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Short-term toxicity to aquatic invertebrates

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Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
January 30 - March 6, 2001
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
In accordance with the OECD Guideline and GLP. Two testconcentrations were analytically monitored using TOC measurements. Missing information on the age of the Daphnids, but expected to be < 24 hours as it is mentioned that neonates are used. All the validity criteria can be verified and are fulfilled.
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
GLP compliance:
yes
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
Not applicable
Analytical monitoring:
yes
Details on sampling:
- Concentrations: 48.4 and 234 mg/L
- Sampling method: no data
- Sample storage conditions before analysis: no data
Vehicle:
no
Details on test solutions:
A homogeneous turbid stock suspension with a nominal concentration of 10 g/l synthetic mineral water was prepared. Appropiate volumes were taken from this constantly stirred suspension and pipetted into the test vessels. Then the volume was filled up to nearly 40 ml with test medium, neonates (age not provided) were transferred and at least the endvolume was adjusted with test medium.
Test organisms (species):
Daphnia magna
Details on test organisms:
- Common name: Daphnids
- Source: Creasel, 98000 Deinze, Belgium
- Age: no data, but it is mentioned that neonates are used, so it is expected < 24 h
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Post exposure observation period:
Not applicable
Hardness:
Not reported
Test temperature:
20 degrees Celsius
pH:
7.8-8.0 (measured after 48 h)
Dissolved oxygen:
96-98% (measured after 48 h)
Salinity:
Not applicable
Nominal and measured concentrations:
Nominal: 0, 10, 22, 48.4, 106, 234, 515 and 1130 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: 100 mL glass beakers, covered with glass plates
- Test volume: 50 mL
- No of daphnids: 20 per concentration in 2 groups of 10
Reference substance (positive control):
yes
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
22 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
8.6 mg/L
Nominal / measured:
nominal
Conc. based on:
other: expressed as solid content
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
46 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
17.9 mg/L
Nominal / measured:
nominal
Conc. based on:
other: expressed as solid content
Basis for effect:
mobility
Details on results:
Because of the good analytical recovery of the test substance (104.5% determined as TOC) during the 48 hours, the nominal concentrations were used for the calculation of the EC values.
Results with reference substance (positive control):
- Potassium dichromate
- 24h-EC50: 0,98 mg/l
Reported statistics and error estimates:
According to the probit-method.

  

Concentrations nominal (mg/L)

Test substance recovery (%)

Product

TOC

0 h

24 h

48 h

Mean

48.4

8.42

116.4

105.7

104.5

108.8

234

40.72

100.9

99.5

100.2

100.2

 

Mean recovery overall: 108.8 + 100.2 / 2 = 104.5 %

Validity criteria fulfilled:
yes
Remarks:
1) Immobility in control < 10% (0%); 2) Oxygen >3 mg/L (8.8 – 9.0 mg/L; reported as 96-98%)
Conclusions:
48h-EC50: 46 mg/L (based on test material)
48h-EC50: 17.9 mg/L (based on solid content)
Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
September 10, 1991 - September 12, 1991
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
no analytical monitoring
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
GLP compliance:
yes (incl. certificate)
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
Not applicable
Analytical monitoring:
no
Details on sampling:
Not applicable
Vehicle:
no
Details on test solutions:
Method of preparation: Direct dispersion in water (dechlorinated and aged laboratory tap water);
1 g of test material was dispersed in 1 litre water to give a stock solution of 1 g/L; 200 mL of this stock solution was dispersed in 2 litres of water to give 100 mg/L stock solution from which serial dilutions were made to give the test series.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Source: Laboratory culture originating from a strain supplied by the Institut National de Recherche Chimique Appliquée, France
- Method of breeding: At 21 degrees Celsius in polypropylene vessels containing 2 litres of dechlorinated and aged tap water. Cultures were fed daily with a suspension of mixed algae
- Selection: Gravid adults were isolated 24 hours prior to initiation of the test. Young daphnids produced overnight were used for testing
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Post exposure observation period:
Not applicable
Hardness:
Test water: 50 mg/L as CaCO3
Test temperature:
22 degrees Celsius (measured)
pH:
7.3 - 7.5 (measured)
Dissolved oxygen:
8.3 - 8.4 mg/L (measured)
Salinity:
Not applicable
Nominal and measured concentrations:
Nominal concentrations: 1.0, 1.8, 3.2, 5.6, 10, 18, 32, 56 and 100 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: glass jar each containing 200 mL test solution
- Type: closed
- Renewal rate of test solution (frequency/flow rate): no
- No. of organisms per vessel: 20
- No. of vessels per concentration (replicates): 2 (10 organisms per replicate)
- No. of vessels per control (replicates): 2 (10 organisms per replicate)
- No. of vessels per vehicle control (replicates): no

