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Long-term toxicity to aquatic invertebrates

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Reference
Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2015-07-06 till 2015-09-30
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: OECD guideline study without restrictions.
Qualifier:
according to
Guideline:
EU Method C.20 (Daphnia magna Reproduction Test)
Version / remarks:
(2008)
Deviations:
no
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 211 (Daphnia magna Reproduction Test)
Version / remarks:
(2012)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Analytical monitoring:
yes
Details on sampling:
- Concentration: 10 mg/L 14H-benz[4,5]isoquino[2,1-a]perimidin-14-one plus control
- Sampling schedule: Control of on day 0, 9 and 19 in the freshly prepared media; on day 2, 12 and 21 in the old media and test concentration of 10 mg/L on day 0, 9 and 19 in the freshly prepared media; on day 2, 12 and 21 in the old media
- 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:
Pre-treatment of 14H-benz[4,5]isoquino[2,1-a]perimidin-14-one
A direct weighing was prepared to produce the only test concentration.
20 mg of 14H-benz[4,5]isoquino[2,1-a]perimidin-14-one were added to 2 litres of dilution water (day 0 – 8 and 16) and 10 mg of 14H-benz[4,5]isoquino[2,1-a]perimidin-14-one were added to 1 litre of dilution water (day 9 – 15 ), treated for 1 hour in an ultrasonic bath and stirred for 24 h on a magnetic stirrer. Finally 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 was measured and ranged between pH 7.7 and 7.8. 100 mL of the prepared solution and one neonate was added to each replicate. For each test item concentration and the control 10 replicates were prepared.
Test organisms (species):
Daphnia magna
Details on test organisms:
- Name : Daphnia magna STRAUS, parthenogenetic females
- Source : Strain of Bundesgesundheitsamt Berlin
- Maintenance and Acclimatisation : A population of parthenogenetic females of synchronized age structure has been maintained for more than 20 years in the test facility under constant temperature conditions (20 +/- 1 °C) at a 16 : 8 hour light-dark photoperiod (light intensity: < 20 μE x m-2 x s-1). The culture water (so-called 'M4 medium') was partly renewed once a week. The Daphnia were exclusively fed unicellular green algae (Desmodesmus subspicatus) 'ad libitum'. Mortalities of parent Daphnia during the culture period were recorded daily in a semi-quantitative way. The neonates were separated from their parent Daphnia by filtration prior to the acute test.
Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
21 d
Hardness:
13.4 °dH to 15.0 °dH (= 239 - 268 mg/L CaCO3) measured at each test vessel at the beginning of the test and at 2, 9, 12, 19 and 21 days
Test temperature:
20.1 - 20.9 °C measured at each test vessel at the beginning of the test and at 2, 9, 12, 19 and 21 days
pH:
7.8 - 8.1 measured at each test vessel at the beginning of the test and at 2, 9, 12, 19 and 21 days
Dissolved oxygen:
8.7 - 9.1 mg O2/L with 102 - 105 % saturation measured at each test vessel at the beginning of the test and at 2, 9, 12, 19 and 21 days
Nominal and measured concentrations:
- Concentration: 10 mg/L 14H-benz[4,5]isoquino[2,1-a]perimidin-14-one plus control
The results are expressed in terms of time-weighted mean measured concentrations. Effective concentrations ranged from 5.3 % to 7.3 % of the nominal values in the freshly prepared media and from 3.8 % to 5.5 % in the media after 48 hours, or 72 hours of exposure.
Details on test conditions:
Exposure conditions
- Test vessels: 150 mL glass beakers covered with watch glasses, holding 1 neonate in 100 mL test medium
- Experimental design: 1 test concentration plus 1 control; 1 neonate per vessel, 10 replicates per concentration/control; Semi static system (renewal of the test media on Monday, Wednesday and Friday)
- Method of initiation- : neonates were placed in prepared media
Photoperiod: 16 h light : 8 h dark
- Light intensity: not exceeding 20 μE x m-2 x s-1
- Temperature of incubation unit: 19.5 to 20.5 °C
- Aeration: none
- Test item concentration: 10 mg/L
- Food source and feeding rate: Desmodesmus subspicatus; day 0-7: 0.1 mg C / Daphnia / day and day 8-21: 0.2 mg C / Daphnia / day
- Duration of exposure: 21 days
-Criteria of effects : The criterion of adverse effects used in this study was the item-induced inhibitory effect on the parthenogenetic reproduction rate and the mobility behaviour/mortality rate of parent Daphnia, all effects being assessed at least three times a week.
Reference substance (positive control):
no
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
>= 0.566 mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
reproduction
Duration:
21 d
Dose descriptor:
LOEC
Effect conc.:
> 0.566 mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
reproduction
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
>= 0.566 mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
21 d
Dose descriptor:
LOEC
Effect conc.:
> 0.566 mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
1st brood on day: 8
Validity criteria fulfilled:
yes
Remarks:
(- The mortality rate in the controls did not exceed 20 % by the end of the test. - The dissolved oxygen concentration remained above 3 mg/L throughout the exposure period.)
Conclusions:
Daphnia magna was exposed under semi-static conditions for 21 days to one concentration (10 mg/L) of 14H-benz[4,5]isoquino[2,1-a]perimidin-14-one. No toxic effects against daphnia were observed at a test concentration of nominally 10 mg/L, corresponding to a time-weighted mean measured concentration of 0.566 mg/L, which reflects the maximum water solubility under exposure conditions.
Executive summary:

A study was performed to assess the effects of 14H-benz[4,5]isoquino[2,1-a]perimidin-14-one on the reproductive output of parthenogenetic Daphnia magna to the test solution of one concentration (10 mg/L) and blank control solution under semi static conditions.

The study was conducted in accordance with Council Regulation (EC) No 440/2008, Method C.20 'Daphnia magna Reproduction Test'' (2008), which equals in all relevant parts to the OECD Test guideline 211 (Daphnia magna Reproduction Test; 2012).

Base on different biological effects observed and endpoints assessed, the following results were obtained at the end of the 21 day exposure of the Daphnia reproduction test:

21d-NOEC (reproduction): ≥ 0.566 mg/L

21d-LOEC (reproduction): > 0.566 mg/L

21d-NOEC (mortality): ≥ 0.566 mg/L

21d-LOEC (mortality): > 0.566 mg/L

1st brood on day : 8

No toxic effects against daphnia were observed at a test concentration of nominally 10 mg/L, corresponding to a time-weighted mean measured concentration of 0.566 mg/L, which reflects the maximum water solubility under exposure conditions.

The results are expressed in terms of time-weighted mean measured concentrations. Effective concentrations ranged from 5.3 % to 7.3 % of the nominal values in the freshly prepared media and from 3.8 % to 5.5 % in the media after 48 hours, or 72 hours of exposure.

Description of key information

Daphnia magna was exposed under semi-static conditions for 21 days to one

concentration (10 mg/L) of 14H-benz[4,5]isoquino[2,1-a]perimidin-14-one. No toxic effects against daphnia were observed at a test concentration of nominally 10 mg/L, corresponding to a time-weighted mean measured concentration of 0.566 mg/L, which reflects the maximum water solubility under exposure conditions.

Key value for chemical safety assessment

EC10, LC10 or NOEC for freshwater invertebrates:
0.057 mg/L

Additional information

The key value should read as >= 0.566 mg/L as there was no effect up to the maximum water solubility of the substance and, thus, the substance has no chronic effects on daphnids.

Discussion

A study was performed to assess the effects of 14H-benz[4,5]isoquino[2,1-a]perimidin-14-one on the reproductive output of parthenogenetic Daphnia magna to the test solution of one concentration (10 mg/L) and blank control solution under semi static conditions.

The study was conducted in accordance with Council Regulation (EC) No 440/2008, Method C.20 'Daphnia magna Reproduction Test'' (2008), which equals in all relevant parts to the OECD Test guideline 211 (Daphnia magna Reproduction Test; 2012).

Base on different biological effects observed and endpoints assessed, the following results were obtained at the end of the 21 day exposure of the Daphnia reproduction test:

21d-NOEC (reproduction): ≥ 0.566 mg/L

21d-LOEC (reproduction): > 0.566 mg/L

21d-NOEC (mortality): ≥ 0.566 mg/L

21d-LOEC (mortality): > 0.566 mg/L

1stbrood on day : 8

No toxic effects against daphnia were observed at a test concentration of nominally 10 mg/L, corresponding to a time-weighted mean measured concentration of 0.566 mg/L, which reflects the maximum water solubility under exposure conditions.

The results are expressed in terms of time-weighted mean measured concentrations. Effective concentrations ranged from 5.3 % to 7.3 % of the nominal values in the freshly prepared media and from 3.8 % to 5.5 % in the media after 48 hours, or 72 hours of exposure.