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Short-term toxicity to aquatic invertebrates

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Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
6 Mar - 26 Apr 2017
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
other: main test not completed
Reference:
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
GLP compliance:
yes
Test material information:
Composition 1
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Appearance: off-white waxy solid
- Source and lot/batch No.of test material: 629582
- Expiration date of the lot/batch: 9 Dec 2017
- Purity: 98.3%
- Purity test date: 14 Jun 2016

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature
- Stability under test conditions: Substance is hydrolytically active
Analytical monitoring:
yes
Details on sampling:
- Concentrations: 2.0 mL
- Sampling method: During the test single 2 mL samples were taken from the center of the test vessels. Samples were taken at the start of the test and at 48 hours.
- Sample storage conditions before analysis: Samples were transferred to the analytical laboratory at the Test Facility and analysed on the day of sampling.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: All glassware used was silanized to minimise adsorption. Preparation of test solutions started with a loading rate of 100 mg/L applying a two-day period of magnetic stirring to ensure maximum dissolution of the test item in medium. This resulted in a hazy mixture with undissolved particles present as floating droplets. The aqueous Saturated Solution (SS) was collected by means of filtration through a 0.45 μm membrane filter (RC55, Whatman) and used as the highest test concentration. Lower test concentrations were prepared by subsequent dilutions of the SS in test medium.
- Controls: no
- Evidence of undissolved material: All test solutions were clear and colorless at the end of the preparation.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Daphnia magna (Straus, 1820), at least third generation, obtained by a cyclical parthenogenesis under specified breeding conditions
- Source: In-house laboratory culture with a known history.
- Age of parental stock: greater than 2 weeks but no more than 4 weeks
- Age at study initiation: < 24 hours
- Feeding during test: No
- Culturing: Adult daphnids were cultured in M7 medium (adjusted ISO medium plus trace elements, micronutrients, and vitamins). Adult daphnids in the cultures were not more than 4 weeks old prior to collection of the juveniles for testing. During the period preceding the test, water temperatures were 18 - 22°C.
Daphnids in the cultures were fed daily a suspension of freshwater algae. The adults were fed prior to test initiation, but neonates were not fed during the test.
- Health: 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.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Hardness:
180 mg CaCO3/L
Test temperature:
20 - 21 °C
pH:
6.9 - 8.1
Dissolved oxygen:
8.5 - 8.9
Nominal and measured concentrations:
Nominal: 1.0, 10, 100% saturated solution
Measured: 0.0042, 0.011, and 0.080 μg/L (at start of test, extrapolated from calibration curve)
Details on test conditions:
TEST SYSTEM
- Test vessel: 100 mL all-glass, silanized
- Type: open
- Fill volume: 80 mL
- Aeration: no
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4 for 100% SS concentration, 2 for 1.0 and 10% SS concentration
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: adjusted ISO medium prepared using reverse-osmosis purified water (GEON Waterbehandeling, Berkel-Enschot, The Netherlands) with the following concentrations:
CaCl2.2H2O 211.5 mg/L
MgSO4.7H2O 88.8 mg/L
NaHCO3 46.7 mg/L
KCl 4.2 mg/L
- Culture medium different from test medium: Yes, culture medium was M7 (adjusted ISO medium with addition of trace elements, micronutrients, and vitamins).

