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Diss Factsheets

Administrative data

Endpoint:
toxicity to terrestrial plants: long-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
23 – 20 Dec 2019
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2019
Report date:
2020

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
other: HJ/T 153-2004, The Guidelines for the Testing of Chemicals [S]. Beijing: State Environmental Protection Administration
Version / remarks:
2004
GLP compliance:
yes
Remarks:
The test was performed in compliance with the requirements of the Principles of Good Laboratory Practices (GB/T 22278-2008).

Test material

Constituent 1
Chemical structure
Reference substance name:
2-nitro-4-(trifluoromethyl)benzonitrile
EC Number:
212-298-1
EC Name:
2-nitro-4-(trifluoromethyl)benzonitrile
Cas Number:
778-94-9
Molecular formula:
C8H3F3N2O2
IUPAC Name:
2-nitro-4-(trifluoromethyl)benzonitrile

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations: 0.0 (Control), 3.125, 6.25, 12.5, 25, 50 and 100 mg/L
- Sampling method: For blank control and 3.125 mg/L: 20 mL stock solution water samples were collected for each treatment group. For 6.25 mg/L, 12.5 mg/L, 25 mg/L: 10 mL stock solution water samples were collected for each treatment group. For 50 mg/L, 100 mg/L: 2 mL stock solution water samples were collected for each treatment group.

Test substrate

Vehicle:
no
Details on preparation and application of test substrate:
- Method of mixing into soil (if used): 30 mL test solution was added to 100 g quartz sand and mixed thoroughly.

Test organisms

open allclose all
Species:
Raphanus sativus
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: radish
- Source of seed: Nanjing Vegetable Science Research Institute
- Seed storage: Test plant seeds were sealed and stored in a refrigerator at 5 °C
Species:
Glycine max (G. soja)
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: soybean
- Source of seed: Nanjing Vegetable Science Research Institute
- Seed storage: Test plant seeds were sealed and stored in a refrigerator at 5 °C
Species:
Lycopersicon esculentum
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: tomato
- Source of seed: Nanjing Vegetable Science Research Institute
- Seed storage: Test plant seeds were sealed and stored in a refrigerator at 5 °C
Species:
other: Capsicum frutescens L.
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: green chili pepper
- Source of seed: Nanjing Vegetable Science Research Institute
- Seed storage: Test plant seeds were sealed and stored in a refrigerator at 5 °C
Species:
Zea mays
Plant group:
Monocotyledonae (monocots)
Details on test organisms:
- Common name: corn
- Source of seed: Nanjing Vegetable Science Research Institute
- Seed storage: Test plant seeds were sealed and stored in a refrigerator at 5 °C
Species:
Cucumis sativus
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: cucumber
- Source of seed: Nanjing Vegetable Science Research Institute
- Seed storage: Test plant seeds were sealed and stored in a refrigerator at 5 °C
Species:
Brassica oleracea var. capitata
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: cabbage
- Source of seed: Nanjing Vegetable Science Research Institute
- Seed storage: Test plant seeds were sealed and stored in a refrigerator at 5 °C
Species:
Oryza sativa
Plant group:
Monocotyledonae (monocots)
Details on test organisms:
- Common name: rice
- Source of seed: Nanjing Vegetable Science Research Institute
- Seed storage: Test plant seeds were sealed and stored in a refrigerator at 5 °C
Species:
Lactuca sativa
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: lettuce
- Source of seed: Nanjing Vegetable Science Research Institute
- Seed storage: Test plant seeds were sealed and stored in a refrigerator at 5 °C
Species:
Brassica campestris var. chinensis
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: pak choi
- Source of seed: Nanjing Vegetable Science Research Institute
- Seed storage: Test plant seeds were sealed and stored in a refrigerator at 5 °C

Study design

Test type:
seed germination/root elongation toxicity test
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
no
Remarks:
When the seed germination rate in the control group reached 65% or higher and when root length was at least 20 mm, the test was terminated.

Test conditions

Test temperature:
24.8 - 25.2 °C
pH:
7.59 - 7.77 (blank control)
6.58 - 7.91 (treatments)
Details on test conditions:
TEST SYSTEM
- Testing facility: Testing laboratory
- Test container (type, material, size): Culture dish (diameter 120 mm, Xuzhou Dahua Glass Products Co., Ltd.); Artificial climate incubator (JM-600, Jiangsu Jiamei Instrument Manufacturing Co., Ltd.)
- Amount of soil: 100 g
- Method of seeding: The plant seeds used in the test were placed on the surface of the test substrate ensuring the direction of the ends of the seed radicles and the direction of growth were in a straight line. These were then sealed using perforated film and placed in an artificial climate box.
- No. of seeds per container: 10 seeds/dish
- No. of replicates per treatment group: 3
- No. of replicates per control: 3

GROWTH CONDITIONS
- Photoperiod: complete darkness
- Day/night temperatures: 24.8 - 25.2 °C
- Relative humidity (%): 79.8 - 80.2%

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
- When the seed germination rate in the control group reached 65% or higher and when root length was at least 20 mm, the test was terminated. When both conditions had been satisfied, mean germination count and mean root length were determined for each treatment (and control) seed

