Quaternary ammonium compounds, benzyl-C16-18-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 27 August, 2012 to 31 August, 2012

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

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Chemical analyses
Samples were taken for chemical analysis from the stock and all test concentrations at the start and end of the test as well as in the old and new test solutions after 48 hours.
Samples of 10 mL were taken and diluted 1:1 in leaching solution. Leaching solution used was 100 g/L of MgCl2.6H20 dissolved in a 50% methanol and 50%
2-propanol solution. Samples were shaken for a short period on a shaking table and further diluted (also in leaching solution) if required to fall within the calibration curve. Chemical analysis was conducted according to the procedure detailed in Annex 1 of the attached background material.

Vehicle:

no

Details on test solutions:

Preparation of the stock solution
0.0121 g of the test substance was weighed out using the analytical balance for preparation of the stock solution. 80 mL of the test medium was added and the solution was stirred at room temperature. The test substance dissolved easily and formed a clear and homogeneous stock solution. The stock solution was then made up to 100 mL the pH was 8.3. The stock solution was then used to generate the chosen test concentrations and waskept stirring when in use.

Preparation of the test solutions
An appropriate amount of the stock solution required to reach the desired concentration in a total volume of 3 L was pipetted into each 3 L aquarium. The aquarium was then brought up to exactly 3 L volume with a volumetric flask and homogenized with a Teflon coated magnetic stirrer. The following test concentrations were prepared:

0.05, 0.11, 0.24, 0.53 and 1.2 mg/L.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebra fish
- Strain: Danio rerio
- Source: “Dierenvriend” in Arnhem, The Netherlands
- Food type: The fish in stock were fed one to three times per day with commercially available dry, deep frozen food, or fresh Artemia salina nauplii /juvenile Daphnia magna neonates. Feeding was stopped 24 hours before the test was started.
- Length at study initiation: >3 cm
- Weight at study initiation (mean and range, SD): 0.3 g
- Feeding during test: No

ACCLIMATION
- A representative number of test fish batch (10 at random) were weighed prior to the test and measured after the test to assess compliance with guideline criteria.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

Dutch Standard Water: 12.3 ºdH (the guideline criteria requires the CaCO3 content to be between 10 and 250 mg/L (approximately 1-14 ºdH)

Test temperature:

Temperature controlled incubator: 22.4-22.8°C

pH:

8.2

Dissolved oxygen:

>8.4 mg/L

Salinity:

Dutch Standard Water: 200 mg of CaCl2.2H2O, 180 mg of MgSO4.7H2O, 100 mg of NaHCO3 and 20 mg of KHCO3.

Nominal and measured concentrations:

Nominal: 0, 0.05, 0.11, 0.24, 0.53 and 1.2 mg/L (based on 98.2% purity of the test substance).

Details on test conditions:

Test vessel:
3-L aquaria (capacity slightly exceeded 3 L) containing 3 L of test solution were used. They were covered with glass plates and aerated gently during the test.

Test room and light regime
The test was carried out in a temperature controlled room. The light regime was 16 h of ambient light per day, provided by fluorescent tubes.

The dissolved oxygen concentrations and conductivity were determined electrochemically using an oxygen electrode / conductivity electrode and meter. The pH was determined with a pH meter. The temperature was measured with a temperature sensor and recorder. Hardness, was measured using a Dr Lange test kit validated against an appropriate standard. TOC measurements were conducted with a Shimatzu TOC apparatus according to laboratory standard operating procedures


Test principle and procedure
The test was performed as a semi static test which means that the test media was replaced after 48 hours to ensure that a worst case scenario was tested. The minimum requirement of 7 fish per test concentration and control were used for ethical reasons.

Under otherwise identical test conditions, the fish were exposed to the chosen concentrations of the test substance and mortality and sub-lethal effects were recorded at least at 30 minutes and approximately 1, 2 and 4 hours after exposure and then and then at approximately 24, 48, 72 and 96 hours. Additional observations after the solution refreshment were made for animal welfare reasons. This data was not used for endpoint determinations.

The fish were considered dead when a lack of opercular movement was observed and touching of the caudal peduncle produced no reaction. Dead fish were removed from the test vessels directly after being observed, which was checked several times up to 4 h on the first day for ethical reasons and at least twice each day on subsequent days. In addition to death, sub-lethal effects such as erratic swimming, loss of reflex, increased excitability, lethargy, changes in physiology, discoloration, pigmentation, excessive mucous production, hyperventilation, opaque eyes, curved spine or hemorrhaging were recorded if observed. Fish that were convulsing or showing other severe forms of distress not considered transient in nature and likely to become more severe before the exposure is terminated, were sacrificed for humane reasons. These fish were treated as having died in the test.

