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Ecotoxicological information

Short-term toxicity to fish

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Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From March to May 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study (OECD, EPA, etc)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report Date:
2012

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 850.1075 (Freshwater and Saltwater Fish Acute Toxicity Test)
Deviations:
no
Qualifier:
according to
Guideline:
other: ASTM Standard E 729-96: Standard Guide for Conducting Acute Toxicity Tests on Test Materials with Fishes, Macroinvertebrates and Amphibians
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): XP-7866
- Physical state: Solid
- Lot/batch No.: 10188-1JM
- Expiration date of the lot/batch: February 28, 2015
- Storage condition of test material: Stored under ambient conditions
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
PHYSICO-CHEMICAL PROPERTIES, refer to chapter 4 for details on test item.

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Sampling method: Samples were collected from newly prepared test solutions in each test chamber of each treatment and control group at the beginning of the test and at 72 hours, and from the 24-hour old solution in each test chamber at 24 hours and at the end of the test. The samples were collected at mid-depth, placed in glass vials.
- Sample storage conditions before analysis: Analysis was proceeded immediately

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method:
1) Primary stock solution in methanol, for a nominal concentration of 100 mg a.i./mL
2) The stock solution was sonicated for approximately 3 minutes and inverted at least 20 times to mix, and appeared clear and colorless with no visible precipitate. Individual test solutions were prepared directly in each clean empty stainless steel aquarium using a glass graduated pipette or glass graduated cylinder.
3) The methanol was evaporated off from the flasks.
4) To each aquarium 10 liters of pH adjusted well water (see characteristic below) was added to prepare the five nominal test concentrations of 20, 40, 80, 160 and 320 μg a.i./L.The test solutions were mixed using a top-down electric mixer for approximately 40 minutes to 1 hour.
- Controls: Dilution water
The dilution water was prepared at the beginning of the test and at each renewal period during the test by adding aliquots of HEPES buffer to the 180-L stainless steel tank filled with well water and mixed with an electric top-down mixer until the final pH of approximately 7 was achieved. The total amount of HEPES buffer used at each preparation was 115, 125 or 130 g.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): methanol for the primary stock solution but evaporated in later stage (refer method above)
- Evidence of undissolved material (e.g. precipitate, surface film, etc): no - All test solutions appeared clear and colorless.
Note: New test solutions were prepared daily and surviving fish were transferred to the new test solutions at approximately 24 hours.

Test organisms

Test organisms (species):
Pimephales promelas
Details on test organisms:
TEST ORGANISM
- Common name: Fathead minnow - Pimephales promelas
- Source: Received as juveniles from Chesapeake Cultures of Hayes, Virginia.
- Age at study initiation (mean and range, SD): juvenile fish
- Length at the end of the study: The average total length of 10 negative control fish measured at the end of the test was 2.6 cm, with a range of 2.5 to 2.9 cm.
- Weight at the end of the study: The average wet weight (blotted dry) of 10 negative control fish measured at the end of the test was 0.16 grams, with a range of 0.14 to 0.21 grams.
- Feeding during test: The fish were not fed for at least two days prior to the test or during the test.

ACCLIMATION
- Acclimation period: At least 14 days prior to the test in water
- Acclimation conditions:
* water temperatures in the cultures ranged from 21.6 to 23.0°C, measured with a liquid-in-glass thermometer.
* pH of the water ranged from 8.3 to 8.6, measured with a Thermo Orion Thermo Orion Benchtop 4 Star Plus pH meter.
* Dissolved oxygen concentrations were > 8.0 mg/L (>92% of saturation), measured with a Thermo Orion Benchtop 3 Star Plus dissolved oxygen meter.
- Type and amount of food: Fed with a commercially-prepared diet supplied by Sera North America, Montgomeryville, Pennsylvania, supplemented with brine shrimp nauplii (Artemia sp.) supplied by Brine Shrimp Direct, Ogden, Utah.
- Feeding frequency: Not reported
- Health during acclimation (any mortality observed): During the 2-week period prior to the test the fish showed no signs of disease or stress.