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light; 8 hours dark

EFFECT PARAMETERS MEASURED: immobilization (including mortality) observed at 24 and 48 hours
Reference substance (positive control):
yes
Remarks:
Potassium dichromate (K2CrO7)
Key result
Duration:
48 h
Dose descriptor:
EC50
Remarks on result:
not measured/tested
Remarks:
Information became available showing that the test item is subject to rapid hydrolysis
Details on results:
Table 1 shows the responses recorded during the range-finding test. No immobility was observed at the test concentration representing 1.0% SS, while all daphnids exposed to 10 and 100% SS were immobilized within 24 hours of exposure; therefore, the expected EC50 was between test groups representing 1.0 and 10% SS. However, the 48-hour EC50 for Daphnia magna exposed to MTDID 15670 was not determined. The project was stopped in consultation with the sponsor. Information became available showing that the test item is subject to rapid hydrolysis. The sponsor decided to perform additional testing on the hydrolysis products instead.
Results with reference substance (positive control):
- Results with reference substance valid? Yes
- Dose-response test: Yes
- ECx: 48-hr EC50 = 0.59 mg/L (95% CI, 0.52 mg/L to 0.66 mg/L)
- Other: The sensitivity of Daphnia magna originating from the batch used in this test, for potassium dichromate fell within the expected range of sensitivity observed during previous years (n > 60; 48-hr EC50 between 0.3 and 1.0 mg/L.) The reference substance test was conducted within three months of the testing of MTDID 15670.
Validity criteria fulfilled:
not applicable
Conclusions:
The 48-hour EC50 for Daphnia magna exposed to MTDID 15670 in a range-finding test according to OECD 202 was not determined. The project was stopped before the range-finding tests were completed. Information became available showing that the test item is subject to rapid hydrolysis.
Executive summary:

A range-finding test preliminary to a full study as per OECD 202 was done. The 48-hour EC50 for Daphnia magna exposed to MTDID 15670 was not determined. The project was stopped before the range-finding test series for all species was completed. Information became available showing that the test item is subject to rapid hydrolysis. Because the test was not completed, the reliability of the results could not be assigned.

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Reference:
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
GLP compliance:
yes
Test material information:
Composition 1
Specific details on test material used for the study:
Dodecyl methyl sulfide is used within this study as a surrogate for dodecyl ethyl sulfide, which is a hydrolysis product of MTDID 15670. Dodecyl ethyl sulfide is not commercially available.
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Sigma-Aldrich, Batch Number MKBW6054V
- Physical state: colorless, liquid
- Quality Release Date: 12JAN2016
- Purity: 97.8%
- Storage condition of test material: room temperature
Analytical monitoring:
yes
Details on sampling:
- Concentrations: negative control and saturated solution
- Volume: 3 mL for the regular samples and 1.5 mL for quality control (QC) samples
- Sampling method: Duplicate samples for analysis, and duplicate reserve samples, were taken from the approximate center of the test vessels. Two extra samples were taken from the limit concentration for analytical QC, and two reserve QC samples were also taken. Samples were taken at test initiation (T = 0 h) and test completion (T= 48 h).
- Sample storage conditions before analysis: Samples were transferred to the analytical laboratory at the test facility on the day of sampling for extraction then stored in a freezer (≤ -15°C) until analyses.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The batch of dodecyl methyl sulfide tested was a colorless liquid which was not completely soluble in test medium at the initially prepared loading rate. Preparation of test solutions started with a loading rate of 100 mg/L (200.1 mg in 2 liters) applying a two-day period of gentle magnetic stirring to ensure maximum dissolution of the test item in medium. The obtained mixture was allowed to settle for an overnight period. Thereafter, the aqueous Saturated Solution (SS) was collected by means of siphoning and used as the highest test concentration.
- Controls: Blank only
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): Test solutions were clear and colorless at the end of the preparation procedure.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Water flea
- Strain/clone: Daphnia magna (Straus, 1820), at least third generation, obtained by a cyclical parthenogenesis under specified breeding conditions.
- Source: In-house laboratory culture with a known history.
- Age of parental stock: > 2 weeks to < 4 weeks
- Age at study initiation: < 24 hrs
- Feeding during test: No
- Culturing: Adult daphnids were cultured in M7 medium (adjusted ISO medium plus trace elements, micronutrients, and vitamins). Adult daphnids in the cultures were held for at least 14 days, and were not more than 4 weeks old prior to collection of the juveniles for testing. During the period preceding the test, water temperatures were 18 - 22°C. Daphnids in the cultures were fed daily a suspension of freshwater algae. The adults were fed prior to test initiation, but neonates were not fed during the test.
- Health: Daphnids originated from a healthy stock, 2nd to 5th brood, showing no signs of stress suchas mortality >20%, presence of males, ephippia or discoloured animals and there was no delay in the production of the first brood.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Hardness:
180 mg/L CaCO3
Test temperature:
19 °C to 21°C
pH:
8.0 to 8.3
Dissolved oxygen:
8.7 mg/L to 9.4 mg/L
Nominal and measured concentrations:
Nominal: 100 mg/L loading level (100% SS), 56% SS, 32% SS, 18% SS, 10% SS, Control (Blank)
Measured: Only the 100 mg/L (100% SS) loading level and the blank were analyzed. At 100 mg/L measured concentrations were estimated in the range of 0.0087-0.011 mg/L at the start of the test, which decreased below the LOD (0.0066 mg/L) after 48 hours. In the control samples, measured concentrations were below the LOD (0.0066 mg/L). Based on the measured initial concentrations in the 100 mg/L loading samples from this test and previous test samples, an estimated actual concentration of 0.01 mg/L was used for expressing effect parameters.
Details on test conditions:
TEST SYSTEM
- Test vessel: 60 mL glass vessels
- Fill volume: 50 mL volume
- Aeration: no
- No. of organisms per vessel: five
- No. of vessels per concentration (replicates): four
- No. of vessels per control (replicates): four
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:adjusted ISO medium prepared using reverse-osmosis purified water (GEON Waterbehandeling, Berkel-Enschot, The Netherlands)
CaCl2.2H2O - 211.5 mg/L
MgSO4.7H2O - 88.8 mg/L
NaHCO3 - 46.7 mg/L
KCl - 4.2 mg/L
- Culture medium different from test medium: Yes, culture medium was M7 (adjusted ISO medium with addition of M7 trace elements, micronutrients, and vitamins).
OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 hours daily
EFFECT PARAMETERS MEASURED: immobilization (including mortality) observed at 24 and 48 hours