Reference substance (positive control):

not specified

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

ca. 0.1 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

act. ingr.

Basis for effect:

other: No lethal effects observed

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

ca. 0.06 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

act. ingr.

Basis for effect:

other: No lethal effects observed

Details on results:

Preliminary results:
The preliminary test results indicated a nominal NOEC of 0.1 mg/L for this substance and gave therefore sufficient information on which to base the definitive test.

- Toxicity:

The results of the test on survival and sub-lethal effects are summarized in Table 1. All effects are based on the nominal test concentrations. Endpoints are based on 96 hours exposure. Toxcalc 5.0.23 outputs are reported as figure 1 for nominal values and as figure 2 for time weighted mean values.

The NOEC for the test substance was determined as 0.06 mg/L based on no significant lethal effects of being observed in this test concentration. (Time weighted mean corrected for both components).

The LC50 calculated using the Spearman Karber Graphical Method was calculated as 0.1 mg/L (Time weighted mean corrected for both components).

Prior to death affected animals displayed reduced activity and a hanging tail in comparison to the control.

The average length of a sample of 7 fish randomly taken from the test animals at the end of the test was 3.39 cm. The mean biomass loading was 0.7 g/L. The results of the length and mass determination are presented in Table 2. The general criterion set for the test animals in the study plan and guideline for biomass loading was met. The criterion for length was exceeded slightly.

- Chemical analyses:
A summary of the analytical results and quality criteria are presented in tables 5, 6, 7 and 8 in the tables and figures section. The analytical results show initial concentrations at T0 close to that of the nominal concentration. The analytical method met the required quality criteria for both the C16 and the C18 component. The concentration of the old solutions after 48 hours declined significantly to approximately 50 % of the nominal concentrations. Time weighted mean values were therefore calculated. See tables 5, 6 and 7.

As was expected the recovery of the C18 was less than that of the C16 in this study, most likely due to stronger adsorption of the C18 component to glass or test organisms. In order to obtain a quantification representative for both of the main test substance components, measured time weighted mean concentrations of each component were corrected for their composition as indicated in a footnote to table 7. These values were used for determination of the endpoints. Additionally the endpoints are also expressed as nominal concentrations.

Parallel samples without fish were also analyzed in the event that a large difference was observed between test solutions with and without test organisms. This was not the case and all calculations were based on in test measurements and the parallel replicate results were not used further.

- Oxygen, pH, Temperature, Conductivity and Hardness:
The oxygen concentrations varied during the test from 7.3 to 8.3 mg/L. The results of the oxygen measurements are summarized in Table 3.
The maximum pH variation during the test was from 7.6 to 8.3. The results of the pH measurements are summarized in Table 4.
The temperature in the test room during the test period ranged from 21.3 to 22.0°C. The conductivity at the start of the test in dilution water was 601 µs/cm .
The hardness of the test water was measured as 12.3°dH, which is equivalent of a maximum of 219 mg/L as calcium carbonate (Ref 4).

All measurements fell within study plan and guideline criteria.

- QUALITY CRITERIA
The following quality criteria have been met in the present study:
• Mortality in the control did not exceed one fish at the end of the test.
• The dissolved oxygen concentration was >60% of the air saturation value throughout the test.
• pH variation did not exceed 1.5 units.

The following quality criterion was not met in this study:
• Test substance recovery (for both components) was not >80% for the entire test duration. Time weighted measured mean calculations were therefore conducted for use in the endpoint determination. The time weighted mean values were then corrected for composition according to the analytical certificate.

- CONCLUSION
Endpoints were calculated or determined as far as possible using the guideline recommended procedures. An LC50 of 0.1mg/L was calculated and a NOEC of 0.06 mg/L were determined. Analytical measurement of the new solutions indicated that the test substance was accurately dosed to the test system but not stable throughout the test period despite a medium renewal to improve the stability of the exposure. As recommended in the guideline time weighted mean values (corrected for each component) were therefore used for endpoint determination. Amendments and Deviations were reported and were not of significant influence to the test outcome. The study can be considered an accurate representation of the effects of the test chemical to D.Rerio in guideline compliant synthetic water.

Reported statistics and error estimates:

The LC50 calculated using the Spearman Karber Graphical Method was calculated as 0.1 mg/L (Time weighted mean corrected for both components).

