Potassium (1R,3R)-3-[(1Z)-2-chloro-3,3,3-trifluoroprop-1-en-1-yl]-2,2-dimethylcyclopropanecarboxylate and Potassium (1S,3S)-3-[(1Z)-2-chloro-3,3,3-trifluoroprop-1-en-1-yl]-2,2-dimethylcyclopropanecarboxylate

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1984-01-10 to 1984-03-31

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1984

Report date:

1984

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Remarks:

based on the guideline issued 1984-04-04. Some newer requirements of the 2004-04-13 issue (water contaminants and hardness) are not met.

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

reaction mass of (1R,3R)-3-[(1Z)-2-chloro-3,3,3-trifluoroprop-1-en-1-yl]-2,2-dimethylcyclopropanecarboxylic acid and (1S,3S)-3-[(1Z)-2-chloro-3,3,3-trifluoroprop-1-en-1-yl]-2,2-dimethylcyclopropanecarboxylic acid

EC Number:

614-283-9

Cas Number:

68127-59-3

Molecular formula:

C9H10ClF3O2

IUPAC Name:

reaction mass of (1R,3R)-3-[(1Z)-2-chloro-3,3,3-trifluoroprop-1-en-1-yl]-2,2-dimethylcyclopropanecarboxylic acid and (1S,3S)-3-[(1Z)-2-chloro-3,3,3-trifluoroprop-1-en-1-yl]-2,2-dimethylcyclopropanecarboxylic acid

Details on test material:

- Physical state: powder
- Colour: white
- Expiration date of the lot/batch: not reported
- Storage condition of test material: not reported (reference sample stored <5°C)

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: A stock solution (200 ppm, equivalent to mg/L) was prepared at the beginning of the experiment, and used as such or appropriately diluted to yield nominal tank concentrations of 30, 50, 80, 130, and 200 ppm.
- Sampling method: Samples were taken from each tank (3 replicates per dose, 2 subsequent studies) at the beginning (0 h) and the end (48 h) of exposure, and diluted with water to yield samples containing 8 - 15 ppm of the test substance.
- Sample storage conditions before analysis: not reported

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Stock solution of test material (500 mg) in dilution water (2.5 L), by mechanical dispersion (probably vigorous); further dilution to yield the lower nominal tank concentrations of 30, 50, 80, and 130 ppm.
- Test solutions prepared once, at the beginning of the experiment (static bioassay)
- Controls: freshwater control
- Evidence of undissolved material (e.g. precipitate, surface film, etc): None. Analytically determined values at 0 hours were practically identical with nominal concentrations (100 +/- 4%), at 48 hours even slightly higher (106 +/- 6%)

Test organisms

Test organisms (species):

Daphnia pulex

Details on test organisms:

TEST ORGANISM
- Common name: daphnia
- Age at study initiation: First instar, less than 24 hours
- Weight and length at study initiation (mean and range, SD): not applicable to daphnia
- Method of breeding: 50 parent Daphnia kept in 2-L holding tank on-site (20+/-1°C, 1000 Lux, 16hr light / 8hr dark period); diet: Selenastrum capricornutum (7.5x10exp8 per tank per day). First instars from 3 - 4 week old parent daphnia used in the test.
- Feeding during test: no

ACCLIMATION
- Acclimation period: none; breeding conditions same as test

QUARANTINE (wild caught): No, daphnia bred on site

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Remarks on exposure duration:

24-h results also reported

Post exposure observation period:

None reported

Test conditions

Hardness:

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

Test temperature:

20 +/- 1°C (min 19.4°C, max 20.3°C)

pH:

7.3 - 7.6 at 0 hours, 6.4 - 7.5 at 48 hours

Dissolved oxygen:

7.8 - 8.4 mg/L at 0 hours, 6.2 - 7.8 mg/L at 48 hours

Salinity:

Not determined (freshwater)

Nominal and measured concentrations:

Nominal: 30, 50, 80, 130, 200 ppm (equivalent to mg/L, assuming water density = 1 kg/L)
Analytical (0 h, 1st / 2nd study, average of 3 tanks each): 31 / 28 ppm, 53 / 51 ppm, 76 / 82 ppm, 130 / 127 ppm, 198 / 196 ppm
Analytical (48 h, 1st / 2nd study, average of 3 tanks each): 33 / 32 ppm, 51 / 48 ppm, 83 / 88 ppm, 136 / 144 ppm, 217 / 220 ppm
Controls: all measurements below LOD (<0.5 ppm)

Details on test conditions:

TEST SYSTEM
- Test vessel: cylindrical aquarium, open
- Material, size, headspace, fill volume: all glass, diameter 7.4 cm, height 10 cm, nominal volume 300 mL, fill volume 250 mL
- Aeration: dilution water aerated before introduction into test tanks, no aeration during test
- Renewal rate of test solution (frequency): 0 (static)
- No. of independent studies: 2 (subsequent)
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 3 per study (total 6)
- No. of vessels per control (replicates): 3 per study (total 6)
- Biomass loading rate: not reported

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: tap water, dechlorinated with activated charcoal and heated to 20°C, then aerated prior to introduction into the test tanks.
- 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: 16 h light and 8 h dark period, 1000 Lux
- Temperature: test tanks kept in a thermo-regulated room (20 +/- 1 °C)

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Daphnia with "inhibited movement" (immobilised or showing only minor movements of their appendages) were recorded after 3, 6, 24, and 48 hours of exposure.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: approx. 1.6
- Justification for using less concentrations than requested by guideline: none (5 concentrations used, including 0% and 100% immobilisation)
- Range finding study (RF): two preliminary tests
- Test concentrations in RF (with % affected daphnia): first study: 10 ppm (0%), 30 ppm (0%), 100 ppm (70%), 300 ppm (100%); second study 30 ppm (0%), 50 ppm (5%), 80 ppm (25%),130 ppm (65%), 200 ppm (100%)
- Results used to determine the conditions for the definitive study: all above

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

3 h

Dose descriptor:

EC50

Effect conc.:

> 200 other: ppm (equivalent to mg/L, with d=1 kg/L)

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

measured: 0 hours: 197 ppm (mg/L); 48 hours: 219 ppm (mg/L)

Basis for effect:

mobility

Remarks on result:

other: no daphnia affected in either study

Effect concentrations 2

Duration:

6 h

Dose descriptor:

EC50

Effect conc.:

> 200 other: ppm (equivalent to mg/L, with d=1 kg/L)

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

measured: 0 hours: 197 ppm (mg/L); 48 hours: 219 ppm (mg/L)

Basis for effect:

mobility

Remarks on result:

other: 3/30 daphnia affected at 200 ppm in second study

Effect concentrations 3

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

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

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

measured (average of all concentrations tested): 0 hours: 100 +/- 4 % of nominal concentration; 48 hours: 106 +/- 6 % of nominal concentration

Basis for effect:

mobility

Remarks on result:

other: 95% confidence interval: 164 - 202 ppm

Effect concentrations 4

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

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

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

measured (average of all concentrations tested): 0 hours: 100 +/- 4 % of nominal concentration; 48 hours: 106 +/- 6 % of nominal concentration

Basis for effect:

mobility

Remarks on result:

other: 95% confidence interval: 95 - 116 ppm

Details on results:

- Behavioural abnormalities: none, except inhibited movement
- Observations on body length and weight: not applicable
- Other biological observations: none reported
- Mortality of control: no immobilisation observed
- Other adverse effects control: none reported
- Abnormal responses: none reported
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: no precipitation observed, measured concentration at 0 hours: 100 +/- 4 % of nominal; at 48 hours: 106 +/- 6 % of nominal
- Effect concentrations exceeding solubility of substance in test medium: Although the saturation concentration of the test material has been reported in a fish toxicity study (Tapp & Caunter 1987) to be about 30 mg/L (using acetone as water-miscible solvent), in the present investigation concentrations up to 200 mg/L (300 mg/L in the pretest) were reached and analytically confirmed (by HPLC). Stock solutions were probably prepared using an Ultra-Turrax rotor/stator high-speed dispersing device, which is known to achieve substantial particle size reduction and create stable emulsions. It is possible that such emulsions were formed from the test substance, and representatively sampled from the test tanks.

