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Administrative data

Endpoint:
toxicity to terrestrial plants: long-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2011-03-24 - 2011-07-25
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2011
Report date:
2011

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 208 (Terrestrial Plants Test: Seedling Emergence and Seedling Growth Test)
Deviations:
yes
Remarks:
: not affecting the outcome of the test
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Test material form:
liquid
Specific details on test material used for the study:
- Name of test material (as cited in study report): XTJ 568
- Physical state: liquid (extremely pale yellow)
- Analytical purity: 97.2%
- Lot/batch No.: DR32630507
- Expiration date of the lot/batch: 31 December 2011
- Storage condition of test material: at room temperature in the dark

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Duplicate samples were taken from deionized reverse osmosis water and the 10 g/L stock solution.
- Samples were stored at ca. -20°C for further analysis if necessary.

Test substrate

Vehicle:
no
Details on preparation and application of test substrate:
- Method of mixing into soil (if used): addition of stock solution (in deionized reverse osmosis water) to soil and mixing with a Hobart A200N mixer
- Controls: idem, but addition of a similar amount of deionized reverse osmosis water without added test substance

Test organisms

open allclose all
Species:
Avena sativa
Plant group:
Monocotyledonae (monocots)
Details on test organisms:
- Common name: oat
- Plant family: Poaceae
- Variety: SW Argyle
- Source of seed: Senova Ltd., Cambridge, UK
- Prior seed treatment/sterilization: none
- Seed storage: cool, dry conditions
- Percentage germination: 98% (data supplied by supplier)
Species:
Lycopersicon esculentum
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: tomato
- Plant family: Solanaceae
- Variety: Orange Pixie
- Source of seed: Herbiseed, Twyford, UK
- Prior seed treatment/sterilization: none
- Seed storage: cool, dry conditions
- Percentage germination: 82% (data supplied by supplier)
Species:
Glycine max (G. soja)
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: soybean
- Plant family: Fabaceae
- Variety: Amsoy
- Source of seed: Herbiseed, Twyford, UK
- Prior seed treatment/sterilization: none
- Seed storage: cool, dry conditions
- Percentage germination: 86% (data supplied by supplier)

Study design

Test type:
seedling emergence toxicity / vegetative vigour test
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
yes
Total exposure duration:
21 d
Remarks:
i.e., 21 days after 50% emergence in the control

Test conditions

Test temperature:
10-42°C - study plan: 10-35°C. Temperatures outside the study plan range were considered not to affect the integrity of the outcome of the study as no morphological abnormalities were observed throughout the test.
pH:
not reported
Moisture:
not reported
Details on test conditions:
TEST SYSTEM
- Testing facility: greenhouse with artificial lighting and heating systems
- Test container (type, material, size): polypropylene pots, depth 9.9 cm, diameter 13 cm, placed in plant pot saucer serving as sub-irrigation reservoir
- Amount of soil: ca. 850 g (dw)
- Method of seeding: planting depth according to recommendations of supplier, initial surface irrigation following sowing to encourage germination
- No. of seeds per container: 5
- No. of replicates per treatment group: 4
- No. of replicates per control: 4

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Artificial sandy loam soil mixed with sand (50/50 w/w) to give a nominal organic carbon content of 0.87% (eq. to 1.5% organic matter, which is within the acceptable range for substrates to be used in plant toxicity tests)
- Supplier: Sporting Surface Supplies Ltd., Reigate, Surrey, UK
- The soil was characterised by NRM Laboratories
- Organic carbon (%): 0.87%
- Dry matter: 88.4% w/w
- Nitrate Nitrogen: 44.74 mg/kg
- Ammonium Nitrogen: 10.08 mg/kg
- Total Nitrogen: 0.14 % w/w
- Total Phosphorus: 461 mg/kg
- Total Potassium: 1010 mg/kg
- pH: 8.0
- Organic matter: 3.8%
- Sand (2.00 - 0.063 mm): 90 % w/w
- Silt (0.063 - 0.002 mm): 5% w/w
- Clay (<0.002 mm): 5 % w/w
- Available Nitrogen: 219.3 kgN/ha
- Available Phosphorus: 72.8 mg/L
- Available Potassium: 991 mg/L
- Available Magnesium: 144 mg/L
- Conductivity: 3009 µS/cm

GROWTH CONDITIONS
- Photoperiod: 16 h light, 8 h dark
- Light source: artificial, triphosphor fluorescent lamps
- Light intensity and quality: 58 wtt, 240 volt triphosphor fluorescent lamps at 5000 lumen
- Day/night temperatures: no details reported
- Relative humidity (%): 16-87%
- Watering regime and schedules: no details reported (top-watering only to stimulate germination, afterwards only sub-irrigation)
- Water source/type: no details reported
- Volume applied: no details reported

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
- Emergence: daily after sowing
- Mortality and/or morphological abnormalities: daily for 21 days following 50% emergence in control
- Shoot dry weight: day 21 (harvested, dried at 60°C)

TEST CONCENTRATIONS
- Limit test (1000 mg/kg)
- Range finding study: yes
- Test concentrations in range finding study: control + 10, 100 and 1000 mg/kg
- Results used to determine the conditions for the definitive study: yes (except the results for soybean due to weak emergence)
Nominal and measured concentrations:
Definitive study (limit test): control + 1000 mg/kg
Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Species:
Lycopersicon esculentum
Duration:
21 d
Dose descriptor:
EC50
Effect conc.:
> 1 000 mg/kg soil ww
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
and emergence
Species:
Lycopersicon esculentum
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
>= 1 000 mg/kg soil ww
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
and emergence
Species:
Glycine max (G. soja)
Duration:
21 d
Dose descriptor:
EC50
Effect conc.:
> 1 000 mg/kg soil ww
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
and emergence
Species:
Glycine max (G. soja)
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
>= 1 000 mg/kg soil ww
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
and emergence
Species:
Avena sativa
Duration:
21 d
Dose descriptor:
EC50
Effect conc.:
> 1 000 mg/kg soil ww
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
and emergence
Species:
Avena sativa
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
>= 1 000 mg/kg soil ww
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
and emergence
Details on results:
No morphological abnormalities were observed for any of the species throughout the test.
Reported statistics and error estimates:
Statistical analysis of emergence and dry weight data was performed using Bartlett's test for homogeneity of variance (Sokal and Rohlf, 1981) and a Students t-test. For comparison of several treatments with the control, Dunnett's multiple comparison procedure was used.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
In this seedling emergence and growth test, the effect of the test substance on emergence and growth (shoot dry weight) was investigated up to 21 days after 50% emergence in the control was reached. Three plant species were included in the study: tomato (Lycopersicon esculentum), soybean (Glycine max), and oat (Avena sativa). Based on the results of the range finding test, the definitive test was decided to be a limit test (control + 1000 mg/kg). At the single concentration of 1000 mg/kg (ww), no statistically significant effects were observed on any of the species. Therefore, all EC50 and NOEC values were > 1000 mg/kg (ww) and >= 1000 mg/kg (ww), respectively.