Cyclopenta[c]cyclopropa[g][1,6]diazacyclotetradecine-12a(1H)-carboxylic acid, 2,3,3a,4,5,6,7,8,9,11a,12,13,14,14a-tetradecahydro-2-[[7-methoxy-8-methyl-2-[4-(1-methylethyl)-2-thiazolyl]-4-quinolinyl]oxy]-5-methyl-4,14-dioxo-, ethyl ester, (2R,3aR,10Z,11aS,12aR,14aR)-

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:

2015-09-07 to 2015-09-16

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

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:

ethyl (1R,4R,6S,7Z,15R,17R)-17-({7-methoxy-8-methyl-2-[4-(propan-2-yl)-1,3-thiazol-2-yl]quinolin-4-yl}oxy)-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^{4,6}]octadec-7-ene-4-carboxylate

EC Number:

618-843-3

Cas Number:

923604-57-3

Molecular formula:

C37H46N4O6S

IUPAC Name:

ethyl (1R,4R,6S,7Z,15R,17R)-17-({7-methoxy-8-methyl-2-[4-(propan-2-yl)-1,3-thiazol-2-yl]quinolin-4-yl}oxy)-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^{4,6}]octadec-7-ene-4-carboxylate

Test material form:

solid

Details on test material:

- Name of test material (as cited in study reports): JNJ-38970191-AAA (T003009)
- Physical state: solid
- Appearance white solid

Specific details on test material used for the study:

- Appearance: White solid (determined by WIL Research Europe at receipt of the test item)
- Batch: I15AB0148
- Purity/Composition: 97.8 % v/v
- Test item storage conditions: At room temperature
- Stable under storage conditions: until 21 January 2017 (retest date)

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Sampling method: Samples for possible analysis were taken from all test concentrations and the control at t=0 and t=48h. 2.0 mL of volume was taken from the approximate centre of the test vessels. At the end of the exposure period, the replicates were pooled at each concentration before sampling.
- Sample storage conditions before analysis: the samples were stored in a freezer. Additionally, reserve samples of 2.0 mL were taken for possible analysis.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Preparation of test solutions started with the highest concentration of 100 mg/L applying 2 days of magnetic stirring to ensure reaching maximum dissolution in test medium. The resulting dispersion was filtered through a 0.45 µm membrane filter (Whatman; RC55) to remove the undissolved test item. The obtained Water Soluble Fraction (WSF) was used as highest test concentration. The lower test concentrations in the combined range-finding test were prepared by subsequent dilutions of the WSF in test medium.
- Evidence of undissolved material (e.g. precipitate, surface film, etc): final solutions were clear and colourless

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Source: in-house laboratory culture with a known history
- Age at study initiation (mean and range, SD): <24h (from parental daphnids of more than 2 weeks old).
- Method of breeding: 250 newborn daphnids are placed into 5 L of medium in an all-glass culture vessel. Medium is M7. After 7 days of cultivation half of the medium is renewed twice a week.
- Feeding during test: no

ACCLIMATION
- Acclimation period: max. 4 weeks
- Acclimation conditions (same as test or not): yes
- Type and amount of food: a suspension of fresh water algae.
- Feeding frequency: daily
- Health during acclimation (any mortality observed): no

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Hardness:

180 mg/L expressed as CaCO3

Test temperature:

19.1-20.7°C

pH:

At t= 0h: 7.7-7.9
At t= 48h: 8.1

Dissolved oxygen:

At t= 0h: 8.1-8.4
At t= 48h: 9.0-9.1

Salinity:

not relevant

Nominal and measured concentrations:

Final test:
Nominal concentrations (mg/L): 0, 100
Measured concentrations (mg/L) at t=0h: 0.000013, 0.000200
Measured concentrations (mg/L) at t=48h: 0.0000285, 0.0000633

Details on test conditions:

TEST SYSTEM
- Test vessel: all-glass test vessels
- Material, size, headspace, fill volume: 100 mL, all-glass vessels filled with 80mL of test solution
- Aeration: no aeration
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

OTHER TEST CONDITIONS
- Photoperiod: 16h light daily

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : immobility

TEST CONCENTRATIONS
- Spacing factor for test concentrations: none (limit test)
- Justification for using less concentrations than requested by guideline:
- Range finding study
- Test concentrations: 0, 100 mg/L
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

yes

Remarks:

Potassium Dichromate

Results and discussion

Effect concentrations

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 other: % v/v

Nominal / measured:

nominal

Conc. based on:

other: saturated solution

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: no data
- Mortality of control: no
- Other adverse effects control: no
- 24h-EC50 > 0.11 µg/L (100 mg/L initial loading rate)

Results with reference substance (positive control):

- Results with reference substance valid? yes (should lie between 0.3 and 1.0 mg/L)
- EC50/LC50: 48h-EC50 = 0.39 mg/L

Reported statistics and error estimates:

Calculations of EC50.
No EC50 could be calculated because the test item proved to be non-toxic (EC50 > maximum soluble concentration)

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of T003009 to Daphnia magna was determined in a 48-hour static test according to the OECD guideline 202. The 48h-EC50 exceeded the 100 % v/v saturated solution prepared at a loading rate of 100 mg/L. This concentration was considered to equal the maximum soluble concentration in test medium. In conclusion, due to the extremely low solubility of the test item in test medium concentrations acutely toxic for Daphnia magna could not be reached.
The results of the test can be considered reliable without restriction.