PIGMENTADDITIV RL

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1989-02-23 to 1989-02-25

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study performed according to relevant guidelines and compliant with GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1989

Report date:

1989

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

(former 84/449/EWG)

Deviations:

no

Remarks:

according to OECD guideline 202, adopted 2004

GLP compliance:

yes

Test material

Reference

Name:

Unnamed

Sampling and analysis

Analytical monitoring:

no

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A stock solution was prepared at 2g/l test item (0.2% solution). The dispersion of the test item occurred through ultrasonication. Test concentrations ranged from 1 to 1000 mg/L test item using a spacing factor of 1.8.
- Eluate: dilution water according to DIN 38412 Teil 11, 8.2
- Controls: yes
- Evidence of undissolved material (e.g. precipitate, surface film, etc): after 24h precipitation of test item particles was observed for all concentration levels.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna STRAUSS
- Source: Bundesgesundheitsamt, Berlin
- Method of breeding: in 2000 ml vessels at 20° C, aerated vessels (0.45 µm filter), water renewal three times a week.
- Food type during breeding: dry food, two times per day
- Age at the beginning of the experiment: not specified.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Test temperature:

Treatment groups: 22.2 °C
Controls: 22.2 °C

pH:

Treatment groups: 7.71 - 7.92
Controls: 7.71

Dissolved oxygen:

Treatment groups: 7.9 - 8.1 mg/l
Controls: 8.1 mg/l

Nominal and measured concentrations:

Nominal test item concentrations were achieved by dilution of a test item stock solution prepared at 2 g/l test item (0.2%).
Range of nominal concentrations: 1.0, 1.8, 3.2, 5.8, 10, 18, 32, 58, 100, 180, 320, 580, 1000 mg/l test item (spacing factor: 1.8)

Details on test conditions:

TEST SYSTEM
- Test vessel: 50 ml vessels (DIN 12 331)
- Fill volume: 20 ml
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: DIN 38412 Teil 11, 8.2
- Conductivity: <5 µS/cm
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 12 h, diffuse
- Light intensity: 10 µE/qm *s,

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : immobilization, after 24h and 48h

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.8
- Range finding study: yes
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 500 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95 % CL could not be determined

Details on results:

No significant dose-response-curve could be generated: precipitation of test item particles occurred with respect to all concentrations after 24h. The particles adsorbed at the cuticle of the test organisms and hindered the daphnid's motility.

- Effect concentrations exceeding solubility of substance in test medium: yes
- Mortality of control: no
- Other adverse effects control: no

Results with reference substance (positive control):

- Results with reference substance valid? Yes.
- EC0: 0.7 mg/l, EC50: 1,2 mg/l, EC100: 2.0 mg/l
- Other: Reference test has been carried out in parallel to the main test for 24h.

Reported statistics and error estimates:

Applied statistics were according to "Ausgewählte Methoden der Wasseruntersuchung", Band II (1982), 379-382, VEB Fischer Verlag.

Any other information on results incl. tables

Test concentrations for the determination of effect concentrations

Conc. (mg/l)	Immobilization (%)				Diss. Ox. (mg/l)	pH	Temp. (°C)
	24h		48h
	1.	2.	1.	2.
1000	40	30	40	40	7.9	7.85	20.2
580	40	50	50	60	7.9	7.87	„
320	40	40	60	60	7.8	7.86	„
180	20	30	30	30	7.9	7.85	„
100	20	30	50	40	8.1	7.84	„
58	40	20	50	30	8.0	7.83	„
32	20	40	20	40	7.9	7.81	„
18	10	0	20	20	8.0	7.80	„
10	10	0	30	10	8.1	7.80	„
5.8	0	10	20	20	7.9	7.92	„
3.2	0	0	20	10	8.0	7.85	„
1.8	0	0	0	0	8.1	7.80	„
1.0	0	0	0	0	8.0	7.77	„
Control	0	0	0	0	8.1	7.71	„

The following effect concentrations have been derived:

EC0-24h: 3.2 mg/l

EC0-48h: 1.8 mg/l

EC50-24h: >1000 mg/l

EC50-48h: >500 mg/l

EC100-24h/48h: >1000 mg/l

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The derived EC50 (48h) for the immobilization of Daphnia magna was >500 mg/l.

Executive summary:

In the present study the immobilization of Daphnia magna has been evaluated after 24h and 48h of exposure to the test item. The concentrations tested were prepared from a super-saturated solution prepared at 2 g/l test item using ultrasonication for dispersion. The nominal concentrations tested were as follows: 1.0, 1.8, 3.2, 5.8, 10, 18, 32, 58, 100, 180, 320, 580, 1000 mg/l test item. No analytical measurements were performed based on the very low solubility of the test item (<1µg/l, see section Physical and chemical properties).

After an exposure of 24h, precipitation of test item particles occurred in all test concentrations, which can be explained by the low water solubility of the test item (<1 µg/l, see section Physical and chemical properties). Furthermore, adsorption of the test item occurred on the cuticle of the test organisms hindering the motility of the organisms. Thus, observed immobility of Daphnia magna may be caused by the physical impact of undissolved test item particles instead of inherent toxicity. Summarizing, the following effect concentrations were determined:

 

EC0-24h: 3.2 mg/l

EC0-48h: 1.8 mg/l

EC50-24h: >1000 mg/l

EC50-48h: >500 mg/l

EC100-24h/48h: >1000 mg/l

 

The study was performed according to 84/449/EWG (later EU Method C.2 (Acute toxicity for Daphnia) and GLP.