RANGE-FINDING STUDY
- Test concentrations: 0 mg/L (control) and saturated solution (100 mg/L loading rate)
- Results used to determine the conditions for the definitive study: yes
Reference substance (positive control):
yes
Remarks:
Potassium dichromate (K2CrO7)
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
Remarks on result:
other:
Remarks:
based on loading level of 100 mg/L, which is above the water solubility of dodecyl methyl sulfide.
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 0.01 mg/L
Nominal / measured:
estimated
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other:
Remarks:
estimated from measured initial concentrations from several tests
Details on results:
Test was conducted twice.
First test:
- Mobility: no immobilization observed (One Daphnid was observed trapped at the surface of the test solutions at 24 hours. This organism were reimmersed into its respective solution before recording of mobility.)
- Mobility of control: no immobilization observed
- Other adverse effects: no other signs of disease or stress
Second Test:
- Mobility: no immobilization observed (Daphnids were observed trapped at the surface of the test solutions at 24 and 48 hours. These organisms were reimmersed into the respective solutions before recording of mobility.)
- Mobility of control: no immobilization observed (At 24 hours one daphnid was observed trapped at the surface of a control vessel. This organism was reimmersed into the solution before recording of mobility.)
- Other adverse effects: no other signs of disease or stress

- Effect concentrations exceeding solubility of substance in test medium: yes
Results with reference substance (positive control):
- Results with reference substance valid? Yes
- Dose-response test: Yes
- ECx: 48-hr EC50 = 0.33 mg/L (95% CI, 0.29 mg/L to 0.38 mg/L)
- Other: The sensitivity of Daphnia magna originating from the batch used in this test, for potassium dichromate fell within the expected range of sensitivity observed during previous years (n > 60; 48-hr EC50 between 0.3 and 1.0 mg/L.) The reference substance test was conducted within one month of the testing of MTDID 22327.
Validity criteria fulfilled:
yes
Remarks:
Control immobilization was < 10% at the end of the test. The dissolved oxygen concentration was ≥ 3 mg/L in the control and test vessels at the end of the test.
Conclusions:
The 48-hour EC50 (immobilization) of dodecyl methyl sulfide to Daphnia magna exceeded a loading rate of 100 mg/L, which exceeded the maximum soluble concentration for dodecyl methyl sulfide in test medium. The test was conducted according to OECD TG 202.
Executive summary:

The 48-hour EC50 of dodecyl methyl sulfide to Daphnia magna was examined in a static test conducted according to OECD TG 202. Dodecyl methyl sulfide is used as a surrogate for dodecyl ethyl sulfide (not commercially available), a hydrolysis product of MTDID 15670. Two tests were conducted in which test solutions were prepared as the control (0 mg/L), a 100 mg/L loading level (water soluble fraction), along with dilutions of the 100 mg/L water soluble fraction. No immobilization of daphnids was observed in the controls, or any of the test solutions in either test. Dodecyl methyl sulfide was measured in initial samples of the 100 mg/L water soluble fraction, but were below the Limit of detection (0.0066 mg/L) at 48 hrs, and an estimated level of 0.01 mg/L dodecyl methyl sulfide was established for expressing the effect parameter (EC50 > 0.01 mg/L). The test was conducted according to internationally accepted test guidelines and was GLP compliant. It is reliable with restriction and suitable for Risk Assessment, Classification and Labeling, and PBT Analysis.

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
27 Feb - 3 Mar 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reference:
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
GLP compliance:
yes
Test material information:
Composition 1
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- State of substance: clear colourless liquid
- Source and lot/batch No.of test material: Sigma-Aldrich Chemie GmbH, Steinheim, Germany, batch: STBD0697V
- Purity: 99.8%, mixture of cis and trans

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature
Analytical monitoring:
yes
Details on sampling:
- Concentrations: control (test medium without test item) and 100 mg/L
- Sampling method: Duplicate samples (6.0 mL) for analysis were taken from both test groups at the start of the test and at 48 hours. In addition, two quality control samples were taken from the limit concentration at the start and the end of the test period. Additionally, duplicate reserve samples of 6.0 mL were taken for possible analysis. If not used, these samples were extracted and stored in a freezer (≤-15°C) for a maximum of three months after delivery of the draft report, pending on the decision of the sponsor for additional analysis.
- Sample storage conditions before analysis: Samples were transferred to the analytical laboratory at the Test Facility on the day of sampling for extraction and then stored in a freezer (≤-15°C) until analyses.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Preparation of test solutions started with the highest concentration of 100 mg/L applying 10 minutes 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.
- Controls: Test medium without test item or other additives
- Evidence of undissolved material: All test solutions were clear and colorless at the end of the preparation procedure.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Daphnia magna (Straus, 1820), at least third generation, obtained by a cyclical parthenogenesis under specified breeding conditions
- Source: In-house laboratory culture with a known history.
- Age of parental stock: > 2 weeks
- Age at study initiation: < 24 hours
- Feeding during test: No
- Culturing: Adult daphnids were cultured in M7 medium (adjusted ISO medium plus trace elements, micronutrients, and vitamins). Adult daphnids in the cultures were at least 2 but not more than 4 weeks old prior to collection of the juveniles for testing. During the period preceding the test, water temperatures were 18 - 22°C. Daphnids in the cultures were fed daily a suspension of freshwater algae. The adults were fed prior to test initiation, but neonates were not fed during the test.
- Health: 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.
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Hardness:
180 mg CaCO3/L
Test temperature:
19 - 21 °C
pH:
7.9 - 8.1
Dissolved oxygen:
9.2 - 9.8 mg/L
Nominal and measured concentrations:
Nominal: 100 mg/L
Measured: 83 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: 60 mL glass vessel
- Type: open
- Fill volume: 50 mL
- Aeration: no
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4 at 100 mg/L (2 at lower concentrations)
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: adjusted ISO medium prepared using reverse-osmosis purified water (GEON Waterbehandeling, Berkel-Enschot, The Netherlands) with the following composition:
CaCl2.2H2O 211.5 mg/L
MgSO4.7H2O 88.8 mg/L
NaHCO3 46.7 mg/L
KCl 4.2 mg/L
- Culture medium different from test medium: Yes, culture medium was M7 (adjusted ISO medium with addition of trace elements, micronutrients, and vitamins).

