4-Amino-N-[3-(diethylamino)propyl]benzamid, hydrochloride

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

30 Apr 2018 - 17 Dec 2018

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

4-amino-N-[3-(diethylamino)propyl]benzamide hydrochloride

EC Number:

814-672-5

Cas Number:

109342-99-6

Molecular formula:

C14H24N3OCl

IUPAC Name:

4-amino-N-[3-(diethylamino)propyl]benzamide hydrochloride

Test material form:

solid

Specific details on test material used for the study:

4-amino-N-[3-(diethylamino)propyl]benzamid, hydrochloride, batch number: Betriebsprobe 3a + 3b, purity analysed: 96.0 % w/w.

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Analytical samples taken at 0 hours (initial value) and 48 hours aged test solutions were analysed from control and all test item concentrations.

Test solutions

Details on test solutions:

The necessary amount of test item for preparing the stock solution S1 was weighed on a weighing scoop and transferred to a volumetric flask. Test medium (Elendt M4) was added up to the bench mark and the solution was homogenised by shaking. Afterwards the solution was clear and transparent. Lower test solutions were prepared by dilution of the appropriate solution with test medium. ~ 50 mL of the prepared solutions were transferred to each test vessel (see Table added in any other information).

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

Daphnia magna Straus, Clone V, was used as the test organism. The animals are continuously bred in the laboratory and were originally purchased in a healthy condition from the Federal Environment Agency in Berlin/Germany.
Daphnia magna was bred as single culture (1 daphnid per 100 mL) in Elendt M4 medium. The pH-value of the aerated water was within a range of 6.0 – 9.0. The dissolved oxygen was above 60 % saturation and the total hardness 140 - 250 mg/L (as CaCO3), corresponding to 7.8 - 14°dH. The animals were fed with single cell green algae (Desmodesmus subspicatus, formerly Scenedesmus subspicatus) at least three times a week.
The daphnids were reared at a temperature of 20 ± 2 °C in a climatic chamber with 16 hours of illumination and 8 hours of darkness. The medium was changed three times per week. A pipette was used to separate the young daphnids from the adults.
Freshly hatched daphnids less than 24 hours old were used for the test.

Study design

Test type:

static

Water media type:

other: Elendt M4

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

13°dH (232 mg/L as CaCO3)

Test temperature:

18.9 – 20.9 °C

pH:

7.62 – 7.92

Dissolved oxygen:

≥ 8.5 mg/L

Salinity:

-

Conductivity:

-

Nominal and measured concentrations:

100, 50.0, 25.0, 12.5, 6.25 mg/L nominal

Details on test conditions:

Static dose-response test with twenty test organisms per treatment (4 replicates of 5 test organisms each) were used. The duration of the test was 48 hours.
The total hardness (as CaCO3) of the test medium (untreated control) was determined to be 13°dH (232 mg/L CaCO3); the mean pH-value of the untreated control was determined to be 7.70 ± 0.07 (Std. Dev.), the mean temperature of the control and all test item concentrations was measured to be 19.7 ± 0.8 °C (Std. Dev.) and the mean oxygen concentration was determined to be 8.7 ± 0.2 mg/L (Std. Dev.).

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 3

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 4

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

97.8 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Control immobilisation: The percentage of immobilisation should be ≤ 10 %. In this study the control immobilisation was 0 %.
Oxygen concentration: The dissolved oxygen concentration at the end of the test should be ≥ 3 mg/L in all test units. In this test, the dissolved oxygen concentration at the end of the test was ≥ 8.5 mg/L.

After 24 hours of exposure no immobilisation was observed in the control and up to and including 100 mg/L.
After 48 hours of exposure no immobilisation was observed in the control and at test item concentrations 12.5 to 50.0 mg/L. No immobilisation higher than the allowed control immobilisation was observed at 6.25 mg/L. At the highest test item concentration of 100 mg/L 60 % of the daphnids were immobile.
No behavioural changes could be observed.

Results with reference substance (positive control):

The results indicate an EC50 (24 h) of the reference item potassium dichromate between 1.00 and 2.00 mg/L. Since the results are in accordance with the requirements of the OECD guideline 202 and fall within the historical data generated with the reference item at the testing facility, the daphnids were suitable for the determination of the toxicological effects of the test item.

Reported statistics and error estimates:

EC50 (24h): >100
EC50 (48h): 97.8 (95 % confidence limits were not calculable)

Any other information on results incl. tables

EC₅₀and NOEC-values of daphnids exposed to the test item evaluated using nominal concentrations

Endpoint	4-amino-N-[3-(diethylamino)propyl]benzamid, hydrochloride[mg/L]
	24 h	48 h
NOEC	100	50.0
EC50	> 100^1)	97.8^2)
95 % confidence limit of EC50	-	n.d.

- not applicable

n.d. not determined due to mathematical reasons or inappropriate data

¹⁾Due to an inhibition below 50% the database was weak for probit analysis which hence was not performed

²⁾Webull analysis using linear max. likelihood regression

Analytical Results

The initial measured content of the test item was between 88 and 99 % of nominal and the aged measured content was between 98 and 100 % of nominal. Since the content of active ingredient in the samples was between 80 and 120 % of nominal the toxicological endpoints were evaluated using nominal concentrations.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

According to the results of the test, the EC50 (48 h) was determined to be 97.8 mg/L (nominal). The corresponding NOEC (48 h) was 50.0 mg/L (nominal).

Executive summary:

Validity criteria: Control immobilisation: The percentage of immobilisation should be ≤ 10 %. In this study the control immobilisation was 0 %.

Oxygen concentration: The dissolved oxygen concentration at the end of the test should be ≥ 3 mg/L in all test units. In this test, the dissolved oxygen concentration at the end of the test was ≥ 8.5 mg/L.

Test conditions: The total hardness (as CaCO₃) of the test medium (untreated control) was determined to be 13°dH (232 mg/L CaCO₃); the mean pH-value of the untreated control was determined to be 7.70±0.07 (Std. Dev.), the mean temperature of the control and all test item concentrations was measured to be 19.7±0.8 °C (Std. Dev.) and the mean oxygen concentration was determined to be 8.7 ± 0.2 mg/L (Std. Dev.).

Analytical Results: The initial measured content of the test item was between 88 and 99 % of nominal and the aged measured content was between 98 and 100 % of nominal. Since the content of active ingredient in the samples was between 80 and 120 % of nominal the toxicological endpoints were evaluated using nominal concentrations.

Statistical Results:  EC₅₀and NOEC-values of daphnids exposed to the test item evaluated using nominal concentrations  

	4-amino-N-[3-(diethylamino)propyl]benzamid, hydrochloride [mg/L] (nominal)
	24 h	48h
NOEC	100	50.0
EC50	> 100^1)	97.8^2)
95 % confidence limit of EC50	-	n.d.

- not applicable

n.d. not determined due to mathematical reasons or inappropriate data

¹⁾Due to an inhibition below 50% the database was weak for probit analysis which hence was not performed

²⁾Weibull analysis using linear max. likelihood regression