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Ecotoxicological information

Short-term toxicity to fish

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Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP Guideline study
Cross-reference
Reason / purpose:
reference to same study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2006
Report Date:
2006

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Chemical name: Amines, coco alkyl

Sampling and analysis

Analytical monitoring:
yes

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Two stock solutions were prepared. The first stock solution could not be analysed due to an
error during sampling. The second stock solution (for test media preparation after 48 h) was
analytically verified.
Dispersion treatment: 6 μL/L test item were applied to 3 L natural river water in a measuring flask. This solution was treated for 30 minutes with ultrasound at 40 °C. After a cooling period of 10 minutes the test item concentrations were prepared from this stock solution. The stock solution concentrations were analytically verified.

Test organisms

Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: Zebrafish
- Strain: Danio rerio
- Source: AQUARIUM AM AEGI, Volgersweg 6, D-30175 Hannover
- Age at study initiation (mean and range, SD):
- Length at study initiation: 2.50 cm
- Weight at study initiation: 0.76 g
- Feeding during test: No


HOUSING:
- Conditions: Holding was performed at the test facility at 23 ± 2 °C, diffuse light (0.1-10 μmol photons · m-2 · s-1, natural photoperiod). The water was changed at least once per week. The dissolved oxygen concentration was more than 80 % of the air saturation value.
- Type and amount of food: The amount of food was 4 % of the fish body weight per feeding day. The test fish were not fed 24 h before test start.
- Feeding frequency: Food was provided 3 times per week.

ACCLIMATION
- Acclimation period: 12 days
- Health during acclimation: Only zebrafish with mortality < 5 % within the last 7 days before the study started were used in the test. No disease treatments were administered throughout holding and testing.


Study design

Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h

Test conditions

Hardness:
116 - 129 mg/L CaCO3
Test temperature:
20 - 24 ± 1 °C
pH:
7.5 +/- 0.4
Dissolved oxygen:
> 90 % of air saturation.
Nominal and measured concentrations:
5 concentrations were tested: 0.20 – 0.36 – 0.63 – 1.12 – 2.00 mg/L.
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Material, fill volume: Glass-aquaria loosely covered by glass tops, fill volume about 3 L.
- Renewal rate of test solution (frequency/flow rate): Renewal of the test media after 48 h.
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1



TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: A natural occurring river water (River Böhme) was used as test medium.
Test water parameter:
pH: 7.68
Conductivity [μS/cm]: 451
DOC [mg C/L]: 6.3
TOC [mg C/L]: 7.1
Ammonium-N [mg N/L]: < 1.94
Nitrate-N [mg N/L]: 3.9
o-Phosphate-P [mg P/L]: 0.029
Total Phosphate [mg P/L]: 0.080
Suspended Matter [mg/L]: 16.9
Total Hardness [mg CO3/L]: 131.4
- Intervals of water quality measurement: pH-value, temperature and oxygen saturation were measured in all vessels at the beginning of the test and every 24 h (from old and new media). Total hardness of the water was determined at the beginning and after 48 h renewal out of the test control. During the test the room temperature was recorded continuously with a thermohygrograph.


OTHER TEST CONDITIONS
- Photoperiod: 12 – 16 h illumination
- Light intensity: 0.1 - 10 μmol photons · m-2 · s-1


EFFECT PARAMETERS MEASURED: Mortality and symptoms were measured. Observations were made after 2, 24, 48, 72 and 96 h.

Reference substance (positive control):
not required

Results and discussion

Effect concentrationsopen allclose all
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
0.63 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
0.84 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
1.12 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality

Any other information on results incl. tables

Table 1: Cumulative Mortality [%] in the test vessels with Amines, coco alkyl

Nominal conc.(mg/l)

Number of fish

Cumulativ Mortality (%) after

2 h

24 h

48 h

72 h

96 h

2.0

7

0

100

100

100

100

1.12

7

0

0

57

100

100

0.63

7

0

0

0

0

0

0.36

7

0

0

0

0

0

0.2

7

0

0

0

0

0

Control

7

0

0

0

0

0

Table 2: Concentrations and Recovery Rates of Amines, coco alkyl

Nominal test item conc.

[mg/L]

Meas. conc. [mg/L]after 0 h (new medium)

Meas. conc. [mg/L]after 48 h (old medium

Meas. conc. [mg/L]after 48 h (new medium)

Meas. conc. [mg/L]after 96 h (old medium

2.0

2.39

1.97

n.d.

n.d.

1.12

1.33

< detection limit

1.32

< detection limit

0.63

0.74

< detection limit

0.72

< detection limit

0.36

0.40

< detection limit

0.43

< detection limit

0.2

0.23

< detection limit

0.26

< detection limit

Control

< detection limit

< detection limit

< detection limit

< detection limit

Applicant's summary and conclusion

Conclusions:
In this study Amines, coco alkyl was found to be toxic to Zebrafish after 96 h at a nominal test item concentration of 1.12 mg/L and higher. Therefore the NOEC was laid down as 0.63 mg/L (nominal test item concentration). The LC50 (96 h) is 0.84 (0.63 - 1.12) mg/L.
Executive summary:

The acute toxicity of the test item Amines, coco alkyl (batch no. S000358) to fish (Zebrafish) was determined according to the principles of OECD 203 from 2006-03-01 to 2006-03-05 at DR.U.NOACK-LABORATORIEN, Sarstedt, Germany. A semi-static test in natural river water with one renewal of the test media after 48 h was conducted with the following nominal concentrations: 0.20 - 0.36 - 0.63 - 1.12 - 2.00 mg/L. Duration of the test was 96 h. 7 test organisms were exposed to each test concentration and control. Water quality parameters temperature, pH-value, O2-saturation measured at 0, 24, 48, 72 and 96 h were determined to be within the acceptable limits. All concentration levels were analytically verified via LC-MS/MS. Freshly prepared test media at sampling time 0 and 48 h and corresponding 48 h old solutions were analysed. The measured initial concentrations of both series were in good accordance. In 48 h old media the test item concentrations were below the limit of quantification (= 10 µg/L). Adsorption to the test vessels after 48 h of exposure, determined from the dosage levels framing the LC50, was found to be negligible (for details of the analytical method and results please refer to chapter 9). The test item has a low water solubility and sorbs to organic and inorganic materials by different mechanisms. The sorption processes are mostly non-linear, means are concentration dependent. Due to these properties the test item is difficult to test in synthetic water (e.g. sorption to the test organism and walls of the test vessel) and results from such tests depend from the test settings applied. Using natural river water which contains particulate as well as dissolved organic carbon to which the test item can sorb partially reduces the difficulties encountered in tests with synthetic water e.g. preventing that the test item settles onto surfaces. The sorbed fraction of the test item is difficult to extract from the test system which normally leads to low analytical recoveries. Due to the short exposure period these low recoveries cannot be associated to biodegradation. This means the test substance is present in the test system and therefore available for exposure (dissolved in water and sorbed also called bulk). Due to the properties of the test item nominal concentrations have to be used instead of measured ones (see Table 1). This so called Bulk Approach is described by ECETOC (2003). Sorption of the test item to the glass ware of the test system was monitored and found to be small.

Test duration [h] LC50 [mg/L] p =95 % [mg/L]
2 > 2.00 not applicable
24 1.50 1.12 - 2.00
48 1.09 0.98 - 1.22
72 0.84 0.63 - 1.12
96 0.84 0.63 - 1.12
LC100
lowest test item concentration with 100 % mortality after 96 h
1.12 mg/L
LC0
highest test item concentration with 0 % mortality after 96 h
0.63 mg/L