2-(1,3-dihydro-3-oxo-2H-indol-2-ylidene)-1,2-dihydro-3H-indol-3-one

Toxicity to terrestrial plants

Administrative data

Endpoint:

toxicity to terrestrial plants: short-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

January to April 2005

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study (OECD, etc.)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2005

Report date:

2005

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 208 (Terrestrial Plants Test: Seedling Emergence and Seedling Growth Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

2-(1,3-dihydro-3-oxo-2H-indol-2-ylidene)-1,2-dihydro-3H-indol-3-one

EC Number:

207-586-9

EC Name:

2-(1,3-dihydro-3-oxo-2H-indol-2-ylidene)-1,2-dihydro-3H-indol-3-one

Cas Number:

482-89-3

Molecular formula:

C16H10N2O2

IUPAC Name:

2-(1,3-dihydro-3-oxo-2H-indol-2-ylidene)-1,2-dihydro-3H-indol-3-one

Details on test material:

- Name of test material: Indigo aus Indigoküpe
- Substance type: active ingredient
- Physical state: solid
- Analytical purity: 99.4%
- Lot/batch No.: V2143/1
- Date of expiry: Oct 2008

Sampling and analysis

Analytical monitoring:

yes

Test substrate

Vehicle:

yes

Details on preparation and application of test substrate:

- Test medium: 2:1 mixture of natural soil (LUFA 2.2, batch: Sp220605) and quartz sand with an organic C-content of 1.32 %.
Soil type: loamy sand (S12, DIN)
Carbon content: 2.29±0.14
pH value: 5.7 ± 0.3

- Method of mixing into soil (if used): The test item was pulverized with a mortar and dispersed in tap water. Directly weighing, each for a soil amount of 5.8 kg soil (5.5 kg DW): The test item solution was mixed with the soil. The soil was mixed carefully with a mixer to insure a homogeneous distribution of the test item.
- Controls: Tap water
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): water
- Concentration of vehicle in test medium (stock solution and final test solution): 62.5 -125 -250 -500 -1000 mg/kg soil dry weight (DW)
- Evaporation of vehicle before use: NA

Test organisms

Species:

Glycine max (G. soja)

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

- Common name: Soybean
- Plant family: Fabaceae / Leguminosae
- Source of seed: SAATBAU LINZ, A-4021 Linz, Schimmerstrasse 19, Austria
- Prior seed treatment/sterilization: no data
- Seed storage: The seeds were stored dark at the test facility at room temperature (20 ± 5°C) and protected from moisture until use

Kingdom: Plantae
Phylum: Magnoliophyta
Class: Magnoliopsida
Order: Fabales
Family: Fabaceae
Subfamily: Faboideae
Genus: Glycine
Species: G. max
Binomial name: Glycine max (L.) Merr.

Study design

Test type:

seedling emergence toxicity test

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

yes

Total exposure duration:

21 d

Post exposure observation period:

no

Test conditions

Test temperature:

13 -21°C

pH:

5.7 ± 0.3

Moisture:

45* -100 %
Deviation down to 45 % due to weather conditions had no impact on quality and integrity of the study

Details on test conditions:

TEST SYSTEM
- Testing facility: DR. U. NOACK-LABORATORIEN Kaethe-Paulus-Str. 1, 31157 Sarstedt, Germany
- Test container (type, material, size): Plastic container (standard flower pots) with a diameter of ca. 12 c
- Amount of soil: 5.8 kg soil (5.5 kg DW)
- Method of seeding: seeding holes were made with a seedling pistil
- No. of seeds per container: 5 seeds were sown per test container
- No. of plants (retained after thinning):
- No. of replicates: 8 replicates per test concentration and control were tested in a randomised block design

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
2:1 mixture of natural soil (LUFA 2.2, batch: Sp220605) and quartz sand with an organic C-content of 1.32 %
- Soil type: loamy sand (S12, DIN)
- Carbon content: 2.29±0.14
- pH value: 5.7 ± 0.3
- Origin: LANDWIRTSCHAFTLlCHE UNTERSUCHUNGS-UND FORSCHUNGS-ANSTALT SPEYER, Obere Langgasse 40, 0-67346 Speyer.
- Quartz sand type 12 a
- Grain size: 0.063 - 0.25 mm
- Origin: DORENTRUP QUARZ GMsH Co. KG, An der Sandgrube 1, 31089 Duingen.
- Storage: The natural soil and the quartz sand have been stored at the test facility at green house conditions until use. The mixture was completed one day prior to test start.

