Quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides

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:

key study

Study period:

From March 21, 1990 to April 14, 1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

No chemical analyses of the test concentrations were performed during the test and all concentrations mentioned are nominal concentrations of the test substance.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

not specified

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

not specified

GLP compliance:

yes

Analytical monitoring:

no

Details on sampling:

- Concentrations: 0.18, 0.23,0.3, 0.4 and 0.51 mg/L

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
The test substance formed a stable, homogeneous dispersion in water. The stock solution used contained 1 g of the test substance/L of deionized water. The test concentrations were prepared by addition of the respective amounts of stock dispersion to the dilution water (DSW), a synthetic water having a pH of approx 8.2 and a hardness of 13 dH.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Source: Local aquarium retailer (batch 15-03-90)
- Age at study initiation (mean and range, SD):
- Length at study initiation (length definition, mean, range and SD): 2.9 cm
- Weight at study initiation (mean and range, SD): 0.3 g( wet weight)
- Feeding during test: No

ACCLIMATION
- Acclimation period: atleast 12 d
- Acclimation conditions (same as test or not): Yes
- Feeding frequency: Fed one to three times each day for 6 d a wk

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

No post observation period.

Hardness:

13⁰dH

Test temperature:

21.7 to 22.3 °C

pH:

7.4 - 8.1

Dissolved oxygen:

Fresh solutions: 8.8 mgO2/L, decreasing to 6.0 mgO2/L at 96h.

Salinity:

DSW: 100 mg of NaHCO3/L
20 mg of KHCO3/L
200 mg of CaCl2.2H2O/L
180 mg of MgSO4.7H2O/L
The deionized water which was used to prepare the DSW contained less than 10 μg of copper/L and a conductivity of less than 5 μS/cm. The TOC content was less than 2 mg/L.

Nominal and measured concentrations:

0 (control), 0.18, 0.23, 0.3, 0.4 and 0.51 mg/L Nominal

Details on test conditions:

TEST SYSTEM
- Test vessel: 5 L glass aquaria
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: 5 L glass aquaria were filled with 3 L of test solution covered with glass plate
- Aeration: During the test the test media were not aerated
- Biomass loading rate: 0.7 g/L

OTHER TEST CONDITIONS
- Adjustment of pH: Yes
- Photoperiod: 12 hours of ambient light per day, provided by fluorescent tubes

Reference substance (positive control):

yes

Remarks:

(Potassium dichromate)

Key result

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

ca. 0.32 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CI: 0.29-0.35 mg/L

Remarks:

equivalent to 0.24 mg a.i./L

Key result

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

ca. 0.3 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CI: 0.27-0.32 mg/L

Remarks:

equivalent to 0.23 mg a.i./L

Key result

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

ca. 0.26 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CI: 0.24-0.29 mg/L

Remarks:

equivalent to 0.20 mg a.i./L

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

ca. 0.26 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CI: 0.24-0.29 mg/L

Remarks:

equivalent to 0.20 mg a.i./L

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

ca. 0.23 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks on result:

other: equivalent to 0.18 mg a.i./L

Details on results:

- The highest concentration causing no mortality after 96 h was 0.23 mg/L.
- The lowest concentration causing 100 % mortality already after 72 h was 0.3 mg/L
- At 0.4 and 0.51 mg/L 100 % mortality was observed after 22 h.
- No other effects, such as deviations in the behaviour or appearance of the fish, were observed, except for the fish at the concentrations of 0.3 mg/L after 22 h and at 0.4 and 0.51 mg/L after 6 and 4 h, respectively, which showed a reduced activity.
The results observed in the definitive study are in good agreement with the results observed in the two preliminary tests.

Results with reference substance (positive control):

The LC50 values for the reference substance are: 215 mg/L at 23 h, 183 mg/L after 48 h, 183 mg/L after 72 h and 183 mg/L after 96 h. The results obtained are in accordance with previous test results generated in an EEC report.

