Mecoprop

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:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

no guideline followed

Principles of method if other than guideline:

Biodegradability of the test material was tested in water and soil, and the toxicity to fish was measured.

GLP compliance:

no

Remarks:

Study pre-dates GLP.

Analytical monitoring:

no

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The test material was dissolved in standard fish water.

Test organisms (species):

Lebistes reticulatus

Details on test organisms:

TEST ORGANISM
- Common name: Guppy
- Age: 3 to 4 weeks old.

ACCLIMATION
- Not specified

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

200 ppm

Nominal and measured concentrations:

0, 560, 1000, 1800, 3200, 5600 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Aquaria
- Fill volume: 1L
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2

TEST MEDIUM / WATER PARAMETERS
- Standard fish water.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- After 96 hours the mortality was recorded and from these data the LC50 (TLM 96) (= concentration in which 50 % mortality occurs) at 96 hours was calculated.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: Several concentrations were made increasing in a geometric progression.
- Test concentrations: 0, 560, 1 000, 1 800, 3 200, 5 600 mg/L

Reference substance (positive control):

yes

Remarks:

2, 4-DP (560, 1 000, 1 800, 3 200, 5 600 mg/L)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 1000 - 1300 mg/L

Details on results:

From these results it appears that the test material is only moderately toxic.

Sublethal observations / clinical signs:

Mortality During the Study

Concentration (mg/L)	Number of Fish Living After 96 Hours
0	10 – 10
560	10 – 10
1 000	8 – 8
1 800	0 – 0
3 200	0 – 0
5 600	0 - 0

Validity criteria fulfilled:

not applicable

Conclusions:

Under the conditions of this study the 96 hour LC50 of the test material to fish was determined to be 1 100 mg/L (1 000 - 1 300 mg/L).

Executive summary:

The short-term toxicity to the gubby by the test material was investigated. Biodegradability of the test material was tested in water and soil and the toxicity to fish was measured over 96 hours. The test material was dissolved in standard fish water with a total hardness of 200 ppm. Several concentrations were made increasing in a geometric progression (each concentration was replicated twice). Guppies (3 to 4 weeks old) were transferred to these solutions in aquaria of 1 litre. To each concentration ten guppies were exposed. After 96 hours the mortality was recorded and from these data the LC50 at 96 hours was calculated. 

Under the conditions of this study the 96 hour LC50 of the test material to fish was determined to be 1 100 mg/L (1 000 - 1 300 mg/L).

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

09 July 1984 to 13 July 1984

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

04 April 1984

Deviations:

no

Principles of method if other than guideline:

It was reported that the test concentrations were determined by UV-Spectroscopic method but no details were provided.

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- The test material was added to the test water without any pretreatment using an ultra-turrax stirrer; after 1 hour the aquarium content was stirred again; subsequently the fish were placed into the aquaria.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Rainbow trout
- Length at study initiation: 4.7 cm (range: 4.5 - 4.9)
- Weight at study initiation: 1.0 g (range: 0.8 - 1.2)

ACCLIMATION
- Acclimation period: 7 weeks
- Acclimation conditions (same as test or not): The fish were kept in a flow-through tank in tap water cleaned and dechlorinated by active carbon and aerated with oil-free air.
- Feeding frequency during acclimation: Ad libitum
- Health during acclimation (any mortality observed): 0 % mortality over 1 week before the beginning of the test.

FEEDING DURING TEST
- Food type: Growing feed and standard feed
- Amount/frequency: Ad libitum

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

2.5 mmol/L

Test temperature:

17 ± 1 °C

pH:

about 7.8

Conductivity:

max. 10 micro MHO

Nominal and measured concentrations:

31.6, 46.4, 68.1, 100.0, 147.0, 215.0 and 316.0 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass aquarium with a stainless steel frame (60 cm x 25 cm x 40 cm)
- Material, size, headspace, fill volume: 50 L
- Aeration: Continuous aeration with oil-free air.
- No. of organisms per test concentration: 10
- Biomass loading rate:0.2 g fish / L test water

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted freshwater according to DIN 38 412, part 11, draft Sept. 1981. Preparation from fully de-mineralised tap water.
- Ca/mg ratio: 4 : 1
- Na/K ratio: 10 : 1
- Culture medium different from test medium: Yes
- Intervals of water quality measurement: Not specified

OTHER TEST CONDITIONS
- Photoperiod: 16 hours light and 8 hours dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of mortality and symptoms.
- Determination of calculation of the median lethal concentration (LC 50) and, if possible, the LC 3 and the LC 95 using the probit analysis after hours (nominal conc.): 1, 4, 24, 48, 72 and 96.