Sublethal observations / clinical signs:

TABLES AND FIGURES

Table1 – Survival and sub lethal effects

Date

27/8

27/8

27/8

27/8

27/8

28/8

28/8

28/8

29/8

29/8

29/8

30/8

31/8

31/8

Time

13:10

13:45

14:10

14:55

17:10

8:10

13:10

16:10

9:30

13:10

16:00

16:00

11:00

13:20

Hours Test

0

24

48

72 (Approx)

96

Control

7

7AN

7AN

7AN

7AN

7AN

7AN

7AN

7AN

7AN

7AN

7AN

7AN

7AN

0.05 mg/L

7

7AN

7AN

7AN

7AN

7AN

7AN

7AN

7AN

7AN

7AN

7AN

7AN

7AN

0.11 mg/L

7

7AN

7AN

7AN

7AN

7AN

7AN

7AN

7AN

7AN

7AN

7AN

7AN

7AN

0.24 mg/L

7

7AN

7AN

5RA 2AN

7RA

2RA

2RA

2RA

0

0.53 mg/L

7

7AN

7RA

7RA

2RA 3D 2#

0

1.2 mg/L

7

7A

4A#

0

AN=Appearing normal, #=Euthanized for Ethical reasons; D=Dead; RA=Reduced activity; A=Surfacing

Table 2: Biomass and weight measurements of fish

 

	Mass (g)	Length (cm)
	0.3	3.57
	0.34	3.56
	0.22	3.32
	0.29	3.29
	0.26	3.52
	0.33	3.08
	0.32	3.41
	0.26
	0.30
	0.34
Average	0.30	3.39

 

Table 3: Oxygen measurements (mg/L)

 

Dilution (mg/L)	0 hours	24 hours	48 Hours O     N		72 hours	96 hours
Control	8.4	8.3	7.6	8.3	N/M	8.1
0.05	8.4	7.8	7.8	8.3	N/M	7.3
0.11	8.4	7.8	7.9			8.3
0.24	8.4	8.3
0.53	8.4	8.6
1.2	8.3

N/M – Not measured

Table 4: pH-measurements

 

Dilution (mg/L)	0 hours	24 hours	48 Hours O      N		72 hours	96 hours
Control	8.3	7.9	7.9	8.0	N/M	7.7
0.05	8.3	7.9	7.9	8.0	N/M	7.6
0.11	8.3	8.0	7.9			7.7
0.24	8.3	8.0
0.53	8.3	8.1
1.2	8.3

 

Table 5: Test concentrations based on compound C16

sample	conc. 0h fresh (mg/L) (C16)	conc. 48h old(mg/L) (C16)	conc. 48h old par. (mg/L) (C16)	conc. 48h fresh (mg/L) (C16)	conc.96h old (mg/L) (C16)	conc. 96h old par. (mg/L) (C16)	TW mean (mg/L)
control	0.00	0.00	--	< LOD	0.00	0.01	--
0.05 mg/L	0.04	0.02	0.02	0.04	0.02	0.02	0.03
0.11 mg/L	0.09	0.05	--	0.09	0.05	0.07	0.07
0.24 mg/L	0.20	0.17	0.18	--	--	--	0.188
0.53 mg/L	0.46	--	--	--	--	--	--
1.2 mg/L	1.02	--	--	--	--	--	--
Stock	122.5	--	--	101.5	--	--	--

 

Conc: Concentration measured --: not measured no living test organisms TW= Time weighted mean

 

 

Table 6 : Test concentrations based on compound C18

sample	conc. 0h fresh (mg/L) (C18)	conc. 48h old (mg/L) (C18)	conc. 48h old par. (mg/L) (C18)	conc. 48h fresh (mg/L) (C18)	conc.96h old (mg/L) (C18)	conc96hold par. (mg/L) (C18)	TW mean (mg/L)
control	0.00	0.00	--	< LOD	0.00	0.00	--
0.05 mg/L	0.03	0.02	0.02	0.04	0.02	0.02	0.02
0.11 mg/L	0.08	0.04	--	0.08	0.04	0.04	0.06
0.24 mg/L	0.20	0.11	0.09	--	--	--	0.148
0.53 mg/L	0.45	--	--	--	--	--	--
1.2 mg/L	1.00	--	--	--	--	--	--
Stock	121.8	--	--	100.8	--	--	--

Conc: Concentration measured --: not measured no living test organisms TW= Time weighted mean

 