Results with reference substance (positive control):

none

Reported statistics and error estimates:

No statistics on analytical concentrations performed in the report. Mean concentrations (test tanks 1-3, first and second study) calculated by reviewer: at 0 hours: 100 +/- 4 % of nominal concentration; at 48 hours: 106 +/- 6 % of nominal concentration.
EC50 and 95% confidence limits·at each time of observation calculated by Litchfield-Wilcoxon method, based on nominal concentrations. The 48-hour LC50 values were 98 (84-114) ppm in the first study, and 117 (99-138) in the second. 24- hour values: 165 (138-198) in the first and 200 (138-290) in the second study. These results were considered to be reproducible, because the 95% confidence intervals overlapped. The acute EC50 values and 95% confidence limits were calculated on the basis of the combined data for both studies.

Any other information on results incl. tables

Number and percentage of affected daphnia

Nominal	Tank	1st study: hours				1st study: hours
conc. ppm	no	3	6	24	48	3	6	24	48
0	1	0	0	0	0	0	0	0	0
control	2	0	0	0	0	0	0	0	0
	3	0	0	0	0	0	0	0	0
Mean	% of total	0.0%	0.0%	0.0%	0.0%	0.0%	0.0%	0.0%	0.0%
30	1	0	0	0	0	0	0	0	0
	2	0	0	0	0	0	0	0	0
	3	0	0	0	0	0	0	0	0
Mean	% of total	0.0%	0.0%	0.0%	0.0%	0.0%	0.0%	0.0%	0.0%
50	1	0	0	0	0	0	0	0	0
	2	0	0	0	1	0	0	0	0
	3	0	0	0	1	0	0	0	0
Mean	% of total	0.0%	0.0%	0.0%	6.7%	0.0%	0.0%	0.0%	0.0%
80	1	0	0	0	2	0	0	0	2
	2	0	0	0	3	0	0	1	2
	3	0	0	0	5	0	0	1	1
Mean	% of total	0.0%	0.0%	0.0%	33.3%	0.0%	0.0%	6.7%	16.7%
130	1	0	0	0	7	0	0	1	5
	2	0	0	2	8	0	0	1	6
	3	0	0	3	7	0	0	2	3
Mean	% of total	0.0%	0.0%	16.7%	73.3%	0.0%	0.0%	13.3%	46.7%
200	1	0	0	7	10	0	1	4	10
	2	0	0	8	10	0	0	5	9
	3	0	0	8	10	0	2	6	10
Mean	% of total	0.0%	0.0%	76.7%	100.0%	0.0%	10.0%	50.0%	96.7%

Analytically determined concentrations

			Analytical concentration ppm
Nominal	Tank		1st study			2nd study
conc. ppm	no	0 hours	48 hours	Mean, SD	0 hours	48 hours	Mean, SD
0	1	<0.5	<0.5	<0.5	<0.5	<0.5
control	2	<0.5	<0.5		<0.5	<0.5
	3	<0.5	<0.5		<0.5	<0.5
30	1	31	33	33	28	32	32
	2	31	34	0.6	28	33	1.0
	3	31	33		28	31
50	1	53	53	51	51	45	48
	2	53	52	2.1	51	49	3.1
	3	53	49		51	51
80	1	76	83	83	82	84	88
	2	76	84	1.5	82	87	4.6
	3	76	81		82	93
130	1	130	135	136	127	145	144
	2	130	133	3.6	127	148	5.1
	3	130	140		127	138
200	1	198	226	217	196	218	220
	2	198	222	11.7	196	242	20.6
	3	198	204		196	201

