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Short-term toxicity to fish

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short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Test performed from 23 April 1990 to 21 May 1990.
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Guideline study: According to the REACH Guidance this study is not suited to evaluate the long-term toxicity on fish since sensitive life-stages (juveniles, fertilised eggs, larvae) were not tested. Nevertheless the study results can be used for the evaluation of the short-term toxicity (Ch. 6.1.2).
according to
OECD Guideline 204 (Fish, Prolonged Toxicity Test: 14-day Study)
GLP compliance:
Analytical monitoring:
Details on sampling:
- Concentrations: all concentrations determined daily
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: 2 mL Tributylamine dissolved in 5 mL ethanol; 2 mL of this solution added under stirring to 10 L test medium in closed glass bottle; mixed for 24 h on magnetic stirrer; set aside for 2 days; aqueous phase drawn off and filtered (folded filter); test substance concentration analytically determined; two solutions were prepared for each occasion of water exchange; theoretical concentration of tributylamine: 44 mg/L
- Controls: 1 negative control, 1 solvent control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): ethanol
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): ethanol in final test solutions: 0.0017 to 0.0773 ml/L; ethanol in control: 0.1 mL/L
- Evidence of undissolved material (e.g. precipitate, surface film, etc): slight turbidity and floccculation in test solutions
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
- Common name: zebra fish
- Strain:
- Source: West Aquarium, Bad Lauterberg, Germany
- Age at study initiation (mean and range, SD): 3 months (hatched: 10 Nov 1989; received 06 Feb 1990)
- Length at study initiation (length definition, mean, range and SD): mean: 3.1 cm; s = 0.24 cm; range = 2.7 to 3.5 cm; n = 10
- Weight at study initiation (mean and range, SD): no data
- Method of breeding: no data
- Feeding during test
- Food type: dried fish food (Tetra Standard Mix; Tetra Werk, Melle, Germany)
- Amount: 60 mg/day/10 fish
- Frequency: 1 to 2 times daily

- Acclimation period: 14 d
- Acclimation conditions (same as test or not):yes
- Type and amount of food: see above
- Feeding frequency: see above
- Health during acclimation (any mortality observed): no data
Test type:
Water media type:
Limit test:
Total exposure duration:
28 d
total hardness: 13-14 °dH (mean = 13.75, s=0.5, n= 4; determined weekly)
carbonate hardness: 2 °dH
Test temperature:
21.0-22.9 °C
Dissolved oxygen:
4.5-9.6 mg/L
Nominal and measured concentrations:
Test concentrations (nominal/analytical; mg/L): 0, 0 (ethanol), 0.315/0.530, 1/1.583, 3.15/4.817, 10/11.936
Recoveries between 67 and 260 % (mean: 119-168 %; only on two occasion below 80 % of nominal: day 17 and 18 at 10 mg/L ); test results evaluated on basis of nominal test item concentrations
Details on test conditions:
- Test vessel:
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: glass (30x22x24 cm), 10 L fill volume
- Aeration: performed when oxygen content below 5.8 mg/L
- Renewal rate of test solution (frequency/flow rate): renewal every 48 or 72 h (except for period of 04 May 1990 and 09 May 1990: 96h and 24 h)
- 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: below 1 g/L

- Source/preparation of dilution water: reconstituted water according to ISO/DIS 7346/1

- Adjustment of pH: no
- Photoperiod: 12:12 h light: dark
- Light intensity: 700 lux directly above water tanks

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
behaviour and general health condition after 2-4, 24, 48, (72 and 96) h after start of an interval

- Spacing factor for test concentrations: 3.15
Reference substance (positive control):
not required
Key result
28 d
Dose descriptor:
Effect conc.:
> 10 mg/L
Nominal / measured:
Conc. based on:
act. ingr.
Basis for effect:
Remarks on result:
other: no mortality occurred during test; highest tested conc.: 10 mg/L
28 d
Dose descriptor:
Effect conc.:
0.315 mg/L
Nominal / measured:
Conc. based on:
act. ingr.
Basis for effect:
28 d
Dose descriptor:
Effect conc.:
1 mg/L
Nominal / measured:
Conc. based on:
act. ingr.
Basis for effect:
Details on results:
- Observed effects at each concentration for each observation time: see "Remarks on results..."
- Concentrations that produce lethal or other effects: effects at 1 mg/L and above (details see below at "Remarks on results..."
- Cumulative mortality at each concentration and for each recommended observation time if possible: No mortality occurred during the test.
- Mortality in the controls: none
- Behavioural observation of the fish:
- Effect concentrations exceeding solubility of substance in test medium: no
- Incidents in the course of the test which might have influenced the results: none

- Control, solvent control, 0.315 mg/L: no effects observed.

- 1 and 3.15 mg/L: delayed feed consumption on first two days; no effects observed afterwards, feeding normalised

- 10 mg/L: feed intake (none at test start, changed to delayed and reduced intake in the course of the test); hypo- or hyperactivity, tonic-clonic convulsions, abnormal swimming behaviour, unregular and forced respiration.

Validity criteria fulfilled:
The 28-d LC50 was determined to be greater than 10 mg/L.
Executive summary:

Tributylamine was tested on zebrafish (Danio rerio, reported as Brachydanio rerio) in a 28 -d semi-static subchronic study according to OECD TG 204. Ten fish were tested per concentration, control and solvent control (ethanol). The test medium was exchanged after 48 or 72 h (on two occasions after 24 h and 96 h). The test concentrations were (nominal/measured): 0, 0 (ethanol), 0.315/0.530, 1/1.583, 3.15/4.817, 10/11.936 mg/L. Recoveries were between 67 and 260 % (mean: 119-168 %; only on two occasions below 80 % of nominal: day 17 and 18 at 10 mg/L ). Since no stable concentrations were established and the test concentrations were - on average - above the nominal level, the test results were evaluated on basis of the nominal test item concentrations. No mortalities were observed up to the highest test concentration of 10 mg/L. Bahvioural abnormalities were observed at 1 mg/L and above. Based on nominal concentrations, the 28 -d LC50 was greater than 10 mg/L.

Description of key information

Danio rerio: 28-d LC50 > 10 mg/L (OECD 204; Hoechst, 1991)
Oryzias latipes: 96-h LC50 = 16.3 mg/L (OECD 203; NITE, 2002)

Key value for chemical safety assessment

LC50 for freshwater fish:
10 mg/L

Additional information

Results are available for several fish species (warm and cold water), but compared to the recent OECD TG 203 - especially regarding exposure period and analytical monitoring - only two studies are valid (Hoechst, 1991; NITE, 2002).


In the BUA report, it was noted that the highest concentration without lethality ranges from 20 to 60 mg/L with an exposure period of 24 to 48 h. This period is not sufficient compared to the recent OECD TG 203, which requires a test period of 96 h. Therefore these values must be considered invalid.




Value [mg/L]




Tributylamine (CAS 102-82-9)

28-d LC50

Danio rerio

> 10


analyt. verified

key study

Hoechst AG (1991)

96-h LC50

Oryzias latipes




supp. study

NITE (2002)


Because the semi-static study on Danio rerio spanned 28 days and was analytically verified, it is regarded to be most relevant and used as the key study. Because only behavioural effects and no lethality occurred up to 10 mg/L, and given that with Oryzias latipes the LC50 was determined as 16.3 mg/L, as a key value for hazard and risk assessment 10 mg/L are used as the acute LC50 for freshwater fish.

Based on these studies, tributylamine is considered acutely harmful to fish.