Reaction products of disodium 3,5-diamino-2-[(2-sulfonatophenyl)diazenyl]benzoate and  diazotised sodium 2-[(4-aminophenyl)sulfonyl]ethyl sulfate

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Experiment start date - 27 January 2003; Experiment end date - 21 March 2003; Study completion date - 09 April 2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identity: FAT 40810/A
Batch: WP 6/02
Purity: approx. 75 %
Appearance: Solid, dark brownish powder
Expiration date: 12 December 2010
Storage: At room temperature at about 20 °C

Analytical monitoring:

yes

Details on sampling:

Specimens of each test concentration were drawn from the approximate centre of the test vessels. They were taken at the beginning and the end of the exposure period.

Vehicle:

no

Details on test solutions:

Identity and concentration of auxiliary solvent for dispersal: No auxillary solvent was used.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Test species Zebra fish (Danio rerio)
Origin Commercial supplier: Universitat Zürich-lrchel, lnstitut für Hirnforschung, 8057 Zurich, Switzerland. Fish supplied in one batch were all of a similar age and size.
Holding: Fish were held in well water of “Lange Erlen” and “Hard” and fed daily (except for weekends) with a commercially prepared zebra fish food diet: Mat Nr. 228221 DE, Tretra Werke 49304 Melle, Germany. Fish used in the test were not fed 48 hours prior to initiation of the test. Quality of the fish was assessed during an acclimatisation period of at least 12 days in well water. Fish were not treated for disease during holding.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

178 mg CaCO3/L

Test temperature:

21.5 - 22.3 °C

pH:

7.8 - 8.3

Dissolved oxygen:

93 - 101 % of saturation

Nominal and measured concentrations:

The analytically determined test item concentrations in the analysed test media varied from 94 % to 92 % of the nominal values between start and end of exposure. The test item FAT 40810/A was sufficiently stable in the test media under the test conditions during the test period of 96 hours. Hence, all biological results are related to the nominal concentration of the test item. As no effects were seen at 100 mg/L concentration, no further doses were tested and the test was terminated as limit test.

Details on test conditions:

The oxygen content (% of the saturation value), pH and temperature (°C) were measured in each test chamber at 0, 24, 48, 72 and 96 hours (as long as any fish remained in the tank). The temperature and the dissolved oxygen were measured with a WT\N OXI 330i dissolved oxygen- and temperature meter. The pH was measured with a Knick Portameter 751. ln addition, total hardness (as CaCO3) and chlorine content were measured monthly in representative specimens of the dilution water. For the determination of the total hardness a complexometric titration (Aquamerck 1 .08011.0001) with Titriplex Ill and a mixed indicator were used, the chlorine content was determined using the Aquamerck kit 1.11160. At 2 - 4, 24, 48, 72 and 96 hours, observations of mortality and sublethal symptoms, such as abnormal behavioural activity and stress were made (swimming behaviour, loss of equilibrium, respiratory function, pigmentation, exophtalmos and other observations). Dead fish, if any, were removed from the test solutions at least at the above mentioned intervals. At test termination all the fish were sacrificed by exposure to tricaine methane sulphonate (MS 222), the fish were retained for measurements of weight and length.
The test item appeared homogeneously distributed in the test vessel over the period of the test. No precipitation was observed in the test solution throughout the duration of the test.

TEST SYSTEM
- Test vessel:
Glass vessel
- Material, size, headspace, fill volume:
Glass, 5 litres with test volume of 3 litre
- Aeration:
Yes
- No. of organisms per vessel:
7 fish per concentration and control.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:
Well water of "Lange Erlen" and "Hard" water used as dilution medium. Prior to its used the water was passed through an activated carbon filter.
- Total organic carbon:
0.4 mg/L
- Chlorine:
(< 0.1 ppm)
- Alkalinity:
277 - 323 mg/L
- Culture medium different from test medium:
No


OTHER TEST CONDITIONS
- Photoperiod:
Fluorescent light, 16 hours daily with 30 min transition period.


EFFECT PARAMETERS MEASURED:
Mortality

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Basis for effect:

mortality (fish)

Details on results:

No mortality was observed in any fish.

Results with reference substance (positive control):

not applicable

Reported statistics and error estimates:

Due to the absence of mortality during the exposure, the median lethal concentration (LC50) was estimated without a statistical program.

Sublethal observations / clinical signs:

The test was considered valid since:

- no fish were dead in the control at the end of the test

- the dissolved oxygen content throughout the test was >60 % of the air saturation value.

Validity criteria fulfilled:

yes

Conclusions:

Acute fish toxicity of FAT 40810/A to Zebrafish (Danio rerio), 96 h exposure: LC50 >100 mg/L, LC100 >100 mg/L, LC0 = 100 mg/L

Executive summary:

The objective of this study was the estimation of the acute toxicity of FAT 40810/A to zebrafish in a 96-hour static test. The toxic effect was determined by  assessing  the mortality of the fish caused by the test item and expressed as the median lethal concentration at which 50 % of the fish population dies (LC50). The study was conducted in accordance with OECD test guideline 203 and EU Method C.1 in a GLP-certified laboratory. The test concentration was 100 mg/L. No mortality of fish was recorded in the control and the nominal test concentration of 100 mg/L FAT 40810/A during the 96-hour exposure period. No sublethal effects such as changes in swimming behaviour, respiratory function, pigmentation, exophtalmos or loss of equilibrium were observed in the control or the treatment concentration of 100 mg/L during the 96 hour test period. The LC values (96h) for FAT 40810/A based on the nominal concentrations were determined as follows: LC50 >100 mg/L, LC100 >100 mg/L, LC0 = 100 mg/L.

Description of key information

Acute fish toxicity of FAT 40810/A to Zebrafish (Danio rerio), 96 h exposure: LC50 >100 mg/L, LC100 >100 mg/L, LC0 = 100 mg/L

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

The objective of this study was the estimation of the acute toxicity of FAT 40810/A to zebrafish in a 96-hour static test. The toxic effect was determined by assessing the mortality of the fish caused by the test item and expressed as the median lethal concentration at which 50 % of the fish population dies (LC50). The study was conducted in accordance with OECD test guideline 203 and EU Method C.1 in a GLP certified laboratory. The test concentration was 100 mg/L. No mortality of fish was recorded in the control and the nominal test concentration of 100 mg/L FAT 40810/A during the 96-hour exposure period. No sublethal effects such as changes in swimming behaviour, respiratory function, pigmentation, exophtalmos or loss of equilibrium were observed in the control or the treatment concentration of 100 mg/L during the 96 hour test period. The LC values (96h) for FAT 40810/A based on the nominal concentrations were determined as follows: LC50 > 100 mg/L, LC100 > 100 mg/L, LC0 = 100 mg/L