TEST CONCENTRATIONS
- Test concentrations: 31.6, 46.4, 681, 100.0, 147.0, 215.0 and 316 mg/L

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 150 - < 220 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

68.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Details on results:

TEST MATERIAL
Undissolved test material was visible in all concentrations 1 hour after beginning of the test. The amount increased with an increase in the concentration. After 24 hours the amount of undissolved test material had decreased and after 72 hours no undissolved test material was visible in the aquaria with surviving fish.

Sublethal observations / clinical signs:

- Symptoms: Convulsions, discolouration, escape reflex, gasping.
- The symptom discolouration is regarded as a sign of stress and not as a sign of intoxication as it was also seen in the control group.

 

Mortality During the Study

Nominal Concentration (mg/L)	Number of Fish at the Beginning	Dead Fish After
		1 h	4 h	24 h	48 h	72 h	96 h
31.6	10	0	0	0	0	0	0
46.4	10	0	0	0	0	0	0
68.1	10	0	0	0	0	0	0
100.0	10	0	0	0	0	0	0
147.0	10	0	0	0	1	1	1
215.0	10	0	0	10	10	10	10
316.0	10	0	0	10	10	10	10
0	10	0	0	0	0	0	0

Validity criteria fulfilled:

not specified

Conclusions:

Under the conditions of this study the LC50 was between 150 and 220 mg/mL. The NOEC was 68.1 mg/L.

Executive summary:

The short-term toxicity of the test material to fish was investigated using rainbow trout in accordance with the standardised guideline OECD 203, under GLP conditions. 

A static test was conducted in freshwater over a period of 96 hours. The test organisms were housed in glass aquarium with a stainless steel frame (60 cm x 35 cm x 40 cm), at 17 ± 1 °C. The test concentrations used were: 31.6, 46.4, 68.1, 100.0, 147.0, 215.0 and 316.0 mg/L. Mortality and symptoms were monitored. 

Undissolved test material was visible in all concentrations 1 hour after beginning of the test. The amount increased with an increase in the concentration. After 24 hours the amount of undissolved test material had decreased and after 72 hours no undissolved test material was visible in the aquaria with surviving fish. Clinical symptoms observed were convulsions, discolouration, escape reflex and gasping. The symptom discolouration is regarded as a sign of stress and not as a sign of intoxication as it was also seen in the control group.

Under the conditions of this study the LC50 was between 150 and 220 mg/mL. The NOEC was 68.1 mg/L.

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

17 February 1986 to 21 February 1986

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Qualifier:

equivalent or similar to guideline

Guideline:

EPA OPP 72-1 (Fish Acute Toxicity Test)

Version / remarks:

October 1982

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- The test material was added to the test water separately for each concentration by mixing the appropriate quantities of the test material in about 1 litre of test water and transferring into the aquaria.

Test organisms (species):

Lepomis macrochirus

Details on test organisms:

TEST ORGANISM
- Common name: Bluegill
- Length at study initiation: 4.2 cm (range: 2.9 - 4.8)
- Weight at study initiation: 0.9 g (range: 0.7 - 1.3)

ACCLIMATION
- Acclimation period: about 3 months
- Acclimation conditions (same as test or not): The fish were kept in a flow-through tank in tap water cleaned by active carbon and aerated with oil-free air.
- Feeding frequency during acclimation: ad libitum
- Health during acclimation (any mortality observed): Mortality during the last 2 weeks of housing: 0 %, mortality during the period of adaptation: 0 %.

