Ethanaminium, N-[4-[[4-(diethylamino)phenyl][4-(ethylamino)-1-naphthalenyl]methylene]-2,5-cyclohexadien-1-ylidene]-N-ethyl-, molybdatetungstatephosphate

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

25Tth June to12th July 2002

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

no guideline followed

Principles of method if other than guideline:

- Principle of test:
- Short description of test conditions: Stock solutions of the test substance were prepared at 10 and 100 mg/L. These were placed in tanks and daphnids (10 per vessel containing 80 mL of test solution) were added. The daphnids were observed for 48 hours
- Parameters analysed / observed: Mobility

GLP compliance:

no

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Source -Sponsor, batch no 0151N/961482
- Expiration date of the lot/batch: 11 June 2003
- Purity test date: Not stated


STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature in the dark
- Stability under test conditions: not indicated
- Solubility and stability of the test substance in the solvent/vehicle:


TREATMENT OF TEST MATERIAL PRIOR TO TESTING
Two stock solutions of 10 and 100 mg/I respectively were stirred overnight. Subsequently the stock solutions, which contained test substance particles, were centrifuged at 3500 x g for 5 minutes. Adequate amounts of the stock solutions were taken from the clear part of the solutions in the centrifuge tubes, to provide for the test concentrations. The final test solutions were all clear and colourless.
FORM AS APPLIED IN THE TEST (if different from that of starting material)

TYPE OF BIOCIDE/PESTICIDE FORMULATION (if applicable)

OTHER SPECIFICS:

Analytical monitoring:

no

Details on sampling:

Not stated

Vehicle:

yes

Details on test solutions:

Two stock solutions of 10 and 100 mg/I respectively were stirred overnight. Subsequently the stock solutions, which contained test substance particles, were centrifuged at 3500 x g for 5 minutes. Adequate amounts of the stock solutions were taken from the clear part of the solutions in the centrifuge tubes, to provide for the test concentrations. The final test solutions were all clear and colourless.

Test organisms (species):

Daphnia magna

Details on test organisms:

No details provided

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

not stated

Test temperature:

19.5 - 20.0 °c

pH:

7.8 - 9.0

Dissolved oxygen:

8.7 - 9.0

Nominal and measured concentrations:

Nominal 1, 10 and 100 mg/L

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Validity criteria fulfilled:

not specified

Conclusions:

Under the conditions of the present study 0151N induced acute immobilisation of Daphnia magna in a WAF loaded at 100 mg/L after 48 hours of exposure. However, the 48h-EC50 was > a WAF loaded at 100 mg 0151N per litre.

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

2-4 June 2004

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

not specified

GLP compliance:

no

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Mo Batch number given, Source - Sponsor
- Expiration date of the lot/batch: 24 May 2005
- Purity test date: Not stated


STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature in the dark
- Stability under test conditions: not stated
- Solubility and stability of the test substance in the solvent/vehicle:
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
Stock solutions were prepared in acetone. The final test solutions in the aqueous test medium were treated as follows: all test solutions were stirred for 15 minutes and then treated with ultrasonic waves for 5 minutes thereafter they were stirred for another 15 minutes. The test substance appeared as completely dissolved at all levels. Those containing 1O and 100 mg/L were transferred to separation funnels for a two-hour settlement period, because due to the opacity of the dark colour at 100 mg/L it was not possible to discern whether all test substance had indeed completely dissolved. When parts of the water phases were taken out of the funnels no test substance particles were present in the remaining part. The appearance of the final test solutions ranged from colourless to dark blue (See Photo 1). The control solution contained the same amount of solvent as in the treated solutions (0.1 ml/L).
FORM AS APPLIED IN THE TEST (if different from that of starting material)

TYPE OF BIOCIDE/PESTICIDE FORMULATION (if applicable)

OTHER SPECIFICS:

Analytical monitoring:

no

Details on sampling:

Not specified

Vehicle:

yes

Details on test solutions:

Stock solutions were prepared in acetone. The final test solutions in the aqueous test medium were treated as follows: all test solutions were stirred for 15 minutes and then treated with ultrasonic waves for 5 minutes thereafter they were stirred for another 15 minutes. The test substance appeared as completely dissolved at all levels. Those containing 1O and 100 mg/L were transferred to separation funnels for a two-hour settlement period, because due to the opacity of the dark colour at 100 mg/L it was not possible to discern whether all test substance had indeed completely dissolved. When parts of the water phases were taken out of the funnels no test substance particles were present in the remaining part. The appearance of the final test solutions ranged from colourless to dark blue. The control solution contained the same amount of solvent as in the treated solutions (0.1 ml/L).