TEST CONCENTRATIONS
- Range finding study
- Test concentrations: 100 mg/L, 500 mg/L and 1000 mg/L test substance
- Results used to determine the conditions for the definitive study: Obvious inhibitory effects on germination and root elongation in the test groups of the 10 seed species exposed to 100 mg/L - 1000 mg/L
Nominal and measured concentrations:
See Table 2 in "Any other information on results incl. tables"
Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Species:
Brassica campestris var. chinensis
Duration:
12 d
Dose descriptor:
EC50
Effect conc.:
> 100 other: mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Remarks on result:
other:
Remarks:
Test was ended when germination rates for each seed species in the blank control group reached 65% or higher and root length exceeded 20 mm.
Species:
Lactuca sativa
Duration:
12 d
Dose descriptor:
EC50
Effect conc.:
> 100 other: mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Remarks on result:
other:
Remarks:
Test was ended when germination rates for each seed species in the blank control group reached 65% or higher and root length exceeded 20 mm.
Species:
Oryza sativa
Duration:
13 d
Dose descriptor:
EC50
Effect conc.:
> 100 other: mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Remarks on result:
other:
Remarks:
Test was ended when germination rates for each seed species in the blank control group reached 65% or higher and root length exceeded 20 mm.
Species:
Brassica oleracea var. capitata
Duration:
12 d
Dose descriptor:
EC50
Effect conc.:
24.1 other: mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Remarks on result:
other:
Remarks:
Test was ended when germination rates for each seed species in the blank control group reached 65% or higher and root length exceeded 20 mm.
Species:
Cucumis sativus
Duration:
6 d
Dose descriptor:
EC50
Effect conc.:
> 100 other: mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Remarks on result:
other:
Remarks:
Test was ended when germination rates for each seed species in the blank control group reached 65% or higher and root length exceeded 20 mm.
Species:
Zea mays
Duration:
13 d
Dose descriptor:
EC50
Effect conc.:
> 100 other: mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Remarks on result:
other:
Remarks:
Test was ended when germination rates for each seed species in the blank control group reached 65% or higher and root length exceeded 20 mm.
Species:
other: Capsicum frutescens L.
Duration:
12 d
Dose descriptor:
EC50
Effect conc.:
> 100 other: mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Remarks on result:
other:
Remarks:
Test was ended when germination rates for each seed species in the blank control group reached 65% or higher and root length exceeded 20 mm.
Species:
Lycopersicon esculentum
Duration:
13 d
Dose descriptor:
EC50
Effect conc.:
29.4 other: mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Remarks on result:
other:
Remarks:
Test was ended when germination rates for each seed species in the blank control group reached 65% or higher and root length exceeded 20 mm.
Species:
Glycine max (G. soja)
Duration:
13 d
Dose descriptor:
EC50
Effect conc.:
> 100 other: mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Remarks on result:
other:
Remarks:
Test was ended when germination rates for each seed species in the blank control group reached 65% or higher and root length exceeded 20 mm.
Species:
Raphanus sativus
Duration:
7 d
Dose descriptor:
EC50
Effect conc.:
> 100 other: mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Remarks on result:
other:
Remarks:
Test was ended when germination rates for each seed species in the blank control group reached 65% or higher and root length exceeded 20 mm.
Species:
Brassica campestris var. chinensis
Duration:
12 d
Dose descriptor:
EC50
Effect conc.:
8.16 other: mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: root elongation
Remarks on result:
other:
Remarks:
Test was ended when germination rates for each seed species in the blank control group reached 65% or higher and root length exceeded 20 mm.
Species:
Lactuca sativa
Duration:
12 d
Dose descriptor:
EC50
Effect conc.:
9.62 other: mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: root elongation
Remarks on result:
other:
Remarks:
Test was ended when germination rates for each seed species in the blank control group reached 65% or higher and root length exceeded 20 mm.
Species:
Oryza sativa
Duration:
13 d
Dose descriptor:
EC50
Effect conc.:
11.7 other: mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: root elongation
Remarks on result:
other:
Remarks:
Test was ended when germination rates for each seed species in the blank control group reached 65% or higher and root length exceeded 20 mm.
Species:
Brassica oleracea var. capitata
Duration:
12 d
Dose descriptor:
EC50
Effect conc.:
6.28 other: mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: root elongation
Remarks on result:
other:
Remarks:
Test was ended when germination rates for each seed species in the blank control group reached 65% or higher and root length exceeded 20 mm.
Species:
Cucumis sativus
Duration:
6 d
Dose descriptor:
EC50
Effect conc.:
5.06 other: mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: root elongation
Remarks on result:
other:
Remarks:
Test was ended when germination rates for each seed species in the blank control group reached 65% or higher and root length exceeded 20 mm.
Species:
Zea mays
Duration:
13 d
Dose descriptor:
EC50
Effect conc.:
53.5 other: mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: root elongation
Remarks on result:
other:
Remarks:
Test was ended when germination rates for each seed species in the blank control group reached 65% or higher and root length exceeded 20 mm.
Species:
other: Capsicum frutescens L.
Duration:
12 d
Dose descriptor:
EC50
Effect conc.:
11.1 other: mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: root elongation
Remarks on result:
other:
Remarks:
Test was ended when germination rates for each seed species in the blank control group reached 65% or higher and root length exceeded 20 mm.
Species:
Lycopersicon esculentum
Duration:
13 d
Dose descriptor:
EC50
Effect conc.:
3.41 other: mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: root elongation
Remarks on result:
other:
Remarks:
Test was ended when germination rates for each seed species in the blank control group reached 65% or higher and root length exceeded 20 mm.
Species:
Glycine max (G. soja)
Duration:
13 d
Dose descriptor:
EC50
Effect conc.:
36.1 other: mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: root elongation
Remarks on result:
other:
Remarks:
Test was ended when germination rates for each seed species in the blank control group reached 65% or higher and root length exceeded 20 mm.
Species:
Raphanus sativus
Duration:
7 d
Dose descriptor:
EC50
Effect conc.:
13.9 other: mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: root elongation
Remarks on result:
other:
Remarks:
Test was ended when germination rates for each seed species in the blank control group reached 65% or higher and root length exceeded 20 mm.
Details on results:
SEED GERMINATION
- Percent seed germination: At the end of the pre-test and the actual test, germination rates for all seed species in the blank control group were 73.3% - 100% (> 65%). At the lowest concentration (100 mg/L), there were already obvious inhibitory effects on germination rates in these 10 seed species, the inhibition rates for all seeds were 6.70% - 84.7%. In the 100 mg/L, 500 mg/L, and 1000 mg/L treatment groups, root elongation was inhibited in all seeds to varying degrees.
- Root length: At the end of the pre-test and the actual test, root lengths exceeded 20 mm.