FEEDING DURING TEST (as applicable)
- Food type: Standard feed and sometimes frozen artemia.
- Amount/frequency: ad libitum

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

2.5 mmol/L

Test temperature:

23 °C

pH:

about 7.8

Conductivity:

max 10 micro MHO

Nominal and measured concentrations:

21.5, 31.6, 46.4, 68.1, 100.0 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass aquarium with a stainless steel frame (60 cm x 35 cm x 40 cm)
- Material, size, headspace, fill volume: 50 litres
- Aeration: Continuous aeration with oil-free air.
- No. of organisms per test concentration: 10
- Biomass loading rate: about 0.2 g fish / L test water.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted freshwater according to DIN 38 412, Par 11, Draft Sept. 1981. Preparation from fully demineralised tap water.
- Ca/mg ratio: 4 : 1
- Na/K ratio: 10 : 1
- Culture medium different from test medium: Yes

OTHER TEST CONDITIONS
- Photoperiod: 16 hours light and 8 hours darkness

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of mortality and symptoms.
- Determination of calculation of the median lethal concentration (LC 50) and, if possible, the LC 3 and the LC 95 using the probit analysis after hours (nominal conc.): 1, 4, 24, 48, 72 and 96.

TEST CONCENTRATIONS
- Range finding study: based on the results of a preceding study with rainbow trout (LC 50 after 96 H: > 100 mg/L) the concentrations, spaced by a factor of about 2.2 were fixed as follows: 21.5, 31.6, 46.4, 68.1 and 100 mg/L.
- Results used to determine the conditions for the definitive study: Yes.

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Details on results:

TEST MATERIAL
Small amounts of undissolved test material were visible at the bottom of the aquaria 24 hours after the beginning. After 96 hours no undissolved test material was visible.

Sublethal observations / clinical signs:

No abnormalities were observed. 

 

Analytical Determination of Test Concentrations

Nominal Concentration (mg/L)	Analytically Detected Concentrations (mg/L)
	1 Hour	96 Hours
21.5	20.47	22.66
31.6	28.54
46.4	39.77	47.15
68.1	59.70
100.0	83.96	102.30

Validity criteria fulfilled:

not specified

Conclusions:

Under the conditions of this study the LC50 was > 100 mg/mL.

Executive summary:

The short-term toxicity to bluegill by the test material was assessed according to the EPA Guideline 72-1 and in compliance with GLP. 

A static test was conducted in freshwater over a period of 96 hours. The test organisms were housed in glass aquarium with a stainless steel frame (60 cm x 35 cm x 40 cm), at 23 °C. Based on the results of a preceding study with rainbow trout (LC 50 after 96 H: > 100 mg/L) the concentrations, spaced by a factor of about 2.2 were fixed as follows: 21.5, 31.6, 46.4, 68.1 and 100 mg/L. Mortality and symptoms were monitored. 

Small amounts of undissolved test material were visible at the bottom of the aquaria 24 hours after the beginning. After 96 hours no undissolved test material was visible. No abnormalities were observed in the test organisms. 

Under the conditions of this study the LC50 was > 100 mg/mL.

Reference 4

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

18 July 2003 to 07 August 2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japanese guideline: 12-Nousan-No 8147 of MAFF

Version / remarks:

24 November 2000. Test guideline: Fish acute toxicity studies § 2-7-1 (JMAFF, 2000)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

At the beginning of the test, before and after each renewal of the solution, a 10 mL aliquot of the test solution was taken from the aquarium to determine the test material concentration. The samples were kept in a refrigerator (approximately 5 °C) until the analysis was performed.

Vehicle:

yes

Remarks:

Acetone

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
Range-finding test:
- The stock solutions for the range-finding test were prepared by dissolving 5,000 and 25,000 mg of test material in 20 and 150 mL of acetone, respectively, and the volume was completed to 1 000 and 5 000 mL, respectively, with dilution water, resulting in two stock solutions with a concentration of 5 000 mg/L. To assist the solubilisation 10 drops of Tween-80 and 500 mL of the first stock solution were diluted into 25 000 mL of dilution water resulting in two test concentrations of 10 and 100 mg/L respectively. 5 000 mL of the second stock solution was diluted into 25 000 mL of dilution water, resulting in a test concentration of 1 000 mg/mL.

Definitive test:
- The stock solution for the definitive test was prepared by dissolving 1 400; 2 500; 4 500; 8 00 and 14 000 mg of test material in 20 mL of acetone. These solutions which presented initially a homogenous aspect, where diluted into 25 000 mL of dilution water.

