1,3,5-Triazine-2,4-diamine, N2-[(1R,2S)-2,3-dihydro-2,6-dimethyl-1H-inden-1-yl]-6-(1-fluoroethyl)-

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1035 (Mysid Acute Toxicity Test)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A primary stock solution was prepared at a nominal concentration of 50 mg a.i./mL by dissolving a calculated amount of AE 1170437 into dimethyl formamide (DMF). The stock solution was mixed for approximately 30 seconds of sonication followed by inversion, and appeared clear and tan in color.
Aliquots of the 50 mg a.i./mL stock solution were proportionally diluted with DMF to prepare secondary stock solutions.
- Chemical name of vehicle: Dimethyl formamide (DMF)

Test organisms (species):

Americamysis bahia (previous name: Mysidopsis bahia)

Details on test organisms:

TEST ORGANISM
- Common name: saltwater mysid
- Source: Cultures maintained by Wildlife International, Ltd., Easton, Maryland. Identification of the species was verified by the supplier of the original culture stock. The original culture was received on April 5, 2007 from Aquatic Bio Systems, Fort Collins, CO 80524.
- Age at study initiation: <24 hours old
- Feeding during test
- Food type: live brine shrimp Artemia nauplii
- Frequency: daily

ACCLIMATION
- Acclimation conditions (same as test or not): Adult mysids were held in water from the same source and at approximately the same
temperature as used during the test.

Test type:

static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

24.0 to 25.3°C

pH:

7.8 to 8.2

Dissolved oxygen:

5.9 to 6.8 mg/L (≥80% of saturation)

Salinity:

19-20 parts per thousand (‰)

Nominal and measured concentrations:

Nominal concentrations: 0.31, 0.63, 1.3, 2.5 and 5.0 mg a.i./L
Measured concentrations: 0.26, 0.52, 1.1, 1.9 and 4.0 mg a.i./L, representing 84, 83, 85, 76 and 80% of nominal concentrations, respectively.

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 2-L glass beakers containing approximately 1.5 L of test solution.
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- No. of vessels per vehicle control (replicates): 2


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: according to guideline (The water used for culturing and testing was natural seawater collected at Indian River Inlet, Delaware that was filtered and diluted to a salinity of approximately 20‰ with well water.)


OTHER TEST CONDITIONS
- Photoperiod: 16 hours of light and 8 hours of darkness
- Light intensity: 205 lux

EFFECT PARAMETERS MEASURED: Observations of mortality and other signs of toxicity were made approximately 4, 24, 48, 72 and 96 hours after test initiation.

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1.5 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

mortality

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

1.1 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

mortality

Saltwater mysids, Americamysis bahia, were exposed under static conditions for 96 hours to five mean measured concentrations of AE 1170437 ranging from 0.26 to 4.0 mg a.i./L. The 96-hour LC50 value was 1.5 mg a.i./L, with conservative 95% confidence limits of 1.1 and 1.9 mg a.i./L. The nomortality concentration and NOEC were 1.1 mg a.i./L.

Results:

Mean Measured				4 Hours	24 Hours		48 Hours		72 Hours		96 Hours		Cumulative Percent Mortality
Concentration (mg a.i./L)	Replicat e	No. Exposed	No. Dead1	Obs.2	No. Dead1Obs.2		No. Dead1Obs.2		No. Dead1Obs.2		No. Dead1Obs.2
Negative Control	A	10	0	10 AN	0	10 AN	0	10 AN	0	10 AN	0	10 AN	0
	B	10	0	10 AN	0	10 AN	0	10 AN	0	10 AN	0	10 AN
Solvent Control	A	10	0	10 AN	0	10 AN	0	10 AN	0	10 AN	0	10 AN	0
	B	10	0	10 AN	0	10 AN	0	10 AN	0	10 AN	0	10 AN
0.26	A	10	0	10 AN	0	10 AN	0	10 AN	0	10 AN	0	10 AN	5
	B	10	0	10 AN	0	10 AN	1	9 AN	1	9 AN	1	9 AN
0.52	A	10	0	10 AN	0	10 AN	0	10 AN	0	10 AN	0	10 AN	0
	B	10	0	10 AN	0	10 AN	0	10 AN	0	10 AN	0	10 AN
1.1	A	10	0	10 AN	0	10 AN	0	10 AN	0	10 AN	0	10 AN	0
	B	10	0	10 AN	0	10 AN	0	10 AN	0	10 AN	0	10 AN
1.9	A	10	0	9 AN; 1 A	1	2 AN; 4 A; 3 E	5	5 E	7	3 E	8	2 E	90
	B	10	0	6 AN; 4 A	0	3 AN; 2 A; 4 E; 1 C	1	9 E	7	3 E	10	-
4.0	A	10	1	4 E; 5 C	10	-	-	-	-	-	-	-	100
	B	10	0	10 E	10	-	-	-	-	-	-	-

