Sodium anisate

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:

15 - 22 Jan 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: OECD 126

Version / remarks:

2010

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Hessisches Ministerium für Umwelt, Klimaschutz, Landwirtschaft und Verbraucherschutz, Wiesbaden, Germany

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Test item concentration and the control were analysed
- Sampling method: At test start and at each test media renewal. For the determination of the stability of the test item, samples from the aged test media were taken in duplicate out of the only test concentration and the control directly before each test medium renewal and at the end of the test. All test medium samples were taken from the approximate centre of the aquaria.
- Sample storage conditions before analysis: All samples were stored in a freezer (≤ -20 °C), protected from light until analysis was performed.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Test item concentration of nominal 100 mg/L was prepared by dissolving 1000 mg test item into 10000 mL test water by intense stirring for 10 min. The test media was prepared just before introduction of the test fish (= start of the test and test medium renewal on day 1, 2 and 3).
- Controls: yes

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Rainbow Trout
- Source: Forellenzuchtbetrieb Störk, 88348 Bad Saulgau, Germany
- Length at study initiation (mean length): 4.48 cm
- Weight at study initiation (mean and range, SD): 0.93 g ± 0.13 g
- Method of breeding: All fish were obtained and held in the laboratory for at least 12 d before the start of the test. They were held in water of the quality to be used in the test for at least seven days immediately before testing under the following conditions.

ACCLIMATION
- Acclimation period: 12 d
- Acclimation conditions (same as test or not): same
- Feeding frequency during acclimation: three times per week or daily until 24 hours before the test was started
- Health during acclimation (any mortality observed): During the last 7 days prior to the start of the test no fish died in the test fish batch. Therefore the mortalities in the fish batch were below 5% and the fish batch was accepted.

FEEDING DURING TEST : no feeding during the test

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

2.5 mmol/L (= 250.0 mg/L) as CaCO3

Test temperature:

13.7 to 14.3 °C

pH:

7.3 to 7.8

Dissolved oxygen:

93 - 100% (Control)
93 - 96% (100 mg test item/L)

Conductivity:

≤ 10 µScm-1

Nominal and measured concentrations:

Control and 100 mg test item/L

Details on test conditions:

TEST SYSTEM
Test vessel:
- Type: open
- Material, size, headspace, fill volume: 12 litre glass aquaria with 10 litre test medium
- Aeration: The test media were slightly aerated during the test
- Renewal rate of test solution (frequency/flow rate): On day 1, 2 and 3
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.93 ± 0.13 g fish/L test water

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Iso medium with deionised water
- Alkalinity: 0.8 mmol/L
- Ca/mg ratio: 4 : 1 (based on molarity)
- Na/K ratio: 10 : 1 (based on molarity)
- Culture medium different from test medium: no
- Intervals of water quality measurement: The water temperature, pH-values and the dissolved oxygen concentrations were determined daily in the freshly prepared and aged test media of each treatment group.

OTHER TEST CONDITIONS
- Photoperiod: 16 h light : 8 h dark (30 min dawn/dusk period was provided)
- Light intensity: 680 - 730 lux

EFFECT PARAMETERS MEASURED: The test fish were observed at test start and after approximately 2, 24, 48, 72 and 96 h test duration for sublethal effects and mortality.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: not necessary due to a limittest
Range finding study : No pre-experiments were performed.

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: no abnormal behaviour was observed
- Mortality of control: No mortality was observed

Reported statistics and error estimates:

No statistical analysis was performed.

Sublethal observations / clinical signs:

Analytical Results:

In the freshly prepared test media at the start of the test and at the renewal of the test media 106% of the nominal test concentrations were found. In the 24 hour old test media after 24 and 96 hours test duration, also a measured concentration of 106% of the nominal value was determined. During the test the test organism were exposed to a mean of 106% of nominal.

 Table 1: Validity criteria of the study

Criterion from the guideline	Outcome	Validity criterion fulfilled
The mortality in the control(s) should not exceed 10% (or one fish if less than ten are used) at the end of the test	No mortality was observed	Yes
The dissolved oxygen concentration must have been at least 60 per cent of the air saturation value throughout the test	>93%	Yes
There must be evidence that the concentration of the substance being tested has been satisfactorily maintained, and preferably it should be at least 80 per cent of the nominal concentration throughout the test. If the deviation from the nominal concentration is greater than 20 per cent, results should be based on the measured concentration.	106%	Yes

Validity criteria fulfilled:

yes

Remarks:

For additional information please see Table 1 within section "Any other information on results incl. tables".

Conclusions:

LC50 (96h) >100 mg/L (Oncorhynchus mykiss, OECD 203 and 126, nominal)

Description of key information

LC50 (96h) >100 mg/L (Oncorhynchus mykiss, OECD 203 and 126, nominal)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

One experimental study is available investigating the toxicity of the test item sodium anisate (CAS 536-45-8) to fish . The study was conducted according to OECD 203 (GLP) using the rainbow trout Oncorhynchus mykiss under semi-static conditions. The test was performed according to the thresholdapproach OECD 126. The absence of mortality at 100 mg/L indicated that fish was not the most sensitive group of test organisms and that the LC50 is greater than the limit test concentration. The measured content of the test item in the fresh as well as in aged test solution was 106% of nominal. Since the content of active ingredient in the samples was between 80 and 120% of nominal, the toxicological endpoints were evaluated using nominal test item concentrations. The LC50 (96 h) of the test item was determined to be > 100 mg/L test item (nominal). The corresponding NOEC (mortality) (96 h) was 100 mg/L test item (nominal).No sublethal effects were observed in the control and test item concentration 100 mg/L after 96 h.