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light; 8 hours dark

EFFECT PARAMETERS MEASURED: immobilization (including mortality) observed at 24 and 48 hours

RANGE-FINDING STUDY
- Test concentrations: 0.1, 1.0, 10, and 100 mg/L, contained within limit test
Reference substance (positive control):
yes
Remarks:
Potassium dichromate (K2CrO7)
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 83 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 10% effect at limit concentration, not considered biologically relevant
Details on results:
- Behavioural abnormalities: At the end of the test, 10% immobility was observed at the limit concentration.
- Other biological observations: In the 10 mg/Land 100 mg/L concentrations, some daphnia were trapped at the surface of the test solution. The organisms were reimmersed into the respective solutions before recording immobility.
- Mortality of control: none
- Other adverse effects control: none
Results with reference substance (positive control):
- Results with reference substance valid? Yes
- Dose-response test: Yes
- ECx: 48-hr EC50 = 0.33 mg/L (95% CI, 0.29 mg/L to 0.38 mg/L)
- Other: The sensitivity of Daphnia magna originating from the batch used in this test, for potassium dichromate fell within the expected range of sensitivity observed during previous years (n > 60; 48-hr EC50 between 0.3 and 1.0 mg/L.) The reference substance test was conducted within one month of the testing of crotononitrile.
Validity criteria fulfilled:
yes
Remarks:
No daphnids became immobilized in the control during the test. The dissolved oxygen concentration was ≥ 3 mg/L in the control and test vessels at the end of the test.
Conclusions:
The 48-hour EC50 of crotononitrile, a hydrolysis product of MTDID 15670, to Daphnia magna was > 83 mg/L (OECD 202).
Executive summary:

The 48-hour EC50 of crotononitrile, a hydrolysis product of MTDID 15670, to Daphnia magna was examined in a range-finding/limit static test conducted according to OECD TG 202. Test solutions were prepared as the control (0 mg/L), 0.1 mg/L, 1 mg/L, 10 mg/L, and 100 mg/L. At the end of the test, 10% immobility was observed at the highest test level, which had geometric mean concentration of 83 mg/L. This immobilization is not considered biologically relevant. The EC50 is therefore > 83 mg/L. The study was well-documented, followed an international standard method, and was GLP compliant; therefore, the test is considered reliable without restriction.

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
04 - 07 Mar 2002
Reliability:
3 (not reliable)
Rationale for reliability incl. deficiencies:
significant methodological deficiencies
Remarks:
The method of making the test solutions, sonication, is expected to produce toxic byproducts
Reference:
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Version / remarks:
1984
Deviations:
no
GLP compliance:
yes
Test material information:
Composition 1
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 114211
- Expiration date of the lot/batch: 30 Apr 2002
- Storage condition of test material: At room temperature away from direct sunlight
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Ultrasonication for 15 minutes, followed by intense stirring for 10 minutes at room temperature
- Controls: Test water without test substance
- Chemical name of vehicle: No vehicle
- Evidence of undissolved material: All test solutions were clear and colorless throughout the test
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Strain/clone: Daphnia magna Straus, Clone 5, originally obtained from the University of Sheffield, UK 10 years prior to the test
- Source: Internal stock
- Age: At the start of the test, organisms were 6-24 hours old and were not first brood progeny
- Feeding during test: no

ACCLIMATION
- Acclimation period: None, organisms were cultured under conditions (reconstituted water quality, light, temperature) identical to that used in the test
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Hardness:
250 mg/L as CaCO3
Test temperature:
21 °C
pH:
7.9 - 8.0
Dissolved oxygen:
8.2 - 9.0 mg/L
Nominal and measured concentrations:
Nominal only: Control, 0.006, 0.019, 0.061, 0.196, 0.629 and 2.0 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: 100 mL covered glass beakers filled with 50 mL test medium
- Type: closed
- Aeration: None
- No. of organisms per vessel: Ten
- No. of vessels per concentration (replicates): Two
- No. of vessels per control (replicates):Two

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted test water made from analytical grade salts and purified water.
CaCl2 x 2H2O: 2.0 mM (= 294.0 mg/L)
MgSO4 X 7H2O: 0.5 mM (= 123.0 mg/L)
NaHCO3: 0.75 mM (= 65.0 mg/L)
KCI: 0.075 mM (= 5.8 mg/L)

- Alkalinity: 0.8 mM
- Ca/Mg ratio: 4:1 (molarity)

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16-h light: 8-h dark with 30 minute transition period.
- Light intensity: 200 - 1200 Lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobilization at 24 and 48 hours