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Common carp
- Fresh weight range: 2.930 to 2.648 g
- Fish fresh total length range (average of 10 fish): 5.16 to 5.41 cm.
- Length at study initiation: 5.0 ± 1.0 cm

ACCLIMATION
- Acclimation period: 7 days
- Acclimation conditions (same as test or not): Seven days before testing, the fish were taken from the maintenance tank and held in the same water quality used in the test, with temperature of 21 to 25 °C, with continuous aeration and with a photoperiod of 12 to 16 hours of light.
- Feeding frequency during acclimation: The fish were daily fed until 24 hours before the test was started.
- Health during acclimation (any mortality observed):The fish were healthy and free from any visible disease and abnormalities during the maintenance and acclimation periods.

FEEDING DURING TEST (as applicable)
- During the exposure period to the test material, the fish were not fed.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

100.98 - 106.08 mg/L in CaCO3

Test temperature:

22.3 and 23.7 °C

pH:

7.65 - 7.70

Dissolved oxygen:

7.08 - 7.32

Conductivity:

297 - 321

Nominal and measured concentrations:

Nominal: 56; 100; 180; 320 and 560 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 28-litre stainless steel beakers
- Material, size, headspace, fill volume: 25 litres of test solution
- Aeration: Continuous
- Renewal rate of test solution (frequency/flow rate): Daily
- No. of organisms per concentration: Range finding study: 7 fish, definitive test: 10 fish.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The dilution water was prepared from water supplied by Water and Sewage Municipal Service (SEMAE). The water was de-chlorinated and the quality characteristic such as hardness was between 100.98 and 106.08 mg/L in CaCO3 and the pH was between 7.65 and 7.70.
- Culture medium different from test medium: No

OTHER TEST CONDITIONS
- Photoperiod: 16 hours of light

EFFECT PARAMETERS MEASURED:
- Mortality and sub-lethal effects were recorded at 24, 48, 72 and 96 hours.

TEST CONCENTRATIONS
- Range finding study: Initially a range-finding test was carried out using seven fish per concentration. The nominal concentrations used were: 10; 100 and 1 000 mg/L and a control and a solvent control groups.
- Test concentrations: 56; 100; 180; 320 and 560 mg/L
- Results used to determine the conditions for the definitive study: Yes, once the toxic range was found, a definitive test was carried out.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate (100, 180, 320, 560 and 1 000 mg/L)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

134.16 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Details on results:

CHEMISTRY ANALYSIS
- In the case of the test solutions with nominal test material concentrations of 180, 320 and 560 mg/L, it was observed that no fish survived for more than 2 hours. For these solutions, there was only one determination of the effective concentration, from the aliquot collected after the initial filling of the tank.
- The deviations for mean effective/mean nominal concentration ratios for the test material in the test solutions range from 2.00 to 14.64 %. The maximum deviation (14.64 %) was less than 20 %, so the LC50 values were calculated using the mean nominal concentration values.

BIOLOGICAL RESULTS
- The values of 24; 48; 72 and 96-hour LC50 of the test material were the same 134.16 mg/L, respectively. The highest concentration that caused no lethal effect on the fish was 100 mg/L and the lowest concentration that killed 100 % of fish was 180 mg/L, in 96 hours of exposure period.

Results with reference substance (positive control):

The value of 24-hour LC50 for the reference substance, potassium dichromate, was 377.90 mg/L (confidence limits at 95 % = 327.36 to 436.24 mg/L), demonstrating, this way, the viability of test system use.

Reported statistics and error estimates:

The LC50 and the respective confidence limits at 95 % were determined using the Trimmec Spearman - Karber Method.

Sublethal observations / clinical signs:

Mortality (10 fish) was observed at test material concentrations of 180 mg/L and above at 24 hours. 

 

Cumulative Mortality of Fish (Cyprinus carpio) During the Exposure Period to Test Material

Nominal Concentration (mg/L)	Number of Dead Fish During the Test Period
	24 Hours	48 Hours	72 Hours	96 Hours
Control	0	0	0	0
Solvent Control	0	0	0	0
56	0	0	0	0
100	0	0	0	0
180	10	10	10	10
320	10	10	10	10
560	10	10	10	10

 

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of this study the 96 hour LC50 was determined to be 134.16 mg/L. The No Observed Concentration (NOEC) was 100 mg/L.