SEEDLING EMERGENCE
- Percent seedling emergence: Rate of inhibition on root elongation for all seeds increased as test substance concentration increased. At the highest concentration, the range of root elongation inhibition rates for all seeds was 54.9% - 97.9%.

Any other information on results incl. tables

Table 1: Newly Prepared Test Medium Actual Measured Concentration and pH Measurement Results 

Batch no.

Prepared concentration (mg/L)

Measured concentration (mg/L)

pH

First batch

Blank control

ND

7.77

3.125

3.13

7.91

6.25

6.20

7.60

12.5

12.1

7.44

25.0

25.4

7.14

50.0

50.1

6.84

100

103

6.59

Second batch

Blank control

ND

7.59

3.125

3.18

7.41

6.25

6.22

7.15

12.5

12.3

7.10

25.0

25.2

6.90

50.0

49.9

6.80

100

101

6.58

 

 

 

Table 2: Effect concentrations with 95% confidence interval for the tested species.

Seed name

EC10 (mg/L)

95% confidence interval (mg/L)

EC50 (mg/L)

95% confidence interval (mg/L)

Germination

Root elongation

Germination

Root elongation

Brassica chinensis pak choi

8.77

2.25

>100

8.16

4.42-13.4

1.65-2.84

/

7.08-9.31

Lactuca sativa lettuce

23.2

1.33

>100

9.62

17.4-28.9

0.806-1.92

/

7.87-11.5

Oryza sativa rice

56.0

2.23

>100

11.7

38.5-93.5

1.56-2.93

/

9.97-13.6

Brassica oleracea cabbage

1.55

0.776

24.1

6.28

0.113-4.02

0-2.77

13.4-51.2

0.535-13.4

Cucumis sativus cucumber

71.5

0.787

>100

5.06

55.4-97.0

0.099-1.89

/

2.25-8.02

Zea mays corn

22.8

2.46

>100

53.5

17.4-28.0

1.21-3.93

/

39.8-79.5

Capsicum frutescens L. green chili pepper

28.4

1.93

>100

11.1

20.5-37.2

1.31-2.61

/

9.37-13.0

Lycopersicon esculentum tomato

2.45

0.636

29.4

3.41

0.369-5.37

0.033-1.70

17.5-60.8

0.996-5.81

Glycine max soybean

6.11

1.96

>100

36.1

1.95-10.9

0.928-3.23

/

27.5-49.5

Raphanus sativus radish

6.83

3.15

>100

13.9

0.904-14.4

0.502-6.35

/

7.23-24.3

 

  

 

Table 3: Validity criteria for OECD 208

Criterion

Outcome

Criterion fulfilled

Seedling emergence in the controls is ≥ 70%.

Germination rates for all seed species in the blank control group were 73.3% - 100%

yes

No visible phytotoxic effects (e.g. chlorosis, necrosis, wilting, deformatons) in control seedlings and the plants of every particular species exhibit normal variation in growth and morphology.

not specified

-

In the controls, the mean survival of emerged control seedlings is ≥ 90% for the duration of the study.

In the controls, the mean survival of emerged control seedlings was 100% for the duration of the study of all species.

yes

The environmental conditions for a particular species are identical and the growing media contain the same amount of soil matrix, support media or substrate from the same source.

The environmental conditions for a particular species were identical and the growing media contain the same amount of soil matrix, support media or substrate from the same source.

yes

 

 

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Remarks:
See Table 3 in "Any other information on results incl. tables".