Reaction mass of N1-benzylpropane-1,2-diamine and N2-benzylpropane-1,2-diamine

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 2017-12-01 to 2018-02-28

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:

2004

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: For the analysis of the test item concentrations, samples of each test concentrations and control were taken from the test solutions.
- Sampling method: Four parallel samples were taken at start and at the end of the test from the test solutions and from the control. Formulation samples were diluted with mobile phase and the control samples were directly injected.
- Sample storage conditions before analysis: ambient conditions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The test solutions used in the test were prepared by mechanical dispersion. An appropriate amount of test item was dissolved in the dilution water (ISO medium) in order to obtain the concentration of 100 mg/L and this stock solution were placed into an ultrasonic bath for appr. 10 minutes. The further test solutions were prepared by appropriate dilution of this stock solution.
- Controls: Untreated control was run parallel in the test. A positive control with the reference substance potassium dichromate was also performed.
- Evidence of undissolved materia: No

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea.
- Source: Laboratory of Hydrobiology (Central Agricultural Office, Directorate of Plant-, and Soil Protection) 2100 Gödöllő, Kotlán S. u. 3. Hungary
- Age at the test start: < 24 hours old
- Feeding during test: No. The test animals were not fed during the exposure.
- Breeding conditions: Daphnia were bred in the Laboratory of TOXI-COOP ZRT. under similar temperature and light conditions as in the test, and in reconstituted water of a similar quality regarding to pH, components of the main ions and total hardness as the test water used in the test. The animals were fed with centrifuged green alga suspension.

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

Remarks on exposure duration:

none

Post exposure observation period:

Not performed

Hardness:

249 mg/L

Test temperature:

20.1 – 20.3 °C

pH:

The pH was in the range of 7.70– 8.93 during the test.

Dissolved oxygen:

The dissolved oxygen concentration was in the range of 6.03 – 7.99 mg/L during the test.

Salinity:

Not applicable

Conductivity:

No data

Nominal and measured concentrations:

- Nominal concentrations: 0 (Control), 6.25, 12.5, 25.0, 50.0 and 100 mg/L
- Measured concentrations: The measured concentration of the test item at the start of the test were 6.02,12.1, 24.8, 47.5 and 95.9 mg/L (corresponding to 95 and 99 % of the nominal value) and 6.56, 12.7, 27.8, 53.0 and 111 mg/L (corresponding to 102 and 111 % of the nominal value) at the end of the test. There is evidence that the concentration of the tested substance was maintained within ± 20 % of the nominal concentrations throughout the test. Therefore, the analysis of the results was based on the nominal concentration values.

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers
- Fill volume: 40 mL
- Aeration: No
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ISO medium (according to OECD 202) was used as dilution water in the experiment.
- Culture medium different from test medium: No
- Intervals of water quality measurement: The test containers were kept during the test in a climate chamber under controlled conditions. The water temperature, dissolved oxygen concentration and pH were measured in each test group at the start (before test solutions distributed into test vessels) and in each test vessel (i.e. each replicate) at the end of the test (after 48 hours). Additionally, the ambient temperature was measured continuously using a min/max thermometer in the climate chamber.

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: The light-dark cycle during the test was 16 hours light (artificial illumination) and 8 hours darkness.

EFFECT PARAMETERS MEASURED: The purpose of this study was to evaluate the influence of the test item on the mobility respectively survival of the test organisms after 24 and 48 hours.

VEHICLE CONTROL PERFORMED: no

RANGE-FINDING STUDY
- Test concentrations: A non-GLP preliminary test was performed at the nominal concentations of 100, 10, 1, 0.1, 0.01 mg/L
- Results used to determine the conditions for the definitive study: The EC50 in the preliminary test was determined to be between 10 and 100 mg/L.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

21.99 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95 % conf. limits: (20.207 - 23.050 mg/L)

Details on results:

- Behavioural abnormalities: None
- Observations on body length and weight: Not observed
- Other biological observations: Not perfomed
- Mortality of control: There was no mortality of the test organisms in the control.
- Other adverse effects control: Not observed
- Abnormal responses: Not observed
- Any observations that might cause a difference between measured and nominal values: No
- Effect concentrations exceeding solubility of substance in test medium: No
- Toxicity results: The 48-h EC50 value was determined to be 21.99 mg/L, the 48-h NOEC was determined to be 12.5 mg/L, the 48-h EC10 was determined to be 17.62 mg/L and the 48-h LOEC was determined to be 25.0 mg/L. The results are based on the nominal concentrations. All validity criteria were met and therefore the study can be considered as valid.

