Reaction products of propane-1,2-diamine with 2-[(2-methylphenoxy)methyl]oxirane

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:

from 2018-01-25 to 2018-05-24

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)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1010 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Four parallel samples were taken from the test solutions and from the control. Formulation samples were diluted with mobile phase and the control samples were directly injected.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- The test solution used in the test was prepared by mechanical dispersion without using of any solubilising agent. An amount of 0.0753 g test item was dissolved in 753 mL dilution water (ISO Medium) to obtain the nominal concentration of 100 mg/L. The test solutions of the further test concentrations were prepared by the appropriate dilution of this stock solution.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Strain/clone: Straus
- Source: Laboratory of Hydrobiology (Central Agricultural Office, Directorate of Plant-, and Soil Protection) 2100 Gödöllő, Kotlán S. u. 3., Hungary
- Feeding during test: no

ACCLIMATION
- Test animals were bred under similar conditions as that used during the exposure period, therefore additional acclimatisation before the test was not necessary.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

249 mg/L CaCO3

Test temperature:

20.8 - 21.3 °C

pH:

7.97 - 8.89

Dissolved oxygen:

7.58 - 8.40 mg/L

Salinity:

NA

Nominal and measured concentrations:

Nominal concentrations: 0, 3.1, 6.3, 12.5, 25, 50 and 100 mg/L
Measured concentrations: 0, 2.87, 5.71, 11.57, 23.45, 47.22 and 99.63 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: beakers
- Material, size, headspace, fill volume: glass, approximately 40 mL test medium
- Aeration: no
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Volume/Organism: 2 mL

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The ISO medium was prepared by adding 25 mL from each of four stock solutions to one litre deionised water proportionally.
Stock solution 1: CaCl2 x 2 H2O 11.76 g/L
Stock solution 2: MgSO4 x 7H2O 4.93 g/L
Stock solution 3: NaHCO3 2.59 g/L
Stock solution 4: KCl 0.23 g/L

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light (artificial illumination) and 8 hours darkness

EFFECT PARAMETERS MEASURED:
The immobility or mortality of the Daphnia was determined by visual observation after 24 and 48 hours of exposure. Those animals not able to swim within 15 seconds after gentle agitation of the test beaker were considered to be immobile.

RANGE-FINDING STUDY
- Results used to determine the conditions for the definitive study: A preliminary test was performed at the nominal concentation of 100 mg/L in order to check the toxicity of the test item on the daphnids up to at least this concentration. In the preliminary test no immobility or any sub-lethal effect was detected in the treated concentration and in the control group during the 48-h exposure period.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

16.595 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

8.614 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

6.3 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

12.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Results with reference substance (positive control):

The date of the last study with reference item Potassium dichromate was: 05 – 06 September 2017.
The 24h EC50: 1.53 mg/L, (95 % confidence limits: 1.30 – 1.78 mg/L)

Validity of the Study

– The immobilisation rate of the introduced Daphnia did not exceed 10 % in the control group. Immobility was not observed in the control and no Daphnia were trapped at the water surface or showed any sign of stress.

– The dissolved oxygen concentration at the end of the test was more than 3 mg/L in control and test vessels. It was 7.75 – 7.84 mg/L.

All validity criteria were within acceptable limits, therefore, the study is considered as valid.

Analytical Results

Samples were taken from the test concentration and the control at the start and at the end of the experiment and analysed by HPLC method with MS detection. The test item concentrations were calculated based on the measured concentrations of the two components: (1-((2-Amino-2-methylethyl)amino)-3-(2-methylphenoxy)propan-2-ol and 3,3’-(Propane-1,2-diylbis(azanediyl))bis(1-(2-methylphenoxy)propan-2-ol)). The measured concentrations of these components were in the range of 91 – 105 % of the nominal during the test. There is evidence that the concentration of the tested substance was maintained within ± 20 % of the nominal concentration throughout the test. Therefore, the analysis of the results was based on the nominal concentration value.

