Reaction mass of sodium hydrogen N,N-dibutyl-10-(sulphonatooxy)octadecanamidate and sodium sulphooctadecanoate

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2012-09-17 to 2012-09-21

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

May 30, 2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

July 17, 1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: OECD Series on Testing and Assessment, No. 23, "Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures"

Version / remarks:

December 15, 2000

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 49, 22.3, 10.1, 4.6 and 2.1 mg test item/L and a control.
- Sampling method: For the determination of the stability of the test item under the test conditions, respectively the maintenance of the test item concentrations during the test period, samples were taken in duplicate out of all test media except the highest test concentration and the control at day 1 and at the end of the test (96 hours). Samples for the highest test concentration were taken after two hours since all fish were dead. Samples from the 22.3 and 10.1 mg test item /L were taken only after day 1 since all fish were dead. All test medium samples were taken from the approximate centre of the aquaria.
- Sample storage conditions before analysis: At room temperature (20 ± 5 °C), under dark and dry conditions

Vehicle:

no

Details on test solutions:

The test media (reconstituted water and test material) were freshly prepared just before introduction of the test fish (= start of the test) and at test medium renewal on day 1, 2 and 3.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebra fish
- Source: Aquaristik-Studio Fröhlich, 63165 Mühlheim, Germany
- Age at study initiation: Juveniles
- Length at study initiation: The mean body length of the fish in the test was 1.80 cm ± 0.07 cm (Mean ± SD)
- Weight at study initiation: the mean body wet weight 33.53 ± 6.88 mg (Mean ± SD)

ACCLIMATION
- Acclimation period: 7
- Acclimation conditions: same as test conditions
- Feeding frequency during acclimation: three times per week or daily
- Health during acclimation: no mortality was observed during acclimation

FEEDING DURING TEST
No feeding during test

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

250 mg CaCO3/L

Test temperature:

23 to 24 °C

pH:

8.1 to 8.2

Dissolved oxygen:

95 to 100 % of the air saturation value

Nominal and measured concentrations:

Nominal concentration: 49, 22.3, 10.1, 4.6 and 2.1 mg test item/L and a control
Measured concentration: average 96 % of the nominal in the two highest concentrations. All other concentrations are below LOD.

Details on test conditions:

TEST SYSTEM
- Test vessel: Aquaria
- Material, size, fill volume: glass, 12 L, 5 L
- Aeration: slightly aerated during test
- Renewal rate of test solution: renewal on days 1, 2 and 3.
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source of dilution water: Reconstituted Water (ISO Medium)

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light: 8 h dark; 30 min dawn/dusk period
- Light intensity: 480 to 880 lux

EFFECT PARAMETERS MEASURED:
Mortality and Sublethal Effects: The test fish were observed after approximately 2, 24, 48, 72 and 96 hours test duration for sublethal effects and mortality. Dead fish were removed at least once daily and discarded.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.19
- Range finding study: yes
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

4.72 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

10.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

2.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: See Table 1 below
- Observations on body length and weight: no
- Other biological observations: no
- Other adverse effects control: no
- Abnormal responses: no
- Any observations that might cause a difference between measured and nominal values: no

Validation criteria:
No fish died in the control until the end of the test. The dissolved oxygen concentration in the test media did not fall below 95 % of air saturation value during the test.

Sublethal observations / clinical signs:

Table 1: Mortality and Sublethal Effects

Mortality and Sublethal Effects
Nominal concentration [mg/L]
Exposure Time [h]		Control	2.1	4.6	10.1	23.3	49
0h	# mort	0	0	0	0	0	0
	# symp	0	0	0	0	0	0
	symp	-	-	-	-	-	-
2 h	# mort	0	0	0	0	2	7
	# symp	0	0	0	0	4	0
	symp	-	-	-	-	MB	-
24 h	# mort	0	0	0	7	7	7
	# symp	0	0	0	0	0	0
	symp	-	-	-	-	-	-
48 h	# mort	0	0	2	7	7	7
	# symp	0	0	0	0	0	0
	symp	-	-	-	-	-	-
72 h	# mort	0	0	2	7	7	7
	# symp	0	0	5	0	0	0
	symp	-	-	MB	-	-	-
96 h	# mort	0	0	3	7	7	7
	# symp	0	0	4	0	0	0
	symp	-	-	MB, TS	-	-	-

# mort: Number of dead fish

# symp: Number of fish with sublethal effects

MB: fish mainly on the bottom

TS: tumbling during swimming

Validity criteria fulfilled:

yes

Conclusions:

Based on the test results, the 96-hour LC50 of the test item for Zebrafish (Danio rerio) was determined to be 4.72 mg test item/L based on nominal test concentrations. The LC0 was determined to be 2.1 mg test item/L and the LC100 was determined to be 10.1 mg test item/L, both values also based on nominal test concentrations.

Executive summary:

The acute toxicity of the test item to Zebrafish (Danio rerio) in a 96-hour semi-static test was determined according to OECD 203. Juvenile Zebrafish (Danio rerio) were exposed in a semi-static test to aqueous test media containing the test item at various concentrations under defined conditions. The recorded effects were mortality and sublethal effects on the fish. This study encompassed 6 treatment groups (5 dose rates of the test item, control) each containing 7 individuals. The acute toxicity to unfed juvenile Zebrafish (Danio rerio) was determined in an aerated, semi-static, 96-hour test. The test fish were observed after approximately 2, 24, 48, 72 and 96 hours of test duration for sublethal effects and mortality. Dead fish were removed at least once daily and discarded.