Reported statistics and error estimates:

The LC50 was determined using a statistical program of Griffioen (RIZA) based on a model of Kooyman (1981)

Sublethal observations / clinical signs:

Table 1. Mortality in the definitive test

mg/L of test substance	Number of surviving animals
	0h	4h	6h	22h	24h	30h	45h	48h	52h	69h	72h	78h	93h	96h
0 (control)	7	7	7	7	7	7	7	7	7	7	7	7	7	7
0.18	7	7	7	7	7	7	7	7	7	7	7	7	7	7
0.23	7	7	7	7	7	7	7	7	7	7	7	7	7	7
0.3	7	7	7	6*	5	5	3	3	3	1	0
0.4	7	7	7*	0
0.51	7	7*	5	0

*reduced activity

Validity criteria fulfilled:

yes

Conclusions:

Under the study conditions, the 96 h LC50 and NOEC value of the test substance were 0.26 mg/L and 0.23 mg/L (equivalent to 0.20 and 0.18 mg a.i./L), respectively.

Executive summary:

A study was conducted to determine the acute toxic effects of test substance, DAQ C12-18 (76.4% active) to Brachydanio rerio (zebra fish), according to the OECD Guideline 203 and EEC method C.1. Based on findings from preliminary test, seven fish per vessel were exposed to nominal concentrations of the test substance at .18, 0.23, 0.3, 0.4 and 0.51 mg/L for 96 h period under static conditions. No chemical analyses of the test concentrations were performed during the test and all concentrations mentioned are nominal concentrations of the test substance. The concentration causing 100% mortality was observed within 72 h at 0.3 mg/L. Sub lethal effects were observed in all groups which also showed mortality. The highest concentration causing no mortality after 96 h was 0.23 mg/L. The lowest concentration causing 100 % mortality already after 72 h was 0.3 mg/L At 0.4 and 0.51 mg/L 100 % mortality was observed after 22 h. No other effects, such as deviations in the behaviour or appearance of the fish, were observed, except for the fish at the concentrations of 0.3 mg/L after 22 h and at 0.4 and 0.51 mg/L after 6 and 4 h, respectively, which showed a reduced activity. Under the study conditions, the 96 h LC50 and NOEC value of the test substance were 0.26 mg/L (95 % CL.: 0.24 - 0.29 mg/L) and 0.23 mg/L (equivalent to 0.20 and 0.18 mg a.i./L), respectively (Mark & Meuwsen, 1990).

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From February 27, 1998 to March 6, 1998

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

No chemical analyses of the test concentrations were performed during the test and all concentrations mentioned are nominal concentrations of the test substance.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

Details not provided

Qualifier:

according to guideline

Guideline:

other: PARCOM Protocols on methods for the testing of chemicals used in the off-shore industry, part B: protocol for a fish acute toxicity test

Deviations:

yes

Remarks:

Details not provided

GLP compliance:

yes

Analytical monitoring:

no

Details on sampling:

No sampling performed

Vehicle:

no

Details on test solutions:

The test material was dispersible in water. A stock solution containing approximately 1 g/L of test substance was prepared as follows: to an accurately measured amount of approximately 1 g of test substance 80 mL of deionized water was added. The test substance dispersed well at room temperature when stirring. Thereafter, deionized water was added up to a final volume of 1000 mL. A homogeneous, clear, foaming stock solution was obtained. This was used for the preparation of the test solutions at the start of the test. For the refreshening of the test solutions on Day 2, a fresh stock solution of approximately 1 g/L was prepared as described above.

The test solutions were prepared by addition of the required amounts of stock solution to the dilution water. The following concentrations were tested: 0.21, 0.47, 1.03, 2.27 and 5.00 mg/L. A control containing only dilution water was included in the test.

The test was performed as a semi-static test with the test media replaced after 48 h.

