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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:
supporting study
Study period:
10 July 2013 to 17 August 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP study conducted in accordance with OECD and EU Guidelines.
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
Remarks:
See below under 'Any other information on materials and methods'
Qualifier:
according to guideline
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Deviations:
no
Remarks:
See below under 'Any other information on materials and methods'
GLP compliance:
yes (incl. QA statement)
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: Main test: 100 mg/L loading rate WAF. Range-finding test: nominal loading rates of 1.0, 10 and 100 mg/L.
- Sampling method: Main test: Water samples were taken from the control and the 100 mg/L loading rate WAF test vessel at 0 and 72 hours (fresh media) and at 24 and 96 hours (old media) for quantitative analysis. Duplicate samples, and samples at 24 (fresh media), 48 (old and fresh media) and 72 hours (old media), were taken for further analysis if necessary.
Range-finding test: A sample of each loading rate WAF was taken for chemical analysis at 0 and 24 hours in order to determine the stability of the test item under test conditions. Only concentrations within the 'range to be used for the definitive test were analyzed.
- Sample storage conditions before analysis: The samples were stored frozen prior to analysis.


Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Prepared as a Water Accommodated Fraction (WAF) of the test item due to low aqueous water solubility. At the request of the Sponsor, a stirring period of 47 hours followed by a 1-Hour standing period was used to prepare the test item.
- Eluate: None
- Differential loading: Based on the results of the range-finding test a "Limit test" was conducted at a single loading rate of 100 mg/L .
- Controls: Yes - without test item
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): No vehicle
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): No vehicle
- Evidence of undissolved material (e.g. precipitate, surface film, etc): Microscopic inspection of the WAFs showed no micro-dispersions or undissolved test item to be present.

Further information:
Range-finding test:
Amounts of test item (21, 210 and 2100 mg) were each separately added to the surface of 21 liters of test water to give the 1.0, 10 and 100 mg/L loading rates respectively. After the addition of the test item, the test water was stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface. The stirring was stopped after 47 hours and the mixtures allowed to stand for 1 hour. A wide bore glass tube, covered at one end with Nescofilm was submerged into the vessel, sealed end down, to a depth of approximately 5 cm from the bottom of the vessel. A length of Tygon tubing was inserted into the glass tube and pushed through the Nescofilm seal. Microscopic inspection of the WAFs showed no micro-dispersions or undissolved test item to be present. The aqueous phase or WAF was removed by mid-depth siphoning (the first 75-100 mL discarded) to give the 1.0, 10 and 100 mg/L loading rate WAFs.

Main test:
An amount of test item (2100 mg) was added to the surface of 21 liters of test water to give the 100 mg/L loading rate. After the addition of the test item, the test water was stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface. The stirring was stopped after 47 hours and the mixture allowed to stand for 1 hour. A wide bore glass tube, covered at one end with Nescofilm was submerged into the vessel, sealed end down, to a depth of approximately 5 cm from the bottom of the vessel. A length of Tygon tubing was inserted into the glass tube and pushed through the Nescofilm seal. Microscopic inspection of the WAF showed no micro-dispersions or undissolved test item to be present. The aqueous phase or WAF was removed by mid-depth siphoning (the first 75-100 mL discarded) to give the 100 mg/L loading rate WAF.

Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
TEST ORGANISM
- Common name: Juvenile rainbow trout
- Strain: Oncorhynchus mykiss
- Source: Brow Well Fisheries Limited, Hebden, near Skipton, Yorkshire, UK and maintained in-house since 20 June 2013. Fish were maintained in a glass fiber tank with a "single pass" water renewal system.
- Age at study initiation (mean and range, SD): Not specified.
- Length at study initiation (length definition, mean, range and SD): Mean standard length of 4.4 cm (sd = 0.4) at the end of the definitive test.
- Weight at study initiation (mean and range, SD): Mean weight of 1.01 g (sd = 0.36) at the end of the definitive test. Based on the mean weight value this gave a loading rate of 0.35 g bodyweight/liter.
- Method of breeding: Not specified.
- Feeding during test: No, the stock fish were fed commercial trout pellets which was discontinued approximately 24 hours prior to the start of the definitive test.
- Food type: N/A
- Amount: N/A
- Frequency: N/A

ACCLIMATION
- Acclimation period: Fish were acclimatized to test conditions from 24 July 2013 to 5 August 2013.
- Acclimation conditions (same as test or not): Yes
- Type and amount of food: commercial trout pellets
- Feeding frequency: Not specified.
- Health during acclimation (any mortality observed): There was no mortality in the 7 days prior to the start of the test.

