Hydroxyprogesterone

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:

2016

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:

yes

Remarks:

Some test fish had a somewhat larger size than the one recommended in the test guideline for the zebra fish. This deviation is not regarded to be relevant to the results.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Remarks:

HPLC analysis and UV/VIS-detection

Details on sampling:

Sampling schedule:
- Control: in the media after 24 hours of exposure.
- Test concentrations: on 2016-02-29 and 2016-03-01 in the freshly prepared media and on 2016-03-01 and 2016-03-02 in the media after 24 hours of exposure

Vehicle:

no

Details on test solutions:

Pre-treatment of test item and preparation of test item concentrations:
- 1000 mg of the test item were added daily to 10 litres of dilution water, treated for 1 h in an ultrasonic bath and stirred for 24 h on a magnetic stirrer
- Undissolved particles were removed by filtration using a folded filter (pore size 7-12 μm)
- The pH was measured to be pH 7.8
- To produce the different test item concentrations appropriate amounts of the stock solution were diluted with dilution water to a volume of 5000 mL

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

- Source: Interaquaristik.de Shop (Germany)
- Date supplied: 2016-01-11
- Acclimatisation: Stock held since 2016-01-11 and acclimatised to the test conditions since then
- Holding water: synthetic freshwater according ISO standard
- 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 = 28): 3.42 cm (S.D. = 0.17 cm)

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

13.2 °dH (= 236 mg/L CaCO3)

Test temperature:

21.1-21.6 °C (concerning the highest test concentration)

pH:

7.6-7.8 (concerning the highest test concentration)

Dissolved oxygen:

8.5-9.0 (mg/L) (concerning the highest test concentration)

Salinity:

n.a.

Conductivity:

No data

Nominal and measured concentrations:

100 mg/L (n): 4.562 mg/L (HPLC-value) after 0 hours of exposure; 3.921 mg/L (HPLC-value) after 48 hours of exposure

Details on test conditions:

EXPOSURE CONDITIONS:
- Test vessels: glass aquaria holding 5 L of test media covered by glass plates
- Experimental design: 3 test concentrations plus 1 control; 7 fish per test concentration
- Feeding: None
- Method of initiation: fish were placed in prepared media
- Loading: 0.71 g body weight (wet weight) per litre
- Photoperiod: 16 h light: 8 h dark
- Temperature: 21.1 to 21.7 °C
- Aeration: gentle aeration via narrow glass tubes
- Test item concentrations: 25, 50 and 100 mg/L
- Method of administration: stock solution
- Medium renewal: daily
- Criteria of effects: Criterion of death (absence of response to physical stimulation): additionally 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

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 4.207 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

2.032 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

> 4.207 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

Effective concentrations ranged from 4.5 % to 4.6 % of nominal values in the freshly prepared media and from 3.4% to 3.9 % of nominal values in the media after 24 hours of exposure according to the semi-static test procedure.

Validity criteria fulfilled:

yes

Remarks:

(mortality in the controls < 10% by the end of the test; dissolved oxygen concentration above 60% of the air-saturation value throughout the exposure period; pH did not vary by more than 1 unit)

Conclusions:

After 96 hours of exposure a LC50 of higher than 4.21 mg/L was determined under semi-static conditions. All results are expressed in terms of geometric mean measured concentrations.

Executive summary:

The acute toxicity of the substance to fish (Danio rerio) was investigated in a semi-static test according to EEC methods for Determination of Ecotoxicity Annex to Directive 92/69/EEC (O.J. No. L383A, 29.12.92) Part C, Method 1 "Acute toxicity for Fish". Groups of seven fish of the recommended size were exposed to a range of concentrations, nominally 25, 50 and 100 mg/L dissolved in dilution water. Auxiliaries used to prepare the test media were an ultrasonic bath, a magnetic stirrer and folded filters . Observations were made on the number of dead fish and the incidence of sub-lethal effects after 2, 24, 48, 72 and 96 hours of exposure. After 96 hours of exposure a LC50 of higher than 4.21 mg/L was determined under semi-static conditions. All results are expressed in terms of geometric mean measured concentrations. Effective concentrations ranged from 4.5 % to 4.6 % of nominal values in the freshly prepared media and from 3.4% to 3.9 % of nominal values in the media after 24 hours of exposure according to the semi-static test procedure. This toxicity study is classified as acceptable and satisfies the guideline requirements for the acute Fish study.

Description of key information

After 96 hours of exposure a LC50 of higher than 4.21 mg/L was determined under semi-static conditions. All results are expressed in terms of geometric mean measured concentrations.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

4.207 mg/L

Additional information

"Should read: > 4.207 mg/L"

Effective concentrations ranged from 4.5 % to 4.6 % of nominal values in the freshly prepared media and from 3.4% to 3.9 % of nominal values in the media after 24 hours of exposure according to the semi-static test procedure.