Test organisms (species):

Cyprinodon variegatus

Details on test organisms:

TEST ORGANISM
- Common name: Sheepshead minnow
- Source: Bio international, Roemond, The Netherlands
- Length at study initiation (length definition, mean, range and SD): 2.4 cm
- Weight at study initiation (mean and range, SD): 0.29 g (wet weight of 5 fish).
- Feeding during test: No, 24 h or longer before the test

ACCLIMATION
- Type and amount of food: Dried or deep-frozen fish food
- Feeding frequency: Once a day
- Health during acclimation (any mortality observed):

Test type:

semi-static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

None

Hardness:

No data

Test temperature:

21.5 to 22.5 ⁰C

pH:

7.8- 8.0

Dissolved oxygen:

6.5 - 7.5 mgO2/L

Salinity:

31.2 to 33.4 g/L

Nominal and measured concentrations:

Nominal: 0.21, 0.47, 1.03, 2.27, 5.00 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 5L aquariums
- Type (delete if not applicable): Closed
- Material, size, headspace, fill volume: 5 L in size and contained 3 L of test solution covered with glass plates
- Renewal rate of test solution (frequency/flow rate): once in 48 h
- No of fishes/dose: 7



OTHER TEST CONDITIONS
- Photoperiod: 16 h of ambient light/d provided by fluorescent tubes

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Justification for using less concentrations than requested by guideline:
- Range finding study: No
- Test concentrations: 0.21, 0.47, 1.03, 2.27, 5.00 mg/L

Reference substance (positive control):

no

Key result

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

ca. 0.696 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 98 % CL: 0.470-1.030 mg/L

Remarks:

equivalent to 0.522 mg a.i./L

Key result

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

ca. 0.696 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 98 % CL: 0.470-1.030 mg/L

Remarks:

equivalent to 0.522 mg a.i./L

Key result

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

ca. 0.696 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 98 % CL: 0.470-1.030 mg/L

Remarks:

equivalent to 0.522 mg a.i./L

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

ca. 0.696 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 98 % CL: 0.470-1.030 mg/L

Remarks:

equivalent to 0.522 mg a.i./L

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

ca. 0.47 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks on result:

other: equivalent to 0.35 mg a.i./L

Details on results:

- Dose groups of 1.03, 2.27 and 5.00 mg/L caused 100 % mortality within 19 h.
- The NOEC was 0.47 mg/L.
- 96h LC50 was 0.696 mg/L.
- Reduced activity was only observed at the concentrations at which mortality occurred. Other sublethal effects, such as deviations in the behaviour or the appearance of the fish, were not observed.

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

The LC50 was calculated with a computer program (Akzo program SKBT, version 1.0) according to SOP L2, using both the trimmed Spearman-Karber method and the binomial test method.

Sublethal observations / clinical signs:

Results

Table 1. Number of surviving animals

Conc (mg/L)	0h	1h	2h	19h	24h	26h	44h	48h	51h	68h	72h	74h	91h	96h
Control	7	7	7	7	7	7	7	7	7	7	7	7	7	7
0.21	7	7	7	7	7	7	7	7	7	7	7	7	7	7
0.47	7	7	7	7	7	7	7	7	7	7	7	7	7	7
1.03	7	7	7	0	-	-	-	-	-	-	-	-	-	-
2.27	7	7	7	0	-	-	-	-	-	-	-	-	-	-
5.00	7	5	0	-	-	-	-	-	-	-	-	-	-	-

Validity criteria fulfilled:

yes

Conclusions:

Under the study conditions, the 96 h LC50 of the test substance was determined to be 0.696 mg/L (equivalent to 0.522 mg a.i./L).