Table 7: Test concentrations corrected for % of each component

sample	conc. 0h (mg/L) (correcte d for A.I.)	conc. 48h old(mg/L) (correcte d for A.I.)	conc.48h old par. (mg/L)(correctedfor A.I.)	conc. 48h fresh (mg/L)(correctedfor A.I.)	conc. 96h old(mg/L) (correct ed for A.I.)	conc.96h old par. (mg/L)(correctedfor A.I.)	TW mean (mg/L) (corrected for A.I.)
control	0.00	0.00	--	< LOD	0.00	0.00	--
0.05 mg/L	0.03	0.02	0.02	0.04	0.02	0.02	0.02
0.11 mg/L	0.08	0.05	--	0.08	0.04	0.05	0.06
0.24 mg/L	0.19	0.13	0.12	--	--	--	0.16
0.53 mg/L	0.45	--	--	--	--	--	--
1.2 mg/L	0.99	--	--	--	--	--	--
Stock	119.8	--	--	99.2	--	--	--

Correction for composition

C16 =30.6 % A.I. was corrected to 31.7% A.I as C16 en C18 are considered representative for 98.2% of active ingredient. C18 =64.2 % A.I.was corrected to 66.5% A.I as C16 en C18 are considered representative 98.2% active ingredient.

Time weighted mean values in tables 5&6 were then corrected for composition as indicated and summed resulting in table 7 above.

A.I. = Active Ingredient.

Time weighted mean formula used

Area =Conc 0 – Conc 1  x Days      Total area / Total days =Timeweighted mean (OECD Guideline 211) Ln(Conc0) 0 Ln(conc1)

 

 

Table 8: Validity criteria of the analytical method

 

Parameter	Limit	Determined
Squared regression coefficient (R^2)	>= 0.98	>= 0.9998
Repeatability (CV (%)) at lowest standard	<= 20	<=3.8
Repeatability (CV (%)) at Highest standard	<= 20	<=0.5
LOQ (µg/L)	10	0.2
LOD (µg/L)	10	0.06
System stability (% of nominal)	<= 10	<= 1.9
Recovery from medium (%)	>= 60 and <= 120	96.0 - 101.2

Validity criteria fulfilled:

yes

Conclusions:

Under the study conditions, the 96 h LC50 was calculated to be 0.1 mg a.i./L (time weighted mean corrected for both components) and the NOEC was 0.06 mg a.i./L based on no significant lethal effects observed at this test concentration (time weighted mean corrected for both components).

Executive summary:

A study was conducted to determine the short-term toxicity of the test substance, C16 -18 ADBAC (98.2% active) to Danio rerio, under the semi-static conditions according to OECD Guideline 203, in compliance with GLP. The substance was tested at nominal concentrations of 0, 0.05, 0.11, 0.24, 0.53 and 1.2 mg/L. Chemical analysis was performed on the stock and all test concentrations at the start and at the end of the test as well as in the old and new test solutions after 48 h. Recoveries did not stay above 80% of the nominal concentrations. As recommended in the guideline, a time weighted mean of the measured concentrations in replicates where fish were surviving, corrected for composition, was therefore calculated used for endpoint determination. The NOEC for the test substance was determined as 0.06 mg a.i./L (time weighted mean corrected for both components) based on no significant lethal effects of being observed in this test concentration. The LC50 calculated using the Spearman Karber Graphical Method was calculated as 0.1 mg a.i./L (time weighted mean corrected for a.i.). Prior to death affected animals displayed reduced activity and a hanging tail in comparison to the control. All validity/quality criteria were fulfilled in the study, except for the >80% recovery for the test substance, which was addressed using time weighted mean values. Under the study conditions, the calculated 96 h LC50 and NOEC values were determined at 0.1 mg a.i./L and 0.06 mg a.i./L (time weighted mean) (Kean, 2012).

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

read-across based on grouping of substances (category approach)

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

Justification for type of information:

Refer to the Quaternary ammonium salts (QAS) category or section 13 of IUCLID for details on the category justification.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: PARCOM 1995 Part B

Deviations:

no

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes

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.:

ca. 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: Calculated

Remarks:

equivalent to 0.8 mg a.i./L

Key result

Duration:

96 h

Dose descriptor:

LC10

Effect conc.:

ca. 1.6 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: Calculated

Remarks:

equivalent to 1.3 mg a.i./L

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

ca. 1.7 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: Calculated

Remarks:

equivalent to 1.36 mg a.i./L

Key result

Duration:

96 h

Dose descriptor:

other: LC90

Effect conc.:

ca. 1.9 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: Calculated

Remarks:

equivalent to 1.5 mg a.i./L

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.

