Registration Dossier

Ecotoxicological information

Long-term toxicity to aquatic invertebrates

Administrative data

Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report date:
2015

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
other: CRC-MEP. The Guidelines for the Testing of Chemicals HJ/T 153-2004, Effects on Biotic Systems
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: GB/T 21828-2008,ChemicaIs-Daphnia magna reproduction test
Deviations:
no
Qualifier:
according to guideline
Guideline:
OECD Guideline 211 (Daphnia magna Reproduction Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method C.20 (Daphnia magna Reproduction Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: EPA OPPTS 850.1010 (Daphnid Chronic Toxicity Test, Freshwater Daphnids)
Deviations:
no
GLP compliance:
yes

Test material

Constituent 1
Test material form:
liquid
Details on test material:
- Purity: >99%

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations: 5.0, 10.0, 20.0, 50.0 and 100.0 mg/L
- Sample storage conditions before analysis: Room temperature
- Stability or Test Solution and Chemical Analysis: Samples were taken from all control and test solutions at the beginning (0 d), immediately prior to the first renewal of the test solution (3 d) and weekly thereafter (9, 21 d). Fifty mL samples at different concentration were diluted to the 100 mL with acetonitrile and then analysed by the above UPLC-PDA method.

Test solutions

Vehicle:
yes
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The stock solution of 10.0 mg/L was prepared by dissolving 0.0100 g test substance into 1000 mL Elendt M7, stirring for 24 hours at 30°C to allow it to fully dissolve. Test solutions at different concentrations were prepared by adding the stock solution and diluting with Elendt M7.
- Controls: Blank control

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Water flea
- Food type: Chlorella vulgaris Beij
- Amount: 0.1-0.2 mg C/Daphnia each time
- Frequency: Once daily

Study design

Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
21 d

Test conditions

Hardness:
221 to 232 mg (CaCO3)/L
Test temperature:
20.5 to 21.2°C
pH:
7.42 ~ 7.81
Dissolved oxygen:
90-101% of air saturation
Nominal and measured concentrations:
10.0 mg/L (Nominal)
10.3 mg/L (Measured)
Details on test conditions:
TEST SYSTEM
- Test vessel: 100 mL glass beakers
- Aeration: None
- No. of organisms per vessel: 10 animals were individually held at each test concentration and in the control.

OTHER TEST CONDITIONS
- Photoperiod: 16-hour light and 8-hour dark cycle daily
- Light intensity: 1000 lux to 1500 lux

EFFECT PARAMETERS MEASURED:
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.

Other parameters: Including time production of first brood; number of broods per animal. Oxygen concentration, temperature, hardness and pH values were measured once a week in the control(s) and the highest test substance concentration.

TEST CONCENTRATIONS
- Range finding study: In the range finding test, animals were exposed to concentrations of 1.00 and 10.0 mg/L (nominal concentrations). 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 was conducted in limit test.

Results and discussion

Effect concentrationsopen allclose all
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
Remarks on result:
other: Measured concentration of 10.3 mg/L
Key result
Duration:
21 d
Dose descriptor:
LOEC
Effect conc.:
> 10 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Survival
Remarks on result:
other: Measured concentration of 10.3 mg/L
Key result
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
10 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Survival
Remarks on result:
other: Measured concentration of 10.3 mg/L
Details on results:
During the test, all animals in the blank control were alive and the number of live offspring produced per parent animal surviving at the end of the test was 66-83. The coefficient of variation around the mean number of living offspring produced per parent animal in the control(s) was 7.45% (<25%). At the end of the test period, no adults died in blank control and 10.0 mg/L treatments. Thus the LOEC for survival of adults was greater than nominal 10.0 mg/L (measured concentration 10.3 mg/L) and the NOEC was 10.0 mg/L (measured concentration 10.3 mg/L). According to the Independent Samples T-test analysis results of the test data (Independent Samples T-test, SPSS 17.0), the significance level of p was more than 0.05. Thus the reproduction rates in the treatments of 10.0 mg/L concentration treatments showed no statistically significant difference from that of the blank control P> 0.05). Thus the LOEC for the Daphnia magna reproduction was greater than nominal 10.0 mg/L (measured concentration 10.3 mg/L) and the NOEC was 10.0 mg/L (measured concentration 10.3 mg/L).

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
NOEC (Reproduction): 10.0 mg/L (measured concentration 10.3 mg/L)
LOEC (Survival): > 10.0 mg/L (measured concentration 10.3 mg/L)
NOEC (Survival): 10.0 mg/L (measured concentration 10.3 mg/L)
Executive summary:

Under semi-static conditions of 72 h renewal, the effect on the reproductive output of Daphnia magna exposed to the test substance for 21 days was conducted according to 'The guidelines for the testing of chemicals-effects on biotic systems" (HJ/T 153); "The Guidelines for the Testing of 2nd edition, 2013); Procedure 211 of the Daphnia magna, Reproduction Test", OECD Guidelines for Testing of Chemicals, 211.

During the test period, the pH values of the control and test media were between 7.42 and 7.81, the Dissolved Oxygen (DO) values varied from 90 to 101% of the air saturation at the test temperature, the hardness values varied from 221 to 232 mg (CaCO3)/L and the temperature of the control and test media was maintained in the range of 20.5 to 21.2°C. All animals in the control were alive and the number of live offspring produced per parent animal surviving at the end of the test was 66~83. The coefficient of variation around the number of living offspring produced per parent animal in the control(s) was 7.45% Therefore, the study met the acceptability criteria and was considered valid.

The concentration of the test substance in test medium was determined by UPLC-PDA to represent the stability of the test substance in test period, A linear regression equation was obtained with the mean peak squares (A) vs. the concentration of the test substance (c, mg/L): A=56710c+42719, with good linearity of r2 = 0.9999. The results show that linearity for the concentration range of 5.00 mg/L to 100 mg/L was good. The minimum detection amount of UPLC-PDA was 0.10 mg/L and the minimum detection concentration for water samples was 0.50 mg/L. The analytical results showed that the concentration of the test substance was consistent in the test medium throughout the test period (deviation within 20% of the initial concentration). Thus a semi-static method of 72h renewal test procedure was reasonable.

The results showed that under valid semi-static test conditions (72 h-renewal), for Daphnia magna reproduction and adult survival, the lowest observed effect concentration (LOEC) was greater than nominal 10.0 mg/L (measured concentration 10.3 mg/L) and the no observed effect concentration (NOEC) was 10.0 mg/L (measured concentration 10.3 mg/L).