Executive summary:

The short-term toxicity to the common carp by the test material was investigated in accordance with the standardised guidelines OECD 203 and the Japanese guideline 12-Nousan-p 8147 of MAFF, under GLP conditions. 

A range-finding test was carried out in order to determine the concentrations that cause 100 and 0 % of mortality to the test organisms. Based on the results of this test, a definitive test was carried out with the following nominal concentrations of the test material: 56, 100, 180, 320 and 560 mg/L and a control group. Ten fish were used at each concentration. Mortality and sub-lethal effects were recorded at 24, 48, 72 and 96 hours. 

Under the conditions of this study the 96 hour LC50 was determined to be 134.16 mg/L. The No Observed Concentration (NOEC) was 100 mg/L.

Reference 5

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

17 January 2005 to 21 January 2005

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), Testing Guidelines for Toxicology Studies, 12 NohSan No.8147 Guideline No. 2-7-1

Version / remarks:

24 November 2000 (partially revised by 13 SeiSan No. 3986 on October 10, 2001).

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Water samples were taken from the control and each replicate test vessel at 0 hours (fresh media), 24 and 96 (old media) hours for quantitative analysis.

Duplicate samples and samples at 24 (fresh media), 48 and 72 hours (old and fresh media) were taken and stored at approximately -20ºC for further analysis if necessary.

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: An amount of test material (2.00 g) was dissolved in dechlorinated tap water with the aid of ultrasonication and vigorous shaking for approximately 2 hours and the volume adjusted to 5 litres to give a 400 mg/L stock solution. The entire volume of the 400 mg/L stock solution was dispersed in a final volume of 20 litres of dechlorinated tap water to give the 100 mg/L test concentration. This method of preparation was conducted in duplicate to give replicates R1 and R2.
- Each of the stock solutions was inverted several times to ensure adequate mixing and homogeneity.

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Common carp
- Length at study initiation: 4.4 cm (sd = 0.2) (at the end of the definitive test)
- Weight at study initiation: 2.23 g (sd = 0.24) (at the end of the definitive test

ACCLIMATION
- Acclimation period: Fish were acclimatised to test conditions from 5 January 2005 to 17 January 2005.
- Type and amount of food during acclimation: The stock fish were fed commercial carp pellets which was discontinued approximately 24 hours prior to the start of the definitive test.
- Health during acclimation (any mortality observed): There was less than 1 % mortality in the 7 days prior to the start of the test.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

100 mg/L as CaCO3

Test temperature:

19.8 to 21.2 °C
At 24 hours exposure (fresh media) the temperatures were measured to be 19.8 to 19.9 °C which were outside the specified range of 21 ± 1 °C. This minor deviation was considered to have no affect on the results of the test given that temperatures at all other times were in the specified range..

pH:

Mean: 7.617

Dissolved oxygen:

Greater than or equal to 8.8 mg O2/L

Conductivity:

Mean: 439.164 μS/cm

Nominal and measured concentrations:

Nominal: 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass exposure vessels (covered)
- Material, size, headspace, fill volume: 20 L
- Aeration: The test vessels were aerated via narrow bore glass tubes.
- Renewal rate of test solution (frequency/flow rate): Daily
- No. of organisms per vessel: 7
- Biomass loading rate: 0.78 g bodyweight/litre

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Laboratory tap water was dechlorinated by passage through an activated carbon filter (Purite Series 500) and partly softened (Elga Nimbus 1248D Duplex Water Softener).
- Total organic carbon: 1.7 mg C/L
- Pesticides: <0.010 μg/L
- Chlorine: 0.221 (mean total) mg/L
- Alkalinity: 113 mg HCO3/L
- Culture medium different from test medium: No

