4-[2-(2-amino-4,7-dihydro-4-oxo-3H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl]benzoic acid

Long-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

3 to 24 December 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP, guideline study was followed

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

1

Reference substance name:

4-[2-(2-amino-4,7-dihydro-4-oxo-3H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl]benzoic acid

EC Number:

429-790-9

EC Name:

4-[2-(2-amino-4,7-dihydro-4-oxo-3H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl]benzoic acid

Cas Number:

137281-39-1

Molecular formula:

Hill formula: C15 H14 N4 O3 CAS formula: C15 H14 N4 O3

IUPAC Name:

4-(2-{2-amino-4-oxo-3H,4H,7H-pyrrolo[2,3-d]pyrimidin-5-yl}ethyl)benzoic acid

Test material form:

solid: particulate/powder

Details on test material:

Name: Compound 202723
Storage condition of test material: Room temperature in dark

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Samples of newly prepared test solutions were taken on Days 0, 2, 6, 8, 13, 15, and 20. Samples of aged solutions were taken on Days 1, 3, 7, 9, 14, 16, and 21. Three QC samples were prepared at each interval and remained with samples.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Test material weighed (weight corrected for purity) and dissolved in dilution water to prepare a 10 mg/L solution (prepared daily)
- Controls: Dilution water only
- No evidence of undissolved material

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Source: On site cultures
- Feeding during test: 200 µL algal suspension (Ankistrodesmus falcatus, 4 x 10+7 cells/mL) and 50 µL of a yeast, cereal leaves and digested flaked fish food suspension
- Feeding frequency: Daily

SOURCE AND AGE OF TEST ORGANISMS:
Test organisms were not from the first brood and were derived from healthy culture stock. Test organisms were < 24 hours old at test initiation

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

21 d

Test conditions

Hardness:

Total hardness range as calcium carbonate (CaCO3) of 170 to 180 mg/L, a total alkalinity range (as CaCO3) of 100 to 110 mg/L

Test temperature:

18 to 21 ºC (continuous monitoring)

pH:

pH range of 7.8 to 8.6

Dissolved oxygen:

7.3 to 10.0 mg/L

Salinity:

Not applicable

Nominal and measured concentrations:

Nominal 10 mg compound 202723/L
Mean Measured: 10 mg compound 202723/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 100 mL open glass beakers (volume 80 mL)
- Aeration: None
- Type of exposure: Static-renewal
- Renewal rate of test solution: 100% solution change daily
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Well-water, autoclaved, fortified with salts and nutrients to prepare hard water, and mixed overnight while aerating
- Total organic carbon: 0.69 mg/L
- Metals: None detected at toxic concentrations
- Pesticides: None detected at toxic concentrations
- Alkalinity: 100 to 110 mg/L as CaCO3
- Conductivity: 500 to 650 µmhos/cm
- Culture medium same as test medium:
- Intervals of water quality measurement: Dissolved oxygen, temperature and pH measured at each renewal period in new and aged solutions. Alkalinity and conductivity measured weekly.

OTHER TEST CONDITIONS
- Photoperiod: 16 h light: 8 h darkness
- Light intensity: 8.6 to 11 µE per m2 per s

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Number of immobilized daphnids measured daily
Numbers of offspring determined upon first brood release in any vessel and daily throughout remainder of test
Number of immobilized offspring recorded

VEHICLE CONTROL PERFORMED: no

TEST CONCENTRATIONS
-Single concentration of 10 mg compound 202723/L (nominal)
-Justification for using less concentrations than requested by guideline: limit test

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

10 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

reproduction

Effect concentrations 2

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

10 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

growth

Effect concentrations 3

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

10 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

mortality

Details on results:

There was acceptable survival (>80%) and reproduction (>60 offspring per female) in the controls in the study. No effects due to exposure to compound
202723 were observed (see Table 1).

Reported statistics and error estimates:

Statistical analysis (t-Test) indicated no significant difference between the control and treatment group for reproduction and length.

Any other information on results incl. tables

Table 1: Mean survival, reproduction and total body lengths of parental daphnids at termination o fthe 21 day static renewal toxicity test exposing daphnids (Daphnia magna) to compound 202723

Mean Measured Conc (mg compound 202723/L)	Mean percent survival	Mean Reproduction (SD)	Total Body Length in mm (SD)
Control	100	128 (18)	4.51 (0.11)
10mg	100	122 (17)	4.61 (0.1)

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Study is considered valid as the performance criteria defined in OECD211 were met. There was acceptable survival (>80%) and reproduction (>60 offspring per female) in the controls in the study.
Results from an acute aquatic study on invertebrates is not available for the test material. The compound is highly likely to be of low acute toxicity to invertebrates based on a lack of toxicity demonstrated in this chronic invertebrate study. The chronic invertebrate test was conducted as a limit test using a test concentration of 10 mg/L and a control group according to OECD 211 guideline.

Executive summary:

Test Substance	Compound 202723
Test organism	Daphnia magna
Test Conditions	21 day duration, daily test solution renewals, temp range of 18 to 21oC, photoperiod of 16 hours light at an illumination range of 8.6 to 11uE.m-2.s-1 and 8 hours dark.
Dilution water	Fortified well water pH 8.1 to 8.5, Dissolved oxygen: 7.4 to 9.4mg/l, Specific conductivity 600umhos/cm, total hardness as Ca CO3: 170 to 180 mg/L, total alkalinity as CaCO3: 100 to 110mg/L
Results	No effects were found on survival, growth or reproduction among daphnids exposed to compound 202723 at 10mg/L. The NOEC for compound 202723 is 10 mg/l