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

Long-term toxicity to fish

Administrative data

Endpoint:

long-term toxicity to fish, other

Type of information:

experimental study

Adequacy of study:

key study

Study period:

11 December 2008 to 12 January 2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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 210 (Fish, Early-Life Stage Toxicity 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:

Water samples were removed from one replicate of test solution and control on test day 0, 4(hatch), 11, 18, 28 and 32. Quality control (QC) samples were prepared at each sampling interval and remained with the exposure solution samples throughout the analytical process.

Test solutions

Vehicle:

no

Details on test solutions:

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

Test organisms

Test organisms (species):

Pimephales promelas

Details on test organisms:

TEST ORGANISM
- Common name: Fathead minnow (Pimephales promelas)
- Source: On site cultures, brood stock originally from US EPA (Duluth)
- Age at study initiation: embryos = 24 hours old
- Feeding during test: Beginning on Day 4, live brine shrimp 3x daily, ad libitum

Study design

Test type:

flow-through

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

32 d

Test conditions

Hardness:

44 to 56 mg/L CaCO3

Test temperature:

24 to 26 ºC (continuous monitoring)

pH:

7.0 to 7.5

Dissolved oxygen:

5.2-9.0 mg/L

Salinity:

Not applicable

Nominal and measured concentrations:

Nominal 10 mg compound 202723/L
Mean Measured: 9.4 mg compound 202723/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Open glass/silicone aquaria (volume 6.5 L). Embryos were incubated glass jars (volume 150 mL) with Nitex screen bottoms which were gently rocked in the vessels.
- Aeration: None
- Type of flow-through: Metering pump
- Renewal rate of test solution: 250 mL per vessel per cycle, 10.2 aquarium volumes per 24 hours, except Days 12 to 29 when it was 6 aquarium volumes per 24 hours
- No. of organisms per vessel: initiated with 30 embryos/vessel; thinned to 10 larvae/vessel after hatch
- No. of vessels per concentration (replicates): 4 vessels/treatment
- No. of vessels per control (replicates): 4 vessels/control
- Biomass loading rate: 0.029 g/L of flowing test solution; 0.049 g/L on Days 12 to 29

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Aerated well water
- Metals: None detected at toxic concentrations
- Pesticides: None detected at toxic concentrations
- Alkalinity: 20 to 22 mg/L as CaCO3
- Conductivity: 250 to 290 µmhos/cm
- Culture medium is the same as test medium:
- Intervals of water quality measurement: Dissolved oxygen, pH and temperature measured daily; hardness, alkalinity, conductivity measured weekly.

OTHER TEST CONDITIONS
- Photoperiod: 16 h light:8 h dark with 30 min transitions
- Light intensity: 1000 to 1400 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
-Dead and live embryos counted daily until hatching complete
-Behavior, appearance, mortality observed daily post-hatch

-Total length and dry weight measured at 28 days post-hatch

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

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

32 d

Dose descriptor:

NOEC

Effect conc.:

9.4 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

number hatched

Effect concentrations 2

Duration:

32 d

Dose descriptor:

NOEC

Effect conc.:

9.4 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

mortality

Effect concentrations 3

Duration:

32 d

Dose descriptor:

NOEC

Effect conc.:

9.4 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

weight

Effect concentrations 4

Duration:

32 d

Dose descriptor:

NOEC

Effect conc.:

9.4 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

length

Details on results:

There was acceptable hatching (>66%) and larval survival (>70%) in the control. No effects due to exposure to compound 202723 were observed (see
Table 1).

Reported statistics and error estimates:

There were no decreases in hatching success, survival, length or dry weight in the deazaguanine treatment compared to the control. Therefore, no statistical analysis was conducted. See table 1

Any other information on results incl. tables

Table 1: Mean percent hatching success and percent normal larvae at completion of hatch (test day 4) and survival, total length and dry weight of fathead minnow (Pimephales promelas) larvae at test termination of the early life-stage (28 days post-hatch) exposure.

MeanMeasuredConc.(mg compound202723/L)	EmbryoHatchingSuccess (SD)(%)	NormalLiveLarvaeat hatch(SD)	28DaysPost-Hatch
			LarvalSurvival (SD)(%)	TotalLength(SD) (mm)	DryWeight (SD)(g)
Control	78(8)	100(0)	88(19)	27.5(0.57)	0.0441(0.0050)
9.4mg	83(11)	100(0)	90(8)	27.5(0.34)	0.0443(0.0030)

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

No statistically significant effects were found on survival, growth (length and dry weight) among fish exposed to a mean measured concentration of 9.4 mg/L of compound 202723. No additonal testing was conducted on compound 202723 as OECD Guideline 210 recommends that concentrations of the substance higher than 10 mg/l need not be tested.

Executive summary:

Test substance: Compound 202723

Experimental Design: Flow through exposure, 4 replicates of 30 eggs each for embryo portion (4 days), 4 replicates of 10 larvae for post-hatch portion (28 days), endpoints of hatching success, larval survival, dry weight and length

Test Organism: Fathead Minnow (Pimephales promelas)

Test Conditions: 32 day duration, 24 to 26oC, illumination of 16 hours light, 8 hours dark at 94 to 120 footcandles (1000 to 1400 lux). Well water pH 6.8 to 7.0, Specific conductivity: 230 to 260 umhos/bm, Total hardness as Ca~CO3: 40 to 58 mg/L, Total alkalinity as CaCO3: 18 to 20mg/L

Mean Measured concentration: 9.4mg compound 202723/L

Results: No effects were obserced on hatching, survival, growth (length and dry wt) among fish exposed to compound 202723 at 9.4 mg/l