OTHER TEST CONDITIONS
- Photoperiod: 16 hours light and 8 hours darkness with 20 minute dawn and dusk transition period.
- The water temperature, pH and dissolved oxygen concentrations were recorded daily throughout
the test. The measurements at 0 hours, and after each test media renewal at 24, 48 and 72 hours,
represent those of the freshly prepared test preparations while the measurements taken prior to
each test media renewal, and on termination of the test after 96 hours, represent those of the used or 24-Hour old test preparations. The pH and the dissolved oxygen concentration were measured using a WTW pH/Oxi 340I pH and dissolved oxygen meter and the temperature was measured using a Hanna Instruments HI93510 digital thermometer.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Any mortalities and sub-lethal effects of exposure were recorded at 3, 6, 24, 48, 72 and 96 hours after the start of exposure. The criteria of death were taken to be the absence of both respiratory movement and response to physical stimulation.
- An estimate of the LC50 values was given by inspection of the mortality data.
- At 24 hours the LC50 value was calculated using the trimmed Spearman-Karber method (Hamilton et al 1977) using the ToxCalc Computer Software Package (ToxCalc 1999) while at 6, 48, 72 and 96 hours the LC50 values and associated confidence limits were calculated using the geometric mean method as follows:

LC50 value = √ C1 x C2

Where:
C1 = concentration showing 0 % mortality
C2 = concentration showing 100 % mortality

- If there are no mortalities between 0 % and 100 % mortality, then the geometric mean of the
highest test concentration showing no lethality and the lowest test concentration showing 100 %
lethality is calculated. The concentrations resulting in 0 % and 100 % mortality will be the 95 %
confidence limits.

TEST CONCENTRATIONS
- Range finding study: In the range-finding test fish were exposed to a series of nominal test concentrations of 1.0, 10 and 100 mg/L. The test material was dissolved directly in water. An amount of test material (2.00 g) was dissolved in dechlorinated tap water with the aid of ultrasonication and vigorous shaking for approximately 2 hours and the volume adjusted to 5 litres to give a 400 mg/L stock solution. The entire volume of the 400 mg/L stock solution was dispersed in a final volume of 20 litres of dechlorinated tap water to give the 100 mg/L test concentration. An amount of test material (400 mg) was dissolved in dechlorinated tap water with the aid of ultrasonication for approximately 2 hours and the volume adjusted to 1 litre to give a 400 mg/L stock solution. Aliquots (50 and 500 mL) of the 400 mg/L stock solution were each dispersed in a final volume of 20 litres of dechlorinated tap water to give the 1.0 and 10 mg/Ltest concentrations respectively. In the range-finding test 3 fish were added to each 20 litre test and control vessel and maintained at approximately 21 °C in a temperature controlled room with a photoperiod of 16 hours light and 8 hours darkness with 20 minute dawn and dusk transition periods for a period of 96 hours under static test conditions. Each vessel was covered to reduce evaporation. After 3, 6, 24, 48, 72 and 96 hours any mortalities or sub-lethal effects of exposure were determined by visual inspection of the test fish.
- Test concentrations: 100 mg/L
- Results used to determine the conditions for the definitive study: Yes, based on the results of the range-finding test a "Limit test" was conducted at a concentration of 100 mg/L to confirm that at the maximum concentration given in the JMAFF Test Guidelines, no mortalities or sub-lethal effects of exposure were observed.

Reference substance (positive control):

yes

Remarks:

Pentachlorophenol, sodium salt (0.10, 0.18, 0.32, 0.56 and 1.0 mg/L)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Details on results:

RANGE-FINDING TEST
- Based on this information, a single test concentration, in duplicate, of 100 mg/L was selected for the definitive test. This experimental design conforms to a "Limit test" to confirm that at the maximum concentration given in the JMAFF Test Guidelines, no mortalities or sub-lethal effects of exposure were observed.

Results with reference substance (positive control):

The results of the positive control showed the highest test concentration resulting in 0 % mortality to be 0.18 mg/L, the lowest test concentration resulting in 100 % mortality to be 0.32 mg/L and the No Observed Effect Concentration (NOEC) to be 0.18 mg/L. The No Observed Effect Concentration is based upon zero mortalities and the absence of any sub-lethal effects of exposure at this concentration.

The results from the positive control with pentachlorophenol, sodium salt were within the normal range for this reference material. The mean 96-Hour LC50 value calculated from all positive control tests is 0.25 mg/L (sd = 0.04).

Reported statistics and error estimates:

DEFINITIVE TEST
- The results of the definitive test showed the highest test concentration resulting in 0 % mortality to be greater than or equal to 100 mg/L, the lowest test concentration resulting in 100 % mortality to be greater than 100 mg/L and the No Observed Effect Concentration (NOEC) to be 100 mg/L. The No Observed Effect Concentration is based upon zero mortalities and the absence of any sublethal effects of exposure at this concentration.