Biological Results

No immobilization was observed in the control during the test. All test animals were observed as immobile at the test concentrations of 50 and 100 mg/L after 48 h. 70 % of the daphnids was observed as immobile at the test concentration of 25 mg/L, while 40 % immobilisation was found at 12.5 mg test item/L after 48 hours. In the test groups of 3.1 and 6.3 mg/L no immobilisation occurred. Accordingly, the NOEC was determined to be 6.3 mg/L.

Table 1: Immobilisation of the Test Animals (Summary Table)

Nominal concentration [mg/L]	No. of Daphnia tested	No. of immobilised Daphnia after		% of immobilised Daphnia after
		24 h	48 h	24 h	48 h
Control	20	0	0	0	0
3.1	20	0	0	0	0
6.3	20	0	0	0	0
12.5	20	2	8	10	40
25	20	13	14	65	70
50	20	20	20	100	100
100	20	20	20	100	100

Validity criteria fulfilled:

yes

Conclusions:

In this 48-hour static acute toxicity test with Daphnia magna, the obtained results showed that the test item had significant toxic effects on the mobility of Daphnia. The 48-h EC50 value was determined to be 16.595 mg/L. The 48-h NOEC was determined to be 6.3 mg/L.

Executive summary:

In the acute immobilisation test with Daphnia magna (STRAUS), the effects of the test item were determined according to OECD 202, Regulation (EC) No. 440/2008 Method C.2 and EPA OPPTS 850.1010. The study was conducted in a static system over a period of 48 hours with a concentration series of 3.1, 6.3, 12.5, 25, 50 and 100 mg/L. 20 daphnids per test concentration and control were tested in 4 replicates. The test item concentration was determined at the start and at the end of the test with HPLC-MS based on its components and it was in the range of 91 - 105 % of the nominal concentration. Therefore, the biological endpoints were based on the nominal concentration. The validity criteria of the test guideline were fulfilled. There was no immobilisation observed in twenty daphnids exposed to the two lowest test concentrations (3.1 and 6.3 mg/L) and the control group. All test animals were observed as immobile at the test concentrations of 50 and 100 mg/L after 48 h. 70 % of the daphnids was observed as immobile at the test concentration of 25 mg/L, while 40 % immobilisation was found at 12.5 mg test item/L after 48 hours. The estimated EC50 value (48 hours) of the test item based on the nominal concentration is 16.595 mg/L. Accordingly, the NOEC was determined to be 6.3 mg/L.

Description of key information

In a 48-hour static acute toxicity test with Daphnia magna, the obtained results showed that the test item had significant toxic effects on the mobility of Daphnia. The 48-h EC50 value was determined to be 16.595 mg/L. The 48-h NOEC was determined to be 6.3 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

16.595 mg/L

Additional information

In the acute immobilisation test with Daphnia magna (STRAUS), the effects of the test item were determined according to OECD 202, Regulation (EC) No. 440/2008 Method C.2 and EPA OPPTS 850.1010. The study was conducted in a static system over a period of 48 hours with a concentration series of 3.1, 6.3, 12.5, 25, 50 and 100 mg/L. 20 daphnids per test concentration and control were tested in 4 replicates. The test item concentration was determined at the start and at the end of the test with HPLC-MS based on its components and it was in the range of 91 - 105 % of the nominal concentration. Therefore, the biological endpoints were based on the nominal concentration. The validity criteria of the test guideline were fulfilled. There was no immobilisation observed in twenty daphnids exposed to the two lowest test concentrations (3.1 and 6.3 mg/L) and the control group. All test animals were observed as immobile at the test concentrations of 50 and 100 mg/L after 48 h. 70 % of the daphnids was observed as immobile at the test concentration of 25 mg/L, while 40 % immobilisation was found at 12.5 mg test item/L after 48 hours. The estimated EC50 value (48 hours) of the test item based on the nominal concentration is 16.595 mg/L. Accordingly, the NOEC was determined to be 6.3 mg/L.