OTHER TEST CONDITIONS
- Photoperiod: 16 h light, 8 h dark
- Aeration: no
Reference substance (positive control):
no
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
3.2 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
1.3 mg/L
Nominal / measured:
nominal
Conc. based on:
other: expressed as solid content
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
6.4 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 5.4-7.6
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
2.5 mg/L
Nominal / measured:
nominal
Conc. based on:
other: expressed as solid content
Basis for effect:
mobility
Remarks on result:
other: 2.1-3
Results with reference substance (positive control):
Not applicable
Reported statistics and error estimates:
The results are obtained by the method of:
Thompson, W.R., (1947) Bact. Reviews 11, p. 115-145
Validity criteria fulfilled:
yes
Remarks:
1) Immobility in control < 10% (0%); 2) Oxygen >3 mg/L (8.3 – 8.4 mg/L)
Conclusions:
48h-EC50: 6.4 mg/L (based on test material)
48h-EC50: 2.5 mg/L (based on solid content)
Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
2007-06-05 to 2007-06-07
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Version / remarks:
1992
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Version / remarks:
2004
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
- Concentrations: Control and 100 mg a.i./L
- Sampling method: Frequency: at t=0 h and t=48 h; Volume: 2 mL from the approximate centre of the test vessels. At the end of the exposure period, the replicates were pooled at each concentration before sampling.
- Sample storage conditions before analysis: Samples were stored in a freezer until analysis.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Preparation of test solutions started with two solutions of 100 mg a.i./L (198 mg/L, based on formulation) applying 16 minutes of magnetic stirring to accelerate the dissolving of the test substance in the test medium, after which the solutions were pooled. Part of the solution was used as highest test concentration and part was used to prepare the lower test concentrations.
- Controls: Test medium without test substance or other additives
- Evidence of undissolved material (e.g. precipitate, surface film, etc):The final test solutions were all clear and colourless.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: water flea
- Source: In-house laboratory culture with a known history.
- Age at study initiation (mean and range, SD): For the test selection of young Daphnia with an age of < 24 hours, from parental daphnids of more than two weeks old.
- Method of breeding:
Start of each batch: with newborn daphnids, i.e. less than 3 days old, by placing about 250 of them into 5 litres of medium in an all-glass culture vessel.
Maximum age of the cultures: 4 weeks
Renewal of the cultures: Alter 7 days of cultivation half of the medium twice a week.
Temperature of medium: 18-22°C
Feeding: Daily, a suspension of fresh water algae.
Medium: M7, as prescribed by Dr. Elendt-Schneider (Elendt, B.-P., 1990: Selenium deficiency in Crustacea. An ultrastructural approach to antenna!
damage in Daphnia magna Straus. Protoplasma 154, 25-33).
- Feeding during test: no feeding
- Validity of batch: Daphnids originated from a healthy stock, 2nd to 5th brood, showing no signs of stress such as mortality >20%, presence of males, ephippia or discoloured animals and there was no delay in the production of the first brood.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Hardness:
180 mg/L expressed as CaCO3 and the pH: 7.7 ± 0.3.
Test temperature:
19.7 - 20.9°C
pH:
control: 7.6 (t=0h); 7.7 (t=48h)
100 mg a.i./L: 7.5 (t=0); 7.7 (t=48h)
Dissolved oxygen:
control: 8.9 (t=0h); 8.9 (t=48h)
100 mg a.i./L: 8.9 (t=0); 8.6 (t=48h)
Nominal and measured concentrations:
nominal: 0, 0.1, 10, 100 mg a.i./L
Analysis of the samples taken during the combined limit/range-finding test showed that measured concentrations were stable and in agreement with nominal throughout the 48-hour test period (94-97%).
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 100 mL, all-glass
- Aeration: no aeration of the test solutions
- No. of organisms per vessel: 5 per vessel containing 80 ml of test solution
- No. of vessels per concentration (replicates): 2 vessels per concentration for 0.1, 1 and 10 mg/L , 4 vessels for 100 mg/L
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of test medium: Adjusted ISO medium (M7, as prescribed by Dr. Elendt-Schneider)
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours photoperiod daily

