5
16:00	Nominal conc (mg/L)	FW*	Solv*	3.2	5.6	7.5	10.0	18.0
	Dead in vessel					1
	Normal behaviour	10	10	10	10	9	10	8
	Respiration rapid							2
16.03.84	Exposure time (h)**	91.5	91.5	87.5	87.5	87.5	87.5	91.5
09:00	Nominal conc (mg/L)	FW*	Solv*	3.2	5.6	7.5	10.0	18.0
	Dead in vessel
	Normal behaviour	10	10	10	10	9	10	7
	Respiration rapid							3

* FW = freshwater, Solv = 100 mg/L acetone in water

** Intermediate concentrations started 4 hours after 18 mg/L and controls

Validity criteria fulfilled:

yes

Conclusions:

The acute LC50 of the test substance to the rainbow trout (Oncorhynchus mykiss, reported as Salmo gairdneri) was estimated to be > 15.8 mg/L at approximately 24, 48, 72, and 96 hr of exposure. Confidence limits could not be calculated. The study report is relevant and reliable with restrictions but not adequate for classification and labeling.

Executive summary:

The acute toxicity of the test material was assessed according to OECD 203 in the rainbow trout (Oncorhynchus mykiss, reported as Salmo gairdneri), at nominal concentrations of 3.2, 5.6, 7.5, 10, and 18 mg/L, in a freshwater semi-static test system. Appropriate amounts of test substance stock solutions in acetone were mixed with water to make up 10 L of test solution, which was changed every 24 hours and analyzed by HPLC before and after exposure. In spite of some transient precipitation, concentrations after each 24-hour exposure period (“OFF”) amounted to 93.9% +/- 3.9% of the nominal values; all results are reported as analytical “OFF” concentrations. Dissolved oxygen (10 - 11.2 mg/L), pH (7.4 - 8.0) and water hardness (60.6 - 68.3 mg/L) were within the recommendations of the guideline; temperature (11.4 – 13.0 °C) and size of fish (31 – 40 mm) were slightly lower than recommended. Dose groups comprised 10 animals each; mortalities and signs of toxicity were recorded twice daily, during the exposure periods of approximately 96 hours (87.5 – 91.5 h, depending on the dose group).

No mortalities in any dose groups (up to 15.8 mg/L) were observed, except one dead fish at 7.3 mg/L found at 70.5 hours (without any forgoing symptoms), so that a statistical calculation of the LC50 could not be performed. The only clinical symptom was rapid respiration, first seen in 5/10 animals of the highest group (15.8 mg/L) at 43.5 hours, gradually declining to 3/10 animals at the end of the study.

The author estimates an acute LC50 of > 15.8 mg/L to Oncorhynchus mykiss at all four time points, 24, 48, 72, and 96 hours. No higher concentrations of the test material were tested, although the limit of solubility was not reached; no rationale is given.

Description of key information

LC50 > 15.8 mg/L, 96-hour, semi-static, rainbow  trout, method equivalent to OECD203, Hill 1984
LC50 > 17 mg/L, 96-hour, static, bluegill sunfish, method equivalent toOECD203, Tapp & Caunter 1987
LC50 > 10 mg/L, 96-hour, semi-static, carp, method equivalent toOECD203, Yamauchi 1984a

Additional information

Rainbow trout (Oncorhynchus mykiss, reported as Salmo gairdneri):

Only a single semi-static toxicity study conducted under GLP and in according to a method equivalent to guideline OECD 203 is available (Hill 1984).

Since none of the five concentrations (nominal up to 18 mg/L) elicited mortalities above 50% (the only fish that died was exposed to the middle concentration of 7.5 mg/L), the LC50 in the rainbow trout (Oncorhynchus mykiss) was extrapolated to be >15.8 mg/L (analytical).

However the concentrations tested were only up to 15.8 mg/L and therefore the conclusions are valid only up to this level. No attempt to reach the limit concentration of 100 mg/L or to determine the LC50 at a concentration up to the limit of solubility. Further minor deviations from guideline OECD 203 were detected. The study is relevant and reliable with restrictions but not adequate for classification and labeling.

Bluegill sunfish (Lepomis macrochirus):

Only a single static toxicity study conducted under GLP and according to a method equivalent to guideline OECD 203 is available (Tapp & Caunter 1987).

Since none of the five concentrations (nominal up to 32 mg/L, greater than the limit of solubility) elicited mortalities above 10% (the only fish that died was exposed to the second-highest concentration of 11 mg/L), the LC50 in the bluegill sunfish (Lepomis macrochirus) was extrapolated (from the highest analytical concentration) to be >17 mg/L (analytical).

A first run, considered less valid due to a test vessel temperature below 21°C, reached a maximum analytical concentration of 14 mg/L (with 10% mortality), thus a lower estimate of the LC50, >14 mg/L, was reported for this run.

Although a preliminary test indicated a solubility limit < 32 mg/L, however in the test with aquatic invertebrates (Yamauchi 1984b) the concentration tested was >200 ppm (equivalent to mg/L assuming water density =1kg/L). No attempt was made to reach the limit concentration of 100 mg/L or to determine the LC50 more precisely.

The study is considered to be relevant and reliable with restrictions but not adequate for classification and labeling.

Carp (Cyprinus carpio):

Only a single semi-static toxicity study conducted under and GLP according to a method equivalent to guideline OECD 203 is available (Yamauchi 1984a).

The intention of the sponsor was to establish the fish toxicity classification 'A' for the test material, according to the Japanese Agricultural Chemicals Control Law (48-h LC50 of >= 10 ppm), so a single concentration level of 10 mg/L was deemed sufficient for the study. Since no mortalities and no symptoms were observed up to 96 gours of exposure, the LC50 in the carp (Cyprinus carpio) was extrapolated to be >10 mg/L (nominal, for the test material as supplied). Since only a single concentration was tested, the study is disregarded and therefore not adequate and reliable for the purpose of classification and labelling.