6'-(dibutylamino)-3'-methyl-2'-(phenylamino)spiro[isobenzofuran-1(3H),9-(9H)-xanthen]-3-one

Long-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

adult fish: sub(lethal) effects

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1991-05-28 to 1991-06-18

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 204 (Fish, Prolonged Toxicity Test: 14-day Study)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Sampling method: Water samples were taken from the solvent control and each exposure level on Days 0, 1,7, 9, 14, 16 and 21.

Vehicle:

yes

Remarks:

1 % Tween 80 -acetone

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Rainbow trout
- Source: Westacre Trout Farm, Norfolk, UKTEST ORGANISM
- Length at study initiation: Mean 5.8 cm, SD 0.4 cm
- Weight at study initiation: Mean 2.64 g, SD = 0.54 g
- Feeding during test : Discontinued 24 h prior to test. Fish were fed commercial trout pellets.
- Amount: approximately 2 % of the bodyweight
- Frequency: daily


ACCLIMATION
- Acclimation period: 10 days
- Acclimation conditions: same as test
- Type of food: Commercial trout pellets
- Feeding frequency during acclimation: daily
- Health during acclimation: 0 % 7 days prior to test, no medication

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Hardness:

160 mg CaCO3/L

Test temperature:

13 ± 1 °C

pH:

8.3 - 8.5

Dissolved oxygen:

> 10.0 mgO2/L

Conductivity:

1000 µS/cm

Nominal and measured concentrations:

Nominal: 0.010, 0.032, 0.10, 0.32, and 1.0 mg/L
Measured: 0.0067, 0.020, 0.078, 0.29, and 0.94 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Aquaria
- Size of vessel: 40 L
- Type: open
- Material: glass
- Aeration: None
- Renewal rate of test solution (frequency): daily
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1
- Biomass loading rate: 0.66 g bw/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Laboratory tap water, dechlorinated by the addition of sodium thiosulphate.
- Total organic carbon: 3.9 mg/L
- Metals:
Magnesium 8.8 mg/L
Sodium 58 mg/L
Potassium 10.0 mg/L
Calcium 130 mg/L
Aluminum 6.4 µg/L
Iron 110 µg/L
Manganese 20 µg/L
Copper 25 µg/L
Zinc 5.3 µg/L
Fluoride 260 µg/L

- Pesticides: 0.27 µg/L
- Chlorine: 83 mg/L
- Alkalinity: 150 mg/L (HCO3)
- Ca/Mg ratio: 14.77
- Culture medium different from test medium: No

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 h light : 8 h dark.

EFFECT PARAMETERS MEASURED: Mortality

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 3.125 / 3.2

Reference substance (positive control):

no

Key result

Duration:

21 d

Dose descriptor:

LC50

Remarks on result:

not determinable

Key result

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 0.94 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

growth rate

Details on results:

- Observed effects at each concentration for each observation time:
There were no adverse reactions to exposure with all fish surviving the 21-day test period at concentrations well in excess of the water solubility value (0.021 mg/L).
- Cumulative mortality at each concentration and for each recommended observation time if possible: No mortality observed.
- Mortality in the controls: No mortality observed.
- Behavioural observation of the fish: No treatment related effects on behaviour were observed.
- Effect concentrations exceeding solubility of substance in test medium: Yes
- Incidents in the course of the test which might have influenced the results: No

Reported statistics and error estimates:

Not applicable

Validity criteria fulfilled:

yes

Conclusions:

No LC50 could be determined in an long-term toxicity test according to OECD guideline 204, since no mortality was observed. A NOEC value of equal to or above 0.94 mg/L was set for the 21 day exposure period.

Executive summary:

The long term aquatic toxicity to fish was examined according to OECD guideline 204 (Huntingdon, 1991). Rainbow trout (Oncorhynchus mykiss) were exposed to the test substance at nominal concentrations of 0.010, 0.032, 0.10, 0.32, and 1.0 mg/L (measured: 0.0067, 0.020, 0.078, 0.29, and 0.94 mg/L) under semi-static conditions. Measured concentrations of expired media have been used for all calculations and estimations. Continuous flow conditions could not be used due to the poor solubility of the test substance in water.