Results with reference substance (positive control):

- Results with reference substance valid?: Yes
- The EC50 for potassium dichromate was determined to be 1.53 mg/L (95% conf. limits: 1.30 – 1.78 mg/L) after 24 hours.

Reported statistics and error estimates:

The Dunnett t-Test was used for the determination of the LOEC and NOEC and the Probit analysis was used for determination of the ECx values.

Validity criteria fulfilled:

yes

Conclusions:

In a static immobilization GLP test Daphnia magna (< 24 h old) were exposed to the test item for 48 hours according to the OECD 202 (1992). The 48h-EC50 was determined to be 21.99 mg/L.

Executive summary:

In a static immobilization GLP test Daphnia magna (< 24 h old) were exposed to the test item for 48 hours according to Directive 92/69/EEC Part C, Method 2 and the OECD 202 (1992). The Daphnia were exposed to a nominally range of concentrations of 0 (Control), 6.25, 12.5, 25.0, 50.0 and 100 mg/L. The test solutions used in the test were prepared by mechanical dispersion. An appropriate amount of test item was dissolved in the dilution water (ISO medium) in order to obtain the concentration of 100 mg/L. The further test solutions were prepared by appropriate dilution of this stock solution. Samples were taken from the test concentrations and the control at the start and at the end of the experiment and analyzed by HPLC method with MS detection. The measured concentration of the test item in the test solutions were 6.02,12.1, 24.8, 47.5 and 95.9 mg/L (corresponding to 95 and 99 % of the nominal value) at the start of the test and 6.56, 12.7, 27.8, 53.0 and 111 mg/L (corresponding to 102 and 111 % of the nominal value) at the end of the test. The concentration of the tested substance was maintained within ± 20 % of the nominal concentrations throughout the test. Therefore, the analysis of the results was based on the nominal concentration values. An untreated control (ISO-Medium without addition of the test item) and a toxic reference control with potassium dichromate were performed. The 24h-EC50 of potassium dichromate was determined to be 1.53 mg/L. The validity criteria were fulfilled. The 48h-EC50 of the test substance was determined to be 21.99 mg/L.

Description of key information

In a static immobilization GLP test Daphnia magna (< 24 h old) were exposed to the test item for 48 hours according to the OECD 202 (1992). The 48h-EC50 was determined to be 21.99 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

21.99 mg/L

Additional information

In a static immobilization GLP test Daphnia magna (< 24 h old) were exposed to the test item for 48 hours according to Directive 92/69/EEC Part C, Method 2 and the OECD 202 (1992). The Daphnia were exposed to a nominally range of concentrations of 0 (Control), 6.25, 12.5, 25.0, 50.0 and 100 mg/L. The test solutions used in the test were prepared by mechanical dispersion. An appropriate amount of test item was dissolved in the dilution water (ISO medium) in order to obtain the concentration of 100 mg/L. The further test solutions were prepared by appropriate dilution of this stock solution. Samples were taken from the test concentrations and the control at the start and at the end of the experiment and analyzed by HPLC method with MS detection. The measured concentration of the test item in the test solutions were 6.02,12.1, 24.8, 47.5 and 95.9 mg/L (corresponding to 95 and 99 % of the nominal value) at the start of the test and 6.56, 12.7, 27.8, 53.0 and 111 mg/L (corresponding to 102 and 111 % of the nominal value) at the end of the test. The concentration of the tested substance was maintained within ± 20 % of the nominal concentrations throughout the test. Therefore, the analysis of the results was based on the nominal concentration values. An untreated control (ISO-Medium without addition of the test item) and a toxic reference control with potassium dichromate were performed. The 24h-EC50 of potassium dichromate was determined to be 1.53 mg/L. The validity criteria were fulfilled. The 48h-EC50 of the test substance was determined to be 21.99 mg/L.