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Short-term toxicity to fish

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Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
from 1993-04-28 to 1993-08-25
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Version / remarks:
1984
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Version / remarks:
1992
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
RCC Umweltchemie GmbH & Co. KG, P.O. Box 1252, D-64374 Rossdorf, Germany
Specific details on test material used for the study:
MCPP-P DMA
MCPP-P content: 602 g/L
Batch: NML/4/93
Analytical monitoring:
yes
Details on sampling:
For the analyses of the test substance concentrations duplicate samples were taken from all test concentrations and the control out of the freshly prepared test media at the start of the test (Day 0) and after 24 hours (Day 1). Additionally, duplicate samples were taken out of the test media of the nominal test concentrations 15 to 148 mg preparation/L and the control at the end of the test (after 96 hours test period). From the highest test concentration of nominal 320 mg preparation/L no sample was taken at this date, since all fish were dead in this concentration already after 24 hours.
All samples were taken from the approximate centre of the aquaria.
Vehicle:
no
Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
TEST ORGANISM
- Common name: Rainbow trout
- Source: Forellenhof Fredelsloh, D-37186 Moringen, Germany
- Age at study initiation: 4 months
- Length at study initiation: 5.1 ± 0.7 cm
- Weight at study initiation: 1.8 ± 0.6 g

ACCLIMATION
- Acclimation period: 7 days
- Acclimation conditions: same as test
- Type of food during acclimation: Brutfutter FB50, Kronen-Fischkraftfutter, D-46483 Wesel, Germany
- Feeding during acclimation: During acclimation until one day before the start of the test fish were fed with a commercial trout diet.
- Health during acclimation: no mortality was observed up to 3 weeks before the start of the test

FEEDING DURING TEST: no
Test type:
flow-through
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Hardness:
225 mg/L as CaCO3
Test temperature:
13 - 15 °C
pH:
7.4 - 8.2
Dissolved oxygen:
7.3 - 9.9 mg/L
Salinity:
NA
Conductivity:
no data
Nominal and measured concentrations:
Nominal concentrations: 0, 15, 32, 69, 148 and 320 mg preparation/L
Measured concentrations varied in the range of 85.5 to 110.1 % of the nominal concentrations
Details on test conditions:
TEST SYSTEM
- Test vessel: aquarium
- Material, size, headspace, fill volume: glass, 48 L
- Renewal rate of test solution: Dosage: 1.88 - 40.0 ml stock solution per hour (20 dosages per hour, 94 - 2000 u.1 per dosage). Test water continuously flowed into the mixing vessels at a rate of 4 1 per hour, regulated by flow-meters. 2 volume additions per day

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: drinking water of the community D-64380 Rossdorf, Germany
- Alkalinity: 4.35 mmol/L

OTHER TEST CONDITIONS
- Photoperiod: 16 h light
- Light intensity: 50 - 300 lux
Reference substance (positive control):
no
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
127 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat. (total fraction)
Remarks:
mg formulation/L
Basis for effect:
mortality (fish)
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
76.4 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Remarks:
corresponding to MCPP-P acid content
Basis for effect:
mortality (fish)
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
69 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat. (total fraction)
Remarks:
mg formulation/L
Basis for effect:
mortality (fish)
Key result
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
41.5 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Remarks:
corresponding to MCPP-P acid content
Basis for effect:
mortality (fish)
Sublethal observations / clinical signs:

In the control and in the test concentrations up to and including nominal 69 mg preparation/L no mortality or intoxication symptoms were observed during the test period of 96 hours. At the nominal test concentration of 148 mg preparation/L all fish showed symptoms of intoxication after 24 hours until the end of the test (a slightly irregular swimming behaviour and/or a changed body colour, and/or the fish stayed mainly at the water surface or mainly at the bottom of the aquarium). The mortality rate of the fish in this test concentration constantly increased from 10 % after 24 hours to 70 % at the end of the test. In the highest test concentration of nominal 320 mg preparation/L all fish were dead after 24 hours test duration.

The 96-hour LC50 of Marks Optica MP n calculated to be 127 mg preparation/L with 95 % confidence limits from 101 to 160 mg preparation/L.

Validity criteria fulfilled:
yes
Conclusions:
For MCPP-P acid, the hydrolysis product of MCPP-P 2-EHE, a 96h-LC50 of 76.4 mg/L and a 96h-NOEC of 41.5 mg/L were determined.
Executive summary:

The acute toxicity of the test substance Marks Optica MP n (MCPP-P DMA) to Rainbow trout was determined in a 96-hour flow-through-test according to the Commission Directive 92/69/EEC, Annex Part C.l, 29.12.1992, and the OECD Guideline for Testing of Chemicals, Section 2, No. 203, adopted April 04, 1984. The test was performed in compliance with Good Laboratory Practice Regulations.

