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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 October 15, 1990 to October 19, 1990
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
EPA OPP 72-1 (Fish Acute Toxicity Test)
Deviations:
no
GLP compliance:
yes
Specific details on test material used for the study:
- Physical state: Yellow liquid (Non-radiolabelled test substance); clear liquid (Radiolabelled test substance)
- Analytical purity: Non-radiolabelled test substance - 30% w/w active substance in aqueous solution; Radiolabelled test substance - 96.5% (by TLC) and 95.5% (by HPLC).
- Lot/batch No.: 05-6K (Non-radiolabelled test substance)
- Stability under test conditions: Non-radiolabelled test substance is hydrolytically and photolytically stable under the conditions of this study and has been shown to be stable in aqueous, alcohol and alcohol/aqueous solutions for extended periods, e.g. at least five years under standard laboratory conditions.
- Storage condition of test material: Radiolabelled and non-radiolabelled test substance was stored refrigerated or frozen in dark for the duration of the study.
- Density: Approximately 0.97 g/mL (Non-radiolabelled test substance)
- Boiling point: Approximately 212°C (Non-radiolabelled test substance)
- Specific activity (if radiolabelling): 25 mCi/mmol
Analytical monitoring:
yes
Details on sampling:
- Concentrations: 0, 0.180, 0.320, 0.490, 0.560 and 0.750 mg/L
- Sampling method: water quality characteristics of the test solutions were measured at test initiation, at the 24, 48 and 72 h renewal events, and at test termination.
Details on test solutions:
Preparation of test solutions:
The test solutions were prepared from a single stock solution of 14C-ADBAC and ADBAC. The stock was prepared by adding 11.70 mg of radiolabelled test substance and 919.5 mg of non-radiolabelled test substance in deionized water and brought to a volume of 250 mL to yield a nominal concentration 3.72 mg/mL test substance. The test solutions were prepared by adding measured aliquots of this stock solution to dilution water at test temperature (22±1°C), and adjusting the final volume to 22L in the test aquaria.
Test organisms (species):
Lepomis macrochirus
Details on test organisms:
Test organism:
- Common name: Bluegill sunfish
- Source: Kurtz’s Fish Hatchery Elverson, PA
- Feeding during test: No
- Length at study initiation: 33-41 mm (Average length 36 mm)
- Weight at study initiation: 0.57-0.98 g (Average weight 0.76 g)
Acclimation:
- Acclimation period: 2 wks
- Food: Commercial trout chow (Purina brand), frozen brine shrimp (Artemia) and Tetra-Min
- Feeding frequency: 5-6 times/wk
- Water (maintenance): dilution water
- Photoperiod: 16h/8h light/dark photoperiod
- Temperature: 22 ± 2.2°C
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Hardness:
48-84 mg/L as CaCO3
Test temperature:
21.5-22.0°C
pH:
6.8-7.2
Dissolved oxygen:
6.9-8.8 mg/L
Nominal and measured concentrations:
Nominal test concentration: 0, 0.180, 0.320, 0.490, 0.560 and 0.750 mg/L
Mean measured test concentrations: 0, 0.1973, 0.3171, 0.4555, 0.515 and 0.638 mg/L
Details on test conditions:
Test system:
- Test vessel: glass aquaria (capacity 25L)
- Material, size, headspace, fill volume: 22L
- Aeration: No aeration was provided during the test
- Renewal rate of test solution (frequency/flow rate): 24h renewals
- No. of organisms per vessel: 10
- No. of vessels per concentration: 2
- No. of vessels per control: 2
- Biomass loading rate: 0.35 g/L

