[No public or meaningful name is available]

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:

2018

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)

Deviations:

no

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

not applicable

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Remarks:

HPLC/MS/MS analysis

Details on sampling:

Sampling schedule:
- Control: at 24 hours (2018-03-02)
- Test concentrations: on 2018-02-26, 2018-02-27, 2018-02-28, and 2018-03-01 freshly prepared media
and on 2018-02-27, 2018-02-28, 2018-03-01 and 2018-03-02 in the media after 24 hours of exposure

Vehicle:

no

Remarks:

Auxiliaries: ultra sonic bath, magnet stirrer folded filter and antiseptic filter

Details on test solutions:

Pre-treatment of test item and preparation of test item concentrations:
- 500 mg of the test item were added to 5 litres of dilution water, treated for 60 seconds at 8000 rpm with an ultra turrax and stirred for 24 h on a magnetic stirrer
- Undissolved particles of the test item were removed by filtration using a folded filter with a pore size of 7-12 µm and an aseptic filter Sartobran sterile capsules (pore size of 0.45 µm + 0.2 µm)

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

- Source: Interaquaristik.de Shop (Germany)
- Date supplied: 2018-01-23
- Acclimatisation: Stock held since 2018-01-23 and acclimatised to the test conditions since then.
- Temperature: 20 - 24 °C
- Dissolved oxygen: > 5 mg/L
- Feeding: Commercial fish food, daily. Feeding discontinued 24 h prior to test start.
- Mortalities during acclimatisation period: < 5 %
- Medication: none
- Mean standard length (n = 14): 3.34 cm (S.D. = 0.12 cm)
- Holding and dilution water: Reconstituted water prepared according to the recommendations of ISO 7346 (ISO water)

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

14.0 °dH (= 250 mg/L CaCO3)

Test temperature:

22.3 °C (after 96 hours)

pH:

7.7 (after 96 hours)

Dissolved oxygen:

8.1 mg/L (after 96 hours)

Salinity:

n.a.

Conductivity:

No data

Nominal and measured concentrations:

- Test concentration (nominal): 100 mg/L
- Test concentration after 96 hours (HPLC/MS/MS values): 90.749 µg/L

Details on test conditions:

EXPOSURE CONDITIONS:
- Test vessels: glass aquaria holding 5 L of test media covered by glass plates
- Experimental design: 1 test concentration plus 1 control
- Number of test organism: 7 fish per test concentration
- Feeding: none during the exposure period
- Method of initiation: fish were placed in prepared media
- Loading: 0.69 g body weight (wet weight) per litre
- Photoperiod: 16 h light : 8 h dark
- Temperature: 21.7 to 22.3 °C
- Aeration: gentle aeration via narrow glass tubes
- Test item concentration/s: 100 mg/L
- Method of administration: direct weighing
- Medium renewal: daily
- Criteria of effects: The criterion of death used in this study was the absence of response to physical stimulation; in addition to observations on mortality at 2, 24, 48, 72 and 96 hours, type and incidence of sub-lethal effects compared with control fish were observed

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

>= 72.061 µg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 72.061 µg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

> 72.061 µg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

All results are expressed in terms of nominal and geometric mean measured concentrations. Effective concentrations ranged from 0.047 % to 0.136 % of nominal values in the freshly prepared medium and from 0.021 % to 0.091 % of nominal values in the medium after 24 hours of exposure.

Validity criteria fulfilled:

yes

Remarks:

Mortality in the controls < 10 % by the end of the test; Dissolved oxygen concentration >60 % of the air-saturation value throughout the exposure period; pH did not vary by more than 1 unit

Conclusions:

No toxic effects against fish were observed at the limit of water solubility under exposure conditions. All results are expressed in terms of nominal and geometric mean measured concentrations. Effective concentrations ranged from 0.047 % to 0.136 % of nominal values in the freshly prepared medium and from 0.021 % to 0.091 % of nominal values in the medium after 24 hours of exposure.

Executive summary:

According to the method described in the Council Regulation (EC) No 440/2008, Method C.1 'Acute toxicity for Fish' (2008) which is in most parts equivalent to the OECD Guideline for Testing of Chemicals No. 203 'Fish, Acute Toxicity Test' (1992) the acute toxic effects (mortality) of various concentrations of a test item to a freshwater fish species were assessed. A range finding test preceded the main test. It provided information about the range of concentrations which were used in the main test. The following nominal concentrations of the test item were tested in the range-finding test: 10 and 100 mg/L. Because of the anticipated low toxicity of the test item, fish were exposed in the main test to a limit test concentration of nominally 100 mg/L for a period of 96 hours. As no toxic effects against fish were observed, no statistical analysis was required to determine the LC 50. Additionally any abnormal behaviour or appearance of the fish was reported every 24 hours. The main test was conducted as semi-static test with daily renewal of test solutions. During the test a temperature range of 20 - 24 °C was maintained in the test vessels, with a maximum temperature fluctuation of +/- 0.6 °C in the test. The temperature, the pH and the oxygen values were measured at the beginning of the test and every 24 hours thereafter. The temperature, the pH and the oxygen values were measured every day in the freshly prepared media and in the media after 24 hours of exposure. The validity criteria of the test were met. No toxic effects against fish were observed at the limit of water solubility under exposure conditions. All results are expressed in terms of nominal and geometric mean measured concentrations. Effective concentrations ranged from 0.047 % to 0.136 % of nominal values in the freshly prepared medium and from 0.021 % to 0.091 % of nominal values in the medium after 24 hours of exposure. This toxicity study is classified as acceptable and satisfies the guideline requirements for the toxicity study to fish.

Description of key information

No toxic effects against fish were observed at the limit of water solubility under exposure conditions (for details see also chapter 4.8). All results are expressed in terms of nominal and geometric mean measured concentrations. Effective concentrations ranged from 0.047 % to 0.136 % of nominal values in the freshly prepared medium and from 0.021 % to 0.091 % of nominal values in the medium after 24 hours of exposure (Woelz, 2018).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

72.061 µg/L

Additional information

"Should read: 96 h-LC50 > 72.061 µg/L"