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1010 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: FIFRA 72-2

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Prior to the exposure numerous solutions were prepared, both with and without the use of a solvent stock, in an effort to determine the functional solubility of AE 1170437 in waters used for aquatic testing. Solutions prepared without a solvent were not soluble (noted visually). After
working with various stock solutions the limit of solubility in dilution water was determined to be approximately 10 mg a.i./L using a solvent load of 0.5 ml/L. The test material did not go into solution at 10 mg a.i./L at solvent loads below 0.5 ml/L.
- Controls: yes
- Chemical name of vehicle: Acetone

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Waterflea
- Source: one breeding; original source was Aquatic Biosystems Inc., Fort Collins, CO
- Age at study initiation: < 24 hours old at initiation

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

172 to 178 mg/L

Test temperature:

20 ± 1.0 °C

pH:

8.1 to 8.4

Dissolved oxygen:

7.9 to 8.4 mg/L (87 to 92 % saturation)

Nominal and measured concentrations:

Nominal Test Concentrations: Control, Solvent Control, 0.63, 1.25, 2.5, 5.0 and 10 mg a.i./L
Mean Measured Test Concentrations: Control (<0.06), Solvent Control (<0.06), 0.62, 1.23, 2.54, 4.86 and 9.88 mg a.i./L

Details on test conditions:

TEST SYSTEM
- Test vessel: 250-mL borosilicate glass beakers
- Fill volume: Approximately 200 ml
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): 4


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: according to guideline

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 hours light, 8 hours dark
- Light intensity: 40 - 70 footcandles, Approximately 430 - 750 lux

EFFECT PARAMETERS MEASURED: Immobilization and sublethal behavioral effects

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 9.88 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

mobility

Remarks on result:

other: (practical limit of solubility in test water)

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

4.86 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

9.88 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

mobility

Results:

Sublethal Observations
	Frequency
Mean Measured Test Concentration (mg a.i./L)	(maximum observed during scheduled observation events)
Control	0
Solvent Control	0
0.62	0
1.23	0
2.54	0
4.86	0
9.88	16
Immobilization
Mean Measured Test Concentration (mg a.i./L)	Percent Immobilized at 48-Hours
Control	0%
Solvent Control	0%
0.62	0%
1.23	0%
2.54	0%
4.86	0%
9.88	35%
Immobilization included organisms that were dead.

CONCLUSION
Based on immobilizations:
48 Hour NOEC	4.86 mg a.i./L
48 Hour LOEC	9.88 mg a.i./L
48 Hour EC50	>9.88 mg a.i./L (practical limit of solubility in test water)

Description of key information

The test substance is toxic to saltwater invertebrates but shows a lower toxicity to freshwater invertebrates.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

9.88 mg/L

Marine water invertebrates

Marine water invertebrates

Effect concentration:

1.5 mg/L

Additional information

Three studies with aquatic invertebrates are available, one with a freshwater and two with a saltwater organism. The short term test with Daphnia magna according to OECD 202 revealed an 48-hour EC50 in Daphnia magna greater than 9.88 mg /L test item (practical limit of solubility). The NOEC was 4.86 mg / a.i. L. Saltwater mysids, Americamysis bahia, were exposed under static conditions for 96 hours and the LC50 was 1.5 mg a.i./L. The NOEC was 1.1 mg a.i./L.

Eastern oysters (Crassostrea virginica) were exposed under flow-through conditions for 96 hours to six mean measured concentrations of the test substance ranging from 0.062 to 1.8 mg a.i./L. The test was conducted at concentrations near the functional limit of solubility of the test substance in a saltwater system. There were no mortalities or clinical signs of toxicity observed at any concentration tested. Based on inhibition of shell deposition, the 96-hour EC50 value was 1.0 mg a.i./L, with a 95% confidence interval of 0.71 to 1.2 mg a.i./L. The NOEC was determined to be 0.47 mg a.i./L.

The test substance is toxic to saltwater invertebrates but shows a lower toxicity to freshwater invertebrates.