D-threo-Pentitol, 2,5-anhydro-1,3,4-trideoxy-2-C-(2,4-difluorophenyl)-4-[[4-[4-[4-[[[1-[(1S,2S)-1-ethyl-2-(phenylmethoxy)propyl]hydrazino]carbonyl]amino]phenyl]-1-piperazinyl]phenoxy]methyl]-1-(1H-1,2,4-triazol-1-yl)-

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:

05 September 2012 to 07 September 2012

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 Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

2008

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

ISO 6341 (Water quality - Determination of the Inhibition of the Mobility of Daphnia magna Straus (Cladocera, Crustacea))

Version / remarks:

1996

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Guidance document onGuidance document on aquatic toxicity testing of difficult substances and mixtures, OECD series on testing and assessment number 23, December 14, 2000.

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

PAZ-D

IUPAC Name:

PAZ-D

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): PAZ-D
- Molecular formula: C43H50O4N8F2
- Molecular weight: 780.9
- Physical state: white powder
- Expiration date of the lot/batch: 13 January 2013 (allocated by NOTOX, 1 year after receipt of the test substance)
- Stability under test conditions: stable
- Storage condition of test material: At room temperature in the dark

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
PHYSICO-CHEMICAL PROPERTIES
- Water solubility: 0.0453 mg/L
- Stability in water: No

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

At the start of the test duplicate samples for possible analysis were taken from all test concentrations and the control. At the end of the test duplicate samples were taken from the control and the two highest concentrations. Sampling was performed according to the schedule below.

Frequency: at t=0 h and t=48 h
Volume: 0.4 ml from the approximate centre of the test vessels
Storage: Not applicable, samples were analysed on the day of sampling.

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

Test solutions

Vehicle:

no

Details on test solutions:

The batch of PAZ-D tested was a white powder with a purity of 99.3% and not completely soluble in test medium at the loading rate initially prepared.

Preparation started with a loading rate of 100 mg/L. An overnight period of magnetic stirring was applied. Thereafter, the obtained mixture was filtered through a 0.45 µm membrane filter (rs 55, Whatman) and used as the highest test concentration. The lower test concentrations were prepared by subsequent dilutions of the filtrate in test medium. The final test solutions were all clear and colourless.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

Species: Daphnia magna (Crustacea, Cladocera) (Straus, 1820), at least third generation, obtained by acyclical parthenogenesis under specified breeding conditions.
Source: In-house laboratory culture with a known history.

Reason for selection: This system has been selected as an internationally accepted invertebrate species.

Validity of batch: Daphnids originated from a healthy stock, 2nd to 5th brood, showing no signs of stress such as mortality >20%, presence of males, ephippia or discoloured animals and there was no delay in the production of the first brood.

Characteristics: For the test selection of young daphnids with an age of < 24 hours, from parental daphnids of more than two weeks old.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Remarks on exposure duration:

none

Post exposure observation period:

no

Test conditions

Hardness:

180 mg/L expressed as CaCO3

Test temperature:

19.0 - 20.0°C

pH:

7.7 - 8.1

Dissolved oxygen:

9.1 - 9.2

Salinity:

-

Nominal and measured concentrations:

Nominal: 100% of the filtrate prepared at 100 mg/L
Average exposure concentration: 1.1 ug/l

Due to possible adsorption to glassware the obtained analytical results should be considered as estimated values.

Details on test conditions:

Test duration: 48 hours

Test type: Static

Test vessels: 100 mL, all-glass

Medium: Adjusted ISO medium

Number of daphnids : 20 each for the control and the highest concentration (limit test), 10 each for lower concentrations (combined range-finding test)

Loading: 5 per vessel containing 80 ml of test solution

Light: 16 hours photoperiod daily

Feeding: No feeding

Aeration: No aeration of the test solutions.

Introduction of daphnids: Within 21 minutes after preparation of the test solutions.

Reference substance (positive control):

yes

Remarks:

potassium dichromate (K2Cr2O7)

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EC50

Remarks:

.

Effect conc.:

> 1.1 µg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Remarks:

.

Basis for effect:

mobility

Remarks:

.

Remarks on result:

other: -

Effect concentrations 2

Duration:

48 h

Dose descriptor:

NOEC

Remarks:

.

Effect conc.:

1.1 µg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Remarks:

.

Basis for effect:

mobility

Remarks:

.

Remarks on result:

other: -

Details on results:

- Behavioural abnormalities: no
- Other biological observations: no
- Immobility in control: 0%
- Immobility in the limit concentration: 5%
- Other adverse effects control: no
- Abnormal responses: no
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: no

Results with reference substance (positive control):

The actual responses in this reference test with K2Cr2O7 were in the lower range of the expected historical responses at the different concentrations, i.e. a 48h-EC50 between 0.3 and 1.0 mg/l. Hence, the sensitivity of this batch of D. magna was in agreement with the historical data collected at NOTOX.

The 24h-EC50 was 0.50 mg/l with a 95% confidence interval between 0.44 and 0.57 mg/l.

The 48h-EC50 was 0.30 mg/l with a 95% confidence interval between 0.28 and 0.35 mg/l.

Reported statistics and error estimates:

Calculation of EC50:
No EC50 could be calculated because the test substance proved to be non-toxic (EC50 > maximum concentration tested).

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of the present study PAZ-D did not induce acute immobilisation of Daphnia magna at 1.1 µg/L after 48 hours of exposure (NOEC).

The 48h-EC50 was beyond the range tested, i.e. exceeded an average exposure concentration of 1.1 µg/L.