The nominal concentrations tested were 15, 32, 69, 148 and 320 mg preparation/L corresponding to 12.4, 26.4, 57.0, 122 and 264 mg/L MCPP-P acid, and a control.

The test results after the 96 hours exposure period:

96-h LC50: 127 mg preparation/L (76.4 mg/L MCPP-P acid)

96-h LC0: 69 mg preparation/L (41.5 mg/L MCPP-P acid)

lowest lethal concentration (LLC): 148 mg preparation/L (89.1 mg/L MCPP-P acid)

96-h LC 100: 320 mg preparation/L (192.6 mg/L MCPP-P acid)

NOEC: 69 mg preparation/L (41.5 mg/L MCPP-P acid)

LOEC: 148 mg preparation/L (89.1 mg/L MCPP-P acid)

The analytically determined concentrations of the active ingredient MCPP-P DMA in the analysed test media varied in the range from 85.5 to 110.1 % of the nominal values during test period. Therefore, all biological results are related to the nominal concentrations of the test substance. The active ingredient MCPP-P DMA was sufficiently stable in the stock solutions during the renewal periods of 24 hours.

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
from 1992-04-13 to 1992-12-30
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
EPA OPP 72-1 (Fish Acute Toxicity Test)
Version / remarks:
1982
Deviations:
no
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Version / remarks:
1990
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Version / remarks:
EEC 84/449
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
- Batch No.: 5007
- Purity: 90.4 %
Analytical monitoring:
yes
Remarks:
RP-HPLC with UV detection
Details on sampling:
- Concentrations: Sampling was done 1 h and 96 h after the start of the test. At the start of the test the mesured concentrations were in the range of 93.1 to 105.1 %, while at the end they were in the range of 43.2 and 85.6 %.
- Sample storage conditions before analysis: in refrigerator
Vehicle:
yes
Remarks:
Polysorbate 80 (Tween 80)
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: For each concentration 100, 147, 215, 316, 464, 681, 1000 and 1470 mg test material were mixed each with 10 g Polysorbate 80 (Tween 80)(0.1 g/L), premixed with small quantities of double-distilled water (Aqua bidest). An ultra turrax stirrer was used to prepare an optimal dispersion and then the solutions were added to the test water.
- Chemical name of vehicle: Polysorbate 80 (Tween 80)
- Concentration of vehicle in test medium: 0.1 g/L (10 g Tween 80 per test concentration)
Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
TEST ORGANISM
- Common name: Rainbow trout
- Source: Forellenhof Fredelsloh, 37186 Moringen, Germany
- Age at study initiation: about 5 months
- Length at study initiation: 6.46 cm (range: 5.5 - 7.2
- Weight at study initiation: 2.81 g (range: 1.9 - 3.9)
- Maintenance of the brood fish: The fish were kept in a flow-through tank in not chlorinated tap water, passed through an active carbon filter and aerated with oil-free air.

ACCLIMATION
- Acclimation period: 17 days
- Acclimation (housing) conditions:
photoperiod: 16 hours light and 8 hours dark;
total harness of the water: about 2.5 mmol/L (250 mg/L CaCO3)
acid capacity: about 5.5 mmol/L
oxygen content: > 60 % of maximum saturation
pH: about 8.0
water temperature: 12 °C;
prophylactical medical treatment: 2 times with 0.05 mg/L Malachite green chloride and 1 time with 10 mg/L Tetracycline hydrochloride
- Type and amount of food during acclimation: Growing feed F/B 50
- Health during acclimation: about 0.6 % mortality observed

FEEDING DURING TEST
No feeding during the exposure period and 1 day before the start of the test.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Hardness:
250 mg/L CaCO3
Test temperature:
12 ± 1 °C
pH:
8.4 - 8.7
Dissolved oxygen:
9.4 - 12.9 mg/L
Salinity:
NA
Conductivity:
350 - 410 µS/cm
Nominal and measured concentrations:
Nominal concentrations: 1.0, 1.47, 2.15, 3.16, 4.64, 6.81, 10.0 and 14.7 mg/L
Measured concentrations at the end of the test: 0.72, 1.19, 1.64, 1.75, 3.68, 6.10, 8.87, 13.73 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: aquarium with stainless steel frame
- Type: open
- Material, size, headspace, fill volume: glass, 100 litres with water depth of 40 cm, 80 x 35 x 46 cm
- Aeration: slight (due to the use of Polysorbate 80)
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 2 (1 negative control and 1 vehicle control)
- Biomass loading rate: 0.28 g fish / L test water