Test medium / water parameters:
- Source/preparation of dilution water: the dilution water consisted of water from two deep wells. The well water was initially treated to remove iron and organic impurities (activated carbon filters). Some of this water was then sequentially passed through reverse osmosis (RO) purifiers to reduce ion concentrations. To obtain test water of proper hardness, the RO water and the treated well water (organic impurity removal and iron removal only) were mixed in a 500 gallon capacity fibreglass storage tank to obtain a hardness of approximately 50 mg/L as CaCO3.
- Total organic carbon: <0.8 mg/L
- Dissolved oxygen (DO) concentration: 6.9-8.8 mg/L, which represented 83-106% of air saturation at 22°C
- Conductivity: 192 µMhos/cm
- Metals: <2 µg/L Zinc, <1 µg/L cadmium, <12 µg/L copper
- Chlorine (residual): <0.05 mg/L
- Alkalinity: 68-128 mg/L as CaCO3
- Culture medium different from test medium: No
- Intervals of water quality measurement: Water quality characteristics of the test solutions were measured at test initiation, at the 24, 48 and 72h renewal events, and at test termination.

Other test conditions:
- Photoperiod: 16h/8h light/dark photoperiod
- Light intensity: 74.2-81.3 foot candles

Effect parameters measured (with observation intervals if applicable):
Observations of symptoms of toxicity and mortality were made at 24, 48, 72 and 96h. Mortality is defined as the lack of movement by the test organism after touching the caudal peduncle. Data from the 96 h static-renewal acute test with bluegill sunfish were used to estimate median lethal concentration (LC50).

Test concentrations:
- Range finding study: A 96 h static acute range-finding test was conducted to determine the concentration range of the test substance to use in the definitive test. The range-finding test was conducted with one replicate test chamber concentration with 10 fish/replicate.
- Test concentrations: 0, 0.100, 0.500, 0.625, 0.825 and 1.000 mg/L
- Results used to determine the conditions for the definitive study: Mortality was ≤10% in the control and the 0.1 mg/L nominal test concentration during the 96 h static exposure. The 96h LC50 value (nominal concentration) was 0.556 mg/L of the test substance. Based on these results, nominal test concentrations were selected for the definitive study.
Reference substance (positive control):
no
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
0.515 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
act. ingr.
Basis for effect:
mortality
Remarks on result:
other: confidence interval: 0.456-0.638 mg/L
Key result
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
0.456 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
act. ingr.
Basis for effect:
mortality
Details on results:
Test substance
- No mortality was observed at the test concentrations of 0.456 mg/L or less during the entire test period. Immobilisation and erratic swimming were noted for some fish in the 0.515 mg/L group. 

Control
- All control fish appeared normal and survived for the duration of the test.

Reported statistics and error estimates:
Estimates of LC50 values were obtained using version 1.61 of the computer program, TOXDAT, cited in Peltier and Weber (Methods for measuring the acute toxicity of effluents to freshwater and marine organisms. 1985, US Environmental Protection Agency).

Table 1. Cumulative mortality data

Concentration (mg/L)

Percent Mortality

 

24 h

48 h

72 h

96 h

0.6384

100

100

100

100

0.515

30

50

50

50

0.4555

0

0

0

0

0.3171

0

0

0

0

0.1973

0

0

0

0

Control

0

0

0

0

 

Table 2. Results based on the mortality data

LC50 (mg/L)

24 h

48 h

72 h

96 h

 

0.540

0.515

0.515

0.515

95% confidence

limits

(0.456-0.638)

(0.456-0.638)

(0.456-0.638)

(0.456-0.638)

 

Validity criteria fulfilled:
yes
Conclusions:
Under the test conditions, the 96h LC50 and NOEC (Lepomis macrochirus) of the test substance were calculated to be 0.515 mg a.i./L and 0.456 mga.i./L respectively.
Executive summary:

A study was conducted to determine the short-term toxicity to fish of the test substance according to US EPA OPP 72 -1, in compliance with GLP. The experiment was carried out in Bluegill sunfish (Lepomis macrochirus) under static conditions. Fish were exposed for 96 h to nominal test substance concentrations of 0, 0.180, 0.320, 0.490, 0.560 and 0.750 mg a.i./L, equivalent to mean measured concentrations of 0, 0.1973, 0.3171, 0.4555, 0.515 and 0.638 mg a.i./L. Analytical dose verification was performed. Twenty fish in two replicates were exposed to each test concentration. The solutions were renewed at 24, 48 and 72 h. Observations of symptoms of toxicity and mortality were conducted at 24, 48, 72 and 96 h. The LC50 was calculated based on measured concentrations following a binomial method. No mortality was observed at the test concentrations of 0.456 mg/L or less during the entire test period. Immobilisation and erratic swimming were noted for some fish in the 0.515 mg/L group. Under the test conditions, the 96 h LC50 and NOEC were calculated to be 0.515 and 0.456 mg a.i./L, respectively (Pate, 1991).