QUARANTINE (wild caught)
- Duration: N/A
- Health/mortality: N/A
Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Post exposure observation period:
None
Hardness:
140 mg/L as CaC03
Test temperature:
The water temperature was controlled at 14 °C.
pH:
7.8 to 8.1
Dissolved oxygen:
9.3 to 10.1 mg O2/L
Salinity:
Not applicable
Nominal and measured concentrations:
Main test: 100 mg/L loading rate WAF.
Analysis of the 100 mg/L loading rate WAF fresh test preparations at 0 and 72 hours showed measured test concentrations of 0.016 and 0.020 mg/L respectively. Analysis of the old or expired media at 24 and 96 hours for the 100 mg/L loading rate WAF test concentration showed measured concentrations of 0.015 and 0.023 mg/L respectively.
Details on test conditions:
TEST SYSTEM
- Test vessel: Glass exposure vessels
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: Glass, 20L
- Aeration: The test vessels were aerated via narrow bore glass tubes.
- Type of flow-through (e.g. peristaltic or proportional diluter): N/A
- Renewal rate of test solution (frequency/flow rate): daily
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): N/A
- Biomass loading rate: loading rate of 0.35 g bodyweight/liter.

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) giving water with a total hardness of approximately 140 mg/L as CaC03. After dechlorination and softening the water was passed through a series of computer controlled plate heat exchangers to achieve the required temperature.
- Total organic carbon: average: 0.92 mg/L
- Particulate matter: Not specified
- Metals: Not specified
- Pesticides: average: 0.009 ug/L
- Chlorine: average free chlorine: 0.207 mg/L, average total chlorine: 0.270 mg/L
- Alkalinity: pH: 7.497
- Ca/mg ratio: Not specified.
- Conductivity: average: 419.0 uS/cm at 20 degrees C
- Culture medium different from test medium:
- Intervals of water quality measurement: The test water used for both the range-finding and definitive tests was the same as that used to maintain the stock fish.

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: Photoperiod of 16 hours light and 8 hours darkness with 20 minute dawn and dusk transition periods for a period of 96 hours.
- Light intensity: Not specified.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Test Organism Observations
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.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: One concentration in main test. Factor of 10 for range-finding test.
- Justification for using less concentrations than requested by guideline: Due to the low aqueous solubility and complex nature of the test item, for the purposes of the study the test medium was prepared as a Water Accommodated Fraction (WAF) of the test item. It was considered unnecessary and unrealistic to test at loading rates in excess of 100 mg/L loading rate WAF.
- Range finding study
- Test concentrations: nominal loading rates of 1.0, 10 and 100 mg/L.
- Results used to determine the conditions for the definitive study: Based on the results of the range-finding test a "Limit test" was conducted at a single loading rate of 100 mg/L to confirm that no mortalities or sub-lethal effects of exposure were observed.
Reference substance (positive control):
no
Duration:
96 h
Dose descriptor:
LL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Details on results:
- Behavioural abnormalities: There were no sub-lethal effects of exposure observed in 7 fish exposed to a 100 mg/L loading rate WAF for a period of 96 hours.
- Observations on body length and weight: Mean standard length of 4.4 cm (sd = 0.4) at the end of the definitive test. Mean weight of 1.01 g (sd = 0.36) at the end of the definitive test. Based on the mean weight value this gave a loading rate of 0.35 g bodyweight/liter.
- Other biological observations: Not specified
- Mortality of control: There were no mortalities.
- Other adverse effects control: No
- Abnormal responses: No
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: At the start of the mixing period the 100 mg/L loading rate was observed to be a clear colorless water column with oily globules of test item floating on the surface. After 47 hours stirring and a 1-Hour standing period the 100 mg/L loading rate was observed to remain a clear colorless water column with oily globules of test item floating on the surface. Microscopic inspection of the WAF showed no micro-dispersions or undissolved test item to be present. For the duration of the test, the 100 mg/L loading rate was observed to be a clear, colorless solution. No media observations were performed at the 0-Hour time-point which is a deviation from the study plan. However, this was considered not to impact the results of the test as the media was observed to be a clear colorless water solution for all other observational time-points.
- Effect concentrations exceeding solubility of substance in test medium: Loading rate WAF reported due to low aqueous water solubility.