Executive summary:

A study was conducted to determine the acute toxicity of test substance, DAQ C12-18 (75% active) with the salt water fish, Cyprinodon variegatus (sheepshead minnow), according to the OECD guideline 203 and the Parcom protocol on methods of the testing of chemicals used in the offshore oil industry, in compliance with GLP. Sen fish per vessel were exposed to the test substance at the concentrations of 0.21, 0.47, 1.03, 2.27 and 5.00 mg/L for 96 h under semi-static conditions. No chemical analyses of the test concentrations were performed during the test and all concentrations mentioned are nominal concentrations of the test substance. Reduced activity was only observed at the concentrations at which mortality occurred. Other sub lethal effects such as deviations in the behaviour and appearance of the fish were not observed. The 96 h NOEC and 19 h LC100 value of the test substance was determined to be 0.47 mg/L and 1.03 mg/L respectively. The 96 h LC50 of the test substance was determined to be 0.696 mg/L (98% CL: 0.470 and 1.030 mg/L). Under the study conditions, the 96 h LC50 of the test substance was determined to be 0.696 mg/L (equivalent to 0.522 mg a.i./L) (Mark, 1998a).

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From February 27, 1998 to March 6, 1998

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

No chemical analyses of the test concentrations were performed during the test and all concentrations mentioned are nominal concentrations of the test substance.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

Details not provided

Qualifier:

according to guideline

Guideline:

other: PARCOM Protocols on methods for the testing of chemicals used in the off-shore industry, part B: protocol for a fish acute toxicity test

Deviations:

yes

Remarks:

Details not provided

GLP compliance:

yes

Analytical monitoring:

no

Details on sampling:

No sampling performed

Vehicle:

no

Details on test solutions:

The test material was dispersible in water. A stock solution containing approximately 1 g/L of test substance was prepared as follows: to an accurately measured amount of approximately 1 g of test substance 80 mL of deionized water was added. The test substance dispersed well at room temperature when stirring. Thereafter, deionized water was added up to a final volume of 1000 mL. A homogeneous, clear, foaming stock solution was obtained. This was used for the preparation of the test solutions at the start of the test. For the refreshening of the test solutions on Day 2, a fresh stock solution of approximately 1 g/L was prepared as described above.

The test solutions were prepared by addition of the required amounts of stock solution to the dilution water. The following concentrations were tested: 0.05, 0.11, 0.24, 0.53 and 1.17 mg/L. A control containing only dilution water was included in the test.

Test organisms (species):

Cyprinodon variegatus

Details on test organisms:

TEST ORGANISM
- Common name: Sheepshead minnow
- Source: Bio international, Roemond, The Netherlands
- Length at study initiation (length definition, mean, range and SD): 2.4 cm
- Weight at study initiation (mean and range, SD): 0.29 g (wet weight of 5 fish).
- Feeding during test: No, 24 h or longer before the test

ACCLIMATION
- Type and amount of food: Dried or deep-frozen fish food
- Feeding frequency: Once a day
- Health during acclimation (any mortality observed):

Test type:

semi-static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

None

Hardness:

No data

Test temperature:

21.0 to 21.5 ⁰C

pH:

7.8- 8.1

Dissolved oxygen:

6.6 - 7.5 mgO2/L

Salinity:

31.2 to 33.4 g/L

Nominal and measured concentrations:

Nominal: 0.05, 0.11, 0.24, 0.53 and 1.17 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 5L aquariums
- Type (delete if not applicable): Closed
- Material, size, headspace, fill volume: 5 L in size and contained 3 L of test solution covered with glass plates
- Renewal rate of test solution (frequency/flow rate): once in 48 h
- No of fishes/dose: 7



OTHER TEST CONDITIONS
- Photoperiod: 16 h of ambient light/d provided by fluorescent tubes


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Justification for using less concentrations than requested by guideline:
- Range finding study: No
- Test concentrations: 0.05, 0.11, 0.24, 0.53 and 1.17 mg/L

Reference substance (positive control):

no

Key result

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

ca. 0.787 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 98 % CL: 0.530- 1.170 mg/L

Remarks:

equivalent to 0.47 mg a.i./L

Key result

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

ca. 0.787 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 98 % CL: 0.530- 1.170 mg/L

Remarks:

equivalent to 0.47 mg a.i./L

Key result

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

ca. 0.787 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 98 % CL: 0.530- 1.170 mg/L

Remarks:

equivalent to 0.47 mg a.i./L

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

ca. 0.787 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 98 % CL: 0.530- 1.170 mg/L

Remarks:

equivalent to 0.47 mg a.i./L

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

ca. 0.53 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks on result:

other: equivalent to 0.40 mg a.i./L

Details on results:

- Dose groups of 1.17 mg/L caused 100 % mortality within 19 h.
- The NOEC was 0.53 mg/L.
- 96h LC50 was 0.787 mg/L.
- Reduced activity was only observed at the concentrations at which mortality occurred. Other sublethal effects such as deviations in the behaviour or the appearance of the fish were not observed.