Sublethal observations / clinical signs:

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

Conclusions:

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.

Executive summary:

A study was conducted to determine the short-term toxicity of the read across substance, C12 -14 ADBAC (60-100% active) to fish, 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 considering 80% as mean active. Analytical dose verification was not performed. The test was performed under semi-static conditions, with the test media changed after 48 h. Mortalities are recorded at 24, 48, 72 and 96 h and the read across substance concentration that are lethal to 50% of the fish were determined where possible. Behaviour was also observed during the test period. The LC50 and 95% confidence intervals were calculated using a Logistic Regression Model. To ensure reproducibility, a single concentration 2.3 mg/L of reference substance 3,5-dichlorophenol was included in the study, which indicated 40% mortality after 96 h. All validity criteria were fulfilled. 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 LC90 were calculated at 1.6 and 1.9 mg/L (i.e. 1.3 and 1.5 mg a.i./L), respectively (Sundfor, 2003). Based on the results of the read across study, similar effects levels are expected for the test substance.

Description of key information

Based on the available information, the freshwater 96 h LC50 and NOEC values of the test substance, C16-18 ADBAC, for toxicity to freshwater fish were determined to be 0.1 mg a.i./L and 0.06 mg a.i./L (time weighted mean) respectively. Further, based on the results of the read across study with C12-14 ADBAC, the marine water 96 h LC50 value of the test substance for acute toxicity in fish is considered to be 1.36 mg a.i./L (nominal).  These respective LC50 values in fresh and marine water have been considered further for hazard/risk assessment.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

0.1 mg/L

Marine water fish

Marine water fish

Dose descriptor:

LC50

Effect concentration:

1.36 mg/L

Additional information

Freshwater

A study was conducted to determine the short-term toxicity of the test substance, C16 -18 ADBAC (98.2% active) to Danio rerio, under the semi-static conditions according to OECD Guideline 203, in compliance with GLP. The substance was tested at nominal concentrations of 0, 0.05, 0.11, 0.24, 0.53 and 1.2 mg/L. Chemical analysis was performed on the stock and all test concentrations at the start and at the end of the test as well as in the old and new test solutions after 48 h. Recoveries did not stay above 80% of the nominal concentrations. As recommended in the guideline, a time weighted mean of the measured concentrations in replicates where fish were surviving, corrected for composition, was therefore calculated used for endpoint determination. The NOEC for the test substance was determined as 0.06 mg a.i./L (time weighted mean corrected for both components) based on no significant lethal effects of being observed in this test concentration. The LC50 calculated using the Spearman Karber Graphical Method was calculated as 0.1 mg a.i./L (time weighted mean corrected for a.i.). Prior to death affected animals displayed reduced activity and a hanging tail in comparison to the control. All validity/quality criteria were fulfilled in the study, except for the >80% recovery for the test substance, which was addressed using time weighted mean values. Under the study conditions, the calculated 96 h LC50 and NOEC values were determined at 0.1 mg a.i./L and 0.06 mg a.i./L (time weighted mean) (Kean, 2012).

Marinewater

A study was conducted to determine the short-term toxicity of the read across substance, C12 -14 ADBAC (60-100% active) to fish, 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 considering 80% as mean active. Analytical dose verification was not performed. The test was performed under semi-static conditions, with the test media changed after 48 h. Mortalities are recorded at 24, 48, 72 and 96 h and the read across substance concentration that are lethal to 50% of the fish were determined where possible. Behaviour was also observed during the test period. The LC50 and 95% confidence intervals were calculated using a Logistic Regression Model. To ensure reproducibility, a single concentration 2.3 mg/L of reference substance 3,5-dichlorophenol was included in the study, which indicated 40% mortality after 96 h. All validity criteria were fulfilled. 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 LC90 were calculated at 1.6 and 1.9 mg/L (i.e. 1.3 and 1.5 mg a.i./L), respectively (Sundfor, 2003). Based on the results of the read across study, similar effects levels are expected for the test substance.

Based on the available information, the freshwater 96 h LC50 and NOEC values of the test substance, C16-18 ADBAC, for toxicity to freshwater fish were determined to be 0.1 mg a.i./L and 0.06 mg a.i./L (time weighted mean) respectively. Further, based on the results of the read across study with C12-14 ADBAC, the marine water 96 h LC50 value of the test substance for acute toxicity in fish is considered to be 1.36 mg a.i./L (nominal).