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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
29 December 2017 - 16 January 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report Date:
2018

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Version / remarks:
13 April 2004
Deviations:
no
Qualifier:
according to
Guideline:
other: Guidance document on aquatic toxicity testing of difficult substances and mixtures, OECD series on testing and assessment number 23
Version / remarks:
2000
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid
Details on test material:
- Appearance: Light tan solid
- Storage conditions: At room temperature protected from light
Specific details on test material used for the study:
- Test item handling: use amber glassware or wrap container in aluminum-foil.
- The test item is soluble in water.

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
Samples were taken from the highest test concentration (100 mg/L) and the control according to the schedule below.
Frequency: at t=0 and t=48 h
Volume: 1.0 and 2.0 mL
Storage: Samples were stored in a freezer (≤ -15°C) until analysis.

At the end of the exposure period, the replicates were pooled at each concentration before sampling.

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION:
- Method: direct application to test medium; weighing of the test item and preparation of the test solution was performed under dimmed light.
The test solution was prepared at a loading rate of 100 mg test item/L applying a 30 minute period of magnetic stirring to accelerate dissolution of the test item in medium. Lower test concentrations were prepared by subsequent dilutions of the highest concentration in test medium.
- Evidence of undissolved material: no, all test solutions were clear and colorless at the end of the preparation procedure.
- Controls: test medium without test item or other additives (blank-control)

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Daphnia magna
- Strain: Straus, 1820
- Source: in-house laboratory culture with a known history. At least third generation, obtained by a cyclical parthenogenesis under specified breeding conditions.
- Age of parental stock: Daphnids originated from a healthy stock, 2nd to 5th brood, showing no signs of stress such as mortality >20%, presence of males, ephippia or discoloured animals and there was no delay in the production of the first brood.
- Age at study initiation: < 24 hours, from parental daphnids of more than two weeks old.
- Feeding during test: no

ACCLIMATION
- Acclimation period: no

BREEDING:
- Method: each batch was started with newborn daphnids, i.e. less than 3 days old, by placing about 250 of them into 5 litres of medium (M7) in an all-glass culture vessel. After 7 days of cultivation half of the medium was renewed twice a week.
- Feeding: daily, a suspension of fresh water algae.

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h

Test conditions

Hardness:
180 mg CaCO3/L
Test temperature:
19-20 °C (throughout the test)
pH:
7.8-8.1 (throughout the test, in the control vessel)
7.2-7.8 (throughout the test, in the test solution)
Dissolved oxygen:
At the start of the test: 9.5-9.7 mg O2/L (in all vessels)
At the end of the test: 8.6-8.7 mg O2/L (in all vessels)
Nominal and measured concentrations:
Nominal concentrations: 0.10, 1.0, 10 and 100 mg/L
Measured concentrations at 100 mg/L (cation): 109 mg/L at t=0; 109 mg/L at t=48
Measured concentrations at 100 mg/L (anion): 115 mg/L at t=0; 114 mg/L at t=48
The measured concentrations at the end of the study were 100% (cation) and 99% (anion) of initial measured concentrations at t=0.
Details on test conditions:
TEST SYSTEM
- Test vessel: 60 mL, all-glass, fill volume: 50 mL
- Aeration: no
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4 for the highest concentration, 2 for the other concentrations
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Standard medium used: yes, adjusted ISO medium
- Source/preparation of dilution water: tap water purified by reverse osmosis
- Culture medium different from test medium: yes, M7 was used as culture medium

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours, daily
- Other: the test was performed in the dark

EFFECT PARAMETERS MEASURED: mobility at 24 and 48 hours.
- Additional measurements: pH and dissolved oxygen at the beginning and at the end of the test, for the highest concentration and the control. Temperature of the medium: continuously in a temperature control vessel.

COMBINED LIMIT/RANGE-FINDING STUDY
- Test concentrations: 0.10, 1.0, 10 and 100 mg/L
- Results used to determine the conditions for the definitive study: as only 1 daphnia was immobilised at the highest concentration after 48 hours, the combined limit/range-finding study was used as the final study and effect parameters were based on the results of this study.
Reference substance (positive control):
yes
Remarks:
potassium dichromate (January 2018)

Results and discussion

Effect concentrations
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
- Analysis showed that test concentrations remained stable during the exposure concentrations: test concentrations after 48 hours were 99-100% of the initial measured concentrations at t=0 (see table 1). Since measured test concentrations were comparable to nominal concentrations (109-115% of nominal), the effect parameter was based on the nominal concentrations.
- At the end of the exposure concentration, only one daphnid was immobilised in the highest concentration and therefore the EC50 was determined to exceed the highest test concentration (i.e. >100 mg/L). No immobilisation was observed at any of the other test concentrations.
- The pH and temperature during the study remained within the limits prescribed by the study plan.
Results with reference substance (positive control):
- Results with reference substance valid? Yes
- Tested concentrations: 0.10, 0.18, 0.32, 0.56, 1.0 and 1.8 mg/L
- 48h-EC50: 0.54 mg/L, 95%-confidence interval: 0.49-0.61 mg/L
- Other: results fell within the historical range.
Reported statistics and error estimates:
No analysis was conducted as the test item proved to be non-toxic at the concentrations tested.

Any other information on results incl. tables

Table 1 Measured concentrations in the test samples - cation

Time of sampling
[hours]

Concentration
[mg/L]

Relative to nominal
[%]

Relative to initial
[%]

Nominal

Analyzed

0

0

n.d.

n.a.

 

 

100

109

109

 

48

0

n.d.

n.a.

 

 

100

109

109

100

n.d. Not detected; n.a. Not applicable

Table 2 Measured concentrations in the test samples - anion

Time of sampling
[hours]

Concentration
[mg/L]

Relative to nominal
[%]

Relative to initial
[%]

Nominal

Analyzed

0

0

n.d.

n.a.

 

 

100

115

115

 

48

0

n.d.

n.a.

 

 

100

114

114

99

n.d. Not detected; n.a. Not applicable

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Remarks:
See 'overall remarks'
Conclusions:
In an acute toxicity study, performed according to OECD 202 and GLP principles, the 48-h EC50 of BMS-482204-03 towards Daphnia magna exceeded the highest tested concentration (i.e. >100 mg/L).