isostearic acid monoisopropanolamide

Long-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

adult fish: sub(lethal) effects

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From February 26, 2002 to June 08, 2002

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 (incl. QA statement)

Analytical monitoring:

yes

Remarks:

HPLC/MS

Details on sampling:

Analytical report 798164 (Biedermann, 2002)
The test substance (stock solution), the test substance solutions and the water (and solvents) (analytical blanks) were analysed. The stock solution (in acetonitrile/purified water) and its dilutions were used to calibrate the HPLC-system. The test substance dissolved in a mixture of acetonitrile/purified water was further diluted with test water and analysed by HPLC. The samples obtained from biological tests (taken on Days 3, 7, 10, 13, 16 and 21 of the exposure period) at different concentration levels were analysed by HPLC with MS detection. Analysis was performed in duplicate.

Vehicle:

no

Details on test solutions:

Aqueous solutions were prepared by dispersing the test substance in test water by ultrasonic treatment. Since the aqueous solutions were not stable for longer time periods, they were renewed regularly (every 2 or 3 days) during the test period.
Aqueous application solutions were first warmed up to obtain a homogeneous liquid and 5 different solutions were prepared by dispersing the test substance in 5 liter purified water each. They were then treated ultrasonically for 45 min and intensively stirred for 15 min to obtain homogeneous dispersions of the test substance in test water. The solutions were prepared at Days 0, 3, 8, 10, 13, 15, 17 and 20. The stock solutions were then added in the aquaria.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

Source: P. Hohler, trout breeding station Zeiningen, CH-4314 Zeiningen, Switzerland.
Acclimatization: at least one week prior to the test
Water temperature: 14 - 15“C
Light conditions: a 16-hour light to 8-hour darkness photoperiod
Diet (prior test): HOKOVIT 502 (adapted to the number of fish)
Number: 10 fish
Mean body length at the start of the test: 5.3 +/- 0.4 cm
Mean body wet weight: 1.5 +/- 0.2 g

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Hardness:

112-182 mg/L as CaCO3

Test temperature:

14-15°C

pH:

7.6-8.2

Dissolved oxygen:

8.6 mg/L or higher (>60% oxygen concentration)

Nominal and measured concentrations:

0, 0.5, 1.6, 5.0, 16 and 50 mg/L (nominal) (chosen based on an acute toxicity test with rainbow trout, a range-finding test and a pre-experiment.

Details on test conditions:

- 60-litre flow-through glass-aquarium with 10 fish

Reference substance (positive control):

no

Key result

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

ca. 1.6 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: body length and wet weight

Remarks on result:

other: 1.2 mg/L (measured)

Key result

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

ca. 5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: body length and wet weight

Remarks on result:

other: 2.5 mg/L (measured)

Key result

Duration:

21 d

Dose descriptor:

other: LLC

Effect conc.:

ca. 50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: 27 mg/L (measured)

Details on results:

Analytical results for test substance concentrations:
- Freshly prepared application solution samples: 93 - 118% of the nominal values (correct preparation)
- Aged application soltuions: 95 - 123% of nominal values (stability)
- In test media: 14 - 121% (inhomogeneous distribution)
- Measured concentrations: 1.2 mg/L (1.6 mg/L nominal), 2.5 mg/L (5.0 mg/L nominal), and 27 mg/L (50 mg/L nominal).
Biological results:
- Mortality: no mortality was observed in the groups 0, 1.6, 5.0 and 16.0 mg/L (nominal). At 50 mg/L, all fish were dead on Day 15
- Visible abnormalities: they were observed in the groups 16.0 and 50.0 mg/L (nominal) (from Day 16 and from Day 6, respectively)
- Measured mean body length and body wet weight: they were significabtly reduced starting at 5.0 mg/L (nominal) at the end of the test period

Reported statistics and error estimates:

Parameters were evaluated on significant differences to the control values by the multivariate statistical Williams-test.

None.

Validity criteria fulfilled:

yes

Conclusions:

Under the study conditions, the nominal 21 d NOEC of the test substance in Rainbow trout was 1.6 mg/L (corresponding to 1.2 mg/L measured).

Executive summary:

A study was conducted to determine the long-term toxicity of the test substance, isoC18 MIPA (96% active), to fish according to OECD Guideline 204, in compliance with GLP. Rainbow trout (Oncorhynchus mykiss) were exposed to the test substance (dispersed in the test medium by ultrasonication) at concentrations of 0, 0.5, 1.6, 5.0, 16.0 and 50.0 mg/L (nominal) for 21 d under flow-through conditions. Samples for analytical confirmation were taken from the freshly prepared solutions, application solutions and test water every 2 or 3 d and analysed by HPLC with MS detection. Mortality, visible parameters and water parameters were recorded throughout the exposure period. The average concentration found in application samples was 103%. Samples diluted in test water showed an average concentration of 92%, so no correction for possible losses during the analytical procedure was necessary. The reported biological results were related to the total mean measured test substance concentrations. These were 1.2 mg/L (1.6 mg/L nominal), 2.5 mg/L (5.0 mg/L nominal) and 27 mg/L (50 mg/L nominal). No mortality was observed up to 16.0 mg/L. At 50 mg/L, all fish were dead on Day 15. Visible abnormalities were observed at 16.0 and 50.0 mg/L. Finally, measured mean body length and wet weight of the body were significantly reduced starting at 5.0 mg/L (nominal) at the end of the test period. Under the study conditions, the nominal 21 d NOEC of the test substance in Rainbow trout was 1.6 mg/L (corresponding to 1.2 mg/L measured) (Peither, 2002).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

1.2 mg/L

Additional information

A study was conducted to determine the long-term toxicity of the test substance, isoC18 MIPA (96% active), to fish according to OECD Guideline 204, in compliance with GLP. Rainbow trout (Oncorhynchus mykiss) were exposed to the test substance (dispersed in the test medium by ultrasonication) at concentrations of 0, 0.5, 1.6, 5.0, 16.0 and 50.0 mg/L (nominal) for 21 d under flow-through conditions. Samples for analytical confirmation were taken from the freshly prepared solutions, application solutions and test water every 2 or 3 d and analysed by HPLC with MS detection. Mortality, visible parameters and water parameters were recorded throughout the exposure period. The average concentration found in application samples was 103%. Samples diluted in test water showed an average concentration of 92%, so no correction for possible losses during the analytical procedure was necessary. The reported biological results were related to the total mean measured test substance concentrations. These were 1.2 mg/L (1.6 mg/L nominal), 2.5 mg/L (5.0 mg/L nominal) and 27 mg/L (50 mg/L nominal). No mortality was observed up to 16.0 mg/L. At 50 mg/L, all fish were dead on Day 15. Visible abnormalities were observed at 16.0 and 50.0 mg/L. Finally, measured mean body length and wet weight of the body were significantly reduced starting at 5.0 mg/L (nominal) at the end of the test period. Under the study conditions, the nominal 21 d NOEC of the test substance in Rainbow trout was 1.6 mg/L (corresponding to 1.2 mg/L measured) (Peither, 2002).

As discussed with ECHA in the frame of a Dossier Improvement Action Plan (DIAP), should the planned new water solubility testing according to OECD Guideline 105 result in a water solubility for the substance <1 mg/L, additional long-term toxicity testing in fish will be conducted with representative short- and a long chain FAA substances.