2-({2-[4-(dimethylamino)benzoyloxy]ethyl}(methyl)amino)ethyl 4-(dimethylamino)benzoate

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:

21 November 2006 to 23 November 2006

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:

October 2000

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

EU Commission Directive 92/69/EEC, 1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: OECD Guidance Document on testing of difficult substances

Version / remarks:

2000

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Test article name LFC 2098
Chemservice sample code : AA26964
Declared purity (according to CoA) 100%
Chemical class: Aminoester
Chemical formula C23H31O4N3
Molecular weight: 413
Preparation date 05/29/2006
Expiry date 05/29/2008
Solubility in water Practically insoluble
Density Not available
Physical form at 20 °C: Solid (powder)
Colour: White
Storage conditions Room temperature, protect from light
Safety precautions Routinary hygienic procedures

Analytical monitoring:

yes

Details on sampling:

Concentrations:
Nominal test concentration 100 mg/L
Actual test concentration: 0.67 mg/L

Sampling method:
Samples taken at 0 and 48 hours

Sample storage conditions before analysis:
The samples were stored in at 4°C in the dark until the analysis day.

Vehicle:

no

Test organisms (species):

Daphnia magna

Details on test organisms:

The study has been performed on the freshwater crustacean Daphnia magna Straus. A strain of this species was originally supplied by the Italian Health Department (lstituto Superiore di Sanita) in September 2004. Since that time, the clone has been bred in the laboratories of Chemservice in reconstituted water of the quality identical to the water quality used in the tests (regarding pH, main ions, and total hardness) and under temperature and light conditions identical to those of the tests (see below). It was daily fed a fixed amount of the laboratory cultured green algae Pseudokirchneriella subcapitata (formerly Selenastrum capricornutum) and of a suspension of the yeast Saccaromices cerevisiae. At the start of the test, the organisms used in the test were less than 24 hours old and were not first brood progeny.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

Not applicable

Hardness:

2.5 mmol/L

Test temperature:

20.6 - 23.7 °C

pH:

7.48

Dissolved oxygen:

≥ 6 mg/L

Salinity:

Not measured

Conductivity:

Not measured

Nominal and measured concentrations:

Nominal test concentration 100 mg/L loading rate
Actual test concentration: 0.67 mg/L

Details on test conditions:

Test water:
CaCl2 x 2H2O : 2.0 mmol/L (=294.0 mg/L)
MgSO4 x 7H20 : 0.5 mmol/L (=123.0 mg/L)
NaHCO3 : 0.75 mmol/L (= 65.0 mg/L)
KCI :0.075 mmoI/L (= 5.8 mg/L)
Alkalinity : 0.8 mmol/L
Ratio of Ca : Mg 4 : 1 (based on molarity)
Ratio of Na : K 10 : 1 (based on molarity)
The test water was aerated prior to the start of the study until oxygen saturation was reached. During the test period, the test water was not aerated.

The test was performed in 50 mL glass beakers filled with 40 mL of test medium. The beakers were covered with glass plates to reduce the loss of water and to avoid the entry of dust into the solutions. The test vessels were labelled with the Chemservice study number and all necessary additional information to ensure unmistakable identification.
For the only test concentration and for the control, 20 daphnids were used divided into four replicates of five daphnids each. The daphnids were randomly distributed to the test vessels at initiation of the test. The daphnid’s loading rate was less than one per 2 mL of test solution.

Temperature: The room temperature has been continuously monitored during the course of the study by the means of a sensor connected to a PC. The incubation temperature was in the range 20. 6 — 237°C.

Light: a 16 hours light and 8 hours darkness daily photoperiod (light intensity during the light period will be within the range of 200—1500 Lux).

Test duration: 48 hours

Feeding: None

Parameter measured: number of immobile daphnids at 24 and 48 hours

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

IC50

Effect conc.:

> 0.67 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

>= 0.67 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOELR

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

The daphnids were observed for immobility after 24 and 48 hours of exposure. The observed immobilization in the test concentrations shows that no adverse effect was observed in the exposed organisms.

Reported statistics and error estimates:

In the control 0% immobilization was observed during the test period at each observation time. This value complies with the validity criteria of the test, according to the mentioned guidelines.

Actual concentration [mg/L]	n. of exposed daphnids	n. immobile daphnids
		24 h	48 h
0 (control)	20	0	0
0.67	20	0	0

Validity criteria fulfilled:

yes

Conclusions:

The test item LFC 2098 caused no immobilization to Daphnia magna after 48-hours exposure at a nominal concentration of 100 mg/L, corresponding to an actual concentration of 0.67 mg/L, therefore the following endpoints have been assessed:

NOEC ≥ 100 mg/L
LOEC > 100 mg/L
IC50 > 100 mg/L.

Corresponding in terms of actual concentration to:
NOEC ≥ 0.67 mg/L
LOEC > 0.67 mg/L
IC50 > 0.67 mg/L

Executive summary:

The acute toxicity of the test item LFC 2098 to Daphnia magna was determined in a 48—hour static test according to the EU Commission Directive 92/69/EEC, Part 0.2 (1992), and the OECD Guideline for Testing of Chemicals, No. 202, revised proposal (2000).

For this purpose, juvenile daphnids of the species Daphnia magna were exposed to an aqueous test medium containing the test item at the only loading rate of 100 mg/L under defined conditions. The exposed organisms were checked for immobilization after 24 and 48 hours from test initiation.

The pH of the test solution at the beginning of the test period was 7.48. Therefore, the pH of the test medium was not adjusted before the start of the test.

The actual test concentration of the test item was analytically measured at the beginning and at the end of the test. The test item concentration has shown a large decrease compared to nominal value short after the test solution preparation (analytical recovery 0.73% of the nominal value at time 0 h), probably due to the low water solubility of the test item, while the concentration was quite stable during the following days (0.60% of the nominal value after 48 h). Therefore, the biological results are not referred to the nominal concentration but to the median of the measured concentrations, according to the OECD Guidance Document on testing of difficult substances (2000).

In the control 0% mortality was observed during the test period of 48 hours. This value complies with the validity criteria of the test, according to the mentioned guidelines.

No mortality was observed in the test medium after 24 and 48 hours, therefore the NOEC (highest concentration tested without toxic effects after the exposure period) of LFC 2098 to Daphnia magna was 0.67 mg/L or higher.

Based on the observed effects, the LOEC and the 48-h |C50 were determined to be greater than 0.67 mg/L, corresponding to 100 mg/L as nominal concentration.

Description of key information

Study conducted to recognised testing guidelines with GLP certification.

Additional information

EC50 96 hour > 100 mg/L