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From June 22, 1987 to July 28, 1987.
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
yes
Remarks:
test media renewal after 48h. Analysis of the test substance was not performed.
Qualifier:
according to
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Deviations:
yes
Remarks:
test media renewal after 48h. Analysis of the test substance was not performed.
Principles of method if other than guideline:
Explanations and other deviations:
No chemical analyses but data from other tests demonstrate the stability and availability of the test substance under test conditions.
The second acclimatisation time is shorter than advised by the guideline but considering the low mortality during the second period. This is considered to have no impact on the final outcome of the test.
GLP compliance:
yes
Specific details on test material used for the study:
- Material containing: ca. 50% Cocobenzyldimethylammonium chloride (CAS no.: 61789-71-7) in water.
- Material is based on full cocoalkylchain , the chain length distribution is somewhat different compared with C12-16 BKC: (chainlength
distribution of full coco: C8: 5%; C10: 6%; C12: 50%; C14: 19%; C16: 10%; C18:10%). The somewhat broader chainlength distribution does not influence the validity for the use of this study for the most commonly used active substance. The results from this substance however are also valid for evaluation of some additional shorter and longer chain lengths present in the test substance.
Analytical monitoring:
no
Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
Species/strain: Salmo gairdneri
Source: Forellenhof Fredesloh, Dr. O. Mueller, 3413 Moringen 1
Wild caught: no
Age/size: 5 ± 1 cm
Food: trout food Pelletgr. S1 (Rheinkrone Kraftfutter)
Pretreatment: during the first time of the adaptation phase (12d) the mortality came to 7% of the fish population. Therefore, the adaptation phase was extended for 5d. During this time the mortality only came to 0.5% of the fish population. 24h before the test started feeding of the fish was stopped.
Feeding of animals during test: no
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Test temperature:
ca. 14°C
pH:
ca. 8
Dissolved oxygen:
ca. 10 mg/L
Nominal and measured concentrations:
Nominal concentrations: 0, 0.5, 0.7, 1.0, 1.4, 2.0, 3.5 and 10.0 mg/L.
Duration:
96 h
Dose descriptor:
LC0
Effect conc.:
0.7 mg/L
Conc. based on:
act. ingr.
Basis for effect:
mortality
Remarks on result:
other: measured
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
0.85 mg/L
Conc. based on:
act. ingr.
Basis for effect:
mortality
Remarks on result:
other: calculated
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
1.75 mg/L
Conc. based on:
act. ingr.
Basis for effect:
mortality
Remarks on result:
other: measured
Details on results:
During the first time of the adaptation phase (12d) the mortality came to 7% of the fish population. Therefore the adaptation phase was extended for 5d. During this time the mortality only came to 0.5% of the fish population. The oxygen content of the water was above 60% during the test and according to analyses in other tests (e.g. algae test), the test substance concentration was above 80% of the initial concentration during the test. Therefore all validity criteria are fulfilled. The concentration-response relationship appears to be typical for this type of substance. The study can be considered as valid.
Validity criteria fulfilled:
yes
Conclusions:
Under the study conditions, the 96h LC0, LC50 and LC100 values (Oncorhynchus mykiss) for the test substances were determined to be 1.4 mg/L (i.e., equivalent to 0.7 mg a.i./L), 1.7 mg/L (i.e., equivalent to 0.85 mg a.i./L) and 3.5 mg/L (i.e., equivalent to 1.75 mg a.i./L).
Executive summary:

A study was conducted to determine the short-term toxicity to fish of the test substance (ca. 50% purity) according to OECD Guideline 203 and EU Method C.1, in compliance with GLP. The experiment was conducted in Rainbow trout (Onchorhynchus mykiss) under static conditions. Fish (5 animals in duplicate) were exposed for 96 h to concentrations of 0, 0.5, 0.7, 1.0, 1.4, 2.0, 3.5 and 10.0 mg/L. No chemical analysis of test concentrations was performed. Abnormal findings during the exposure period were recorded in the swimming behaviour, visible physiological functions and outward appearance of the fish. Under the study conditions, the 96 h LC0, LC50 and LC100 values were determined to be 1.4 mg/L (i.e., equivalent to 0.7 mg a.i./L), 1.7 mg/L (i.e., equivalent to 0.85 mg a.i./L) and 3.5 mg/L (i.e., equivalent to 1.75 mg a.i./L) (Dommrose, 1987).

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From May 12, 2003 to May 16, 2003
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
RA study
Justification for type of information:
Refer to the section 13 for details on the read across justification. The fish study with the read across substance is considered sufficient to fulfil the information requirements as further explained in the provided endpoint summary.
Qualifier:
according to
Guideline:
other: PARCOM 1995 Part B
Deviations:
no
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
GLP compliance:
yes
Specific details on test material used for the study:
- Composition of test material, percentage of components: ADBAC C12-14 content of 79.75%
- Appearance: A clear light yellow liquid
- Stability: Stable
- Solubility: Soluble in water
- Specific gravity: 0.976 g/mL at 15°C
- Date received: May 05, 2003
- Storage: 20°C
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
- Preparation of the test solutions:
The test substance was thoroughly homogenized before use. The test substance was soluble in water, so the sample preparation method followed for water soluble test substances was used. The test substance was added to a beaker containing 0.5L of seawater. The amount of test substance added was 3 times the final concentration (i.e., 3x5 mg/L = 15 mg of test substance to the beaker). The solution was mixed on magnetic stirrers for >30 minutes after covering the top of the beaker to minimize volatilization. The content of beaker was added to 2 L of clean seawater and the beaker was rinsed with 0.5 L of seawater. This was also added to the above solution to make 3 L of stock solution. Required test concentrations were prepared from this solution.
- Reference seawater:
Reference seawater used for testing was bought from Bjolsen Akvarium AS and was stored for no longer than 4 weeks at temperatures between 18 and 22°C.
Test organisms (species):
Cyprinodon variegatus
Details on test organisms:
Test organism:
- Common name: Sheepshead minnow
- Source: Seaplantation Engineering Incorporation
- Date of arrival: April 02, 2003
- Length at study initiation: Control fish (Average of 10 fish): 1.9 cm
- Weight at study initiation: Control fish (Average of 10 fish): 95.1 mg
Acclimation
On arrival at the laboratory, the fish were transferred to a temperature room to allow temperature acclimatization. Temperature in the fish containers and the acclimatization tank were measured. The fish were held in a seawater flow-through system and pH, salinity, O2 saturation and temperature were measured within one hour of arrival and subsequently on every working day. Aeration was supplied and the fish were fed at least once per working day. Mortality during the first 9 days after arrival was 0.8%. This was within the valid range.
Holding conditions:
- Synthetic seawater
- Temperature: 17.6 - 22.4°C
- Salinity: 3.3%
- Dissolved oxygen: >60%
- Condition: Semi-static
Holding period:
- From April 30, 2003 to May 12, 2003
Test type:
semi-static
Water media type:
saltwater
Limit test:
no
Total exposure duration:
96 h
Test temperature:
Temperatures are reported in °C
Control: 18.9 - 19.9
1 mg/L: 19.3 - 20.6
3 mg/L: 19.4 - 19.6
10 mg/L: 19.5 - 19.6
30 mg/L: 19.4 - 19.6
100 mg/L: 19.4 - 19.6
pH:
Control: 8.00 - 8.20
1 mg/L: 7.89 - 8.18
3 mg/L: 8.09 - 8.19
10 mg/L: 8.07 - 8.19
30 mg/L: 8.11 - 8.19
100 mg/L: 8.12 - 8.20
Dissolved oxygen:
Dissolved oxygen levels are reported in % saturation
Control: 95 -100
1 mg/L: 95 -100
3 mg/L: 98 - 100
10 mg/L: 99 - 100
30 mg/L: 99 -100
100 mg/L: 100
Salinity:
Salinity is reported in %
Control: 3.2 - 3.3
1 mg/L: 3.3 - 3.3
3 mg/L: 3.3
10 mg/L: 3.3
30 mg/L: 3.3 - 3.3
100 mg/L: 3.3 - 3.3
Nominal and measured concentrations:
- Test substances concentrations (nominal concentrations of test substance) used for the definitive test: 1, 3, 10, 30 and 100 mg/L.
- Reference substance (3,5-dichlorophenol) concentrations (nominal): 2.3 mg/L
Details on test conditions:
Test system:
- Renewal rate of test solution: 48 h
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 320 mg/L
Effects parameters measured: Mortality
Reference substance (positive control):
yes
Remarks:
3,5-dichlorophenol
Key result
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
1 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: Calculated
Duration:
96 h
Dose descriptor:
LC10
Effect conc.:
1.6 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: Calculated
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
1.7 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: Calculated
Duration:
96 h
Dose descriptor:
other: LC90
Effect conc.:
1.9 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: Calculated
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
1.28 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
mortality
Remarks on result:
other: Calculated
Details on results:
- Mortality of control: 0%
- Mortality with test substance: Refer to Table 1
Results with reference substance (positive control):
- Results with reference substance valid? yes
- Mortality: 40% after 96h
Reported statistics and error estimates:
The LC50 and 95% confidence intervals were calculated using a Logistic Regression Model.