Test organisms (species):

Daphnia magna

Details on test organisms:

Not specified

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

Not specified

Test temperature:

19.7 - 20.6 °C

pH:

7.7 - 8.0

Dissolved oxygen:

>9 mg/L

Nominal and measured concentrations:

Nominal 0, 0.01, 0.1, 1, 10 and 100 mg/L

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

LC0

Effect conc.:

0.01 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LC100

Effect conc.:

0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Validity criteria fulfilled:

not specified

Conclusions:

Based on these results the visible solubility of PASTE VON PB1 - 310304 appeared to be> 100 mg/L, while the 48h-EC50 was between 0.01 and 0.1 mg/L.

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Remarks:

screening test non-GLP

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2-4 June 2004

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

not specified

Principles of method if other than guideline:

- Principle of test: The purpose of the study was to evaluate the test substance for its ability to generate acute toxic effects on the mobility of Daphnia magna during an exposure period of 48 hours and, if possible, to determine the EC50 at 24 and 48 hours of exposure.

- Short description of test conditions: Three solutions with respective initial loadings of 1, 10 and 100 mg/L were prepared and loaded into tanks, daphnids were added and observed for 48 hours
- Parameters analysed / observed:Immobilisation of Daphnids

GLP compliance:

no

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Batch not stated, source Sponsor
- Expiration date of the lot/batch: 24 May 2005
- Purity test date: 35% - substance provided as a liquid - 36 % Pigment PB 1, 0.2% (BIT/MIT Combination) Preservative, <0.5% (Polyether) Antifoaming agent, 63 % Water


STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature in the dark
- Stability under test conditions: stable for at least 96h in vehicle
- Solubility and stability of the test substance in the solvent/vehicle:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The sponsor requested to apply a preparation procedure that is used for products that are more hydrophobic. In this procedure the test substance solutions were stirred for ca. 24 hours. Subsequently all solutions were then centrifuged at 3500 x g for 10 minutes. In the case of PASTE VON PB1 - 310304, the test solutions were completely clear after the 24-hour stirring period and no deposits were observed after the centrifuging step.
The resulting solutions were clear and ranged from nearly colourless to dark blue. The medium used in the control received the same treatment but without test substance.

Analytical monitoring:

no

Vehicle:

yes

Details on test solutions:

Nominal concentrations tested were 1, 10 and 100 mg/L. The test substance was a liquid containing 63% water. Hence it was expected that the product would be easily soluble at these levels in the aqueous test medium. However, the sponsor requested to apply a preparation procedure that is used for products that are more hydrophobic. In this procedure the test substance solutions were stirred for ca. 24 hours. Subsequently all solutions were then centrifuged at 3500 x g for 10 minutes. In the case of PASTE VON PB1 - 310304, the test solutions were completely clear after the 24-hour stirring period and no deposits were observed after the centrifuging step.
The resulting solutions were clear and ranged from nearly colourless to dark blue. The medium used in the control received the same treatment but without test substance.

Test organisms (species):

Daphnia magna

Details on test organisms:

No details given

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

Not specified

Test temperature:

19.7 - 20.6°C,

pH:

7.8-8.0

Dissolved oxygen:

>9 mg/L

Nominal and measured concentrations:

Nominal concentrations 0, 1, 10 and 100 mg/L

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

LC100

Effect conc.:

1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Validity criteria fulfilled:

not applicable

Conclusions:

In conclusion: the 48h-EC50 can be quantified as being below the soluble fraction of PASTE VON PB1 - 310304 in water prepared at a loading of 1 mg/L.

Description of key information

Three non-GLP screening test studies have been performed to assess the possible toxicity of the test substance to fresh water crustaceans.

No analytical information is available to verify the concentration of the test substance in solution.

As the test substance is very poorly water soluble it is proposed to conduct a long term aquatic toxicity test to fulfil this endpoint as indicated in the REACH guidance