Butanedioic acid, sulfo-, 1-C12-18-alkyl esters, disodium salts

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Guideline compliant GPL study with good documentation.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Control, 0.15, 0.47, 1.5, 4.7 and 15 mg/L
- Sampling method: For the determination of the actual test item concentrations in this semi-static test, duplicate samples were taken from each treatment just before the start of the first and of the last renewal periods (Days 0 and 3). At the end of these renewal periods (Days 1 and 4), additional samples were taken for determination of the stability of the test item during the renewal periods of 24 hours.

- Sample storage conditions before analysis: All samples were taken from the approximate center of each aquarium without mixing the test medium, and were stored deep-frozen (at about -20 °C) immediately after sampling. Test samples and control samples were thawed at room temperature for 1.5 hours and shaken manually to obtain homogeneous sample solutions. If necessary, they were further diluted into the calibration range with test water before they were analyzed.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method:Prior to the start of the test and prior to each test medium renewal, a stock solution of nominal 150 mg/L was freshly prepared by completely dissolving 150 mg of the test item (dosed in the range of 150.2 to 153.1 mg) in 1000 mL of test water (except for day 3 where 75 mg of test item was dissolved in 500 mL of test water) using ultrasonic treatment for 15 minutes and stirring for 15 minutes at room temperature. Then, adequate volumes of the intensively stirred stock solution were diluted with test water to obtain the desired test concentrations. The test media were freshly prepared just before introduction of the fish (= start of the exposure) and before each test medium renewal.

- Controls: dilution water only
- Evidence of undissolved material (e.g. precipitate, surface film, etc): No remarkable observations were made concerning the appearance of the test media. All test media were clear solutions throughout the entire test duration.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebra fish
- Strain: not provided
- Source: The fish were obtained from a breeding culture at Harlan Laboratories.
- Age at study initiation (mean and range, SD): not provided
- Length at study initiation (length definition, mean, range and SD): From the acclimated test fish batch, 10 fish were measured at the start of the test. The mean total body length (measured from the mouth to the end of the tail fin) of the fish was 2.8 ± 0.22 cm (Mean ± SD).
- Weight at study initiation (mean and range, SD): the mean body wet weight was 0.20 ± 0.02 g (Mean ± SD).
- Method of breeding: similar to test conditions
- Feeding during test: no

ACCLIMATION
- Acclimation period: Prior to test start, they were acclimated for one week to the test water and temperature.
- Acclimation conditions (same as test or not): similar
- Type and amount of food: During holding and acclimatization until one day before the start of the test, the fish were fed with a commercial fish diet (Tetra Min® Hauptfutter, supplied by TETRA-Werke, 49304 Melle / Germany).
- Feeding frequency: not reported
- Health during acclimation (any mortality observed):During holding and acclimatization, no fish died in the test fish batch and all fish were healthy.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

15 mg/L as CaCO3

Test temperature:

21-22 °C

pH:

7.2 to 7.5

Dissolved oxygen:

The oxygen concentration was at least 7.9 mg/L, corresponding to at least 88% oxygen saturation.

Nominal and measured concentrations:

The nominal test item concentrations tested were 0.15, 0.47, 1.5, 4.7 and 15 mg/L. Additionally, a control group was tested in parallel. The measured concentrations of the test item in the test media of the test concentrations of 0.15 to 15 mg/L were between 81 and 112% of the nominal values at the start of the test and between below Limit of Quantification (LOQ) and 109% of the nominal concentrations at the end of each renewal period. Thus, the correct dosage of the test item was confirmed, but the test item concentrations declined below 80% at the lower concentrations in the test media over the renewal period of 24 hours. Therefore, the biological results were related to the mean measured test item concentrations, i.e., control, 0.090, 0.33, 1.6, 4.8 and 16 mg test item/L.

Details on test conditions:

TEST SYSTEM
- Test vessel: glass vessels filled with 5 L test solution each
- Aeration: not reported, but DO concentration was within the validity range
- Renewal rate of test solution (frequency/flow rate): 24 h renewal interval.
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.28 g/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted test water (AAP Medium) prepared according to the test guideline was used for this study.
- Culture medium different from test medium: no
- Intervals of water quality measurement: at every renewal of the freshly prepared and aged solutions

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: A 16 hour light to 8-hour dark photoperiod, with a 30-minute transition period was used
- Light intensity: light intensity during the light period was approximately within the range of 140 to 480 Lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
EC50 96 hours

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 3.2
- Justification for using less concentrations than requested by guideline: NA
- Range finding study
- Test concentrations: not performed
- Results used to determine the conditions for the definitive study: not performed

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

0.09 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% C.I.: 0.8 - 3.3 mg test item/L

Details on results:

- Behavioural abnormalities:
- Observations on body length and weight: NA
- Other biological observations: In the control and at the test concentrations up to and including 0.090 mg/L, all fish survived until the end of the test and no visible abnormalities were observed in the test fish.
- Mortality of control: none
- Other adverse effects control: none
- Abnormal responses: none
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No remarkable observations were made concerning the appearance of the test media. All test media were clear throughout the entire test duration.
- Effect concentrations exceeding solubility of substance in test medium: no

Reported statistics and error estimates:

The LC50 and the 95%-confidence interval at the observation dates were calculated Weibull Analysis. The NOEC, LOEC, LC0 and LC100 were determined directly from the raw data.

