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Ecotoxicological information

Short-term toxicity to fish

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Reference
Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2002-02-25 / 2002-09-26
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Validity criteria can be considered as fulfilled. Mortality of control animals was <10%, the concentration of dissolved oxygen in all test vessels was > 60% saturation and the concentration of test substance >= 80% of initial concentration during test. Although quantification of the applied concentrations was not possible due to the absence of a suitable analytical method. Therefore only the two stock solutions used to prepare the test concentrations were analyzed. These analyses showed that the stock solutions were stable even after storage for almost 7 months in the refrigerator. Considering the high water solubility of the test substance and its stability it is reasonable to assume that the actual concentrations were identical to the nominal concentrations during the entire test. A dose-response related increase in mortality was observed. Study peformed according to standard protocol and under GLP.
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
GLP compliance:
yes
Analytical monitoring:
yes
Remarks:
Concentration of the stock solution was quantified
Details on sampling:
Quantification of the applied concentrations was not possible due to the absence of a suitable analytical method. The currently available analytical method is capable of analyzing down to only 10 mg/L. Therefore only the two stocksolutions used to prepare the test concentrations were analyzed. The analysis showed that the stocksolutions were stable even after storage for almost 7 months in the refrigerator. All concentrations mentioned in the report are therefore based on nominal concentrations. Considering the high water solubility of the test substance and its stability it is reasonable to assume that the actual concentrations were identical to the nominal concentration during the entire test.
Vehicle:
no
Details on test solutions:
STOCK AND TEST SOLUTION AND THEIR PREPARATION:
- Dispersion: A stock solution was prepared by dissolving approximately 1 g of test substance in 120 mL deionized water at 50 °C under stirring. After cooling pH was adjusted to 8.5 and deionized water was added up to a volume of 1000mL (at day 2 a fresh solution was prepared to change the medium). The test solutions were prepared by addition of the required amounts of stock solution to the test medium (Dutch Standard Water). - Vehicle, solvent: deionized water - Concentration of vehicle/ solvent: 1 g/L -
Other procedures: - STABILITY OF THE TEST CHEMICAL SOLUTIONS: REFERENCE SUBSTANCE: DILUTION WATER - Source: Dutch Standard Water; Containing per liter of dionized water 100 mg Na HCO3, 20 mg KHCO3, 200 mg CaCl2.2H2O and 180 mg MgSO4.7H2O - Aeration: test solutions were aerated with water-saturated air purified by an active coal filter and cotton filter. - Alkalinity: - - Hardness: 12°dH - Salinity: - - TOC: - - TSS: - - pH: 8.2 - Oxygen content: 8.2-8.7 mg/L - Conductance: < 5uS/cm - Holding water: DSW
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISMS - Strain: Brachydanio rerio (zebra fish) - Supplier: Retailer (fish were accepted according to the quality control criteria of the guidelines and SOP) - Wild caught: no - Age/size/loading: Juvenile/<= 3 cm/1g/L (wet weight) - Feeding: Commercially available dry or deep-frozen food - Pretreatment: 1-3 times a day - Feeding during test: No; feeding stopped 24 hrs before study
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Post exposure observation period:
no
Hardness:
Hardness: 12°dH
Test temperature:
19.4-21.4 °C
pH:
8.2
Dissolved oxygen:
Oxygen content: 8.2-8.7 mg/L
Salinity:
DSW was used which is a freshwater
Nominal and measured concentrations:
Concentrations: 0.1 - 0.18 - 0.32 - 0.58 and 1.05 mg/L
Details on test conditions:
TEST SYSTEM - Test type: semi static - Concentrations: 0.1 - 0.18 - 0.32 - 0.58 and 1.05 mg/L - Dosing rate: - - Renewal of test solution: day 2 - Exposure vessel type: 5L aquaria containing 3L of test solution - Number of replicates, fish per replicate: one aquarium per dose with 7 fish - Test temperature: 19.4-21.4 °C - Dissolved oxygen: 8.2-8.7 mg/L - pH: 7.6-8.2 - Adjustment of pH: the pH of the stock solution (1.0031 g in 120 ml and 1.0004 g in 120 ml) of the chemical prepared in deionized water of approximately 50 °C after cooling down the pH was adjusted to 8.5. Deionized water was added up to a volume of 1000 mL. - Intensity of irradiation: Ambient light provided by fluorescent tubes - Photoperiod: 16 h photoperiod daily
DURATION OF THE TEST: 96 hrs
TEST PARAMETER: The fish were considered dead when a lack of opercular movement was observed and touching of the caudal peduncle produced no reaction. Dead fish were removed from the test vessels directly after being observed, which was checked at least twice each day. In addition to death, sub-lethal effects such as erratic swimming, loss of reflex, increased excitability, lethargy, changes in physiology, discoloration, pigmentation, excessive mucous production, hyperventilation, opaque eyes, curved spine or hemorrhaging were recorded. Fish that were convulsing or showing other severe forms of distress not considered transient in nature and likely to become more severe before the exposure is terminated, were sacrificed for humane reasons. These fish were treated as having died in the test.
SAMPLING: Not performed
MONITORING OF TEST SUBSTANCE CONCENTRATION: Not performed
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
1.05 mg/L
Remarks on result:
other: IUCLID4 note: "m" (measured/nominal)
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
0.18 mg/L
Remarks on result:
other: IUCLID4 note: "m" (measured/nominal)
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
0.431 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks on result:
other: IUCLID4 note: "c" (calculated)
Duration:
96 h
Dose descriptor:
LC0
Effect conc.:
0.18 mg/L
Remarks on result:
other: IUCLID4 note: "m" (measured/nominal)
Sublethal observations / clinical signs:
RESULTS: EXPOSED
- Nominal/measured concentrations: 0, 0.1, 0.18, 0.32, 0.58,1.05 mg/L (nominal)
- Effect data (Mortality):
    24h    48h 72h 96h
Control 0 0 0 0
0.1 0 0 0 0
0.18 0 0 0 0
0.32 0 0 0 1
0.58 0* 1* 5 6
1.05 1** 7
* reduced acitivity
** very reduced activity
For ethical reasons two fish were euthanised at the
concentration of 0.58 ml/L after 53 hours.

