Registration Dossier

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
16 Apr 2007 - 29 Jun 2007
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Guideline study under GLP, test substance could not be maintained in solution

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2007
Report Date:
2007

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:
other: ISO International Standard 7346-2
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, December 14, 2000.
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): MTDID 7145
- Substance type: Multiconstituent substance
- Physical state: Clear, colourless liquid
- Analytical purity: 94.5%
- Lot/batch no.: 142072:43
- Expiration date of the lot/batch: 30 Dec 2007
- Stability under test conditions: Stable
- Storage condition of test material: At room temperature in the dark

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations: WAFs prepared at loading rates of 1.0, 10 and 100 mg/L for the rangefinding test, and 100 mg/L only for the limit test.
- Sampling method: Aliquots (10 mL) were taken from the appoximate center of the test vessels. Samples were taken of freshly-prepared test solution at start of test and at 72 hours, and of spent solution (before renewal) at 24 hours and at test completion.
- Sample storage conditions before analysis: Samples were analyzed day of sampling

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Water accomodated fraction. Test solutions were prepared daily. Loading rates of 1.0, 10 and 100 mg/I were stirred for approximately 1 hour in air-tight vessels. After a stabilisation period of 5-25 minutes, the Water Accommodated Fraction (WAF) was collected by siphoning and used for the test. The final test solutions were all clear and colourless.
- Controls: Blank
- Evidence of undissolved material (e.g. precipitate, surface film, etc): None

Test organisms

Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: Danio rerio (Zebrafish)
- Source: Hubrecht lab, Utrecht, Netherlands. All fish were first generation from a single pair in UV-treated water.
- Length at study initiation (length definition, mean, range and SD): rangefinder, 3.2 ± 0.2 cm. Limit test, 2.8 ± 0.2 cm
- Weight at study initiation (mean and range, SD): rangefinder, 0.57 ± 0.09 g. Limit test, 0.43 ± 0.10 g
- Feeding during test: No

ACCLIMATION
- Acclimation period: More than 12 days
- Acclimation conditions (same as test or not): Same as test
- Type and amount of food: Pelleted fish food (Nutra 3.0 T, TROUW NUTRITION, Putten, The Netherlands)
- Feeding frequency: Daily until 24 hours prior to the start of the test.
- Health during acclimation (any mortality observed): less than 5% mortaliy in the seven days prior to the test

Study design

Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h

Test conditions

Hardness:
180 mg CaCO3 / L
Test temperature:
21.4 °C - 22.2 °C
pH:
7.3 - 8.0
Dissolved oxygen:
7.0 - 8.9 mg/L
Salinity:
NA
Nominal and measured concentrations:
WAF based on 100 mg/L loading rate
Details on test conditions:
TEST SYSTEM
- Test vessel: 10-L sealed glass containers. Fill volume was 9.5 L.
- Aeration: None
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): one
- No. of vessels per control (replicates): one
- Biomass loading rate: 0.32 g/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Water used to make growth medium was purified by reverse osmosis.
- Culture medium different from test medium:no, standard ISO medium in both cases

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 h:8 h light:dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Mortality and other effects: Daily

TEST CONCENTRATIONS
- Spacing factor for test concentrations: One concentration tested
- Justification for using less concentrations than requested by guideline: Limit test
- Range finding study: Yes
- Test concentrations: WAFs based on 0.1 mg/L, 1 mg/L and 10 mg/L loading rates
- Results used to determine the conditions for the definitive study: Yes. No mortality at any concentration tested.
Reference substance (positive control):
yes
Remarks:
Pentachlorophenol, reference test done 1 week prior to FC-770 study

Results and discussion

Effect concentrations
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 0.096 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
act. ingr.
Basis for effect:
mortality
Remarks on result:
other: No mortality. Analytical result using 100 mg/L loading rate
Details on results:
No mortality or other clinical effects were observed during the test period.

- Any observations e.g. precipitation) that might cause a difference between measured and nominal values: Concentration of test substance was < LOQ in all blanks and in 24-hour-old test solutions sampled before replacement (see Figures 1 and 2, attached). Test substance concentration is the arithmetic mean of the 0-24 hour and 72-96 hour geometric mean concentrations. The 0-24 hour and 72-96-hour concentrations are the geometric means of initial and final concentrations. Half the limit of quantitation (0.100 mg/L) was used as the final concentration at 24 and 96 hours. Concentrations of fresh solutions were 0.159 mg/L and 0.214 mg/L at 0 and 72 hours, respectively.

- Effect concentrations exceeding solubility of substance in test medium: Yes
Results with reference substance (positive control):
In a test of pentachlorophenol conducted 1 week prior to the FC-770 test, an LC50 of 0.22 mg/L was observed. The range of 96-h LC50 values for zebrafish is generally between 0.10 mg/l and 0.46 mg/l based on historical data of reference tests performed by the testing lab.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Remarks:
No control mortality, DOC > 60% of saturation, measured concentrations used.
Conclusions:
The LC50 of FC-770 to Danio rerio (zebrafish) was >0.096 mg/L. The NOEC was 0.096 mg/L
Executive summary:

The toxicity of FC-770 to Danio rerio (zebrafish) was assessed in a semi-static test conducted according to OECD TG203. The test was performed as a limit test after a rangefinding test indicated no toxicity in water accomodated fractions (WAFs) made using loading rates of 0.10, 1, and 10 mg/L. The limit test used a WAF made with a loading rate of 100 mg/L. FC-770 concentrations in 24 hour-old test solutions were below the limit of quantitation. Because concentrations could not be stably maintained, the average of the geometric means of the fresh and 24-hour old solutions at 0-24 hours and 72-96 hours was used to establish a concentration of 0.096 mg/L in the exposed fish. No toxicity was observed in the exposed fish. Due to the low solubility of the test substance in water combined with its volatility, concentration levels that might be acutely toxic for zebrafish could not be reached. The LC50s of FC-770 to Danio rerio was >0.096 mg/L. The NOEC was 0.096 mg/L.

The study was conducted under accepted guidelines and according to GLP criteria. The concentration of FC-770 could not be maintained above the limit of quantitation. However, reasonable efforts were made to obtain and maintain the highest possible concentration over the whole test duration. Therefore, the study is classified as reliable with restrictions.

Results Synopsis

Test organism weight/length: 0.43 g / 3.2 cm

Test Type: Semi-Static

LC50:  >0.096 mg/L                        95% C.I.:  NA

NOEC:  0.096 mg/L                 Probit Slope:  NA