Tetrasodium 5-[4-chloro-6-(N-ethyl-anilino)-1,3,5-triazin-2-ylamino]-4-hydroxy-3-(1,5-disulfonatonaphthalen-2-ylazo)-naphthalene-2,7-disulfonate

Toxicity to soil macroorganisms except arthropods

Link to relevant study record(s)

Reference

Endpoint:

toxicity to soil macroorganisms except arthropods: short-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

June 14,1993 to August 23,1993

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 207 (Earthworm, Acute Toxicity Tests)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: European community official journal No. L 133,87/302/EEC, Part C (May 30, 1988)

Qualifier:

according to guideline

Guideline:

other: Guideline of Biologische Bundesanstalt (Berlin-Dahlem) and Umweltbundesamt to perform tests according to § 9 Abs. 1 No.8 of the Chemikaliengesetz (Chemicals Act, FRG, March 1990)

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No. of test material:
SR 6947/14
- Expiration date of the lot/batch:
July 31 1995

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material:
at room temperature in dark
- Stability under test conditions:
in solvent, DMSO, DMF >48 h
- Solubility of the test substance in the solvent/vehicle:
water: >100 g/l
org. solvents: DMSO, DMF >10 g/l

Analytical monitoring:

no

Vehicle:

no

Details on preparation and application of test substrate:

A stock solution of 5 mg/mL of the test substance was prepared with bidestilled water. According to the results of the range finding test, the stock solution was diluted with bidestilled water to prepare the following nominal concentrations of the test substance in the artificial soil:
- 0.1 mg/kg,
- 1.0 mg/kg
- 10.0 mg/kg
- 100.0 mg/kg
- 1000.0 mg/kg
All test concentrations and the control were tested in four replicates.

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

The study was performed with earthworms Eisenia foetida foetida bred by RCC under standardised conditions (see OECD 207). Only adult worms (older than two month) with clitellum and body weight of 300-600 mg were used; they could acclimate to artificial soil for 24 h before test start.

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Total exposure duration:

14 d

Post exposure observation period:

No

Test temperature:

The test vials were exposed in an air conditioned room at 20 ± 2 °C.

pH:

5.6-5.7

Moisture:

The water content was 35.8 % - 36.7 % (dry weight) at start and in a range from 33.9 % to 35.5 % at the end of the study.

Details on test conditions:

Artificial soil for tests was prepared according to OECD 207 before the study started and stored until use in a closed plastic-container. Ultimately before starting the experiments, the test substance was dissolved in bidistilled water and then added to the artificial soil (laboratory mixer). In course of applying the test substance, the soil was ventilated and moistened to ca. 35% of dry weight with demineralised (mQ) water. Water content and pH were controlled at start and finish of the study for each concentration. Test vials were normal bottling jars (1 L), which were loosely covered by glass-lids to enable exchange of air and to minimise evaporation of the artificial soil.
At test start, ten worms were placed on the surface of the artificial soil of each test vial (four replicates per concentration/control). The continuous illumination was measured to be 400 - 700 Lux during the entire study. The vials were controlled at days 7 and 14 respectively.
At test start, ten worms were placed on the surface of the artificial soil of each test vial (four replicates per concentration/control). The test vials were exposed in an air conditioned room at 20 ± 2 °C. The continuous illumination was measured to be 400 - 700 lux during the entire study. The vials were controlled at days 7 and 14 respectively.

Nominal and measured concentrations:

Values are based on nominal concentrations: 0.1, 1, 10, 100 and 1000 mg/kg of dry weight of soil.

Reference substance (positive control):

yes

Remarks:

Chloracetamide, Merck, D- 64293 Darmstadt, FRG, Art.No. 802412.

Duration:

7 d

Dose descriptor:

LC50

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: None

Duration:

14 d

Dose descriptor:

LC50

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: None

Details on results:

The results of this study did not reveal any toxic effects of FAT 40406/A to Eisenia foetida. Only two of the 240 worms involved in the experiment died within the 14 days of experimental time and this mortality could not be related to an influence of the test substance. According to these results it was not possible to calculate the LC50 (14 d) of FAT 40406/A to Eisenia foetida, which in artificial soil clearly must be more than 1000 mg/kg. In all test concentrations, the body weights of the surviving worms decreased slightly due to the starvation of the animals within the test period. No behavioural abnormalities could be observed. Summarizing the results it can be concluded that FAT 40406/A did not cause any acute toxic effects to Eisenia foetida in artificial soil up to a concentration of 1000 mg/kg, the highest tested concentration in this study.

Results with reference substance (positive control):

The last check conducted in November 1992 of the whole test system with standard compound (Chloracetarnide, Merck, D- 64293 Darmstadt, FRG, Art.No. 802412) resulted in a LC50 (14 days) of 15.2 mg/kg artificial soil (95 % confidence range: 12.5- 18.5 mg/kg).

Reported statistics and error estimates:

As no results were observed up to the highest concentration tested, no statistics was performed.

Validity criteria fulfilled:

yes

Conclusions:

LC50 of FAT 40406/A for 7 and 14 days was determined to be >1000 mg/kg of dry weight of soil.

Executive summary:

In a 14 days lasting study the mortality of Eisenia foetida under the impact of the test substance was determined according to OECD 207 guideline and in compliance with GLP regulations (including certificate). The results of this study did not reveal any acute toxic effect of the test substance to E. foetida, i.e. LC50 after 7 and 14 days could not be quantified as they were both >1000 mg/kg. The body weights of the worms decreased slightly in all concentrations and controls. No behavioural abnormalities of the surviving worms were observed. During the entire study, water content and pH of the artificial soil were within the ranges given by the guideline.

Description of key information

The 14-day LC50 of Reactive Red 245 is >1000 mg/kg soil dw in earthworms.

Key value for chemical safety assessment

Short-term EC50 or LC50 for soil macroorganisms:

1 000 mg/kg soil dw

Additional information

In a 14 days lasting study the mortality of Eisenia foetida under the impact of the test substance was determined according to OECD 207 guideline and in compliance with GLP regulations (including certificate). The results of this study did not reveal any acute toxic effect of the test substance to E. foetida, i.e. LC₅₀ after 7 and 14 days could not be quantified as they were both >1000 mg/kg. The body weights of the worms decreased slightly in all concentrations and controls. No behavioural abnormalities of the surviving worms were observed. During the entire study, water content and pH of the artificial soil were within the ranges given by the guideline