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Municipal water of the city of Frankenthal, not clorinated and passed through an active carbon filter
- Ca/mg ratio: 9:1 (90 mg/L Ca : 10 mg/L Mg)
- Culture medium different from test medium: no difference
- Intervals of water quality measurement: regularly

OTHER TEST CONDITIONS
- Adjustment of pH: no adjustment
- Photoperiod: 16/8 hours light/dark period

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.5
- Range finding study: yes, LC50 after 96h: between 1 and 10 mg/L
- Test concentrations: 1.0, 1.47, 2.15, 3.16, 4.64, 6.81, 10.0 and 14.7 mg/L
Reference substance (positive control):
no
Key result
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
3.16 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
< 3.68 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
< 4.6 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Key result
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
14.7 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
72 h
Dose descriptor:
other: LC5
Effect conc.:
4.38 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
72 h
Dose descriptor:
other: LC95
Effect conc.:
18.09 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
72 h
Dose descriptor:
LC50
Effect conc.:
8.9 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Details on results:
- Behavioural abnormalities: apathy, discolouration, gasping, narcotic-like state, tumbling
- Any observations that might cause a difference between measured and nominal values: The test solutions were slightly cloudy.
Validity criteria fulfilled:
yes
Conclusions:
In a static acute toxicity test to fish, Oncorhynchus mykiss (rainbow trout) were exposed for 96 hours according to OECD guideline 203 (1990). The LC50 was determined to be smaller than 4.64 mg/L (nominal concentration) and smaller than 3.68 mg/L (measured concentration).
Executive summary:

The acute toxicity of MCPP-P 2-EHE to the Oncorhynchus mykiss (rainbow trout) was investigated in a 96-hour static test according to the OECD Guideline 203 (1990), The EEC Directive 84/449, C.1 and the EPA OPP 72 -1 (1982) under GLP compliance. The test substance was tested in a nominal concentration from 1.0 to 14.7 mg/L with a spacing factor of 1.5 between concentrations. The test concentrations were prepared with the assistance of an emulsifier - Polysorbate 80. The concentration in the test vessels was determined with an HPLC analysis after 1 and 96 h from the start of the test. From concentrations 4.64 mg/L to 14.70 mg/L behavioural abnormalities like apathy, gasping, discolouration, narcotic-like state and tumbling were observed. The 96-h LC50 was determined to be smaller than 4.64 mg/L based on nominal concentrations and smaller than 3.68 mg/L based on measured concentrations. The 96-h NOEC was determined to be 3.16 mg/L. The validity criteria were fulfilled by the test.

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
from 1992-04-13 to 1992-12-30
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
EPA OPP 72-1 (Fish Acute Toxicity Test)
Version / remarks:
1982
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Version / remarks:
EEC 84/449
Deviations:
no
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Version / remarks:
1990
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
- Batch No.: 5007
- Purity: 90.4 %
Analytical monitoring:
yes
Remarks:
RP-HPLC with Uv detection
Details on sampling:
- Concentrations: Sampling was done 1 h and 96 h after the start of the test. At the start of the test the mesured concentrations were in the range of 86.7 to 101.3 %, while at the end they were in the range of 93.1 and 96.3 %.
- Sample storage conditions before analysis: in refrigerator
Vehicle:
yes
Remarks:
Polysorbate 80 (Tween 80)
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: For each concentration 0.147, 0.215, 0.316, 0.464, 0.681, 1.0, 1.47, 2.15, 3.16, 4.64, 6.81 and 10 g test material were mixed each with 10 g Polysorbate 80 (Tween 80), premixed with small quantities of double-distilled water (Aqua bidest). An ultra turrax stirrer was used to prepare an optimal dispersion and then the solutions were added to the test water.
- Chemical name of vehicle: Polysorbate 80 (Tween 80)
- Concentration of vehicle in test medium: 0.1 g/L (10 g Tween 80 per test concentration)
Test organisms (species):
Lepomis macrochirus
Details on test organisms:
TEST ORGANISM
- Common name: Bluegill
- Source: OSAGE Catfisheries, INC., Missouri, USA
- Age at study initiation: about 6 - 10 months
- Length at study initiation: 5.49 cm (range: 4.9 - 6.0
- Weight at study initiation: 1.9 g (range: 1.2 - 2.7)
- Maintenance of the brood fish: The fish were kept in a flow-through tank in not chlorinated tap water, passed through an active carbon filter and aerated with oil-free air.