No mortality or adverse effects were observed at any concentration up to and including 0.94 mg/L, the maximum attainable concentration due to the limited solubility of the test substance in water and having regard to the limited amount of auxiliary solvent permitted in the test. Length and weight measurements made on all surviving fish at the end of the exposure period indicated that no adverse effects on growth occurred. Further there were no adverse reactions to exposure with all fish surviving the 21 -day test period at the concentrations well in excess of the water solubility.

The NOEC was set to equal to or above 0.94 mg/L. An LC50 could not be determined.

Reference 2

Endpoint:

adult fish: sub(lethal) effects

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Qualifier:

according to guideline

Guideline:

OECD Guideline 204 (Fish, Prolonged Toxicity Test: 14-day Study)

GLP compliance:

yes

Details on test solutions:

Identity and concentration of auxiliary solvent for dispersal: Dimethylformamide at 0.6 g/L as auxiliary solvent.

Test organisms (species):

Oryzias latipes

Test type:

static

Water media type:

freshwater

Limit test:

no

Hardness:

80 mg CaCO3/L

Details on test conditions:

Run using nominal concentrations of 1 to 1000 µg/L.

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 10 mg/L

Basis for effect:

mortality

Key result

Duration:

14 d

Dose descriptor:

LC50

Effect conc.:

> 0.922 mg/L

Basis for effect:

mortality

Key result

Duration:

21 d

Dose descriptor:

LC50

Effect conc.:

> 0.922 mg/L

Basis for effect:

mortality

Key result

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

>= 0.922 mg/L

Details on results:

%Concentration loss over test: 7 - 11

Validity criteria fulfilled:

not specified

Conclusions:

A 14 d-NOEC of 0.922 mg/L was determined in a prolonged toxicity study in fish.

Executive summary:

A prolonged toxicity study according to OECD guideline 204 was performed (NIPPON SODA, ECHA RSS, 2017). Oryzias latipes were exposed to nominal concentrations of 1 to 1000 µg/L. 0.6 g/L dimethylformaide were used as auxiliary sovlent for dispersal. No adverse effects or treatment-related mortalities observed up to the highest concentration tested. No statistical differences between control and test fish weights were observed. An LC50 of above 0.922 µg/L was determined. The NOEC was set to equal or above 0.922 µg/L.

Description of key information

A NOEC of equal to or above 0.94 mg test item/L was observed after 21 days in Oncorhynchus mykiss in the key study.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

0.94 mg/L

Additional information

Key study

The long term aquatic toxicity to fish was examined according to OECD guideline 204 (Huntingdon, 1991). Rainbow trout (Oncorhynchus mykiss) were exposed to the test substance at nominal concentrations of 0.010, 0.032, 0.10, 0.32, and 1.0 mg/L (measured: 0.0067, 0.020, 0.078, 0.29, and 0.94 mg/L) under semi-static conditions. Measured concentrations of expired media have been used for all calculations and estimations. Continuous flow conditions could not be used due to the poor solubility of the test substance in water.

No mortality or adverse effects were observed at any concentration up to 0.94 mg/L, the maximum attainable concentration due to the limited solubility of the test substance in water and having regard to the limited amount of auxiliary solvent permitted in the test. Length and weight measurements made on all surviving fish at the end of the exposure period indicated that no adverse effects on growth occurred. Further there were no adverse reactions to exposure with all fish surviving the 21 -day test period at the concentrations well in excess of the water solubility (0.021 mg/L).

The NOEC was set to equal to or above 0.94 mg/L. An LC50 could not be determined.

In accordance with Article 25(3) of the REACH Regulation the robust study summaries submitted 12 years previously, are additionally used for the purpose of the registration.

Supporting Study

A prolonged toxicity study according to OECD guideline 204 was performed (NIPPON SODA, ECHA RSS, 2017). Oryzias latipes were exposed to nominal concentrations of 1 to 1000 µg/L. 0.6 g/L dimethylformamide were used as auxiliary solvent for dispersal. No adverse effects or treatment-related mortalities observed up to the highest concentration tested. No statistical differences between control and test fish weights were observed. An LC50 of above 0.922 µg/L was determined. The NOEC was set to equal or above 0.922 µg/L.