3-[(2-acetamidoacetyl)amino]propanoic acid

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

January to February 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

adopted April 13, 2004

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

Commission Regulation (EC) No 440/2008, C.2., 2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Sampling: Duplicate samples from the freshly prepared test media of all test concentrations and the controls were taken at the start of the test.
For the determination of the stability of the test item under the test conditions and of the maintenance of the test item concentrations during the test period, duplicate samples from the test media of all
test concentrations and the controls were collected at the end of the test (after 48 hours) by pouring together the contents of the test beakers of each treatment.
Additional samples of the control were taken at test start and test end.
Analyses: The concentrations of the test item GenoWhite were analysed in the duplicate test media samples from all test concentrations, and in the duplicate control samples, from both sampling times (0 and
48 hours).

Vehicle:

yes

Remarks:

Test item dissolved in test water.

Details on test solutions:

The test medium of the highest test concentration of nominal 100 mg test item/L was prepared by dissolving 60 mg test item into 600 mL test water by intense stirring for 15 minutes. Adequate volumes of
this test medium were diluted with test water to prepare the test media.
The test media were prepared just before introduction of the Daphnia (= start of the test).

Test organisms (species):

Daphnia magna

Details on test organisms:

Daphnia magna (Straus), clone5; 2.75 to 18.75 hours old.
Sex: Female.
Origin: Daphnia introduced in the test were taken from ibacon's in-house laboratory culture.
Breeding Conditions: The Daphnia were bred in the laboratories of ibacon under similar temperature and light conditions as used in the test. The cultivation of the parental Daphnia was performed in Elendt M4 medium (see 6.5). The test organisms were not first brood progeny. The Daphnia in the stock culture were fed at least on all working days with green algae (Desmodesmus subspicatus) freshly grown in the
laboratories of ibacon.
Reference Item: For the evaluation of the quality of the Daphnia clone and the consistency of the experimental conditions, the reference item potassium dichromate is tested at least twice a year to demonstrate
satisfactory test conditions. The results of the most recent reference test are included in Table 10.
Acclimatisation: Was not necessary, since the test was performed in the same medium as the culturing (see 6.5).

Test type:

static

Water media type:

freshwater

Remarks:

Reconstituted Water (Elendt "M4")

Limit test:

no

Total exposure duration:

48 h

Hardness:

2.5 mmol/L as CaCO3

Test temperature:

20-19 ºC

pH:

7.8 to 8.0

Dissolved oxygen:

9.8 to 10.0 mg/L at test start
9.6 to 9.9 mg/L at test end

Salinity:

Alcalinity: 0.9 mmol/L

Nominal and measured concentrations:

At the start of the test 94% of the nominal test concentrations were found (average of all test concentrations). After 48 hours test duration, 92% of the nominal value was determined (average of all test concentrations). During the test the daphnids were exposed to a mean of 93% of nominal. Therefore, all reported results refer to nominal concentrations.

Details on test conditions:

Test Units
Type and Size: Glass beakers of 100 mL volume containing approximately 60 mL of test medium. The test vessels were covered with a lid to reduce the loss of water due to evaporation and to avoid the entry of dust into the solutions.
Identification: Each test unit was uniquely identified with serial number, treatment and replicate number.
Test Conditions
Test Environment: Controlled environment room
Measurement of pH, Dissolved Oxygen and Water Temperature: The water temperature, pH-values and dissolved oxygen concentrations were determined at test start and test end in each treatment group.
Water Temperature: 20 to 20 °C at test start; 19 to 19 °C at test end
pH-Values: 7.8 to 8.0 at test start; 7.8 to 7.9 at test end; and thus the pH-value did not vary by more than 1.5 units
Dissolved Oxygen Concentration: 9.8 to 10.0 mg/L at test start;9.6 to 9.9 mg/L at test end (see Table 6)
Light Regime: 16 h light : 8 h dark
Light Intensity: The light intensity was 210 to 670 lux (measured once during the test).
Recording: Test conditions were recorded with suitable instruments and documented in the raw data.

Key result

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 100 ng/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 ng/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

other: EC20

Effect conc.:

> 100 ng/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

other: EC20

Effect conc.:

> 100 ng/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC10

Effect conc.:

> 100 ng/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

> 100 ng/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

24 h

Dose descriptor:

NOEC

Effect conc.:

>= 100 ng/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

> 100 ng/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Validity Criteria of the Study
Control Immobilisation Rate: Was 0 % and furthermore no daphnid showed signs of disease or stress; thus the validity criterion was met.
Dissolved Oxygen Concentration: Was ≥ 9.6 mg O2/L in the control and test vessels at the end of the test; thus validity criterion was met.

Biological test results: After 48 hours of exposure no immobilisation of the test animals was observed in the control and up to and including the highest test item concentration of 100 mg test item/L.

Validity criteria fulfilled:

yes

Conclusions:

The toxic effect of the test item to Daphnia magna was assessed in a static dose-response test. The 48-hour NOEC was determined to be ≥ 100 mg test item/L.
The 48-hour LOEC was determined to be > 100 mg test item/L and the 48-hour EC50 value was determined to be >100 mg test item/L.

Executive summary:

The toxic effect of the test item to Daphnia magna was assessed in a static dose-response test (OECD 202: "Daphnia sp., Acute Immobilisation Test").

After 48 hours of exposure no immobilisation of the test animals was observed in the control and up to and including the highest test item concentration of 100 mg test item/L.

The 48-hour NOEC was determined to be ≥ 100 mg test item/L.

The 48-hour LOEC was determined to be > 100 mg test item/L and the 48-hour EC50 value was determined to be >100 mg test item/L.

Description of key information

The toxic effect of the test item to Daphnia magna was assessed in a static dose-response test (OECD 202: "Daphnia sp. Acute Immobilisation Test").

The 48-hour NOEC was determined to be ≥ 100 mg test item/L.

The 48-hour LOEC was determined to be > 100 mg test item/L and the 48-hour EC50 value was determined to be >100 mg test item/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 ng/L