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Short-term toxicity to fish

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Reference
Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2014-11-13 till 2015-01-29
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: OECD guideline study without restrictions.
Qualifier:
according to
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Version / remarks:
(2008)
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Version / remarks:
(1992)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Analytical monitoring:
yes
Details on sampling:
- Concentrations: Concentration of 10 mg/L was measured at 0, 2, 24, 48, 72 and 96 hours; and a concentration of 0 mg/L at 96 hours.
- Sample storage: Routinely, the samples were analysed immediately. Only in exceptional cases, they were stored overnight deep frozen and protected from light.
Vehicle:
no
Details on test solutions:
Direct weighings were prepared daily to produce 14H-benz[4,5]isoquino[2,1-a]perimidin-14-one concentration. To achieve this 50 mg of the test item were added daily to 5 litres of dilution water, treated for 1 h in an ultrasonic bath and stirred for 24 h on a magnetic stirrer. Undissolved particles of 14H-benz[4,5]isoquino[2,1-a]perimidin-14-one were removed by filtration using a folded filter with a pore size of 7-12 μm.
The pH value was measured and ranged between pH 7.6 and 7.7 in the freshly prepared solution.
Finally 7 fish were given to all 14H-benz[4,5]isoquino[2,1-a]perimidin-14-one concentrations and the control.
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
- Name: Zebra fish (Danio rerio)
- Source: Interaquaristik.de Shop
- Date supplied: 2014-10-31
- Acclimatisation: Stock held since 2014-10-31 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): 3.26 cm (S.D. = 0.17 cm)
Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Hardness:
14.4 °dH (= 257 mg/L CaCO3)
Test temperature:
21.2 -22.0 °C measured at each test vessel at the beginning and the end of test
pH:
7.6 - 7.8 measured at each test vessel at the beginning and the end of test
Dissolved oxygen:
8.5 -9.0 mg O2/L with 95.0 - 100.8 % O2 saturation measured at each test vessel at the beginning and the end of test
Nominal and measured concentrations:
10 mg/L (nominal) plus control
All results are expressed in terms of geometric mean measured concentrations. Effective concentrations ranged from 0.4 % to 2.0 % of nominal values in the freshly prepared medium and from 0.1 % to 0.8 % of nominal values in the medium after 24 hours of exposure.
Details on test conditions:
An algal growth inhibition test preceded the fish test. It provided information about the concentration which was used in the fish test.
The following nominal concentrations of 14H-benz[4,5]isoquino[2,1-a]perimidin-14-one were tested in the algal growth inhibition range-finding test: 1.0 and 100.0 mg/L.

A group of seven fish of the recommended size was exposed to a limit test concentration of nominally 10.0 mg/L of 14H-benz[4,5]isoquino[2,1-a]perimidin-14-one dissolved in dilution water. Auxiliaries used to prepare the test media were an ultrasonic bath, a magnetic stirrer and a folded filter.

- Test vessels: glass aquaria holding 5 L of test media covered by glass plates
- Experimental design: 1 test concentration plus 1 control; 7 fish per test concentration; no feeding during the exposure period; semi-static
- Method of initiation: fish were placed in prepared media
- Loading: 0.48 g body weight (wet weight) per litre
- Photoperiod: 16 h light: 8 h dark
- Temperature: 21.2 to 22.0 °C
- Aeration: gentle aeration via narrow glass tubes
- Test item concentration: 10 mg/L
- Method of administration: direct weighing
- Medium renewal: daily
- Duration of exposure: 96 hours
- 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:
2 h
Dose descriptor:
LC0
Effect conc.:
>= 0.041 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
24 h
Dose descriptor:
LC0
Effect conc.:
>= 0.024 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
48 h
Dose descriptor:
LC0
Effect conc.:
>= 0.044 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
72 h
Dose descriptor:
LC0
Effect conc.:
>= 0.063 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LC0
Effect conc.:
>= 0.069 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
2 h
Dose descriptor:
LC100
Effect conc.:
> 0.041 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
24 h
Dose descriptor:
LC100
Effect conc.:
> 0.024 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
48 h
Dose descriptor:
LC100
Effect conc.:
> 0.044 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
72 h
Dose descriptor:
LC100
Effect conc.:
> 0.063 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
> 0.069 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
2 h
Dose descriptor:
LC50
Effect conc.:
> 0.041 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
24 h
Dose descriptor:
LC50
Effect conc.:
> 0.024 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
> 0.044 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
72 h
Dose descriptor:
LC50
Effect conc.:
> 0.063 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 0.069 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality

No toxic effects against fish were observed at the limit of water solubility under exposure conditions.

All results are expressed in terms of geometric mean measured concentrations. Effective concentrations ranged from 0.4 % to 2.0 % of nominal values in the freshly prepared medium and from 0.1 % to 0.8 % of nominal values in the medium after 24 hours of exposure.

Validity criteria fulfilled:
yes
Remarks:
(- The mortality in the controls did not exceed 10 % by the end of the test. - The dissolved oxygen concentration remained above 60 % of the air-saturation value throughout the exposure period. - The pH did not vary by more than 1 unit.)
Conclusions:
The acute toxicity to fish (danio rerio) was tested in a semi-static test. After 96 hours of exposure no effects against fish were observed at the limit of water solubility under exposure conditions.
Executive summary:

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) assesses the acute toxic effects (mortality) of various concentrations of a 14H-benz[4,5]isoquino[2,1-a]perimidin-14-one to a freshwater fish species.

The purpose of this method was to determine that concentration which causes a 50 % mortality rate (= LC 50) or, if conducted as a limit test, to determine the acute toxic effects at a maximum test concentration of 10.0 mg/L or at the limit of water solubility.

An algal growth inhibition test preceded the fish test. It provided information about the concentration which was used in the fish test. The following nominal concentrations of 14H-benz[4,5]isoquino[2,1-a]perimidin-14-one were tested in the algal growth inhibition range-finding test: 1.0 and 100.0 mg/L.

Because of the low solubility of 14H-benz[4,5]isoquino[2,1-a]perimidin-14-one, fish were exposed in the main test to a limit test concentration of nominally 10 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.

After 96 hours of exposure a LC50 of > 0.069 mg/L was determined.

No toxic effects against fish were observed at the limit of water solubility under exposure conditions.

This toxicity study is classified acceptable and satisfies the guideline requirement for the acute fish.

Description of key information

The acute toxicity to fish (danio rerio) was tested in a semi-static test. After 96 hours of exposure no effects against fish were observed at the limit of water solubility under exposure conditions.

Key value for chemical safety assessment

LC50 for freshwater fish:
0.069 mg/L

Additional information

The key value should read as LC50 > 0.069 mg/L as there was no effect up to the limit of water solubility.