In the control and the nominal test concentration of 2.1 mg test item/L, all fish survived until the end of the experiment and no signs of intoxication occurred. At the nominal test concentration of 4.6 mg test item/L 2 fish were dead after 48 hours of test duration and 5 fish were observed mainly at the bottom of the aquarium after 72 hours in addition. At test end after 96 hours of exposure 3 fish were dead at nominal 4.6 mg test item/L and the remaining 4 fish were observed mainly at the bottom of the aquarium showing tumbling during swimming. At the nominal test concentration of 10.1 mg test item/L all fish were dead after 24 hours of test duration. At the nominal test concentration of 22.3 mg test item/L 4 fish were observed mainly at the bottom of the aquarium and 2 fish were dead after 2 hours of test duration. After 24 hours all fish were dead at nominal 22.3 mg test item/L. At the highest test concentration of nominal 49 mg test item/L all fish were dead after 2 hours of test duration.

The quantification of the test item was performed using the determination of the Total Organic Carbon (TOC-method). The purpose of the analytical part of this study was to verify the concentrations of the test item in the test medium. At the start of the test just before introduction of the fish 94 % of the nominal test concentrations were found (average of the two highest test concentrations; all other values were below the Limit of Quantification). In the aged test media 98 % of the nominal values were determined (average of the two highest test concentrations; all other values were below the Limit of Quantification). Thus, during the test period of 96 hours the fish were exposed to a mean of 96 % of nominal. Therefore, all reported results are related to nominal concentrations of the test item.

Based on the test results, the 96-hour LC₅₀ of the test item for Zebrafish (Danio rerio) was determined to be 4.72 mg test item/L based on nominal test concentrations. The LC₀ was determined to be 2.1 mg test item/L and the LC₁₀₀ was determined to be 10.1 mg test item/L, both values also based on nominal test concentrations.

Description of key information

Based on the test results, the 96-hour LC₅₀of the test item for Zebrafish (Danio rerio) was determined to be 4.72 mg test item/L based on nominal test concentrations. The LC₀was determined to be 2.1 mg test item/L and the LC₁₀₀was determined to be 10.1 mg test item/L, both values also based on nominal test concentrations

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

4.72 mg/L

Additional information

The acute toxicity of the test item to Zebrafish (Danio rerio) in a 96-hour semi-static test was determined according to OECD 203. Juvenile Zebrafish (Danio rerio) were exposed in a semi-static test to aqueous test media containing the test item at various concentrations under defined conditions. The recorded effects were mortality and sublethal effects on the fish. This study encompassed 6 treatment groups (5 dose rates of the test item, control) each containing 7 individuals. The acute toxicity to unfed juvenile Zebrafish (Danio rerio) was determined in an aerated, semi-static, 96-hour test. The test fish were observed after approximately 2, 24, 48, 72 and 96 hours of test duration for sublethal effects and mortality. Dead fish were removed at least once daily and discarded.

In the control and the nominal test concentration of 2.1 mg test item/L, all fish survived until the end of the experiment and no signs of intoxication occurred. At the nominal test concentration of 4.6 mg test item/L 2 fish were dead after 48 hours of test duration and 5 fish were observed mainly at the bottom of the aquarium after 72 hours in addition. At test end after 96 hours of exposure 3 fish were dead at nominal 4.6 mg test item/L and the remaining 4 fish were observed mainly at the bottom of the aquarium showing tumbling during swimming. At the nominal test concentration of 10.1 mg test item/L all fish were dead after 24 hours of test duration. At the nominal test concentration of 22.3 mg test item/L 4 fish were observed mainly at the bottom of the aquarium and 2 fish were dead after 2 hours of test duration. After 24 hours all fish were dead at nominal 22.3 mg test item/L. At the highest test concentration of nominal 49 mg test item/L all fish were dead after 2 hours of test duration.

The quantification of the test item was performed using the determination of the Total Organic Carbon (TOC-method). The purpose of the analytical part of this study was to verify the concentrations of the test item in the test medium. At the start of the test just before introduction of the fish 94 % of the nominal test concentrations were found (average of the two highest test concentrations; all other values were below the Limit of Quantification). In the aged test media 98 % of the nominal values were determined (average of the two highest test concentrations; all other values were below the Limit of Quantification). Thus, during the test period of 96 hours the fish were exposed to a mean of 96 % of nominal. Therefore, all reported results are related to nominal concentrations of the test item.

Based on the test results, the 96-hour LC₅₀ of the test item for Zebrafish (Danio rerio) was determined to be 4.72 mg test item/L based on nominal test concentrations. The LC₀ was determined to be 2.1 mg test item/L and the LC₁₀₀ was determined to be 10.1 mg test item/L, both values also based on nominal test concentrations.

The reported effect concentrations refer to the commercial formulation containing 46.2% (w/w) of the REACH registration substance which reflects the test item used in the study.