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
lmmobility (including mortality): At 24 hours and at 48 hours.
pH and dissolved oxygen: At the beginning and at the end of the test, in the highest concentration and the control.
Temperature of medium: Continuously in a temperature control vessel, beginning at the start of the test.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 10
- Justification for using less concentrations than requested by guideline:
- Range finding study: Limit test with 100 mg a.i./L
- Test concentrations: 0, 0.1, 1, 10 and 100 mg a.i./L
Reference substance (positive control):
yes
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
mobility
Details on results:
No significant immobilisation was observed in any of the concentrations tested.
Results with reference substance (positive control):
- Results with reference substance valid: Yes (potassium dichromate)
- EC50:
24 h EC50 = 0.58 mg/L (95% ci 0.53 - 0.68 mg/L)
48 h EC50 = 0.37 mg/L (95% ci 0.33 -0.42 mg/L)
results were within the ranges of historical data

Acceptability of the test

1. In the control, no daphnia became immobilised or trapped at the surface of the water.

2. The oxygen concentration at the end of the test was ≥ 3 mg/L in control and test vessels.

Table 1: Acute immobilisation of daphnia after 48 hours

Concentration Amphopropionate C8 (mg a.i./L)

Vessel number

Number Daphnia exposed

Response at 48 h

Number

Total %

Control

A

B

C

D

5

5

5

5

0

0

0

0

0

0.10

A

B

5

5

0

0

0

1.0

A

B

5

5

0

0

0

10

A

B

5

5

0

0

0

100

A

B

C

D

5

5

5

5

1

0

0

0

5

Validity criteria fulfilled:
yes
Conclusions:
No significant immobilisation was observed in any of the concentrations tested. The 48 h EC50 of Amphopropionate C8 in Daphnia magna was > 100 mg a.i./L (nominal concentration).
Executive summary:

The 48-hr-acute toxicity of Amphopropionate C8 (50.6% a.i.) to Daphnia magna was studied under static conditions.  Daphnids were exposed to control and test chemical at nominal concentrations of 0 (control), 0.10, 1.0, 10, 100 mg a.i./L for 48 h.  Mortality and immobilisation were observed daily. 

Analysis of the samples taken during the combined limit/range-finding test showed that measured concentrations were stable and in agreement with nominal throughout the 48-hour test period (94-97%). Under the conditions of the present study Amphopropionate did not induce acute immobilisation of Daphnia magna at or below 100 mg a.i./L, after 48 hours of exposure.

 

Results Synopsis

Test Organism Age: Daphnia magna, <24 h

Test Type: Static

 

EC50: > 100 mg a.i./L

Endpoint(s) Effected:  immobilisation

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
2016-01-05 to 2016-01-06
Reliability:
3 (not reliable)
Rationale for reliability incl. deficiencies:
significant methodological deficiencies
Remarks:
non-GLP concentration range finding study
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Version / remarks:
(2004)
Deviations:
yes
Remarks:
for detaisl see "Principles of method if other than guideline"
Principles of method if other than guideline:
Screening Test with only three concentrations with a separation factor of 10 instead of at least five test concentrations that should be arranged in a geometric series with a separation factor preferably not exceeding 2.2.
Analytical verification (TOC) of test concentrations only for the highest concentration level.
GLP compliance:
no
Remarks:
performed as concentration range finding study
Analytical monitoring:
yes
Details on sampling:
At the start of the exposure (0 hours), sampling was carried out after preparation of the test item concentration.
At the end of the exposure (48 hours), samples were taken from additional replicates, which were prepared with test media, but without daphnids, and incubated under test conditions until sampling.
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A stock solution (100 mg/L of the test item were weighed out) was freshly prepared with dilution water (ISO test water, acc. to OECD 202,
Annex 3) before the start of the exposure (0 hours).The stock solution was mixed thoroughly by manual agitation.
- Controls: Dilution water without test item tested under the same conditions as the test groups
- Evidence of undissolved material (e.g. precipitate, surface film, etc): All concentration levels were visually clear throughout the exposure intervals. The stock solution was foaming at the start of the exposure (0 hours).
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: water flea
- Strain: STRAUS (Clone 5)
- Source: Institut für Wasser-, Boden- und Lufthygiene (WaBoLu), 14195 Berlin, Germany
- Age at study initiation (mean and range, SD): 2 - 24 h
- Method of breeding:
In glass vessels (2 - 3 L capacity) with approximately 1.8 L culture medium, at 20 ± 2°C, in an incubator, 16 h illumination; light intensity of max. 20 μE*m-2*s-1 (max. 1340 lx)
Elendt M4, according to ELENDT ( 1990), modified to a total hardness of 160 to 180 mg CaCO3/L, is used.
The culture daphnids were fed at least 5 times per week ad libitum with a mix of unicellular green algae, e.g. Pseudokirchneriella subcapitata and Desmodesmus subspicatus, with an algae cell density of > 1E06 cells/mL. The algae were cultured at the test facility.
- Feeding during test: no