ACCLIMATION
- Acclimation period: 4 months
- Acclimation (housing) conditions:
photoperiod: 16 hours light and 8 hours dark;
total harness of the water: about 2.5 mmol/L (250 mg/L CaCO3)
acid capacity: about 5.5 mmol/L
oxygen content: > 60 % of maximum saturation
pH: about 8.0
water temperature: 18 - 22 °C;
medical treatment: 4 times with 0.05 mg/L Malachite green chloride and 2 times with 10 mg/L Tetracycline hydrochloride
- Type and amount of food during acclimation: "Tetra Min" standard feed for auarium fish, additionally live and frozen artemia
- Health during acclimation: 0.4 - 0.5 % mortality observed

FEEDING DURING TEST
No feeding during the exposure period and 1 day before the start of the test.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Hardness:
250 mg/L CaCO3
Test temperature:
21 ± 1 °C
pH:
8.2 - 8.8
Dissolved oxygen:
4.1 - 8.8 mg/L
Salinity:
NA
Conductivity:
330 - 470 µS/cm
Nominal and measured concentrations:
Nominal concentrations: 1.47, 2.15, 3.16, 4.64, 6.81, 10.0, 14.7, 21.5, 31.6, 46.4, 68.1 and 100 mg/L
Measured concentrations at the end of the test: 0.98, 1.78, 2.44, 2.40, 4.30, 5.49, 12.10 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: aquarium with stainless steel frame
- Type: open
- Material, size, headspace, fill volume: glass, 100 litres with water depth of 40 cm, 80 x 35 x 46 cm
- Aeration: slight (due to the use of Polysorbate 80)
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 2 (1 negative control and 1 vehicle control)
- Biomass loading rate: 0.19 g fish / L test water

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Municipal water of the city of Frankenthal, not clorinated and passed through an active carbon filter
- Ca/mg ratio: 9:1 (90 mg/L Ca : 10 mg/L Mg)
- Culture medium different from test medium: no difference
- Intervals of water quality measurement: regularly

OTHER TEST CONDITIONS
- Adjustment of pH: no adjustment
- Photoperiod: 16/8 hours light/dark period

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.5
- Range finding study: yes, LC50 after 96h: between 10 and 100 mg/L
- Test concentrations: 1.47, 2.15, 3.16, 4.64, 6.81, 10.0, 14.7, 21.5, 31.6, 46.4, 68.1 and 100 mg/L
Reference substance (positive control):
no
Key result
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
3.16 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
< 2.4 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
< 4.6 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Key result
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
14.7 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Details on results:
- Behavioural abnormalities: apathy, narcotic-like state, tumbling
- Any observations that might cause a difference between measured and nominal values: from concentration 4.64 to 31.6 mg/L the test solutions were cloudy; from 46.4 mg/L upwards were non-transparent
Validity criteria fulfilled:
yes
Conclusions:
In a static acute toxicity test to fish, Lepomis macrochirus (bluegill) were exposed for 96 hours according to OECD guideline 203 (1990). The LC50 was determined to be smaller than 4.64 mg/L (nominal concentration) and smaller than 2.40 mg/L (measured concentration).
Executive summary:

The acute toxicity of MCPP-P 2-EHE to the Lepomis macrochirus (bluegill) was investigated in a 96-hour static test according to the OECD Guideline 203 (1990), The EEC Directive 84/449, C.1 and the EPA OPP 72 -1 (1982) under GLP compliance. The test substance was tested in a nominal concentration from 1.47 to 100 mg/L with a spacing factor of 1.5 between concentrations. The test concentrations were prepared with the assistance of an emulsifier - Polysorbate 80. The concentration in the test vessels was determined with an HPLC analysis after 1 and 96 h from the start of the test. From concentrations 4.64 mg/L to 100 mg/L behavioural abnormalities like apathy, narcotic-like state and tumbling were observed. The 96-h LC50 was determined to be smaller than 4.64 mg/L based on nominal concentrations and smaller than 2.40 mg/L based on measured concentrations. The 96-h NOEC was determined to be 3.16 mg/L. The validity criteria were fulfilled by the test.