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

The LC50 was calculated with a computer program (Akzo program SKBT, version 1.0) according to SOP L2, using both the trimmed Spearman-Karber method and the binomial test method.

Sublethal observations / clinical signs:

Results

Table 1. Survival of fish

Nominal Concentration (mg/L)	Hours of test duration
	0	1	2	19	24	26	44	48	51	68	72	74	91	96
	Number of living animals
Control	7	7	7	7	7	7	7	7	7	7	7	7	7	7
0.21	7	7	7	7	7	7	7	7	7	7	7	7	7	7
0.47	7	7	7	7	7	7	7	7	7	7	7	7	7	7
1.03	7	7^c	7^c	0
2.27	7	7^b	7^b	0
5.00	7	5^a	0

^aalmost dead;^bvery reduced activity;^cone fish almost dead

Table 2 Statistical evaluation

Hour of test duration	LC50 values (mg/L)	98% confidence intervals (mg/L)
24	0.787	0.530	1.170
48	0.787	0.530	1.170
72	0.787	0.530	1.170
96	0.787	0.530	1.170

Validity criteria fulfilled:

yes

Conclusions:

Under the study conditions, the 96 h LC50 of the test substance was determined to be 0.787 mg/L (equivalent to 0.47 mg a.i./L).

Executive summary:

A study was conducted to determine the acute toxicity of test substance, DAQ C12-18 (60% active) with the salt water fish, Cyprinodon variegates (sheepshead minnow), according to the OECD guideline 203 and the Parcom protocol on methods of the testing of chemicals used in the offshore oil industry, in compliance with GLP. Seven fish per vessel were exposed to the test substance at the concentrations of 0.05, 0.11, 0.24, 0.53 and 1.17 mg/L for 96 h under semi-static conditions. No chemical analyses of the test concentrations were performed during the test and all concentrations mentioned are nominal concentrations of the test substance. Reduced activity was only observed at the concentrations at which mortality occurred. Other sub lethal effects such as deviations in the behaviour and appearance of the fish were not observed. The 96 h NOEC and 19 h LC100 of the test substance was determined to be 0.53 mg/L and 1.17 mg/L respectively. The 96 h LC50 of the test substance was determined to be 0.787 mg/L (98% CL: 0.470 and 1.170 mg/L). Under the study conditions, the 96 h LC50 of the test substance was determined to be 0.787 mg/L (equivalent to 0.47 mg a.i./L) (Mark, 1998b).

Reference 4

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

No chemical analyses of the test concentrations were performed during the test and all concentrations mentioned are nominal concentrations of the test substance.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

not specified

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Details on sampling:

Sampling not performed

Vehicle:

no

Details on test solutions:

Stock solution: 20 mg/L
0, 0.05, 0.1, 0.25, 0.5 and 1 mg/L. Dilutions were prepared with tapwater.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Source: Zoo Stumpe Hildesheim
- Length at study initiation (length definition, mean, range and SD): 3.2 cm
- Weight at study initiation (mean and range, SD): 0.42 g

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

No post exposure observation period.