ACCLIMATION
- Acclimation period:
- Acclimation conditions (same as test or not): same
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Test temperature:
19-20°C
pH:
Dilution water (0 hours): 7.68
highest test concentration (0 hours): 7.32
Dissolved oxygen:
Dilution water: 9.13 mg/L at start of experiment
Nominal and measured concentrations:
nominal: 1, 10, 100 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: Glass beakers (4 (ID) x 7 (H) cm), 50 mL capacity
- Type (delete if not applicable): loosely covered with watch glasses
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ISO Test water, according to OECD 202, Annex 3
- Conductivity: 648 µS/cm
- Culture medium different from test medium: The CaCO3 content of the culture medium was reduced to 176 mg/L to achieve a total water hardness of 160 to 180 mg CaCO3
- Intervals of water quality measurement: prior to the start of the exposure

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16/8 hours light/dark cycle
- Light intensity: Diffuse light, light intensity of max. 20 µE x m-2 x s-1 (max. 1340 Ix)

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
lmmobilisation was determined in all groups after 24 and 48 hours. An animal was considered to be immobile, if it was not able to swim in the water phase within 15 seconds after gentle agitation of the test vessel. Other observations were not made.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 10
- Test concentrations: 1, 10 and 100 mg/L
- Justification for using less concentrations than requested by guideline: designed as dose range finding study
Reference substance (positive control):
yes
Remarks:
potassium dichromate
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
76.8 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 95% CL 10-100 mg/L
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
10 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
30.7 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
mobility
Remarks on result:
other: 95% CL 4-40 mg/L
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
4 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
mobility
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
13.2 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
act. ingr.
Basis for effect:
mobility
Remarks on result:
other: 95% c.i.: 1.72 - 17.22 mg/L
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
1.72 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
act. ingr.
Basis for effect:
mobility
Results with reference substance (positive control):
- Results with reference substance valid? yes
- 24 h EC50 = 1.50 mg/L (CI: 1.75 - 2.04 mg/L)
- validity ranges: 24 h EC50: historical range 0.6 - 2.4 mg/L; recommended range according to OECD-Guideline 202: 0.6 - 2.1 mg/L

Table: Immobilisation Rates in the test (n=20, divided into 2 replicates with 10 daphnids each)

Nominal test item concentration [mg/L]

IMMOBILISATION [%]

24 hours

48 hours

Replicates

Replicates

1

2

MV

1

2

MV

100

80

80

80

100

100

100

10.0

0

0

0

0

0

0

1.0

0

0

0

0

0

0

Control

0

0

0

0

0

0

The total carbon content (TOC, according to DIN EN 1484) was 29.7 mg/L at the start (0 hours) and 28.0 mg/L at the end of the exposure (48 hours) in the highest concentration level (below the limit of quantification in both control samples). No analytical quantification of the other two lower concentrations levels was performed.

TOC content was determined within the OECD TG 301 study to be 67%. This value was used to recalculat TOC to active ingredient.

Conclusions:
Based on measured concentrations, the 48 h EC50 of Amphopropionates C12-18 to Daphnia magna was 13.2 mg/L (95% c.i. 1.72 - 17.22 mg/L).
Executive summary:

The 48–hr-acute toxicity of Amphopropionates C12-18 (40% in water) to Daphnia magna was studied in a screening test comparable to OECD Guideline 202 (2004) with deviations under static conditions.  Daphnids were exposed to control and test item at nominal concentrations of 1, 10 and 100 mg/L for 48 hr.  Immobilization was observed daily.  The 48 hr EC50 was 76.8 mg/L (nominal). Based on measured concentrations, the 48 h EC50 was 13.2 mg/L (95% c.i. 1.72 - 17.22 mg/L).

