Formaldehyde, reaction products with 5,12-dihydroquino[2,3-b]acridine-7,14-dione and 3,5-dimethyl-1H-pyrazole, sulfonated

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2007-12-31 to 2008-02-09

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline Study (OECD 203) including analytical monitoring.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: JMAFF Test Guidelines, 2-7-1-1, Acute fish toxicity, 2005

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: limit concentration and controls
- Sampling method: The water temperature, pH and dissolved oxygen concentrations were measured at the start of the test and once every day during the test at the single test concentration and in the control. At the same dates the appearance of the test medium was recorded. For the analysis of the actual test item concentrations the following samples were taken:
Just before test start:
- duplicate samples from the test medium
- one of the duplicate samples from the control
after 48 and 96 hours:
- duplicate samples from the test medium
- duplicate samples from the control at 48 hours (one of the duplicate control samples at 96 hours)
- Sample storage conditions before analysis: All samples were taken from the approximate center of the aquaria without mixing the test media, and were deep-frozen (at about -20 °C) immediately after sampling. Based on pre-experiments for investigation of the storage stability (without GLP) the test item is sufficiently stable in the test water under the storage conditions.

Test solutions

Vehicle:

no

Details on test solutions:

For the preparation of the test medium, a dispersion of the test item with the loading rate of 100 mg/L was prepared. The dispersion was stirred for 3 hours to dissolve a maximum amount of the different compounds of the test item in the dispersion. Then, the dispersion was filtered through a membrane filter (0.45 μm pore size) and the undiluted filtrate was tested as WAF.

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- the study was performed with zebra fish (Brachydanio rerio).
- the test fish were raised in the RCC laboratories without any medication.
- From the acclimated test fish batch, 10 fish were measured at the start of the test.
- The mean body length of the fish was 2.9 ± 0.14 cm (Mean ± SD), the mean body wet weight was 0.23 ± 0.06 g (Mean ± SD).

ACCLIMATION
- Prior to test start, they were acclimated for one week to the test water and temperature.
- During holding and acclimatization until one day before the start of the test the fish were fed with a commercial fish diet .
- During the last two weeks prior to the test no fish died in the test fish batch and all fish were healthy.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Test conditions

Hardness:

125 mg/L as CaCO3

Test temperature:

21 +/- 0 °C

pH:

7.2 - 7.4

Dissolved oxygen:

8.0 - 8.4 mg/L

Nominal and measured concentrations:

Since a WAF of the test item with sparingly soluble components was tested, all reported biological results are based on the loading rate of 100 mg/L. The analytically measured test item concentration in the test medium was in the range of 0.97 to 1.3 μg/L during the test period of 96 hours.

Details on test conditions:

- One glass test vessel with 5 liters of test medium was used for each treatment (the single test concentration and the control).
- At the start of the test 7 fish were introduced into each test vessel in a random order.
- The loading rate was less than 1 g fish wet weight per liter test medium.
- The test medium and the control were slightly aerated during the test period.
- The fish were not fed during the test.

A static test without test medium renewal was performed under the following conditions:
- Test duration: 96 hours
- Water temperature: 21 °C during the test period.
- Light conditions: A 16-hour light to 8-hour dark photoperiod, with a 30-minute transition period (light intensity during the light period was approximately within the range of 100 to 560 Lux).
- Reconstituted test water was used in the study.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: WAF (Water Accomodated Fraction); no acute toxic effects within the range of solubility.

Effect concentrations 2

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: LC50 was estimated; no acute toxic effects within the range of solubility.

Details on results:

A limit test was performed in accordance with the test guidelines to demonstrate that water accommodated fractions (WAFs) of the test item have no toxic effect on the test fish up to the loading rate of 100 mg/L. Thus, the only treatment tested was a WAF with the loading rate of nominal 100 mg/L in parallel to a control.

Results with reference substance (positive control):

NA

Reported statistics and error estimates:

NA

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

With high probability not acutely harmful to fish. No acute toxic effects within the range of solubility.