Amines, N-C12–14 (even numbered)-alkyltrimethylenedi-, reaction products with chloroacetic acid and diethylenetriamine, N-C12–14 (even numbered)-alkyl derivs.

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

The limit concentration and the control were analytically verified from freshly prepared media at after 0 and 72 h and from the corresponding 24 h aged test media after 24 and 96 h.
All original samples were stored at room temperature before preparation. Prepared samples were stored in an autosampler at room temperature until analysis.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A stock solution of 100 mg/L was freshly prepared in demineralized tap water. The dispersion was agitated until the stock solution was clear. After transferring an appropriate volume of the stock solution into the test vessel (filled with river water), the test medium was mixed with an ultraturrax (1 min, at least 17000 rpm). This procedure was repeated daily for the preparation of the stock solution and the water renewal of the threshold concentration.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebrafish
- Source: All fish used in the test was gained at Noack Laboratorien GmbH from a single brood stock (supplier: Umweltbundesamt, Schichauweg 58, D-12307 Berlin)
- Length at study initiation (length definition, mean, range): 2.3 cm (2.0 – 2.5 cm)
- Weight at study initiation (mean): 0.11 g

ACCLIMATION
- Acclimation period: 7 d
- Acclimation conditions (same as test or not): same
- Type and amount of food during acclimation: Food (Sera Vipan; SERA GMBH, 52518 Heinsberg, Germany) was provided 3 times per week. The amount of food was 4 % of the fish body weight per feeding day.
- Health during acclimation (any mortality observed): No mortality was observed. No disease treatments were administered throughout holding and testing.


FEEDING DURING TEST
- The test fish were not fed 24 h before test start.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

67 mg/L as CaCO3

Test temperature:

22.3 - 22.8

pH:

7.15 - 7.64

Dissolved oxygen:

saturation 96-99%

Nominal and measured concentrations:

146 µg/L, corresponding to 38.8 µg a.i./L

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass aquaria of 3 L, filled with about 2.5 L of natural river water (dimensions: 11.5/13/20 cm, depth of water: about 16.5 cm), and covered with glass plates.
- Aeration: yes
- Renewal rate of test solution: daily renewal
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.30 g fish per litre test solution

TEST MEDIUM / WATER PARAMETERS
- see Any other information on material and methods

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 12/12
- Light intensity: 7 – 750 Lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
Observations were made after 3, 24, 48, 72 and 96 h
Fish were considered dead if there was no visible movement (e.g. gill covers movement) and if touching of the caudal peduncle produced no reaction. Records were kept of visible abnormalities (e.g. loss of equilibrium, swimming behavior, respiratory function, pigmentation, etc.).


TEST CONCENTRATIONS
- limit test; selection of the test concentration was based on the derivation of a threshold concentration (TC) from the results of an alga toxicity test (ErC50 (0-72 h) = 146 µg/L (nominal test item concentration) and a daphnia acute immobilization test
(EC50 (0-48 h) = 609 µg/L (nominal test item concentration))

Reference substance (positive control):

not required

Key result

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

>= 38.8 µg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

>= 146 µg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: none observed
- Mortality of control: no mortality observed in the control

Sublethal observations / clinical signs:

Validity criteria:

·     The O2 saturation should be >/=60% (this study: >/=96%).

·     The mortality in the control group should not exceed 1/7 fish (this study: 0/7 fish).

·     The pH-value in the control group should not deviate by more than one unit over the test period (this study: 0.47 units).

Observations in the Definitive Test

(n = 7)

 

Nominal dry content concentration of the test item [µg/L]	Observation*)	Number of fish effected after different exposure periods [hours]
		3	24	48	72	96
38.8	(1)	7	7	7	7	7
Control	(1)	7	7	7	7	7

*)The number in brackets corresponds to the following observation: 

(1) = Normal behavior

 

Cumulative Mortality [%] in the Definitive Test

 

Nominal dry content concentration of the test item [µg/L]	Cumulative Mortality [%] after different exposure periods [hours]
	3	24	48	72	96
38.8	0	0	0	0	0
Control	0	0	0	0	0

 

Validity criteria fulfilled:

yes

Conclusions:

The LC0 after 96 hours was ≥ 146 µg test item/L, corresponding to ≥ 38.8 µg/L of the dry content of the test item.

Executive summary:

The acute toxicity of Reaction product of lauryl-PDA/lauryl-DETA with chloroacetic acid (26.6% a.i.) to fish (zebrafish) was determined according to OECD Guideline 203 (1992) and EU Method C.1 (2008). An acute toxicity test under semi-static conditions with natural river water and daily renewal of the test media was conducted with the limit concentration of 146 µg/L of the test item, corresponding to a nominal dry content concentration of 38.8 µg/L. Duration of the test was 96 hours. Seven test organisms were exposed to the limit concentration and the control. Water quality parameters (temperature, pH-value and oxygen-saturation) measured at 0, 24, 48, 72 and 96 hours were determined to be within the acceptable limits.

 

The limit concentration of the test item and the control were analytically verified via LC-MS/MS from freshly prepared media after 0 and 72 h and from corresponding 24 h aged test media after 24 and 96 h. The measured concentrations at the start of the exposure intervals after 0 and 72 hours were 94 and 100% of the nominal value. At the end the exposure intervals after 24 to 96 hours the measured test item concentrations were 41and 39% of the nominal value.

Since the test is an UVCB substance all effect values given are based on the nominal concentration of the test item and its dry content. All validity criteria of the test guideline were met.

The LC0 was >/=146 µg/L in terms of test material, corresponding to >/=38.8 µg a.i./L.

Results Synopsis

Test organism size/age: Danio rerio (mean length: 2.3 cm; mean weight: 0.11 g)

Test Type: semi-static

LC50:  >/= 38.8 µg a.i./L                       

Endpoint(s) Effected: mortality and sublethal effects

Description of key information

LC0  >/= 38.8 µg a.i./L   

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

38.8 µg/L

Additional information

The acute toxicity of Reaction product of lauryl-PDA/lauryl-DETA with chloroacetic acid (26.6% a.i.) to fish (zebrafish) was determined according to OECD Guideline 203 (1992) and EU Method C.1 (2008). An acute toxicity test under semi-static conditions with natural river water and daily renewal of the test media was conducted with the limit concentration of 146 µg/L of the test item, corresponding to a nominal dry content concentration of 38.8 µg/L. Duration of the test was 96 hours. Seven test organisms were exposed to the limit concentration and the control. Water quality parameters (temperature, pH-value and oxygen-saturation) measured at 0, 24, 48, 72 and 96 hours were determined to be within the acceptable limits.

The limit concentration of the test item and the control were analytically verified via LC-MS/MS from freshly prepared media after 0 and 72 h and from corresponding 24 h aged test media after 24 and 96 h. The measured concentrations at the start of the exposure intervals after 0 and 72 hours were 94 and 100% of the nominal value. At the end the exposure intervals after 24 to 96 hours the measured test item concentrations were 41 and 39% of the nominal value.

Since the test is an UVCB substance all effect values given are based on the nominal concentration of the test item and its dry content. All validity criteria of the test guideline were met.

The LC0 was >/=146 µg/L in terms of test material, corresponding to >/=38.8 µg a.i./L.

The 96 h LC50 of the source substance DOPA-Glycinate in rainbow trout (Oncorhynchus mykiss) was 207.4 µg a.i./L (CL: 157.5–273.2 µg a.i./L).