3-cyclohexylaminopropylamine

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:

2019-05-06 to 2019-06-24

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)

Version / remarks:

(2004)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Remarks:

LC-MS/MS method

Details on sampling:

- Concentrations: 6.25 - 12.5 - 25.0 - 50.0 - 100 mg/L
- Sampling method: Pipetting from test vessels
- Sample storage conditions before analysis: Samples were analyzed on the day of sampling as far as possible. If necessary, the samples were stored at room temperature until the start of the analysis

Vehicle:

no

Details on test solutions:

Preparation of the stock solution A stock solution with a nominal concentration of 100 mg/L of the test item was prepared with dilution water directly prior
to start of the exposure (at day 0).
An appropriate amount of the test item was weighed out and transferred into a measuring flask with an appropriate
amount of dilution water. This stock solution was mixed by manual agitation until it was visually clear.
The stock solution was used as the highest test concentration and for the preparation of further concentration levels by dilution with dilution water.

Test item concentrations Five concentration levels 6.25 - 12.5 - 25.0 - 50.0 - 100 mg/L were prepared from the stock solution 100 mg/L in a
geometric series with a separation factor of 2 by dilution of the stock solution with dilution water.
The test item concentrations were selected based on the results of a non-GLP preliminary range finding test.

Control Dilution water without test item incubated under the same conditions as the test groups.

Test organisms (species):

Daphnia magna

Details on test organisms:

Test system: Daphnia magna STRAUS, obtained from continuous laboratory cultures.
Origin: Institut für Wasser-, Boden- und Lufthygiene (WaBoLu), 14195 Berlin, Germany
Breeder: Noack Laboratorien GmbH, Käthe-Paulus-Str. 1, 31157 Sarstedt, Germany
Culture: In glass vessels (2 - 3 L capacity) with approximately 1.8 L culture medium, at 20  2
°C, in an incubator, 16 hours illumination; light intensity of max. 1500 lx
Culture medium: Elendt M4, according to OECD 202, Annex 3 (2004) is used.
Culture feeding: The culture daphnids were fed at least 5 times per week ad libitum with a mix of unicellular green algae, e.g. Pseudokirchneriella subcapitata and Desmodesmus subspicatus. The algae were cultured at the test facility.
Age of parental stock (mean and range, SD): Not specified. Continuous laboratory culture.
Feeding during test: The daphnids were not fed during the study.
Acclimation period: Acclimation of the daphnids was not necessary, because the dilution water was equivalent to the culture medium.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

Dilution water: 282 mg CaCO3/L (0 h: start of test)

Test temperature:

19 - 20 °C

pH:

Control
- t(0h) = 7.41
- t(48h) = 7.63
100 mg/L
- t(0h) = 9.45
- t(48h) = 7.89

Dissolved oxygen:

Dilution water:
8.00 mg/L
Control
- t(0h) = 8.00 mg/l
- t(48h) = 7.88 mg/L
100 mg/L
- t(0h) = 8.08 mg/l
- t(48h) = 7.38 mg/L

Conductivity:

Dilution water
588 [µS/cm]

Nominal and measured concentrations:

Nominal concentration measured concentration
0 h:
Control < LOQ < LOQ (600 µg/L)
100 mg/L 116 mg/L
6.25 mg/L 7.05 mg/L
48 h:
Control < LOQ < LOQ
100 mg/L 100 mg/L
6.25 mg/L 6.16 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers (4 (ID) x 7 (H) cm), 50 mL
- Type (delete if not applicable): Loosely covered with watch glasses
- Material, size, headspace, fill volume: 20 mL
- Aeration: No
- No. of organisms per vessel: 20 daphnids
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: According to OECD202
- Total organic carbon: 282 [mg CaCO3/L]
- Alkalinity: pH= 7.41
- Conductivity: 588 [µS/cm]
- Culture medium different from test medium: No
- Intervals of water quality measurement: At the start (t= 0h) and at the end (t= 48h) of the test.
OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16/8 hours light/dark cycle
- Light intensity: Diffuse light, light intensity of max. 1500 lx