The test was considered to be valid given that none of the control fish died or showed signs of stress during the test and that the oxygen concentration at the end of the test was 60% of ASV (>6.2 mg O/L) in the control and test vessels.

Temperature was maintained at approximately 14 °C throughout the test, while there were no treatment related differences for oxygen concentration or pH.

The vortex depth was recorded at the start and end of each mixing period and was observed to be a dimple at the water surface on each occasion.
Results with reference substance (positive control):
Not applicable
Sublethal observations / clinical signs:

Cumulative Mortality Data in the Range-finding Test:

 

Nominal Loading Rate (mg/L)

Cumulative Mortality (Initial Population=3)

3 Hours

6 Hours

24 Hours

48 Hours

72 Hours

96 Hours

Control

0

0

0

0

0

0

1.0

0

0

0

0

0

0

10

0

0

0

0

0

0

100

0

0

0

0

0

0

Cumulative Mortality Data in the Definitive Test:

 

Nominal LoadingRate(mg/L)

Cumulative Mortality (Initial Population=7)

%

Mortality

3

Hours

6

Hours

24

Hours

48

Hours

72

Hours

96

Hours

96

Hours

Control

0

0

0

0

0

0

0

100

0

0

0

0

0

0

0

Water Quality Measurements:

 

Nominal Loading Rate (mg/L)

Time (Hours)

0 Hours (Fresh Media)

24 Hours (Old Media)

24 Hours (Fresh Media)

pH

mg 02/L

T (degrees C)

pH

mg 02/L

T (degrees C)

pH

mg 02/L

T (degrees C)

Control

7.9

9.9

15

8.0

9.5

14

7.9

9.8

15

100

7.8

9.8

15

8.0

9.3

14

7.8

9.8

15

 

 

Nominal Loading Rate (mg/L)

Time (Hours)

48 Hours (Old Media)

48 Hours (Fresh Media)

72 Hours (Old Media)

pH

mg 02/L

T (degrees C)

pH

mg 02/L

T (degrees C)

pH

mg 02/L

T (degrees C)

Control

7.9

10.1

14

7.9

9.8

15

7.9

9.7

14

100

7.9

9.6

14

7.8

9.7

15

7.9

9.5

14

 

 

Nominal Loading Rate (mg/L)

Time (Hours)

72 Hours (Fresh Media)

96 Hours (Old Media)

pH

mg 02/L

T (degrees C)

pH

mg 02/L

T (degrees C)

Control

7.9

9.8

14

8.1

9.9

14

100

7.8

9.7

15

8.0

9.5

14

Validity criteria fulfilled:
yes
Conclusions:
The acute toxicity of the test item to the freshwater fish rainbow trout (Oncorhynchus mykiss) has been investigated and gave a 96-Hour LL50 value of greater than 100 mg/L loading rate WAF. The No Observed Effect Loading rate was 100 mg/L loading rate WAF.
Executive summary:
Introduction

A study was performed to assess the acute toxicity of the test item to rainbow trout (Oncorhynchus mykiss). The method followed was designed to be compatible with the OECD Guidelines for Testing of Chemicals (1992) No 203, "Fish, Acute Toxicity Test" referenced as Method C.1 of Commission Regulation (EC) No. 440/2008.

 

Methods

Due to the low aqueous solubility and complex nature of the test item, for the purposes of the test, the test medium was prepared as a Water Accommodated Fraction (WAF).

 

Following a preliminary range-finding test, seven fish were exposed to a Water Accommodated Fraction (WAF) of the test item, at a single nominal loading rate of 100 mg/L for a period of 96 hours at a temperature of approximately 14 °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.

 

Results

Analysis of the 100 mg/L loading rate WAF fresh test preparations at 0 and 72 hours showed measured test concentrations of 0.016 and 0.020 mg/L respectively. Analysis of the old or expired media at 24 and 96 hours for the 100 mg/L loading rate WAF test concentration showed measured concentrations of 0.015 and 0.023 mg/L respectively.