Table 1. Log test substance concentrations and mortality after 24, 48, 72and 96h

Test concentration (mg/L)

Log test concentration

Number of fish per vessel

Mortality (24 h)

Mortality (48 h)

Mortality (72 h)

Mortality (96 h)

Cumulative mortality %

Control

---

10

0

0

0

0

0

1

0

10

0

0

0

0

0

3

0.48

10

10

10

10

10

100

10

1.00

10

10

10

10

10

100

30

1.48

10

10

10

10

10

100

100

2.00

10

10

10

10

10

100

 

Table2. LC50 values and their 95% confidence limits after 24, 48, 72 and 96h for the test substance (The 95% confidence range could not be calculated)

Duration (h)

LC50 (mg/L)

24

1.7

48

1.7

72

1.7

96

1.7

Summary of the results:

The LC50 value for the test substance was found to be 1.7 mg/L after 96 h.

The 95% confidence range could not be calculated.

The NOEC value was estimated to 1 mg/L.

LC50 values after 24, 48 and 72 h were all 1.7 mg/L.

 

Table 3: Calculation of LC10, LC50 and LC90-values after 24, 48, 72 and 96h (The 95% confidence range could not be calculated)

24, 48, 72 and 96 h

Effect level

mean (log)

LC10

0.21

LC50

0.24

LC90

0.27

Effect level

mean (inverse log)

LC10

1.6

LC50

1.7

LC90

1.9

 

Reference substance:

To ensure reproducibility, each batch of C.variegatus was tested with a reference substance. A reference test was conducted using a single concentration 2.3 mg/L of 3,5-dichlorophenol. The recorded mortality was 40 % after 96h.

Validity criteria fulfilled:
yes
Executive summary:

A study was conducted to determine the short term toxicity to fish of the read across substance, benzyl-C12-14-alkyldimethylammonium chlorides (C12-14 ADBAC), according to PARCOM 1995 Part B which is equivalent or similar to OECD Guideline 203, in compliance with GLP. Juvenile marine Sheepshead minnow (Cyprinodon variegatus) were exposed for 96 h at nominal concentrations of 0, 1, 3, 10, 30 and 100 mg/L, equivalent to 0, 0.8, 2.4, 8.0, 24 and 80 mg a.i./L based on a purity of 79.75%. The reference substance 3,5-dichlorophenol was included in the study. Under the study conditions, the 96 h LC50 was 1.7 mg/L (i.e. 1.36 mg a.i./L) and the NOEC was 1 mg/L (i.e. 0.8 mg a.i./L) . Further, the 96 h LC10 and C90 were calculated at 1.6 and 1.9 mg/L (i.e. 1.3 and 1.5 mg a.i./L) , respectively (Sundfor, 2003).