Results Synopsis

 

Test Organism Age: 2-24 hours

Test Type: static

NOEC:  1.72 mg a.i./L(meas.)                       

EC50: 13.2 mg a.i./L                       95% C.I.: 1.72 - 17.22 mg a.i./L

Endpoint(s) Effected:  Mobility

Description of key information

48 h EC50 = 2.5 mg/L, nominal (Daphnia magna, OECD TG 202; RL2; GLP) read-across: Amphoacetates C8-C18 

Key value for chemical safety assessment

EC50/LC50 for freshwater invertebrates:
2.5 mg/L

Additional information

For the assessment of acute toxicity of Amphopropionates C12-18 to aquatic invertebrates a dose range finding study in Daphnia magna is available. However, as this study was designed only as dose range finding study, it is not fully in compliance with the testing guideline. Thus, it is assigned a Klimisch score of 3. Nevertheless, it is used in a weight-of-evidence-approach together with data from the structurally related source substances Amphoacetates C8-C18 and Amphopropionate C8. A justification for read-across is given in the Endpoint summary Aquatic toxicity.

 

The 48–hr-acute toxicity of Amphopropionates C12-18 (40% in water) to Daphnia magna  was studied in a screening test according to OECD Guideline 202 (2004) with deviations under static conditions.  Daphnids were exposed to control and test item at nominal concentrations of 1, 10 and 100 mg/L for 48 hr.  Immobilization was observed daily.  The 48 hr EC50 was 76.8 mg/L (nominal). Based on measured concentrations, the 48 h EC50 was 13.2 mg/L (95% c.i. 1.72 - 17.22 mg/L).

 

The 48-hr-acute toxicity of Amphoacetates C8-C18 to Daphnia magna was studied according to OECD Guideline 202 under static conditions. Daphnids were exposed to test chemical at nominal concentrations of 0 (control), 1.0, 1.8, 3.2, 5.6, 10, 18, 32, 56 and 100 mg/L for 48 h without analytical monitoring.  Mortality and immobilisation were observed daily. 

The 48 h EC50 based on test material was 6.4 mg/L. The 48 h EC50 based on solids content was 2.5 mg/L.

 

The 48-hr-acute toxicity of Amphoacetates C8-C18 to Daphnia magna was studied according to OECD Guideline 202 under static conditions. Daphnids were exposed to test chemical at nominal concentrations of 0 (control), 10, 22, 48.4, 106, 234, 515 and 1130 mg/L for 48 h.  Mortality and immobilisation were observed daily. 

Because of the good analytical recovery of the test substance (104.5% determined as TOC) during the 48 hours, the nominal concentrations were used for the calculation of the EC values.

The 48 h EC50 based on test material was 46 mg/L. The 48 h EC50 based on solids content was 17.9 mg/L.

 

The 48-hr-acute toxicity of Amphopropionate C8 (50.6% a.i.) to Daphnia magna was studied according to OECD Guideline 202 under static conditions.  Daphnids were exposed to test chemical at nominal concentrations of 0 (control), 0.10, 1.0, 10, 100 mg a.i./L for 48 h.  Mortality and immobilisation were observed daily. 

Analysis of the samples taken during the combined limit/range-finding test showed that measured concentrations were stable and in agreement with nominal throughout the 48-hour test period (94-97%). Under the conditions of the present study Amphopropionate C8 did not induce acute immobilisation of Daphnia magna at or below 100 mg a.i./L, after 48 hours of exposure.

 

Supporting data are available for Acrylic acid: The 48 h EC50 of acrylic acid to Daphnia magna was 47 mg/L.

 

The 48 h EC50 obtained in the dose range finding study conducted with the target substance Amphopropionates C12-18 was 13.2 mg/L (95% c.i. 1.72 - 17.22 mg/L) and thus higher than the effect concentrations obtained with the source substance Amphoacetates C8-C18. Also Acrylic acid did not show considerable toxicity to aquatic invertebrates. Therefore, the value of the source substance Amphoacetates C8-C18 is also protective for the target substance Amphopropionates C12-18.

For chemical safety assessment the lowest EC50 of 2.5 mg/L obtained with Amphoacetates C8-C18 will be used.