Hardness:

7.6-7.8 mg/L

Test temperature:

23 ±2⁰C

pH:

7.9 - 8.2

Dissolved oxygen:

7.6 - 8.9

Salinity:

-

Nominal and measured concentrations:

Nominal: 0, 0.05, 0.1, 0.25, 0.5 and 1 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 16 L aquarium
- Material, size, headspace, fill volume: 16 L aquarium
- No. of organisms per vessel: 10 fish/tank
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Tap water

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

ca. 0.66 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CI: 0.57 - 0.77

Remarks:

equivalent to mg 0.50 mg a.i./L

Key result

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

ca. 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: equivalent to mg 0.75 mg a.i./L

Key result

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

ca. 0.25 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: equivalent to mg 0.19 mg a.i./L

Details on results:

- The % mortality was determined at 24, 48 , 72 and 96 h. The data shows that the mortality rate was the same from 24 to 96 h.
- Death caused by the test substance must occur in the first 24 h.
-The LC50 at 24 h was 0.63 mg/L. At 48, 72 and 96 h the LC50 was 0.66 mg/L

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

Not reported

Sublethal observations / clinical signs:

Results

Table 1: Mortality rates

Concentration mg/l	% Mortality
	24 h	48 h	72 h	96 h
1	100	100	100	100
0.50	10	20	20	20
0.25	0	0	0	0
0.10	10	10	10	10
0.05	0	0	0	0
control	0	0	0	0

Table 2: Estimated LC50 values.

Time (h)	LC50 (mg/L)	P 95%
24	0.63	0.25-1.00
48	0.66	0.57-0.77
72	0.66	0.57-0.77
96	0.66	0.57-0.77

Validity criteria fulfilled:

yes

Conclusions:

Under the study conditions, the 96 h LC50 of the test substance was determined to be 0.66 mg/L (equivalent to 0.50 mg a.i./L).

Executive summary:

A study was conducted to determine the acute toxic effects of test substance, DAQ C12-18 (75% active) to the fresh water fish, Brachydanio rerio (new name: Danio rerio), according to the OECD guideline 203, in compliance with GLP. Ten fish per vessel were exposed to nominal concentrations of the test substance at 0, 0.05, 0.1, 0.25, 0.5 and 1 mg/L for 96 h period under static conditions. The % mortality was determined at 24, 48 , 72 and 96 h. Based on the results, the mortality rate was found to be same from 24 to 96 h. The LC50 at 24 h was 0.63 mg/L. At 48, 72 and 96 h the LC50 was 0.66 mg/L. The lowest concentration which caused 100 % mortality was 1.00 mg/L. The lowest concentration without mortalities was 0.25 mg/L. Under the study conditions, the 96 h LC50 of the test substance was determined to be 0.66 mg/L (equivalent to 0.50 mg a.i./L) (Noack, 1990).

Description of key information

Based on the study results, the 96 h LC50 in freshwater fish were determined to be 0.20 to 0.50 mg a.i./L (nominal) and the 96 h LC50 in marine water fish were determined to be 0.47 to 0.522 mg a.i./L (nominal).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

0.2 mg/L

Marine water fish

Marine water fish

Dose descriptor:

LC50

Effect concentration:

0.47 mg/L

Additional information

Fresh water:

Study 1:A study was conducted to determine the acute toxic effects of the test substance, C12-18 DAQ (76.4% active) toBrachydanio rerio(zebra fish), according to OECD guideline 203 and EEC method C.1. Based on findings from preliminary test, seven fish per vessel were exposed to nominal concentrations of the test substance at .18, 0.23, 0.3, 0.4 and 0.51 mg/L for 96 h period under static conditions. No chemical analyses of the test concentrations were performed during the test and all concentrations mentioned are nominal concentrations of the test substance. The concentration causing 100% mortality was observed within 72 h at 0.3 mg/L. Sub lethal effects were observed in all groups which also showed mortality. The highest concentration causing no mortality after 96 h was 0.23 mg/L. The lowest concentration causing 100% mortality already after 72 h was 0.3 mg/L At 0.4 and 0.51 mg/L 100% mortality was observed after 22 h. No other effects, such as deviations in the behaviour or appearance of the fish, were observed, except for the fish at the concentrations of 0.3 mg/L after 22 h and at 0.4 and 0.51 mg/L after 6 and 4 h, respectively, which showed a reduced activity. Under the study conditions, the 96 h LC50 and NOEC value of the test substance were 0.26 mg/L (95% CL.: 0.24 - 0.29 mg/L) and 0.23 mg/L (equivalent to 0.20 and 0.18 mg a.i./L), respectively (Mark & Meuwsen, 1990).  

