Reaction mass of lithium sodium hydrogen 4-amino-6-({5-[(5-chloro-2,6-difluoropyrimidin-4-yl)amino]-2-sulfonatophenyl}diazenyl)-5-hydroxy-3-[(4-{[2-(sulfonatooxy)ethyl]sulfonyl}phenyl)diazenyl]naphthalene-2,7-disulfonate and lithium sodium hydrogen 4-amino-6-({5-[(5-chloro-2,6-difluoropyrimidin-4-yl)amino]-2-sulfonatophenyl}diazenyl)-5-hydroxy-3-{[4-(vinylsulfonyl)phenyl]diazenyl}naphthalene-2,7-disulfonate

Long-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

fish early-life stage toxicity

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 210 (Fish, Early-Life Stage Toxicity Test)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 0, 1, 3.16 and 10 mg/L
- Days of sampling: -7, 0, 6, 12, 20, 27 and 31
- Sampling method: Not specified
- Sample storage conditions before analysis: Not specified

Vehicle:

no

Test organisms (species):

Pimephales promelas

Details on test organisms:

TEST ORGANISM
- Common name: Fathead minnow
- Strain: Rafinesque
- Source: Bio International B.V., Netherlands
- Feeding during test: yes
- Food type: Tetramin fish food
- Amount: 2% of body weight per day
- Frequency: three times daily

ACCLIMATION
- Acclimation period: eight days
- Acclimation conditions (same as test or not): same
- Type and amount of food: Tetramin fish food, 2% of body weight per day
- Feeding frequency during acclimation: three times daily
- Health during acclimation (any mortality observed): <5%

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

32 d

Hardness:

215.3 mg/L CaCO3

Test temperature:

23-27°C

pH:

7.7-8.1

Dissolved oxygen:

6.1-8.7 mg/L

Nominal and measured concentrations:

Nominal: 0, 1, 3.16 and 10 mg/L
Measured: 0, 1.17, 3.62 and 11.6 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass vessels, diameter 14 cm x height 7.5 cm and glass aquarium holding test medium
- Size of vessel: glass aquarium 5L
- Aeration: gentle aeration via narrow glass tubes
- Renewal rate of test solution: continuous
- No. of organisms per vessel: 30 fertilised eggs
- No. of vessels per concentration: two
- No. of vessels per control: two

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: freshly prepared reconstituted water
- Culture medium different from test medium: no
- Intervals of water quality measurement: five times during the test

OTHER TEST CONDITIONS
- Photoperiod: 16 h light:8 h dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): hatching success, mortality, body abnormalities, type and incidence of abnormalities and length and weight of juvenile fish at the end of the test

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 3.16

Key result

Duration:

32 d

Dose descriptor:

NOEC

Effect conc.:

>= 11.6 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr. (total fraction)

Basis for effect:

number hatched

Details on results:

The hatching success was not affected by the test substance exposure in any of the concentrations compared to the control. Further, there were no adverse behavioural and appearance abnormalities in any of the tested concentrations.

Validity criteria fulfilled:

yes

Conclusions:

Under the study conditions, the nominal 32 d NOEC of the test substance to the early-life stages of fathead minnow was determined to be >=10 mg/L, equivalent to >=11.6 mg a.i./L.

Executive summary:

A study was conducted to determine the toxicity of the test substance (in the form of aqueous dispersion of 64% purity) to the early-life stages of fathead minnow (Pimephales promelas) according to OECD Guideline 210, in compliance with GLP. Batches of 60 fertilised eggs, evenly distributed in two replicates were exposed to the substance dispersed in test medium at nominal concentrations of 0, 1, 3.16 and 10 mg/L (equivalent to 0, 1.17, 3.62 and 11.6 mg a.i./L respectively). The test concentrations were prepared through a flow-through device. Observations were made daily on the numbers of fertilised eggs and/or juvenile fish to record the hatching success, behavioural and morphological abnormalities during the exposure period of 32 d. Analytical validation of test concentrations was conducted in regular intervals. There was no significant difference in mortality among the test and control groups during the study and no behavioural abnormalities were noted. Under the study conditions, the nominal 32 d NOEC of the test substance to the early-life stages of fathead minnow was determined to be >=10 mg/L, equivalent to >=11.6 mg a.i./L (Bruns, 2001a).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

11.6 mg/L

Additional information

A study was conducted to determine the toxicity of the test substance (in the form of aqueous dispersion of 64% purity) to the early-life stages of fathead minnow (Pimephales promelas) according to OECD guideline 210. Batches of 60 fertilised eggs, evenly distributed in two replicates were exposed to the substance dispersed in test medium at nominal concentrations of 0, 1, 3.16 and 10 mg/L (equivalent to 0, 1.17, 3.62 and 11.6 mg a.i./L respectively). The test concentrations were prepared through a flow-through device. Observations were made daily on the numbers of fertilised eggs and/or juvenile fish to record the hatching success, behavioural and morphological abnormalities during the exposure period of 32 d. Analytical validation of test concentrations was conducted in regular intervals. There was no significant difference in mortality among the test and control groups during the study and no behavioural abnormalities were noted. Under the study conditions, the nominal 32 d NOEC of the test substance to the early-life stages of fathead minnow was determined to be >10 mg/L, equivalent to >11.6 mg a.i./L (Bruns, 2001a).