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Immobility/mortality; after 24 and 48 hours.
VEHICLE CONTROL PERFORMED: No vehicle was involved
RANGE-FINDING STUDY: Yes
- Test concentrations: 1; 10 and 100 mg/L prepared with dilution water
- Results used to determine the conditions for the definitive study: Yes

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

17.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Key result

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

7.97 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Key result

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

- Behavioural abnormalities: No
- Observations on body length and weight: No
- Other biological observations: No
- Mortality of control: No
- Other adverse effects control: No
- Abnormal responses: No
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No
- Effect concentrations exceeding solubility of substance in test medium: No

Results with reference substance (positive control):

- Results with reference substance valid: Yes
- Relevant effect levels: EC50 (24 h)
- Limit test: no
- Dose-response test: yes
- EC50 (24h): 1.89 mg/l (CI; 1.5 – 2.40)
- Other: Valid range according to AQS P9/2 (02/2000), clone 5 = 0.6 – 2.4 mg/l
-Study period: 2019-05-14 to 2019-05-15

Validity criteria fulfilled:

yes

Conclusions:

Based on the nominal concentrations of the test item, the 48-hour EC50 for Daphnia magna was 17.1 mg/L (95% confidence limits: 14.3 – 20.0 mg/L).

Executive summary:

The toxicity of the test item was determined according to OECD 202 (2004).

The study was conducted under static conditions over a period of 48 hours with five nominal test concentrations in the range of 6.25 to 100 mg/L, prepared in a geometric series with a separation factor of 2. All test item concentrations and the control were colorless and visually clear throughout the exposure period.

Twenty daphnids (divided into 4 replicates with 5 daphnids each) were exposed to each concentration level and the control.

The concentrations of the test item were analytically verified via LC-MS/MS in the fresh media at the start of the exposure (0 hours) and in the old media at the end of the exposure (48 hours) in all concentration levels and in the control.

The measured concentrations of the test item at the start of the exposure (0 hours) were in the range of 111 to 117% of the nominal values. At the end of the exposure period (48 hours), the measured concentrations were in the range of 96 to 102% of the nominal values.

The measured test item concentrations remained within ±20% of the nominal concentrations over the test period. This indicates that the test item concentrations were stable during the duration of the test.Therefore, the effect levels given are based on the nominal concentrations of the test item. The 48 hour-EC₅₀ value of the test item on Daphnia magna was 17.1 mg/L (95% confidence limits: 14.3 – 20.0 mg/L).

The validity criteria of the test guideline were fulfilled.

Description of key information

The toxicity of the test item was determined according to OECD 202 (2004).

The study was conducted under static conditions over a period of 48 hours with five nominal test concentrations in the range of 6.25 to 100 mg/L, prepared in a geometric series with a separation factor of 2. All test item concentrations and the control were colorless and visually clear throughout the exposure period.

Twenty daphnids (divided into 4 replicates with 5 daphnids each) were exposed to each concentration level and the control.

The concentrations of the test item were analytically verified via LC-MS/MS in the fresh media at the start of the exposure (0 hours) and in the old media at the end of the exposure (48 hours) in all concentration levels and in the control.

The measured concentrations of the test item at the start of the exposure (0 hours) were in the range of 111 to 117% of the nominal values. At the end of the exposure period (48 hours), the measured concentrations were in the range of 96 to 102% of the nominal values.

The measured test item concentrations remained within ±20% of the nominal concentrations over the test period. This indicates that the test item concentrations were stable during the duration of the test.Therefore, the effect levels given are based on the nominal concentrations of the test item. The 48 hour-EC₅₀ value of the test item on Daphnia magna was 17.1 mg/L (95% confidence limits: 14.3 – 20.0 mg/L).

The validity criteria of the test guideline were fulfilled.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

17.1 mg/L