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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:
13 March 2014 to 8 August 2014
Reliability:
1 (reliable without restriction)
Qualifier:
according to
Guideline:
OECD Guideline 211 (Daphnia magna Reproduction Test)
GLP compliance:
yes
Specific details on test material used for the study:
Identifier: EXP1200078
Appearance: Very dark brown (almost black) viscous liquid
Batch: E00275-350
Sample Expiration Date: end-2014
Purity:100%
Analytical monitoring:
yes
Vehicle:
no
Details on test solutions:
WAFs (water accommodated fractions) solutions with a nominal concentration of 10.0 mg/L can be used by adding 0.0200 g
of the test substance to the 2000mL Elendt M7 test medium in glass aspirator bottles with appropriate size. The mixing
aspirator bottle was stoppered and the aqueous test substance mixture was stirred for 24 hrs at room temperature (about 23°C ~ 25°C). The vortex height was set at least 10% of the static liquid depth. At the end of the 24 h, stirring was stopped and
the test substance was allowed to keep afloat. The mixture was allowed to settle for 1 hour at room temperature. Then the
aqueous portion removed through the outlet of the aspirator stirring bottle was the WAFs solutions of the test substance without further filtration to remove the remaining undissolved test substance.

WAFs (water accommodated fractions) solutions with a nominal concentration of 1.00 mg/L can be used by adding 0.0020 g
of the test substance to the 2000mL Elendt M7 test medium in glass aspirator bottles with appropriate size. The mixing
aspirator bottle was stoppered and the aqueous test substance mixture was stirred for 24 hrs at room temperature (about 23°C ~ 25°C). The vortex height was set at least 10% of the static liquid depth. At the end of the 24 h, stirring was stopped and
the test substance was allowed to keep afloat. The mixture was allowed to settle for 1 hour at room temperature.
Test organisms (species):
Daphnia magna
Details on test organisms:
Daphnia magna, not being the first brood progeny, was the test species which were 6 to 24 hours old at the start of the test. All organisms used for a particular test originated from cultures established from the same healthy stock of daphnia. The stock animals were maintained in culture conditions (light, temperature, and medium) similar to those to be used in the test.
Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
21 d
Post exposure observation period:
none
Hardness:
221 to 232 mg CaCO3/L
Test temperature:
20C to 21C
pH:
7.4 to 8.2
Dissolved oxygen:
90% to 101% of air saturation
Salinity:
no data
Nominal and measured concentrations:
In the range-finding test, animals were exposed to concentrations of 1.00 and 10.0 mg/L (nominal concentration). A blank control was tested in parallel (dilution medium without test substance).

Based on the results of the range-finding test, nominal concentration of 10.0 mg/L WAFs were conducted in limit test. Additionally, one control group was tested in parallel (dilution medium without test substance).
Details on test conditions:
Conditions of Exposure
Depend on the stability of the test solution, the concentration of the test substance was demonstrated to be maintained
within ±20 percent of the measured initial concentration throughout the test under a 72h-renewal condition (Table 2).
When the medium was renewed, a second series of test vessels were prepared and the parent animals were then be
transferred to them by a glass pipette of suitable diameter. All the beakers lids were put on.The volume of medium
transferred with the Daphnia was minimized.

10 animals individually were held at each test concentration and 10 animals individually in the control series. Treatments
were allocated to the test vessels and all subsequent handling of the test vessels were done in a random fashion. The test
conditions were maintained as follows:
-Light: 16-hour light and 8-hour dark cycle daily (light intensity: 1000 to 1500 lux);
-Temperature: (20±1) oC;
-Oxygen concentration: 90% to 101%; no aerating;
-pH: 7.4~8.2;
-Feeding: Chlorella vulgaris Beij. was given once every day, 0.1 - 0.2 mg C/Daphnia each time.
-Aeration: None.

Observations & Determinations
(1) Observations: the number of living offspring was counted for each vessel daily. The offspring produced by each parent
animal were preferably removed and counted daily from the appearance of the first brood to prevent them consuming food
intended for the adult.
(2) Concentration: all test concentrations were analyzed at the beginning (0 d), immediately prior to the first renewal of the
test solution (3 d) and weekly thereafter (9, 21 d). In all cases, determination of test substance concentrations prior to
renewal was performed on one seperate vessel at each test concentration.
(3) Other parameters: including time to production of first brood; number of broods per animal.Besides, oxygen
concentration, temperature, hardness and pH values were measured once a week, in the control(s) and the highest test
substance concentration.
Reference substance (positive control):
no
Key result
Duration:
21 d
Dose descriptor:
LOEC
Effect conc.:
> 10 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Key result
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
10 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Key result
Duration:
21 d
Dose descriptor:
LOEC
Effect conc.:
> 10 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Key result
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
10 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Validity criteria fulfilled:
yes
Remarks:
All animals in the control were alive and the mean number of live offspring produced per parent animal surviving at the end of the test was > 60. So the study met the acceptability criteria.
Conclusions:
Under valid semi-static test conditions (72h-renewal), for the daphnia reproduction the LOEC was more than 10.0 mg/L
WAFs (measured concentration 2.20 mg/L), the NOEC was 10.0 mg/L WAFs (measured concentration 2.20 mg/L). For
survival of adults the LOEC was more than 10.0 mg/L WAFs (measured concentration 2.20 mg/L), the NOEC was 10.0
mg/L WAFs (measured concentration 2.20 mg/L). i.e.:
LOEC (Reproduction) > 10.0 mg/L (measured concentration 2.20 mg/L);
NOEC (Reproduction) =10.0 mg/L(measured concentration 2.20 mg/L).
LOEC (Survival) > 10.0 mg/L (measured concentration 2.20 mg/L);
NOEC (Survival) = 10.0 mg/L(measured concentration 2.20 mg/L).
Executive summary:

A well conducted study using OECD 211 determined that the NOEC concentration for both survival and reproduction was > 10 mg/L which was the solubility limit in this study.

Description of key information

Under valid semi-static test conditions (72h-renewal), for the daphnia reproduction the LOEC was more than 10.0 mg/L WAFs (measured concentration 2.20 mg/L), the NOEC was 10.0 mg/L WAFs (measured concentration 2.20 mg/L).

 

For survival of adults the LOEC was more than 10.0 mg/L WAFs (measured concentration 2.20 mg/L), the NOEC was 10.0 mg/L WAFs (measured concentration 2.20 mg/L). i.e.:

 

LOEC (Reproduction) > 10.0 mg/L (measured concentration 2.20 mg/L);

NOEC (Reproduction) =10.0 mg/L(measured concentration 2.20 mg/L).

LOEC (Survival) > 10.0 mg/L (measured concentration 2.20 mg/L);

NOEC (Survival) = 10.0 mg/L(measured concentration 2.20 mg/L).

Key value for chemical safety assessment

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

Additional information

A well conducted study using OECD 211 determined that the NOEC concentration for both survival and reproduction was > 10 mg/L which was the solubility limit in this study.