 

The results show that test item concentrations in the region of 3-4 ppb were detected in the control sample

s at each of the study sampling time points. On inspection of the chromatograms it is clearly evident the peaks detected in the control samples correspond to the characteristic profile of the test item. The analytical method used LC MS-MS, which was very specific for test item mass fragments and the chromatographic profile seen consisted of several unresolved peaks in similar ratios to that of the standard calibration solutions. No peaks were detected in the solvent matrix or in the synthetic control prepared at the same time of the test sample analysis. However, whilst test item concentrations above that of the LOQ have been detected in the control samples, the concentrations are extremely low and have only been detected due to the superior detection limits of the LC MS-MS analytical instrumentation used in the study.

 

The dissolved test item may have been one or several components of the test item. Given that toxicity cannot be attributed to a single component or mixture of components but to the test item as a whole, the results were based on nominal loading rates only.

 

Exposure of rainbow trout to the test item gave LL50 values of greater than 100 mg/L loading rate WAF. The No Observed Effect Loading Rate was 100 mg/L loading rate WAF. It was considered unnecessary and unrealistic to test at loading rates in excess of 100 mg/L loading rate WAF.

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
from 1988-07-11 to 1988-07-15
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: No full study report, only summarised results
Principles of method if other than guideline:
No data concerning used method.
GLP compliance:
not specified
Analytical monitoring:
not specified
Vehicle:
not specified
Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
TEST ORGANISM
- Common name: rainbow trout
- Weight at study initiation: 0.50 g (mean weight)

No further details mentioned
Test type:
static
Water media type:
not specified
Limit test:
yes
Total exposure duration:
96 h
Test temperature:
15 +/- 1 °C
Nominal and measured concentrations:
1 mg/L
Details on test conditions:
10 fish/concentration were used, no further details mentioned
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 1 mg/L
Nominal / measured:
not specified
Conc. based on:
not specified
Basis for effect:
mortality (fish)
Sublethal observations / clinical signs:
concentration [mg/L]  % survivors         
   24 h  48 h  72 h  96 h
 0 (control)  100  100  100  100
 1  100  100  90  90
Validity criteria fulfilled:
not specified
Conclusions:
The LC50 of the test substance was greater than 1 mg/L under the conditions described.
Executive summary:

The LC50 of 1,2,4 -Benzenetricarboxylic acid, mixed decyl and octyl triesters was greater than 1 mg/L, tested with Salmo gaidneri.

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1994-07-04 to 1994-07-08
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Guideline study with acceptable restrictions
Qualifier:
according to guideline
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
- Concentrations: WSF, measured every day (days 0 - 3)
- Sampling method: One sample was taken directly from the stock solution and analyzed, a second sample was taken out of an aquarium which was set up with test solution but without fish after 24h and analyzed to determine the test item's stability under test conditions.
- Sample storage conditions before analysis: not mentioned
Vehicle:
no
Details on test solutions:
As the substance is poorly soluble in water, an aqueous filtrated stock solution was prepared. The content of dissolved organic carbon in the water was 3 mg/L in average.
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: zebrafish
- Strain: Brachydanio rerio
- Source: West Aquarium, Bad Lauterberg, Germany
- Age at study initiation (mean and range, SD): not mentioned
- Length at study initiation (length definition, mean, range and SD): 3 ± 0.5 cm, no further details mentioned
- Weight at study initiation (mean and range, SD): approx. 0.13 g, no further details mentioned
- Method of breeding: not mentioned
- Feeding during test: none


ACCLIMATION
- Acclimation period: 14 days
- Acclimation conditions (same as test or not): The fish were kept in 200 L earthenware basins, or in 300 L glass aquariums in flow rate.
- Type and amount of food: TetraMin, administration in an amount of approx. 1 % of body weight
- Feeding frequency: daily
- Health during acclimation (any mortality observed): The animals showed a normal behaviour pattern at the beginning of the test and were free from visible diseases. The mortality rate during the holding phase was <5% seven days before the beginning of the test.
Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Post exposure observation period:
none
Hardness:
12.3 °d.H. (German hardness)
Test temperature:
21 ± 1 °C
pH:
7.8 - 8.4
Dissolved oxygen:
92 - 105 % relative to saturation
Salinity:
not mentioned
Nominal and measured concentrations:
Nominal: WAF
Measured: 3.0 mg/L (geometric mean of measurement of freshly prepared and after 24 hours solutions)
Details on test conditions:
TEST SYSTEM
- Test vessel: 30 L aquarium
- Type (delete if not applicable): not mentioned
- Material, size, headspace, fill volume: 30 L total, 10 L fill volume, no further details mentioned
- Aeration: continously, no further details mentioned
- Renewal rate of test solution (frequency/flow rate): daily renewal
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.13 g/L


