1H-Isoindole-1,3(2H)-dione, 2-[(2R)-2-hydroxy-3-[[4-(3-oxo-4-morpholinyl)phenyl]amino]propyl]-

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

other information

Study period:

2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

adopted April 13, 2004

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

1H-Isoindole-1,3(2H)-dione, 2-[(2R)-2-hydroxy-3-[[4-(3-oxo-4-morpholinyl)phenyl]amino]propyl]-

IUPAC Name:

1H-Isoindole-1,3(2H)-dione, 2-[(2R)-2-hydroxy-3-[[4-(3-oxo-4-morpholinyl)phenyl]amino]propyl]-

Constituent 2

Reference substance name:

2-[(2R)-2-hydroxy-3-{[4-(3-oxomorpholin-4-yl)phenyl]amino}propyl]-1H-isoindole-1,3(2H)-dione

EC Number:

610-200-5

Cas Number:

446292-07-5

Molecular formula:

C21 H21 N3 O5

IUPAC Name:

2-[(2R)-2-hydroxy-3-{[4-(3-oxomorpholin-4-yl)phenyl]amino}propyl]-1H-isoindole-1,3(2H)-dione

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Concentrations: 25 mg/L; 50 mg/L; 100 mg/L were measured at 24 and 48 h only.
Sample storage conditions before analysis: Routinely, the sample were analysed immediately. Only in exceptional cases, there were stored overnight deep frozen and protect from light.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Stock solutions were prepared to give the desired series of test concentrations. To achieve this 120.2 mg on Jan. 23, 2007 and 120.3 mg on Jan. 24, 2007 of the test item were added each to 1 L of dilution water treated for 1 h in an ultrasonic bath and afterwards stirred for 24 h on a magnetic stirrer. Finally undissolved particles of the test item were removed by filtration using a folded filter of pore size 7-12 µm.
- Eluate: no
- Differential loading: no
- Controls: yes, test medium control

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Age at study initiation (mean and range, SD): < 24 h
- Method of breeding: under constant temperature conditions (20 ± 1 °C) at a 16:8 hour light-dark photoperiod (illumination: < 1000 lux). The culture water (so-called 'M4 medium') is partly renewed once a week. The Daphnia are exclusively fed with unicellular green algae (Desmodesmus subspicatus) 'ad libitum'.
- Source: Strain of Bundesgesundheitsamt Berlin, Germany
- Feeding during test: none

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

15.5 °dH (= 276.68 mg/L CaCO3)

Test temperature:

20.3 - 21.2 °C

pH:

7.7 - 7.8

Dissolved oxygen:

8.6 - 8.8 mg O2/L (94-98% saturation)

Nominal and measured concentrations:

Nominal: control, 25, 50 and 100 mg/L
Measured: see "Any other information on results incl. tables".

Details on test conditions:

Test system:
-Test vessels: 50 ml glass beakers holding 10 neonates in 20 ml of test medium
-3 test concentration plus 1control
-10 neonates per vessel, 2 replicates per concentration/control
-no feeding during the exposure period semi-static system
-Temperatur: mean +/- 1°C (in the temperature range 18-22 °C)
-Photoperiod: 16 h light: 8 h dark
-Aeration: none
-Medium renewal: daily
-Duration of exposure: 48h
-Criteria of effects:The criterion of adverse effects used in this study was the item-induced alteration of the normal mobility behaviour and the loss of locomotory actions of the neonates, observed at 24 and 48 hours

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 12.79 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 4.36 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Any other information on results incl. tables

Table 1: Measured concentrations

Nominal concentration (mg/L)	Geometric mean measured concentration (mg/L)
	0 h	24 h (old)	24 h (fresh)	48 h
Control	< 0.103	< 0.103	< 0.103	< 0.103
25	4.15	0.256	3.236	0.367
50	7.127	0.495	6.011	0.798
100	12.792	1.221	12.088	1.916

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

(The immobilisation in the controls must not exceed 10% by the end of the test. The dissolved oxygen concentration should remain above 3 mg/l throughout the exposure period)

Conclusions:

EC50 (48 h) > 4.36 mg/L (geometric mean measured)

Executive summary:

In order to test acute toxicity to invertebrates of the substance, Daphnia magna was exposed to three nominal concentrations of the test substance (25, 50, 100 mg/L) and a blank control solution for a period of 24 and 48 h under semi-static conditions. Mobility and visible abnormalities were recorded at 24 and 48 h. This toxicity study is classified as acceptable and satisfies the guideline requirements for the acute Daphnia study. Hydroxyaminophthalimid does not show any toxic effects on the mobility of Daphnia, EC50 (24 h) >12.79 mg/L (initially measured) and EC50 (48 h) >4.36 mg/L (geometric mean measured).