Sublethal observations / clinical signs:

In the control and at the test concentrations up to and including 0.090 mg/L, all fish survived until the end of the test and no visible abnormalities were observed in the test fish. At the next higher test concentration of 0.33 mg/L, three test fish showed one or several visible abnormalities. One of the seven test fish died at this test concentration. At the test concentration of 4.8 mg/L all test fish were dead after 48 hours of test duration while at the highest test concentration of 16 mg/L, all fish were dead already after 24 hours of test duration.

Validity criteria fulfilled:

yes

Executive summary:

In the Klimisch 1 GLP study from Kimmel (2013) the acute toxicity of Sulfosuccinic (FA C12-18)E, 2Na to zebra fish (Danio rerio) was determined in a 96‑hour semi-static test with daily test medium renewal according to the EU Commission Directive 92/69/, Part C.1, the Commission Regulation (EC) No 440/2008, Part C.1 and the OECD Guideline for Testing of Chemicals No. 203 (1992). The following nominal concentrations were tested: 0.15, 0.47, 1.5, 4.7 and 15 mg/L mg test item/L. Additionally, a control (test water without test item) was tested in parallel.

The measured concentrations of the test item in the test media of the test concentrations of 0.15 to 15 mg/L were between 81 and 112% of the nominal values at the start of the test and between below Limit of Quantification (LOQ) and 109% of the nominal concentrations at the end of each renewal period. Thus, the correct dosage of the test item was confirmed, but the test item concentrations of the two lowest concentrations declined below 80% over the renewal period of 24 hours. Therefore the biological results were related to the following mean measured test item concentrations: control, 0.090, 0.33, 1.6, 4.8 and 16 mg test item/L.

In the control and at the test concentrations up to and including 0.090 mg/L, all fish survived until the end of the test and no visible abnormalities were observed in the test fish. At the next higher test concentration of 0.33 mg/L, three test fish showed one or several visible abnormalities. One of the seven test fish died at this test concentration. At the test concentration of 4.8 mg/L all test fish were dead after 48 hours of test duration while at the highest test concentration of 16 mg/L, all fish were dead already after 24 hours of test duration. Thus, the 96‑hour NOEC and LC₀of Sulfosuccinic (FA C12-18)E, 2Na to zebra fish were both determined to be 0.090 mg/L. The 96‑hour LC₅₀of Sulfosuccinic (FA C12-18)E, 2Na was calculated to be 2.0 mg/L with a 95% confidence interval of 0.8-3.3 mg/L. The 96‑hour LC₁₀₀was 4.8 mg/L.

The results of this study are considered relevant and reliable for the risk assessment.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

2 mg/L

Additional information

In the Klimisch 1 GLP study from Kimmel (2013) the acute toxicity of Sulfosuccinic (FA C12-18)E, 2Na to zebra fish (Danio rerio) was determined in a 96‑hour semi-static test with daily test medium renewal according to the EU Commission Directive 92/69/, Part C.1, the Commission Regulation (EC) No 440/2008, Part C.1 and the OECD Guideline for Testing of Chemicals No. 203 (1992). The following nominal concentrations were tested: 0.15, 0.47, 1.5, 4.7 and 15 mg/L mg test item/L. Additionally, a control (test water without test item) was tested in parallel.

The measured concentrations of the test item in the test media of the test concentrations of 0.15 to 15 mg/L were between 81 and 112% of the nominal values at the start of the test and between below Limit of Quantification (LOQ) and 109% of the nominal concentrations at the end of each renewal period. Thus, the correct dosage of the test item was confirmed, but the test item concentrations of the two lowest concentrations declined below 80% over the renewal period of 24 hours. Therefore the biological results were related to the following mean measured test item concentrations: control, 0.090, 0.33, 1.6, 4.8 and 16 mg test item/L.

In the control and at the test concentrations up to and including 0.090 mg/L, all fish survived until the end of the test and no visible abnormalities were observed in the test fish. At the next higher test concentration of 0.33 mg/L, three test fish showed one or several visible abnormalities. One of the seven test fish died at this test concentration. At the test concentration of 4.8 mg/L all test fish were dead after 48 hours of test duration while at the highest test concentration of 16 mg/L, all fish were dead already after 24 hours of test duration. Thus, the 96‑hour NOEC and LC₀of Sulfosuccinic (FA C12-18)E, 2Na to zebra fish were both determined to be 0.090 mg/L. The 96‑hour LC₅₀of Sulfosuccinic (FA C12-18)E, 2Na was calculated to be 2.0 mg/L with a 95% confidence interval of 0.8-3.3 mg/L. The 96‑hour LC₁₀₀was 4.8 mg/L.

The results of this study are considered relevant and reliable for the risk assessment.