5-[4-Carboxy-4-(1,3-dioxo-1,3-dihydro-isoindol-2-yl)-butyrylamino]-2-nitro-benzoic acid, L-

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:

15-September 2015 - 17 September 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: This study has been performed according to OECD and/or EC guidelines and according to GLP principles

Data source

Reference

Reference 1

Reference Type:

other: study amendment

Title:

Unnamed

Year:

2021

Report date:

2021

Reference 2

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

5-[(S)-4-Carboxy-4-(1,3-dioxo-1,3-dihydro-isoindol-2-yl)-butyrylamino]-2-nitro-benzoic acid

EC Number:

926-197-1

Cas Number:

2561176-48-3

Molecular formula:

C20 H15 N3 O9

IUPAC Name:

5-[(S)-4-Carboxy-4-(1,3-dioxo-1,3-dihydro-isoindol-2-yl)-butyrylamino]-2-nitro-benzoic acid

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

Identification: P-Glupa-C
Appearance: Yellow powder
Batch: 11328700
Purity/Composition: 86.2 weight-% (average content)
Test substance storage: At room temperature
Stable under storage conditions until: 15 May 2016 (expiry date)

Purity/composition correction factor: No correction factor required
Test substance handling: No specific handling conditions required
Volatile: Not indicated
Solubility in water:Not available
Stability in water: Not available

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Samples for possible analysis were taken from all test concentrations and the control according to the schedule below.

Frequency: at t=0 h and t=48 h
Volume: 4.0 ml from the approximate centre of the test vessels
Storage : Samples were stored in a freezer until analysis.

At the end of the exposure period, the replicates were pooled at each concentration before sampling.

Additionally, reserve samples of 4.0 ml were taken for possible analysis. If not used, these samples were stored in a freezer for a maximum of three months after delivery of the draft report, pending on the decision of the sponsor for additional analysis

Test solutions

Vehicle:

no

Details on test solutions:

The batch of P-Glupa-C tested was a yellow with a purity of 86.2 weight-% (average content) and completely soluble in test medium at the concentrations tested.

Preparation of test solutions started with a loading rate of 100 mg/l applying a 13 minute treatment period with ultrasonic waves to accelerate the dissolving of the test substance in the test medium. The lower test concentrations were prepared by subsequent dilutions of the highest concentration in test medium. The final test solutions were all clear and colourless, except the highest concentration which was clear and slightly yellow.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna (Crustacea, Cladocera) (Straus, 1820)
- Source: In-house laboratory culture with a known history.
- Method of cultivation: With newborn daphnids, i.e. less than 3 days old, by placing about 250 of them into 10 litres of medium in an all-glass culture vessel
- Age at study initiation: < 24 hours
- Feeding during test: No

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Test temperature:

Between 20 and 22°C

pH:

Start of the test between 6.4 and78
End of the test between 8.0 and 8.1

Dissolved oxygen:

Start of the test between 8.5 and 8.7 mg/l
End of the test 8.6 mg/l

Nominal and measured concentrations:

Nominal concentration:
0.10, 1.0, 10 and 100 mg/l

Measured concentration:
The actual concentrations at 100 mg/l was at the level of nominal (97% of nominal) at the start of the test and remained stable during the test (98% of initial, 95% of nominal). Consequently, the effects parameters were reported in terms of analytically confirmed nominal concentration.

Details on test conditions:

COMBINED LIMIT/RANGE-FINDING TEST
Since, no significant effects on immobility were observed at the highest concentration the test was used as a limit test.

TEST SYSTEM
- Type: static, open
- Material, size, headspace, fill volume: glass, 100 ml, normal headspace, 80 ml
- Aeration: no
- No. of organisms per vessel: 5

replicates
- No. of vessels per concentration (replicates): 4 for the highest concentration and 2 for the lower concentrations
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Adjusted ISO medium: the following chemicals (analytical grade) are dissolved in tap water purified by Reverse Osmosis (RO-water, GEON Waterbehandeling, Berkel-Enschot, The Netherlands):
Macro salts:
CaCl2.2H2O: 211.5 mg/l
MgSO4.7H2O: 88.8 mg/l
NaHCO3 : 46.7 mg/l
KCl : 4.2 mg/l
The hardness: 180 mg/l expressed as CaCO3 and the pH: 7.7 ± 0.3 after aeration.
- Culture medium different from test medium: yes, for culturing M7 medium is used

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours photoperiod daily

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : EC50 t=24 and 48 h

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 10
- Test concentrations:control and 0.10, 1.0, 10 and 100 mg/l

Reference substance (positive control):

yes

Remarks:

(potassium dichromate)

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

meas. (initial)

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:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

- Behavioural abnormalities: no
- Immobility of control: no immobility was observed.
- Other adverse effects control: no

Results with reference substance (positive control):

The 24h-EC50 was estimated to correspond with 0.80 mg/l.
The 48h-EC50 was estimated to correspond with 0.39 mg/l.

Reported statistics and error estimates:

The 24 / 48h-EC50 could not be determined because the observed effects were below 50%.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The 48h-EC50 was beyond the range tested, i.e. exceeded an analytically confirmed nominal concentration of 100 mg/l.