Fyroflex SOL-DP

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP and appropriate guidelines

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD 203 and EC Directive 79/831, Annex V, Part C.1 with deviations (Water Accomodated Fraction (WAF) solution)

Deviations:

no

Principles of method if other than guideline:

The test item is poorly soluble in water and therefore Water Accomodated Fraction (WAF) of the test substance was tested. A toxicity test was performed using 100 and 1000 mg/l test substance and a control.

GLP compliance:

yes (incl. QA statement)

Test material

Reference

Name:

Unnamed

Type:

Constituent

Details on test material:

E-AF098T (Batch No. T# 176-3), a white powder. Stored at room temp, dry, closed container.

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
Not relevant

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: WAF 100 mg/l and 1000 mg/l
At the beginning of the test, samples of 10 ml were taken in triplicate from the WAF (100 and 1000 mg/l) and from the control solution, and transfered into 10 ml HPLC vials by pipette just before adding the test animals and at the end of the test. All sample bottles were stored at room temperature in the dark prior to chemical analysis.

Test solutions

Vehicle:

no

Details on test solutions:

The test solutions were prepared as water accomodated fractions (WAF) prepared from 0.5002 g test substance in 5000 ml of Dutch Standard Water (DSW) to make the 100 mg/l loading and from 4.9998 g test substance in 5000 ml of DSW to make the 1000 mg/l loading. The test substance was weighed into a beaker and rinsed into 5000 ml volumetric flask then filled up to the mark with DSW. Then the solution was mechanically stirred for 40.5 h. The prepared WAF was then allowed to settle overnight and, as particles in the 100 and 1000 mg/l WAFs were found to be both floating at the top of the solution and lying on the bottom of the volumetric flask, the test solutions were carefully siphoned off from the middle of the volumetric flasks into their respective test aquaria. The pH of the control and both WAF solutions were checked and found to be between 7.9 and 8.2. No adjustment was considered necessary.
No further dilutions of the stock solution were prepared during the test.

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

The fish were obtained from the ERCA in-house breeding facillity and were accepted according to the quality control criteria of the guidelines and Standard Operation Procedure E2 (8.3). The fish in stock were fed one to three times per day with commeciallly available dry, deep frozen food, or fresh nauplii. The feeding was stopped at 24h before the test was started, and animals were not fed during the test. A number of representative fish was measured for their length and weight to assess the biomass load. Average length: 1.82. Average weight: 0.098 g. The mean biomass at the limit test concentration of 100 mg/l: 0.22 g/l.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

no abnormalities observed

Test conditions

Hardness:

200 mg CaCl2*H2O

Test temperature:

22.9-23.1 °C

pH:

Control: 7.6- 8.2
Treatment (WAF 100 mg/L): 7.6- 7.9
Treatment (WAF 1000 mg/L): 7.8- 8.2

Dissolved oxygen:

Control: 8.3- 8.5 mg/l
Treatment (WAF 100 mg/L): 8.4- 8.6 mg/l
Treatment (WAF 1000 mg/L): 8.4- 8.6 mg/l

Salinity:

Not relevant

Nominal and measured concentrations:

Samples were quantified using a calibration curve of external standards. More details on analysis, see attched report, pp. 17-19.

Details on test conditions:

The test was performed as a limit test with 100 mg/l and a highest recommended concentration of 1000 mg/l based on nominal concentration of the product to demonstrate that the LC50 is higher than the tested concentration. A control containing only test medium was also included in the test.

The test medium contained per liter of de-ionized water (DSW): 100 mg of NaHCO3, 20 mg of KHCO3, 200 mg of CaCl2*2H2O and 180 mg of MgSO4*7H2O.

The fish were randomly placed in the test vessels which were positioned in the test room in a random manner. During the test the vessels were covered with glass plates. The test solutions present in the test vessels were aerated with water-saturated air purified by an active coal filter and a cotton filter during testing.
7 fish were used at the limit test concentration (100 mg/l) and control, and 3 fish were used at 1000 mg/l. The fish were exposed to the chosen concentrations of the test substance and sublethal effects were recorded at least within the first six hours and then at approximately 24, 48, 72 and 96 hours. In addition to death, sub-lethal effects such as erratic swimming, loss of reflex, increased excitability, lethargy, changes in physiology, discoloration, pigmentation, excessive mucus production, hyperventilation, opaque eyes, curved spine or hemorrhaging were recorded.
Since the test was performed as a limit test, the results on toxicity were directly read from the test observations.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

dissolved

Basis for effect:

other: mortality, appearance and behaivor

Effect concentrations 2

Duration:

24 h

Dose descriptor:

EC100

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

dissolved

Basis for effect:

other: mortality, appearance and behaivor

Effect concentrations 3

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

dissolved

Basis for effect:

other: mortality, appearance and behaivor

Effect concentrations 4

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

dissolved

Basis for effect:

other: mortality, appearance and behaivor

Effect concentrations 5

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

dissolved

Basis for effect:

other: mortality, appearance and behaivor

Effect concentrations 6

Duration:

72 h

Dose descriptor:

EC100

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

dissolved

Basis for effect:

other: mortality, appearance and behaivor

Effect concentrations 7

Duration:

96 h

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

dissolved

Basis for effect:

other: mortality, appearance and behaivor

Effect concentrations 8

Duration:

96 h

Dose descriptor:

EC100

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

dissolved

Basis for effect:

other: mortality, appearance and behaivor

Effect concentrations 9

Duration:

24 h

Dose descriptor:

NOEC

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

dissolved

Basis for effect:

other: mortality, appearance and behaivor

Effect concentrations 10

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

dissolved

Basis for effect:

other: mortality, appearance and behaivor

Effect concentrations 11

Duration:

72

Dose descriptor:

NOEC

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

dissolved

Basis for effect:

other: mortality, appearance and behaivor

Effect concentrations 12

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

dissolved

Basis for effect:

other: mortality, appearance and behaivor

Details on results:

Since the test was performed as a limit test, the results on toxicity were directly read from the test observations.
The results (see attached report, pp. 15-16) are based on the performed definitive test.
The results show that WAF of the test substance had no lethal effect on the fish at either the test concentration. No other sub-lethal effects were observed in the WAF.
The chemical analyses performed on the WAF of the test substance showed that only one of the two peaks selected for quantification was observed in the WAF and the retention time of this peak was identical to the retention time of triphenylphosphate (The test item contains <1% triphenylphosphate as a reaction product). The chemical analyses performed showed that the concentration of test substance in the WAF at 100 mg/l and at 1000 mg/l were all

Results with reference substance (positive control):

Not relevant

Reported statistics and error estimates:

See pages 17-19 in the attached report.

Any other information on results incl. tables

Sublethal observations / clinical signs:

No further information

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The results show that WAF of the test substance had no lethal effect on the fish at either the test concentration. No other sub-lethal effects were observed in the WAF.

Executive summary:

The purpose of the test was to determine the acute lethal toxicity of the Water Accommodated Fraction (WAF) of E-AF098T to the freshwater fish Danio rerio (Zebra fish) under continious exposure of 96 hours in static test. The results show that WAF of the test substance had no lethal effect on the fish at either the test concentration. No other sub-lethal effects were observed in the WAF.