Sublethal observations / clinical signs:

Range-finding test

There were no sub-lethal effects of exposure during the range-finding test.

 

Definitive test

There were no mortalities in 7 fish exposed to a test concentration of 100 mg/L for a period of 96 hours. 

There were no sub-lethal effects of exposure observed in 14 fish exposed to a test concentration of 100 mg/L for a period of 96 hours.

 

Positive control

Sub-lethal effects of exposure were observed at the test concentrations of 0.32 and 1.0 mg/L. These responses were swimming at the surface and the presence of moribund fish. After approximately 5 hours exposure seven out of seven fish at 1.0 mg/L were observed to be moribund. Due to the approach of the substantial severity limit (Animals (Scientific Procedures) Act 1986) these fish were killed and classed as mortalities at the 6-Hour time point.

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of this study the 96 hour LC50 was > 100 mg/L. Correspondingly the No Observed Effect Concentration was 100 mg/L.

Executive summary:

The short-term toxicity to the common carp by the test material was assessed in accordance with the standardised guidelines OECD 203, EU Method C.1 and the Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), Testing Guidelines for Toxicology Studies, 12 NohSan No.8147 Guideline No. 2-7-1, under GLP conditions. 

Following a preliminary range-finding test fish were exposed, in two groups of seven, to an aqueous solution of the test material at a single concentration of 100 mg/L for a period of 96 hours at a temperature of 19.8 to 21.2 °C under semi-static test conditions. The number of mortalities and any sub-lethal effects of exposure in each test and control vessel were determined 3 and 6 hours after the start of exposure and then daily throughout the test until termination after 96 hours.

A positive control conducted approximately every six months used pentachlorophenol, sodium salt as the reference material. In the most recent test, fish were exposed, in groups of seven, to an aqueous solution of the reference material at a range of concentrations of 0.10, 0.18, 0.32, 0.56 and 1.0 mg/L for 96 hours at a temperature of 19.7 to 20.6 ºC under semi-static test conditions. The number of mortalities and any sub-lethal effects of exposure in each vessel were determined 3 and 6 hours after the start of exposure and then daily throughout the test until termination after 96 hours.

Under the conditions of this study the 96 hour LC50 was > 100 mg/L. Correspondingly the No Observed Effect Concentration was 100 mg/L.

Description of key information

Key Study: Wetton & McKenzie (2005)

Under the conditions of this study the 96 hour LC50 was > 100 mg/L. Correspondingly the No Observed Effect Concentration was 100 mg/L.

Supporting Study: Munk (1984)

Under the conditions of this study the LC50 was between 150 and 220 mg/mL. The NOEC was 68.1 mg/L.

 

Spporting Study: Munk (1986)

Under the conditions of this study the LC50 was > 100 mg/mL. The NOEC was 100 mg/L.

 

Supporting Study: Bidinotto (2004)

Under the conditions of this study the 96 hour LC50 was determined to be 134.16 mg/L. The No Observed Concentration (NOEC) was 100 mg/L.

 

Supporting Study: ten Berge (1978)

Under the conditions of this study the 96 hour LC50 of the test material to fish was determined to be 1 100 mg/L (1 000 - 1300 mg/L).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

134.16 mg/L

Additional information

Key Study: Wetton & McKenzie (2005)

The short-term toxicity to the common carp by the test material was assessed in accordance with the standardised guidelines OECD 203, EU Method C.1 and the Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), Testing Guidelines for Toxicology Studies, 12 NohSan No.8147 Guideline No. 2-7-1, under GLP conditions. The study was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).

Following a preliminary range-finding test fish were exposed, in two groups of seven, to an aqueous solution of the test material at a single concentration of 100 mg/L for a period of 96 hours at a temperature of 19.8 to 21.2 °C under semi-static test conditions. The number of mortalities and any sub-lethal effects of exposure in each test and control vessel were determined 3 and 6 hours after the start of exposure and then daily throughout the test until termination after 96 hours.