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: drinking water, Gelsenwasser AG, Germany
no further details mentioned
- Intervals of water quality measurement: not mentioned

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light, 8 hours darkness
- Light intensity: not mentioned


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality after 24, 48, 72 and 96 hours; pH, temperature, O2 saturation at the end of the test


TEST CONCENTRATIONS
- Spacing factor for test concentrations: not applicable (limit test, test concentration limited by test item's water solubility)
- Range finding study: not applicable (see above)
Reference substance (positive control):
no
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 3 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
dissolved
Basis for effect:
mortality (fish)
Remarks on result:
other: no mortality at limit of water solubility
Duration:
96 h
Dose descriptor:
LC0
Effect conc.:
>= 3 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
dissolved
Basis for effect:
mortality (fish)
Remarks on result:
other: no mortality at limit of water solubility
Results with reference substance (positive control):
no reference substance tested
Reported statistics and error estimates:
no statisitcs performed, as no mortality occured
Sublethal observations / clinical signs:

 concentration [mg/L]  No. of fish at start of study  observation period                     
     24 h  48 h 72 h 96 h
     No. dead  mortality [%]  No. dead  mortality[%]  No. dead  mortality[%]  No. dead  mortality[%]
control   10  0  0  0  0  0  0  0
 3  10  0  0  0  0  0  0  0  0
Validity criteria fulfilled:
yes
Conclusions:
Not toxic under test conditions in the range of water solubility.
Executive summary:

The acute toxicity of 1,2,4-Benzenetricarboxylic acid, mixed decyl and octyl triesters to Zebra fish (Brachydanio rerio) was determined in a 96 -hour static test according to the Commision Directive 92/96/EEC, Annec Part C.1, dated December 29, 1992 and the OECD Guideline for Testing of Chemicals No. 203, adopted July 17, 1992. The test was performed in compliance with Good Laboratory Practice Regulations. Due to the low water solubility limit of the test substance a filtrate of a 1 g/L in tap water (stirring for 18 hours) was tested. Analytical verification was done by means of DOC determination. The measured values of the test solutions after 24 hours deviated more than 20% from the values of the freshly prepared solutions. Therefore the geometric mean (3.0 mg/L) of the these solutions were used for evaluation. Additionally a control was tested in parallel.


No lethality of Brachydanio rerio was observed at the tested concentration after 96 hours exposure.

Description of key information

In all three available studies the test substance was not toxic under test conditions in the range of water solubility.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Dose descriptor:
LC50
Effect concentration:
> 3 mg/L

Additional information

Three studies were conducted with the test substances with different fish species.


 


In the key study the acute toxicity of 1,2,4 -Benzenetricarboxylic acid, mixed decyl and octyl triesters to Zebra fish (Brachydanio rerio) was determined in a 96 -hour static test according to the Commision Directive 92/96/EEC, Annex Part C.1, dated December 29, 1992 and the OECD Guideline for Testing of Chemicals No. 203, adopted July 17, 1992. The test was performed in compliance with Good Laboratory Practice Regulations. Due to the low water solubility limit of the test substance a filtrate of a 1 g/L in tap water (stirring for 18 hours) was tested. Analytical verification was done by means of DOC determination. The measured values of the test solutions after 24 hours deviated more than 20% from the values of the freshly prepared solutions. Therefore the geometric mean (3.0 mg/L) of the these solutions were used for evaluation. Additionally a control was tested in parallel. No lethality of Brachydanio rerio was observed at the tested concentration after 96 hours exposure.


 


In a second study the LC50 of 1,2,4 -Benzenetricarboxylic acid, mixed decyl and octyl triesters was greater than 1 mg/L, tested with Salmo gardneri.


 


In a supporting study the acute toxicity of the test item to the freshwater fish rainbow trout (Oncorhynchus mykiss) has been investigated and gave a 96-Hour LL50 value of greater than 100 mg/L loading rate WAF. The No Observed Effect Loading rate was 100 mg/L loading rate WAF.


 


Therefore the test substance was not toxic under test conditions in the range of water solubility.