Pentanedioic acid, 2-(1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl)-, (2S)-

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:

24 June 2015 to 26 June 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

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

Data source

Reference

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:

(2S)-2-(1,3-DIOXO-2,3-DIHYDRO-1H-ISOINDOL-2-YL)PENTANEDIOIC ACID

EC Number:

608-945-6

Cas Number:

340-90-9

Molecular formula:

C13H11NO6

IUPAC Name:

(2S)-2-(1,3-DIOXO-2,3-DIHYDRO-1H-ISOINDOL-2-YL)PENTANEDIOIC ACID

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

Identification : Phthalyl-L-Glu
Appearance: White powder
Batch: 10844200 (taken from label)
Purity/Composition: 99.2 weight-% (average content)
Test substance storage: At room temperature
Stable under storage conditions until: Not indicated

Purity/composition correction factor: No correction factor required
Test substance handling: No specific handling conditions required
CAS Number: 340-90-9
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:

Preparation of test solutions started with a the highest concentration of 100 mg/l applying 42 minutes of magnetic stirring to accelerate the dissolving of the test substance in the test medium and ensure homogeneity. 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.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

- Common name: Daphnia magna (Crustacea, Cladocera) (Straus, 1820)
- Source: In-house laboratory culture with a known history.
- Age at study initiation: < 24 hours
- 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.
- Feeding during test: No

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

none

Test conditions

Test temperature:

between 20 and 21°C

pH:

between 6.5 and 8.0

Dissolved oxygen:

between 8.9 and 9.4 mg/l

Nominal and measured concentrations:

- Nominal concentrations: 0.10, 1.0, 10 and 100 mg/l

- Measured concentrations:
The samples from 100 mg/l were analyzed. The actual concentrations at the start was 98 mg/l and remained stable and within nominal during the test (94% of initial, 92% of nominal). Consequently, the parameters were reported in terms of the analytical confirmed nominal concentration.

Details on test conditions:

COMBINED LIMIT/RANGE-FINDING TEST:

Since, no effects on mobbility were observed at the highest concentration the test was used as a limit test.

TEST SYSTEM:

- Type: 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: yes of the highest concentration
- Photoperiod: 16 hours photoperiod daily

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
- EC50 t=24, 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:

24 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: analytical confirmed nominal concentration

Effect concentrations 2

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: analytical confirmed nominal concentration

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 0.72 mg/l with a 95% confidence interval between 0.64 and 0.82 mg/l.
The 48h-EC50 was estimated to correspond with 0.36 mg/l with 10% effect at 0.32 mg/l and 100% effect at 0.56 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.