A positive control conducted approximately every six months used pentachlorophenol, sodium salt as the reference material. In the most recent test, fish were exposed, in groups of seven, to an aqueous solution of the reference material at a range of concentrations of 0.10, 0.18, 0.32, 0.56 and 1.0 mg/L for 96 hours at a temperature of 19.7 to 20.6 ºC under semi-static test conditions. The number of mortalities and any sub-lethal effects of exposure in each vessel were determined 3 and 6 hours after the start of exposure and then daily throughout the test until termination after 96 hours.

Under the conditions of this study the 96 hour LC50 was > 100 mg/L. Correspondingly the No Observed Effect Concentration was 100 mg/L.

Supporting Study: Munk (1984)

The short-term toxicity of the test material to fish was investigated using rainbow trout in accordance with the standardised guideline OECD 203, under GLP conditions. The study was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).

A static test was conducted in freshwater over a period of 96 hours. The test organisms were housed in glass aquarium with a stainless steel frame (60 cm x 35 cm x 40 cm), at 17 ± 1 °C. The test concentrations used were: 31.6, 46.4, 68.1, 100.0, 147.0, 215.0 and 316.0 mg/L. Mortality and symptoms were monitored. 

Undissolved test material was visible in all concentrations 1 hour after beginning of the test. The amount increased with an increase in the concentration. After 24 hours the amount of undissolved test material had decreased and after 72 hours no undissolved test material was visible in the aquaria with surviving fish. Clinical symptoms observed were convulsions, discolouration, escape reflex and gasping. The symptom discolouration is regarded as a sign of stress and not as a sign of intoxication as it was also seen in the control group.

Under the conditions of this study the LC50 was between 150 and 220 mg/mL. The NOEC was 68.1 mg/L.

 

Supporting Study: Munk (1986)

The short-term toxicity of the test material to fish was investigated using bluegills, in a study closely similar to the standardised Guideline EPA 72-1, under GLP conditions. The study was awarded a reliability score of 2 in accordance with the criteria set forth by Klimisch et al. (1997).

A static test was conducted in freshwater over a period of 96 hours. The test organisms were housed in glass aquarium with a stainless steel frame (60 cm x 35 cm x 40 cm), at 23 °C. Based on the results of a preceding study with rainbow trout (LC 50 after 96 H: > 100 mg/L) the concentrations, spaced by a factor of about 2.2 were fixed as follows: 21.5, 31.6, 46.4, 68.1 and 100 mg/L. Mortality and symptoms were monitored. 

Small amounts of undissolved test material were visible at the bottom of the aquaria 24 hours after the beginning. After 96 hours no undissolved test material was visible. No abnormalities were observed in the test organisms. 

Under the conditions of this study the LC50 was > 100 mg/mL. The NOEC was 100 mg/L.

 

Supporting Study: Bidinotto (2004)

The short-term toxicity to the common carp by the test material was assessed in accordance with the standardised guidelines OECD 203 and the Japanese guideline 12-Nousan-p 8147 of MAFF, under GLP conditions. The study was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).

A range-finding test was carried out in order to determine the concentrations that cause 100 and 0 % of mortality to the test organisms. Based on the results of this test, a definitive test was carried out with the following nominal concentrations of the test material: 56; 100; 180; 320 and 560 mg/mL and a control group. Ten fish were used at each concentration. Mortality and sub-lethal effects were recorded at 24, 48, 72 and 96 hours. 

Under the conditions of this study the 96 hour LC50 was determined to be 134.16 mg/L. The No Observed Concentration (NOEC) was 100 mg/L.

 

Supporting Study: ten Berge (1978)

The short-term toxicity to the gubby by the test material was investigated. The study was awarded a reliability score of 2 in accordance with the criteria set forth by Klimisch et al. (1997).

Biodegradability of the test material was tested in water and soil and the toxicity to fish was measured over 96 hours. The test material was dissolved in standard fish water with a total hardness of 200 ppm. Several concentrations were made increasing in a geometric progression (each concentration was replicated twice). Guppies (3 to 4 weeks old) were transferred to these solutions in aquaria of 1 litre. To each concentration ten guppies were exposed. After 96 hours the mortality was recorded and from these data the LC50 at 96 hours was calculated. 

Under the conditions of this study the 96 hour LC50 of the test material to fish was determined to be 1100 mg/L (1000 - 1300 mg/L).