Test water: pH, dissolved oxygen, and temperature

Nominal	Tank		pH			Dissolved oxygen mg/L				Temperature °C
		1st study		2nd study		1st study		2nd study		1st study		2nd study
concentration	no	0 h	48 h	0 h	48 h	0 h	48 h	0 h	48 h	0 h	48 h	0 h	48 h
0 ppm	1	7.6	7.4	7.4	7.5	8.3	7.4	8.2	7.0	20.3	19.4	19.5	19.6
(control)	2	7.6	7.4	7.4	7.5	8.3	7.6	8.2	6.8	20.3	19.4	19.5	19.6
	3	7.6	7.4	7.4	7.5	8.3	7.7	8.2	6.7	20.3	19.4	19.5	19.6
30 ppm	1	7.6	7.4	7.6	7.3	8.1	7.8	8.3	7.0	20.3	19.4	19.5	19.6
	2	7.6	7.5	7.6	7.4	8.1	7.7	8.3	6.2	20.3	19.4	19.5	19.6
	3	7.6	7.4	7.6	7.4	8.1	7.7	8.3	6.4	20.3	19.4	19.5	19.6
50 ppm	1	7.6	7.3	7.6	7.5	8.0	7.1	8.3	6.9	20.3	19.4	19.5	19.6
	2	7.6	7.2	7.6	7.5	8.0	7.1	8.3	6.3	20.3	19.4	19.5	19.6
	3	7.6	7.2	7.6	7.5	8.0	7.2	8.3	6.5	20.3	19.4	19.5	19.6
80 ppm	1	7.6	7.1	7.6	7.4	8.1	7.4	8.1	6.9	20.3	19.4	19.5	19.6
	2	7.6	7.1	7.6	7.3	8.1	6.9	8.1	6.7	20.3	19.4	19.5	19.6
	3	7.6	7.0	7.6	7.4	8.1	6.6	8.1	6.9	20.3	19.4	19.5	19.6
130 ppm	1	7.5	7.0	7.5	7.3	8.4	7.5	7.8	7.1	20.3	19.4	19.5	19.6
	2	7.5	7.0	7.5	7.3	8.4	7.5	7.8	6.9	20.3	19.4	19.5	19.6
	3	7.5	6.9	7.5	7.3	8.4	7.0	7.8	7.0	20.3	19.4	19.5	19.6
200 ppm	1	7.4	6.6	7.3	6.9	8.4	7.0	8.2	7.0	20.3	19.4	19.5	19.6
	2	7.4	6.7	7.3	6.9	8.4	7.3	8.2	6.9	20.3	19.4	19.5	19.6
	3	7.4	6.4	7.3	6.9	8.4	6.6	8.2	7.2	20.3	19.4	19.5	19.6

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The acute EC50 of the test substance to the daphnia (Daphnia pulex) was estimated to be 182 (164 - 202) mg/L at 24 hours and 105 (95 - 116) mg/L at 48 hours of exposure. The study report is relevant, adequate and reliable for risk assessment, classification and labeling.

Executive summary:

The acute toxicity of the test material was assessed in the daphnia (Daphnia pulex), at five nominal concentrations, 30, 50, 80, 130, and 200 ppm (equivalent to mg/L), in a freshwater static test system. The test substance (500 mg) was mechanically dispersed in 2.5 L water for the top concentration, and further diluted to make up the other test solutions. Concentrations in all tanks were analyzed by HPLC before and after exposure (0 and 48 hours); analytical concentrations averaged 100% +/- 4% of the nominal concentrations at 0 hours, and 106% +/- 6% at the end of exposure. Dissolved oxygen levels always remained >= 6.2 mg/L; pH (6.4 - 7.6) and temperature (19.4 – 20.3 °C) were within the recommendations, whereas water hardness (48.8 mg/L) was lower than the requirement of the guideline issued in 2002 (not part of the 1984 issue). The dose groups comprised 30 first instar daphnia (less than 24 hours old) evenly distributed to 3 tanks (250 ml each) per concentration (for each of the two independent studies). Immobilisation and signs of toxicity were recorded at 3, 6, 24, and 48 hours.

Concentration-dependent immobilisation between 0 and 100% was observed (0% in freshwater controls). A statistical calculation of the concentration inhibiting normal swimming in 50% of the animals (EC50) was performed, and confidence limits (CL) estimated; the reproducibility of the two independent studies was good. The author estimates an EC50 of 182 (CL: 164 - 202) ppm (mg/L) to first instars of Daphnia pulex at 24 hours, and 105 (CL: 95 - 116) ppm at 48 hours, on the combined data of both experiments.