Study 2:A study was conducted to determine the acute toxic effects of the test substance, C12-18 DAQ (75% active) to the fresh water fish,Brachydanio rerio(new name:Danio rerio), according to OECD guideline 203, in compliance with GLP. Ten fish per vessel were exposed to nominal concentrations of the test substance at 0, 0.05, 0.1, 0.25, 0.5 and 1 mg/L for 96 h period under static conditions. The% mortality was determined at 24, 48, 72 and 96 h. Based on the results, the mortality rate was found to be same from 24 to 96 h. The LC50 at 24 h was 0.63 mg/L. At 48, 72 and 96 h the LC50 was 0.66 mg/L. The lowest concentration which caused 100% mortality was 1.00 mg/L. The lowest concentration without mortalities was 0.25 mg/L. Under the study conditions, the 96 h LC50 of the test substance was determined to be 0.66 mg/L (equivalent to 0.50 mg a.i./L) (Noack, 1990).  

Based on the available study results, the lower 96 h LC50 value of 0.2 mg/L (nominal) for freshwater fish has been considered further for hazard/risk assessment, as a conservative approach.  

Salt water: 

Study 1:A study was conducted to determine the acute toxicity of the test substance, C12-18 DAQ (75% active) with the salt water fish, Cyprinodon variegatus (sheepshead minnow), according to OECD Guideline 203 and the Parcom protocol on methods of the testing of chemicals used in the offshore oil industry, in compliance with GLP. Sen fish per vessel were exposed to the test substance at the concentrations of 0.21, 0.47, 1.03, 2.27 and 5.00 mg/L for 96 h under semi-static conditions. No chemical analyses of the test concentrations were performed during the test and all concentrations mentioned are nominal concentrations of the test substance. Reduced activity was only observed at the concentrations at which mortality occurred. Other sub lethal effects such as deviations in the behaviour and appearance of the fish were not observed. The 96 h NOEC and 19 h LC100 value of the test substance was determined to be 0.47 mg/L and 1.03 mg/L respectively. The 96 h LC50 of the test substance was determined to be 0.696 mg/L (98% CL: 0.470 and 1.030 mg/L). Under the study conditions, the 96 h LC50 of the test substance was determined to be 0.696 mg/L (equivalent to 0.522 mg a.i./L) (Mark, 1998a).  

Study 2:A study was conducted to determine the acute toxicity of the test substance, C12-18 DAQ (60% active) with the salt water fish,Cyprinodon variegates (sheepshead minnow), according to OECD Guideline 203 and the Parcom protocol on methods of the testing of chemicals used in the offshore oil industry, in compliance with GLP. Seven fish per vessel were exposed to the test substance at the concentrations of 0.05, 0.11, 0.24, 0.53 and 1.17 mg/L for 96 h under semi-static conditions. No chemical analyses of the test concentrations were performed during the test and all concentrations mentioned are nominal concentrations of the test substance. Reduced activity was only observed at the concentrations at which mortality occurred. Other sub lethal effects such as deviations in the behaviour and appearance of the fish were not observed. The 96 h NOEC and 19 h LC100 of the test substance was determined to be 0.53 mg/L and 1.17 mg/L respectively. The 96 h LC50 of the test substance was determined to be 0.787 mg/L (98% CL: 0.470 and 1.170 mg/L). Under the study conditions, the 96 h LC50 of the test substance was determined to be 0.787 mg/L (equivalent to 0.47 mg a.i./L) (Mark, 1998b).  

Based on the available study results, the lower 96 h LC50 value of 0.47 mg/L (nominal) for marine water fish has been considered further for hazard/risk assessment, as a conservative approach. ​