Study design

Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h

Test conditions

Hardness:
144 Expressed in mg/L as CaCO3 - range: 140 - 148
(N = 4)
Test temperature:
22 +/-1°C
pH:
ranged from 7.3 to 7.9
Dissolved oxygen:
>= 6.4 mg/L (>= 74% of saturation)
Nominal and measured concentrations:
- Nominal concentrations: 20, 40, 80, 160 and 320 µg a.i./L
- Measured concentrations average: 11, 23, 34, 75 and 153 µg a.i./L, reprensenting 55, 58, 43, 47 and 48 % of nominal concentration.
Note: Measured concentrations of the samples collected from the newly prepared test solutions at 0 and 72 hours ranged from approximately 20.2 to 70.7% of nominal. Measured concentrations of the samples collected from the old test solutions at 24 and 96 hours ranged from approximately 37.7 to 76.9% of nominal.
Details on test conditions:
TEST SYSTEM
- Test vessel: Test chambers were 25 or 38-L stainless steel aquaria filled with 10 liters of test water.
- Material, size, headspace, fill volume: The depth of the test water in representative 25 and 38-L test chambers was 12 and 7.5 cm, respectively.
- Aeration: Not reported
- Type of flow-through (e.g. peristaltic or proportional diluter): semi-static condition
- Renewal rate of test solution (frequency/flow rate): 24 hours intervals
- No. of organisms per vessel: 10 fish in each test chamber
- No. of vessels per concentration (replicates): two replicates
- No. of vessels per control (replicates): two replicates
- No. of vessels per vehicle control (replicates): No vehicle control

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Obtained from a well approximately 40 meters deep located on the Wildlife International, Ltd. site. The well water was passed through a sand filter to remove particles greater than approximately 25 µm, and pumped into a 37,800-L storage tank where the water was aerated with spray nozzles. Prior to use, the water was filtered to 0.45 µm to remove fine particles.
- Analyses of pesticides, organics and Metals of water were conducted by Lancaster Laboratories on samples collected on December, 28, 2011. Details are available in the original report.

OTHER TEST CONDITIONS
- Adjustment of pH: the ph of the dilution water was adjusted to pH neutral
- Photoperiod: 16 hours of light and 8 hours of darkness. A 30-minute transition period of low light intensity was provided when lights went on and off to avoid sudden changes in lighting.
- Light intensity: ambient laboratory light Fluorescent = 106 lux at the surface of the water - Light bulbs that emit wavelengths similar to natural sunlight. Light intensity was measured at the water surface of one representative test chamber at test initiation using a SPERScientific Model 840006C light meter.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
* mortality
* signs of toxicity
were made approximately 1, 24, 48, 72 and 96 hours (±1 hour) after test initiation.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: Geometric series of 5 test concentration and a negative control (dilution water)
- Justification for using less concentrations than requested by guideline: No justification
- Test concentrations: Nominal concentrations: 20, 40, 80, 160 and 320 µg a.i./L
Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Duration:
24 h
Dose descriptor:
LC50
Effect conc.:
> 153 µg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
act. ingr.
Basis for effect:
mortality
Remarks on result:
other: could not be calculated with the mortality data obtained
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
> 153 µg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
act. ingr.
Basis for effect:
mortality
Remarks on result:
other: could not be calculated with the mortality data obtained
Duration:
72 h
Dose descriptor:
LC50
Effect conc.:
> 153 µg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
act. ingr.
Basis for effect:
mortality
Remarks on result:
other: could not be calculated with the mortality data obtained
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 153 µg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
act. ingr.
Basis for effect:
mortality
Remarks on result:
other: could not be calculated with the mortality data obtained
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
> 153 µg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
act. ingr.
Basis for effect:
mortality
Remarks on result:
other: could not be calculated with the mortality data obtained
Details on results:
Fathead minnows exposed to XP-7866 at concentrations ≤153 μg a.i./L appeared normal with no mortality or overt signs of toxicity.
Results with reference substance (positive control):
No reference substance
Reported statistics and error estimates:
There was less than 50% mortality in any of the XP-7866 treatment groups during the test which precluded the statistical calculation of LC50 values at 24, 48, 72 and 96 hours. Therefore, the LC50 values were estimated to be greater than the highest concentration tested. The no-mortality concentration and the no-observed-effect concentration (NOEC) were determined by visual interpretation of the mortality and observation data.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
Fathead minnows (Pimephales promelas) were exposed for 96 hours under static-renewal conditions to five mean measured concentrations of XP-7866 ranging from 11 to 153 μg a.i./L. The 96-hour LC50 value was >153 μg a.i./L, the highest concentration tested. The 96-hour no-mortality concentration and the NOEC were 153 μg a.i./L.