REAKTIV-GELB F-68 072 FW

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

21 April 1993 to 23 April 1993

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

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

Analytical monitoring:

yes

Details on sampling:

In order to determine the substance concentration, water samples were taken from the middle of the test chamber by pipetting ca. 20 ml into a screw-neck glass bottle in the case of each preparation of the test after 0 and 24 hours

Vehicle:

no

Details on test solutions:

The test concentrations were prepared by diluting a stock batch (1000 mg/L). For this purpose, the test substance was weighed into a graduated flask, filled up with test water and stirred for about 5 minutes with a magnetic stirrer. The amounts corresponding to the test concentrations were pipetted into a graduated flask, made up to a total volume of 200 mL with test water, and then transferred to the test chambers. The test batches were present as clear orange solutions .

No separation or formation of deposits was observed with any of the concentrations tested.

In order to determine the substance concentration, water samples were taken from the middle of the test chamber by pipetting ca. 20 mL into a screw-neck glass bottle in the case of each preparation of the test after 0 and 24 hours.

Test organisms (species):

Daphnia magna

Details on test organisms:

Species: Daphnia magna - Clone 5 (IRChA, France)
Origin: Stockculture of the Department of Toxicology (Hoechst AG)
Breeding conditions: 100 ml beakers (height 90 mm, diameter 50 mm), filled with 70 ml test water (treated fresh water) served as culture vessels. One daphnid was used per culture vessel. The animals were used for breeding up to an age of 42 days. The reproduction rate and the state of health of the animals were monitored daily apart from weekends. The animals were fed three times a week with monocellular green algae (Scenedesmus subspicatus).

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

No post exposure observation period

Hardness:

No data

Test temperature:

20.2 - 21.8 deg C

pH:

6.7 - 8.3

Dissolved oxygen:

8.3 - 9.7 mg/l

Salinity:

Not applicable.

Nominal and measured concentrations:

Test concentrations 0 / 100 / 180 / 320 / 560 / 1000 mg/L

Details on test conditions:

Study groups
20 daphnids, divided into two parallel groups of 10 animals, were used for each concentration. The test concentrations, which were selected on the basis of range finding tests, were as follows:

Test chamber No. Concentration (mg/L)
1/2/3/4/5/6 (a,b) 0/100/180/320/560/1000

Water for dilution
Reconstituted fresh water was used as water for dilution.
The dilution water, prepared as described, was aerated until saturated with oxygen. The pH, oxygen content, conductance and total hardness were measured and recorded once a week.

Test conditions
The study was conducted in a semistatic system. The daphnids were transferred to new test chambers every 24 hours. The test chambers were 250 ml beakers (height 110 mm, diameter 70 mm, water level about 75 mm), each filled with 200 ml test water. The room temperature was about 21 ⁰C. The room was illuminated by fluorescent tubes in a day/night cycle, with each period lasting 12 hours.

Study conduct
After the test concentrations had been prepared and water parameters recorded, 10 daphnids were used in each test chamber. The daphnids received no feed for the entire study period. Inspection of the daphnids took place after 24 and 48 hours and involved recording the immobility. Immobile daphnids were removed from the chambers. Daphnids were considered immobile when there was a lack of movement 15 sec. after touching the test chamber. The water parameters were measured after adding the substance and at the end of the test.

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

180 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

>= 180 - <= 320 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

560 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Immobility was observed at 320 - 1000 mg/L. The 100 and 180 mg/L groups showed no effect in comparison to the negative control.

All tested concentrations were present as clear orange solutions.

The substance concentration was determined in the 100 - 1000 mg/L groups. All values determined were in a range of + 20 % of the theoretical value with the exception of the 24 hour values in the 180 mg/L group (73 and 76 % of the nominal concentration).

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

On the basis of the immobility rate, the EC50 values were determined after 24 hours by means of the probit analysis.

Probit analysis could not be carried out after 48 hours because of the immobility rate. As the factor for the concentration steps was 1.8, no further concentrations were tested. The EC50 range is given.

Immobility

 

Under the given study conditions, the following immobility rates of Reaktiv-Gelb F-68 072 FW were recorded after 24 and 48 hours:

Concentration (mg/l)	Immobility
	24h		48h
	Absolute	Relative (%)	Absolute	Relative (%)
0	0/20	0	0/20	0
100	0/20	0	0/20	0
180	0/20	0	0/20	0
320	9/20	45	12/20	60
560	17/20	85	20/20	100
1000	20/20	100	20/20	100
EC0 (mg/l) highest tested concentration without immobility	180		180
EC50 (mg/l) Confidence interval (p = 0.05)	361 (308 – 422.7)		180 – 320
EC100 (mg/l) lowest tested concentration with 100% immobility	1000		560

Validity criteria fulfilled:

yes

Conclusions:

EC50 > 180 mg/L < 320 mg/L

Executive summary:

The acute toxicity of Reaktiv-Gelb F-68 072 FW to aquatic invertebrates was determined in compliance with the requirements of EEC-Guideline 79/831 Annex V Methods for the Determination of Ecotoxicity 5.1.2 Acute Toxicity of Daphnia and the OECD-Guidelinefor testing of chemicals, Section 2 - Effects on Biotic Systems Testguideline 202 Daphnia sp., 14-day Reproduction Test (including an Acute Immobilisation Test). Study conducted in compliance with the Principles of Good Laboratory Practice (GLP).

 

The 48-hour acute toxicity study of Reaktiv-Gelb F-68 072 FW in Daphnia magna (water flea) was conducted in a semistatic system, because in a static system the substance was not analytically detectable to a sufficient degree over 48 hours.

 

Immobility was observed at -320 - 1000 mg/L. The 100 and 180 mg/L groups showed no effect in comparison to the negative control.

 

The concentrations tested were 100, 180, 320, 560, 1000 mg/L and a negative control (0 mg/L).

 

All tested concentrations were present as clear orange solutions.

 

The substance concentration was determined in the 100 - 1000 mg/L groups. All values determined were in a range of + 20 % of the theoretical value with the exception of the 24 hour values in the 180 mg/L group (73 and 76 % of the nominal concentration).

 

The substance is not classified as harmful to Daphnia.

Description of key information

The EC50 after 48h was determined to be: > 180 mg/L< 320 mg/L (nominal conc.).

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Remarks:

(48h)

Effect concentration:

> 180 - < 320 mg/L

Additional information

The acute toxicity of Reaktiv-Gelb F-68 072 FW to aquatic invertebrates was determined in compliance with the requirements of EEC-Guideline 79/831 Annex V Methods for the Determination of Ecotoxicity 5.1.2 Acute Toxicity of Daphnia and the OECD-Guidelinefor testing of chemicals, Section 2 - Effects on Biotic Systems Testguideline 202 Daphnia sp., 14-day Reproduction Test (including an Acute Immobilisation Test). Study conducted in compliance with the Principles of Good Laboratory Practice (GLP).

The 48-hour acute toxicity study of Reaktiv-Gelb F-68 072 FW in Daphnia magna (water flea) was conducted in a semistatic system, because in a static system the substance was not analytically detectable to a sufficient degree over 48 hours.

All tested concentrations were present as clear orange solutions.

The substance concentration was determined in the 100 - 1000 mg/L groups.Allvalues determined were in a range of + 20 % of the theoretical value with the exception of the 24 hour values in the 180 mg/L group (73 and 76 % of the nominal concentration).

The substance is not classified as harmful to Daphnia.