Description of key information

The short-term toxicity of MCPP-P 2-EHE to two fish species - Lepomis macrochirus (bluegill) and Oncorhynchus mykiss (rainbow trout), in a 96-h static studies was determined to be the same - LC50 values smaller than 4.64 mg/L (nominal concentration). The results of these studies are considered invalid, as a solubilising agent is used to prepare test solution above the water solubility of MCPP-P 2-EHE. As the ester hysrolyses rapidly to MCPP-P acid, its acute toxicity is used to descibe the effect of the ester in a short-term exposure to fish. The 96h-LC50 for MCPP-P acid, was determined in a flow-through studies with the corresponding MCPP-P DMA salt. The 96 h-LC50 was determined to be 127 mg formulation/L or corresponding to the MCPP-P acid content: 76.4 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Effect concentration:
76.4 mg/L

Additional information

In a key study, the acute toxicity of the test substance Marks Optica MP n (MCPP-P DMA) to Rainbow trout was determined in a 96-hour flow-through-test according to the Commission Directive 92/69/EEC, Annex Part C.l, 29.12.1992, and the OECD Guideline for Testing of Chemicals, Section 2, No. 203, adopted April 04, 1984. The test was performed in compliance with Good Laboratory Practice Regulations.

The nominal concentrations tested were 15, 32, 69, 148 and 320 mg preparation/L corresponding to 12.4, 26.4, 57.0, 122 and 264 mg/L MCPP-P acid, and a control.

The test results after the 96 hours exposure period:

96-h LC50: 127 mg preparation/L (76.4 mg/L MCPP-P acid)

96-h LC0: 69 mg preparation/L (41.5 mg/L MCPP-P acid)

lowest lethal concentration (LLC): 148 mg preparation/L (89.1 mg/L MCPP-P acid)

96-h LC 100: 320 mg preparation/L (192.6 mg/L MCPP-P acid)

NOEC: 69 mg preparation/L (41.5 mg/L MCPP-P acid)

LOEC: 148 mg preparation/L (89.1 mg/L MCPP-P acid)

The analytically determined concentrations of the active ingredient MCPP-P DMA in the analysed test media varied in the range from 85.5 to 110.1 % of the nominal values during test period. Therefore, all biological results are related to the nominal concentrations of the test substance. The active ingredient MCPP-P DMA was sufficiently stable in the stock solutions during the renewal periods of 24 hours.

In a supporting study, the acute toxicity of MCPP-P 2-EHE to the Lepomis macrochirus (bluegill) was investigated in a 96-hour static test according to the OECD Guideline 203 (1990), The EEC Directive 84/449, C.1 and the EPA OPP 72 -1 (1982) under GLP compliance [MCPP Task Force, 1992]. The test substance was tested in a nominal concentration from 1.47 to 100 mg/L with a spacing factor of 1.5 between concentrations. The test concentrations were prepared with the assistance of an emulsifier - Polysorbate 80. The concentration in the test vessels was determined with an HPLC analysis after 1 and 96 h from the start of the test. From concentrations 4.64 mg/L to 100 mg/L behavioural abnormalities like apathy, narcotic-like state and tumbling were observed. The 96-h LC50 was determined to be smaller than 4.64 mg/L based on nominal concentrations and smaller than 2.40 mg/L based on measured concentrations. The 96-h NOEC was determined to be 3.16 mg/L. The validity criteria were fulfilled by the test.

In another supporting study, the acute toxicity of MCPP-P 2-EHE to the Oncorhynchus mykiss (rainbow trout) was investigated in a 96-hour static test according to the OECD Guideline 203 (1990), The EEC Directive 84/449, C.1 and the EPA OPP 72 -1 (1982) under GLP compliance [MCPP Task Force, 1992]. The test substance was tested in a nominal concentration from 1.0 to 14.7 mg/L with a spacing factor of 1.5 between concentrations. The test concentrations were prepared with the assistance of an emulsifier - Polysorbate 80. The concentration in the test vessels was determined with an HPLC analysis after 1 and 96 h from the start of the test. From concentrations 4.64 mg/L to 14.70 mg/L behavioural abnormalities like apathy, gasping, discolouration, narcotic-like state and tumbling were observed. The 96-h LC50 was determined to be smaller than 4.64 mg/L based on nominal concentrations and smaller than 3.68 mg/L based on measured concentrations. The 96-h NOEC was determined to be 3.16 mg/L. The validity criteria were fulfilled by the test.

In conclusion, both supporting studies are of equivalent status.