The LC50 (96 h) was calculated to be 0.431 mg/L with 95% confidence limits of 0.346 and 0.538 m/L. The highest
concentration causing no mortality (no observed effect concentration, NOEC) after 96 h amounted to 0.18 mg/L
whereas 0.32 m/L was the lowest tested concentration (LOEC,96 h) at which lethality was observed as compared to the
control. The LC100 was 1.05 m/L, which was observed after 29 hours.

- Concentration / response curve: -
- Effect concentration vs. test substance solubility: -
- Other effects: see mortality data * and **
RESULTS: CONTROL
- Number/percentage of animals showing adverse effects: None
- Nature of adverse effects: -
Validity criteria fulfilled:
yes
Conclusions:
The LC50 (96 h) was calculated to be 0.431 mg/L with 95% confidence limits of 0.346 and 0.538 m/L. The highest concentration causing no mortality (no observed effect concentration, NOEC) after 96 h amounted to 0.18 mg/L whereas 0.32 m/L was the lowest tested concentration (LOEC,96 h) at which lethality was observed as compared to the control. The LC100 was 1.05 m/L, which was observed after 29 hours.
Executive summary:

The acute toxicity of Triameen Y12D to the fresh water fish Brachydanio rerio (zebra fish) was tested under semi-static conditions in accordance with OECD and EEC guidelines for testing of chemicals. The test was performed in compliance with Good Laboratory Practice (GLP). The fish were exposed to the following nominal test concentrations: 0.1- 0.18 – 0.32 – 0.58 and 1.05 mg/l. The LC50(96 h) was calculated to be 0.43 mg/l with 95% confidence limits of 0.346 and 0.538 mg/l. The highest concentration causing no mortality (no observed effect concentration, NOEC) after 96 h amounted to 0.18 mg/l whereas 0.32 mg/l was the lowest tested concentration (LOEC, 96 h) at which lethality was observed as compared to the control. The LC100was 1.05 mg/l, which was observed after 29 hours.

At the concentrations where lethality was established, sublethal effects, such as reduced activity, deviations in the behavior or the appearance of the fish, were observed. For ethical reasons two fish were euthanised at the concentration of 0.58 mg/l after 53 hours.

Quantification of the applied concentrations was not possible due to the absence of a suitable analytical method. The currently available analytical method is capable of analyzing down to only 10 mg/l. Therefore only the two stock solutions used to prepare the test concentrations were analyzed. These analyses showed that the stock solutions were stable even after storage for almost 7 months in the refrigerator. All concentrations mentioned in this report are therefore based on nominal concentrations. Considering the high water solubility of the test substance and its stability it is reasonable to assume that the actual concentrations were identical to the nominal concentrations during the entire test.

Description of key information

Only a limit number of short-term toxicity to fish studies are available for the substances under consideration. Therefore the test results for dodecyltriamine Y was added to the dossier. Tri- and tetramines contain respectively about 22 – 11% diamines and for this reason also the data for the diamines was added to the overview.  

It is considered reasonable to add dodecyltriamine Y as a worst-case as this substance is similar as the linear triamine and considered a the most toxic triamine. Dodecyltriamine Y is therefore selected as the Key study as this study is a Klimisch 1. The LC50 of this study is therefore used as the Key value for chemical safety assessment as a reasonable worst-case. In the final risk assessment this value will not be critical as two chronic end points are available.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Effect concentration:
0.431 mg/L

Additional information

Most of the short-term toxicity fish tests were conducted in a period when no reliable specific method of analyses was available. The concentrations were therefore not analytically verified, the reliability of the results is limited because of the poor solubility of the test substances and partial sorption onto the walls of test vessels. Hence, most of the tests were considered to be valid with restrictions. Only one acute fish test study is available for N-(3-aminopropyl)-N-dodecylpropane-1,3-diamine (2372-82-9; dodecyltriamine Y) which is used as a biocide and this substance is considered as the most ecotoxic substance for the alkyl polyamines (>2N).The observed LC50 for this substance of 0.43 mg/L, suggests that the alkyl-1,3-diaminopropanes (2N) are more ecotoxic than the alkyl polyamines (>2N). Tri- and tetramines contain respectively about 22 – 11% diamines and for this reason the data for the diamines was added to the overview. 

For oleyl tetramine (4N) an LC50 (96 h) has been observed of 0.13 mg/L. This value is slightly lower than the value observed for dodecyltriamine Y. In the presence of humic acid however the toxicity of oleyl tetramine is reduced to an LC50 (96 h) of 2.01 mg/L demonstrating that under environmental conditions the toxicity is significantly reduced by river water constituents.