Description of key information

The 96 h LC50 value for mortality to Bluegill sunfish (freshwater) and Sheepshead minnow ( marine water) due to the test substance were determined to be 0.515 mg a.i./L (measured) and 1.28 mg a.i./L (nominal) respectively.

Key value for chemical safety assessment

LC50 for freshwater fish:
0.515 mg/L
LC50 for marine water fish:
1.28 mg/L

Additional information

Study 1. A study was conducted to determine the short-term toxicity to fish of the test substance according to US EPA OPP 72 -1, in compliance with GLP. The experiment was carried out in Bluegill sunfish (Lepomis macrochirus) under static conditions. Fish were exposed for 96 h to nominal test substance concentrations of 0, 0.180, 0.320, 0.490, 0.560 and 0.750 mg a.i./L, equivalent to mean measured concentrations of 0, 0.1973, 0.3171, 0.4555, 0.515 and 0.638 mg a.i./L. Analytical dose verification was performed. Twenty fish in two replicates were exposed to each test concentration. The solutions were renewed at 24, 48 and 72 h. Observations of symptoms of toxicity and mortality were conducted at 24, 48, 72 and 96 h. The LC50 was calculated based on measured concentrations following a binomial method. No mortality was observed at the test concentrations of 0.456 mg/L or less during the entire test period. Immobilisation and erratic swimming were noted for some fish in the 0.515 mg/L group. Under the test conditions, the 96 h LC50 and NOEC were calculated to be 0.515 and 0.456 mg a.i./L, respectively (Pate, 1991).

Study 2. A study was conducted to determine the short-term toxicity to fish of the test substance (ca. 50% purity) according to OECD Guideline 203 and EU Method C.1, in compliance with GLP. The experiment was conducted in Rainbow trout (Onchorhynchus mykiss) under static conditions. Fish (5 animals in duplicate) were exposed for 96 h to concentrations of 0, 0.5, 0.7, 1.0, 1.4, 2.0, 3.5 and 10.0 mg/L. No chemical analysis of test concentrations was performed. Abnormal findings during the exposure period were recorded in the swimming behaviour, visible physiological functions and outward appearance of the fish. Under the study conditions, the 96 h LC0, LC50 and LC100 values were determined to be 1.4 mg/L (i.e., equivalent to 0.7 mg a.i./L), 1.7 mg/L (i.e., equivalent to 0.85 mg a.i./L) and 3.5 mg/L (i.e., equivalent to 1.75 mg a.i./L) (Dommrose, 1987).

Study 3.

A study was conducted to determine the short term toxicity to fish of the read across substance, benzyl-C12-14-alkyldimethylammonium chlorides (C12-14 ADBAC), according to PARCOM 1995 Part B which is equivalent or similar to OECD Guideline 203, in compliance with GLP. Juvenile marine Sheepshead minnow (Cyprinodon variegatus) were exposed for 96 h at nominal concentrations of 0, 1, 3, 10, 30 and 100 mg/L, equivalent to 0, 0.8, 2.4, 8.0, 24 and 80 mg a.i./L based on a purity of 79.75%. The reference substance 3,5-dichlorophenol was included in the study. Under the study conditions, the 96 h LC50 was 1.7 mg/L (i.e. 1.36 mg a.i./L) and the NOEC was 1 mg/L (i.e. 0.8 mg a.i./L) . Further, the 96 h LC10 and C90 were calculated at 1.6 and 1.9 mg/L (i.e. 1.3 and 1.5 mg a.i./L) , respectively (Sundfor, 2003).