RANGE-FINDING STUDY
- Results used to determine the conditions for the definitive study: Test concentrations based on rangefinder, results not in study report
Reference substance (positive control):
no
Remarks:
Potassium dichromate is tested at least annually to verify system
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
0.11 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 95% CI: 0.08 - 0.16 mg/L
Details on results:
- Mortality of control: None
- Other adverse effects control: None
- Abnormal responses: None
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: None. Test solutions were clear and colorless throughout the test.
- Effect concentrations exceeding solubility of substance in test medium: No

Table 1, Effects during the Daphnia acute toxicity test

Nominal concentration (mg/L)

Number organisms tested

Number immobile at 24 h

Percent immobile at 24 h

Number immobile at 48 h

Percent immobile at 48 h

Control (0)

20

0

0

0

0

0.006

20

0

0

0

0

0.019

20

0

0

0

0

0.06

20

0

0

6

30

0.196

20

0

0

16

80

0.629

20

17

85

18

90

2.0

20

17

85

20

100

Validity criteria fulfilled:
yes
Remarks:
<10% immobilization in controls (0%), ≥3 mg/L DO in all test vessels (≥8.2 mg/L)
Conclusions:
48-h EC50 (immobilization) of MTDID 15670 to D. magna was 0.11 mg/L (OECD 202).
Executive summary:

The 48-h EC50 (immobilization) of MTDID 15670 to D. magna was determined according to OECD 202.  Test substance concentration was not confirmed analytically. An EC50 of 0.11 mg/L was determined using nominal concentrations. The test substance is hydrolytically active and is expected to have formed toxic byproducts during sonication. Without analytical confirmation, actual exposures cannot be determined. Therefore, this study is considered not reliable.

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
test procedure in accordance with national standard methods with acceptable restrictions
Reference:
Composition 0
Qualifier:
equivalent or similar to
Guideline:
other: EPA 600/4-90/027F, Methods for measuring the acute toxicity of effluents and receiving waters to freshwater and marine organisms.
Deviations:
not specified
GLP compliance:
no
Test material information:
Composition 1
Specific details on test material used for the study:
Sodium tetrafluoroborate obtained from Sigma-Aldrich, St. Louis, MO, USA
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
Stock solutions were prepared in water. Test concentrations (1 mg/L to 10,000 mg/L) were prepared by pipetting required volumes of stock solutions into known volumes of test water.
Test organisms (species):
Daphnia magna
Details on test organisms:
Daphnia magna neonates (age, 24 h) were born from parthogenic females grown in batch cultures maintained in groundwater-filled laboratory aquaria at 20 ± 2°C and a 16:8-h light:dark photoperiod with a 15-min transition period. Stock animals were originally obtained from Carolina Biological Supply (Burlington, NC, USA). Cultures were fed a mixture of pond water and Spirulina Dry Powder Microalga® (Petaluma, CA, USA).
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Test temperature:
20 ± 1 °C in laboratory
pH:
recorded, but not reported
Dissolved oxygen:
recorded, but not reported
Nominal and measured concentrations:
Nominal concentrations ranged from 1 mg/L to 10,000 mg/L, but individual solution concentrations were not reported.
Details on test conditions:
TEST SYSTEM
- Test vessel: 250 mL glass beakers
- No. of organisms per vessel: eight
- No. of vessels per concentration (replicates): five
- No. of vessels per control (replicates): five
OTHER TEST CONDITIONS
- Photoperiod: 16:8 h light:dark
EFFECT PARAMETERS MEASURED: The number of living and dead neonates was noted at 24 and 48 h after the initiation of the test. Neonates observed as motionless and without a discernable heartbeat were considered to be dead.

Key result
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
4 766 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other:
Remarks:
95% confidence interval, 4228 mg/L to 5323 mg/L
Reported statistics and error estimates:
The LC50 value as well as the associated 95% confidence intervals (CIs) were obtained by fitting the dose–response curves to a normal model using the maximum-likelihood method. All analyses were performed using SAS® Version 8.02 statistical software (SAS, Cary, NC, USA).
Conclusions:
The 48-hour LC50 of sodium tetrafluoroborate (the sodium salt of tetetrafluoroboric acid) to Daphnia magna is 4766 mg/L. Tetrafluoroborate is the anion of MTDID 15670, and forms immediately on dissociation.
Executive summary:

The toxicity of sodium tetrafluoroborate (the sodium salt of tetrafluoroboric acid), to Daphnia magna was examined in a static test reported in a publication. Tetrafluoroborate is the anion of MTDID 15670, and forms immediately on dissociation. The test was reported to have been run according to United States Environmental Protection Agency (U.S. EPA) protocols, and the study seemed to have been conducted appropriately, although some details of the study were not reported. A 48-hr LC50 of 4766 mg/L (95% CI, 4228 - 5323 mg/L) was reported. Although some details of the study were not reported, the study was conducted according to national standard methods, and is considered reliable with acceptable restrictions. The result may be used in a weight of evidence for risk assessment, Classification & Labelling, and PBT analysis.

Description of key information

MTDID 15670 hydrolyzes rapidly (T1/2 < 12 hrs) in water. Results from short-term toxicity tests (OECD TG 202) with the hydrolysis products of MTDID 15670 (or a surrogate chemical) all indicate that with Daphnia magna the 48-hour EC50 is > 100 mg/L.

Key value for chemical safety assessment

Additional information

A preliminary short-term toxicity to aquatic invertebrate study (OECD 202) with MTDID 15670 for Daphnia magna was initiated, but stopped after consultation with the sponsor, after encountering phase separation (precipitation) during the range-finding test. Some immobility was observed in the range-finding test, however the EC50 was not determined. Subsequently, information became available showing that MTDID 15670 is subject to rapid hydrolysis. Because the test was not completed, the reliability of the results could not be assigned. Analysis after hydrolysis revealed three products; two chemicals with moderate water solubility, crotononitrile (CAS# 4786-20-3) and tetrafluoroborate anion (tetrafluoroboric acid, CAS# 16872-11-0), along with the poorly soluble chemical, dodecyl ethyl sulfide (No CAS#). Short-term toxicity testing results for these hydrolysis products follow.

The 48-hour EC50 of crotononitrile, to Daphnia magna was examined in a rangefinding/static limit test conducted according to OECD TG 202. Test solutions were prepared as the control (0 mg/L), 0.1 mg/L, 1 mg/L, 10 mg/L, and at 100 mg/L. Ten percent immobilization was seen at the 100 mg/L loading rate, which is not considered biologically relevant. The measured concentration at the highest loading rate was 83 mg/L. The study was well-documented, followed an international standard method, and was GLP compliant; therefore, the test is considered reliable without restriction.

The 48-hour EC50 of dodecyl methyl sulfide (CAS# 3698-89-3) to Daphnia magna was examined in duplicate static, limit tests conducted according to OECD TG 202. Dodecyl methyl sulfide is a surrogate for dodecyl ethyl sulfide (not commercially available). Test solutions were prepared as the control (0 mg/L) and as a water soluble fraction at a loading rate of 100 mg/L. No immobilization was observed at the 100 mg/L loading rate or in the control. Dodecyl methyl sulfide was measured in initial samples of the 100 mg/L water soluble fraction, but were below the limit of detection (0.0066 mg/L) at 48 hrs, and an estimated level of 0.01 mg/L dodecyl methyl sulfide was established for expressing the effect parameter (EC50 > 0.01 mg/L). The study was well-documented, followed an international standard method, and was GLP compliant. The study is considered reliable with restrictions. The results from this study are considered suitable for Risk Assessment, Classification and Labeling, and PBT Analysis.

The toxicity of tetrafluoroboric acid, to Daphnia magna was examined as sodium tetrafluoroborate in a test conducted according USEPA protocols. The study seemed to have been conducted appropriately, although some details of the study were not reported. A 48-hr LC50 of 4766 mg/L (95% CI, 4228 - 5323 mg/L) was reported. Although some details of the study were not reported, the study was conducted according to national standard methods, and is considered reliable with acceptable restrictions. The result may be used in a weight of evidence for risk assessment, Classification & Labelling, and PBT analysis.

In addition, an earlier test on the parent compound showed unexpectedly high toxicity. However, the method of making test solutions, ultrasonication, is believed to have created toxic thiol byproducts that were responsible for the observed effects. The test lacked analytical confirmation of dose, and the actual exposures could not be determined. The study was deemed unreliable and is not considered further.