Reaction mass of 3,7-bis(diethylamino)phenoxazin-5-ium chloride and 3,7-bis(diethylamino)phenoxazin-5-ium trichlorozincate

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

other: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Unchanged samples
- All test medium samples for the analytical determinations were taken from the approximate center of the test vessels (after mixing the medium) and kept at-18°C to -25°C until analysis.

Details on test solutions:

STOCK SOLUTION
A stock solution of 219.9 mg test substance was mixed in a beaker with 200 ml test water and filled up to 1000 ml with test water. This stock solution was diluted 1:10 with test water to get a final stock solution.
The test concentrations were prepared with this final stock solution.

PREPARATION AND APPLICATION OF TEST SOLUTION:
- the test medium used had the following composition (concentration in test)
NaHCO3 (65 mg/l)
MgSO4 * 7 H2O (123.3 mg/l)
KCl (5.8 mg/l)
CaCI2 * 2 H2O (294 mg/l)
Na2SiO3 * 9 H2O 4.3 (mg/l)
NaNO3 274 (μg/l)
KH2PO4 1430 (μg/l)
K2HPO4 184 (μg/l)
- the test water medium was prepared with bidistilled water
- the medium was oxygen saturated at the beginning of the test

REFERENCE SUBSTANCE: potassium dichromate

Test organisms (species):

Daphnia magna

Details on test organisms:

The organism used in this test system is the species Daphnia magna Strauss. The Daphnia were bred in the laboratories of the Product Ecology under standardised conditions.

Breeding:
- Cultures of Daphnia magna are held in glass vessels containing approx. 3.0 litre of reconstituted water at 18 - 22 °C. The water is renewed partially three times per week.The Daphnia are fed with a suspension of green algae (Scenedesmus subspicatus) supplemented by a Tetramin-extract in such quantites that the food is consumed within 24 hours.

Pretreatment:
- 24 hours before the start of the exposure reproductive Daphnia were separated from the young Daphnia by sieving all individuals through a 1000 μm sieve.

For each test concentration 20 Daphnia divided into two groups of 10 animals were used.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

282 mg/l CaCO3

Test temperature:

21 °C

pH:

in a range of 7.6 - 8.0

Dissolved oxygen:

saturated

Nominal and measured concentrations:

0.038, 0.083, 0.18, 0.40, 0.9 and 1.9 mg test article/l (nominal)

Details on test conditions:

TEST SYSTEM
- a range finding test was carried out at concentration from 0.01 - 100 mg/l to determine the concentrations for the main test.
- the Daphnia were exposed to the test article in 250 ml beakers containing 60 - 100 ml test medium with 10 animals per beaker
- all tests were running in duplicate

OTHER TEST CONDITIONS
- Light conditions: 12 hours light / 12 hours dark- alkalinity: 0.8 mmol/l

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

0.6 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: 95 %confidence limit: 0.499 - 0.720 mg/l

Duration:

24 h

Dose descriptor:

EC0

Effect conc.:

>= 0.18 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

24 h

Dose descriptor:

EC100

Effect conc.:

1.9 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.2 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: 95 % confidence limit: 0.16 - 0.252 mg/l

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

0.038 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

0.9 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

In all test vessels no visible alteration of the test substance in the test medium was observed after 48 hours. During the test the actual test concentrations in the analysed test media were in the range of 89 - 108 % of the nominal values. Therefore, the reported results are related to the nominal values of the test concentrations of the test article. No immobilization and abnormal behaviour of the Daphnia was observed in the blank control.

Measurements at the end of the test:

Nominal conc. (mg/l)	blank	0.038	0.083	0.18	0.40	0.9	1.9
oxygen (%)	97	97	97	97	98	98	96
pH	8.0	7.9	8.0	7.9	8.0	8.0	8.0
temperature (°C)	21	21	21	21	21	21	21

Validity criteria fulfilled:

yes

Conclusions:

EC50 (48h) = 0.20 mg/l

Executive summary:

A study was conducted to determine the acute toxicity of the test substance to Daphnia magnain a 48 h static test according to the OECD Guideline 202, in according to the GLP Principles.

 

Daphnia were exposed to nominal test substance concentrations of0.038, 0.083, 0.18, 0.40, 0.9 and 1.9mg/l. Analytical determination of the test substance concentration was carried out at 0 and 48 h by spectrophotometry, leading to test substance concentrations in the range of 89 - 108 % of the nominal values.

 

Results:

EC0 (48h) = 0.038 mg/l

EC50 (48h) = 0.20 mg/l

EC100 (48h) = 0.9 mg/l

Description of key information

EC50 (48h) = 0.20 mg/l

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

0.2 mg/L