NUTRIENT MEDIUM (if used)
- Watering / Fertilisation: Bottom watering of the test containers with nutrient solution as necessary. The watering interval was based on visual observations of soil surface and plants.
- Nutrient solution: Flory 9 hydro, Origin: GARTNEREIBEDARF ASPERG E.G., Niederlassung Hannover, Ringstrasse 7, 30457 Hannover

GROWTH CONDITIONS
- Photoperiod: 16 h
- Light source: Special-fluorescent tube FLUORA (Art-No. 25414925) Special-fluorescent tube DAYLIGHT (Art.-No. 25414922) Origin: HOKLATHERM GMBH, Postfach 206, 26687 Apen
- Light intensity and quality: 4475 ± 865 Lux
- Day/night temperatures: 13 -21°C
- Relative humidity (%): 45% to 100%
- Method of application: The test item solution was mixed with the soil. The soil was mixed carefully with a mixer to insure a homogeneous distribution of the test item. The amount of stock solution and tap water volume, respectively, was the same for the test concentration and the control (test concentration: 250 mL stock solution + 250 mL tap water, control: 500 mL tap water). The treated soil was given in the test container and the seeds were sown.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Day 7, 14, 21: The plants were observed for number of emerged seedlings, visual phytotoxic effects and number of dead plants.
Day 21: Final shoot heights and fresh weight measurements


TEST CONCENTRATIONS
- Test concentrations: 62.5 -125 -250 -500 -1000 mg/kg soil dry weight (DW)

Nominal and measured concentrations:

62.5 -125 -250 -500 -1000 mg/kg soil dry weight (DW)
Concentration mg/kg 62.5 125 250 500 1000
content nominal 343.8 687.5 1375.0 2750.0 5500.0
[g] actual 343.9 687.7 1375.0 2750.3 5500.3

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Species:

Glycine max (G. soja)

Duration:

21 d

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/kg soil ww

Effect concentrations 2

Species:

Glycine max (G. soja)

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

1 000 mg/kg soil ww

Details on results:

Method of recording the values for growth (units used): Seedling emergence test

Signs of toxicity:
No test item related phytotoxic effects.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

After 21 days no test item related phytotoxic effects were observed for soybean at the tested concentration.
The test item did not cause test item related inhibitory effects> 10 % of shoot height to soybean.
The test item did not cause test item related inhibitory effects> 10 % of fresh weight (biomass) to soybean
The test item did not cause test item related inhibitory effects> 10 % of rate of emergence to soybean

Executive summary:

The phytotoxicity of the test item Indigo aus Indigo-Küpe (batch numberV2143/1) to the terrestrial plant species soybean was determined in a seedling emergence test over a period of 21 days, from March 01, 2005 to March 22, 2005 according to the guideline OECD 208 (2003). The test was conducted with the concentrations 62.5 -125 -250 -500 -1000 mg/kg soil dry weight (DW). The test item was incorporated into the soil in which the seeds were sown. The test container were bottom watered with nutrient solution as needed. The toxic effects of the test item were determined on day 7, 14 and 21 by visual observations (number of emerged seedlings and phytotoxic effects) and on day 21 by shoot fresh weight and shoot height determination. The inhibition of shoot height,shoot fresh weight and rate of emergence was calculated in relation to the untreated control.

The NOEC's of the test item, as well as the statistically calculated EC25-and EC50-values for Shoot Height, Shoot Fresh Weight, and Number of Emerged Seedlingsare:

 

NOEC:     1000 mg/kg

EC25:    > 1000 mg/kg

EC50:    > 1000 mg/kg