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Toxicity to terrestrial plants

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Reference
Endpoint:
toxicity to terrestrial plants: short-term
Remarks:
The study on toxicity to terrestrial plants was conducted solely to comply with a non-EU national registration requirement, and has been provided here in accordance with REACH, Article 22(1)e.
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
The study was conducted under a National quality assurance scheme (HJ/T 155-2004) similar to OECD GLP
Qualifier:
according to guideline
Guideline:
other: SEPA HJ/T 153-2004 The guidelines for the testing of chemicals [S]. Beijing: SEPA, 2004
Version / remarks:
2004
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: CRC-MEP The Guidelines for the Testing of Chemicals-Effects on biotic systems [M]. 2nd edition Beijing: China Environmental Science Press 2013, 459 - 463
Version / remarks:
2013
Deviations:
no
GLP compliance:
no
Remarks:
The study was conducted under a National quality assurance scheme (HJ/T 155-2004) similar to OECD GLP
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Expiration date of the lot/batch: 31 May 2018
- Purity test date: 13 July 2015

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: < 30 °C
Analytical monitoring:
yes
Details on sampling:
- Concentrations: Control and 100, 500 and 1000 mg/L test solutions
- Sampling method: A sample was taken from an additional treatment at the start of the test, dissolved in acetonitrile and filtered by 0.22 µm membrane.
- Sample storage conditions before analysis: Not reported
Vehicle:
no
Details on preparation and application of test substrate:
- Method of mixing into soil: The appropriate test solution and deionised water were added to 10 g quartz sand and then 90 g quartz sand was thoroughly mixed.
- Controls: Blank control
Species:
Raphanus sativus
Details on test organisms:
- Common name: Radish
- Prior seed treatment/sterilization: Only untreated seed (not treated with fungicides, repellents, etc.) taken from the same lot and year or season of collection was used in the test
- Seed storage: Seeds were held at 5 °C in moisture-proof containers with a seed moisture content of less than 10 %
Species:
other: Capsicum annuum
Details on test organisms:
- Common name: Pepper
- Prior seed treatment/sterilization: Only untreated seed (not treated with fungicides, repellents, etc.) taken from the same lot and year or season of collection was used in the test
- Seed storage: Seeds were held at 5 °C in moisture-proof containers with a seed moisture content of less than 10 %
Species:
Lycopersicon esculentum
Details on test organisms:
- Common name: Tomato
- Prior seed treatment/sterilization: Only untreated seed (not treated with fungicides, repellents, etc.) taken from the same lot and year or season of collection was used in the test
- Seed storage: Seeds were held at 5 °C in moisture-proof containers with a seed moisture content of less than 10 %
Species:
other: Sesamum indicum
Details on test organisms:
- Common name: Sesame
- Prior seed treatment/sterilization: Only untreated seed (not treated with fungicides, repellents, etc.) taken from the same lot and year or season of collection was used in the test
- Seed storage: Seeds were held at 5 °C in moisture-proof containers with a seed moisture content of less than 10 %
Species:
Zea mays
Details on test organisms:
- Common name: Corn
- Prior seed treatment/sterilization: Only untreated seed (not treated with fungicides, repellents, etc.) taken from the same lot and year or season of collection was used in the test
- Seed storage: Seeds were held at 5 °C in moisture-proof containers with a seed moisture content of less than 10 %
Species:
Cucumis sativus
Details on test organisms:
- Common name: Cucumber
- Prior seed treatment/sterilization: Only untreated seed (not treated with fungicides, repellents, etc.) taken from the same lot and year or season of collection was used in the test
- Seed storage: Seeds were held at 5 °C in moisture-proof containers with a seed moisture content of less than 10 %
Species:
other: Brassica oleracea
Details on test organisms:
- Common name: Kohlrabi
- Prior seed treatment/sterilization: Only untreated seed (not treated with fungicides, repellents, etc.) taken from the same lot and year or season of collection was used in the test
- Seed storage: Seeds were held at 5 °C in moisture-proof containers with a seed moisture content of less than 10 %
Species:
Oryza sativa
Details on test organisms:
- Common name: Rice
- Prior seed treatment/sterilization: Only untreated seed (not treated with fungicides, repellents, etc.) taken from the same lot and year or season of collection was used in the test
- Seed storage: Seeds were held at 5 °C in moisture-proof containers with a seed moisture content of less than 10 %
Species:
Lactuca sativa
Details on test organisms:
- Common name: Lettuce
- Prior seed treatment/sterilization: Only untreated seed (not treated with fungicides, repellents, etc.) taken from the same lot and year or season of collection was used in the test
- Seed storage: Seeds were held at 5 °C in moisture-proof containers with a seed moisture content of less than 10 %
Species:
other: Brassica chinensis L.
Details on test organisms:
- Common name: Cabbage
- Prior seed treatment/sterilization: Only untreated seed (not treated with fungicides, repellents, etc.) taken from the same lot and year or season of collection was used in the test
- Seed storage: Seeds were held at 5 °C in moisture-proof containers with a seed moisture content of less than 10 %
Test type:
seed germination/root elongation toxicity test
Study type:
laboratory study
Substrate type:
other: Quartz sand
Limit test:
no
Total exposure duration:
6 d
Post exposure observation period:
None
Test temperature:
25.0 - 25.1 °C
pH:
6.85 - 7.03
Moisture:
80 - 81 % humidity
Details on test conditions:
TEST SYSTEM
- Testing facility: Climatic chamber
- Test container (type, material, size): Glass dishes sealed with preservative film containing holes
- Amount of soil: 100 g
- Method of seeding: The seed was positioned on the substrate allowing adequate room for anticipated growth. The radicle end of the seed was aligned in the direction of growth.
- No. of seeds per container: 15
- No. of replicates per treatment group: 1
- No. of replicates per control: 1

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- % sand: 100 % quartz sand

GROWTH CONDITIONS
- Photoperiod: Complete darkness
- Relative humidity (%): 80 ± 5 %

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Number of germinated seeds were counted and root lengths were measured at the end of the study. The test period ended when at least 65 % of the control seed had germinated and developed roots as least 20 mm long.

Temperature in the germination facility was reported at the start and end of the test. Test solution pH was recorded at the start of the test.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2 - 5
- Justification for using less concentrations than requested by guideline: Range-finding study conducted only
- Range finding study: Yes
- Test concentrations: 100, 500 and 1000 mg/L
- Results used to determine the conditions for the definitive study: No significant phytotoxicity was observed in the range-finding test, therefore no definitive testing was carried out
Nominal and measured concentrations:
Nominal concentrations: 100, 500 and 1000 mg/L
Measured concentrations: 100, 519 and 1012 mg/L
Reference substance (positive control):
no
Key result
Species:
other: Brassica chinensis L.
Duration:
6 d
Dose descriptor:
NOEC
Effect conc.:
1 012 other: mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
germination
Key result
Species:
Lactuca sativa
Duration:
6 d
Dose descriptor:
NOEC
Effect conc.:
1 012 other: mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
germination
Key result
Species:
Oryza sativa
Duration:
6 d
Dose descriptor:
NOEC
Effect conc.:
1 012 other: mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
germination
Key result
Species:
other: Brassica oleracea
Duration:
6 d
Dose descriptor:
NOEC
Effect conc.:
1 012 other: mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
germination
Key result
Species:
Cucumis sativus
Duration:
6 d
Dose descriptor:
NOEC
Effect conc.:
1 012 other: mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
germination
Key result
Species:
Zea mays
Duration:
6 d
Dose descriptor:
NOEC
Effect conc.:
1 012 other: mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
germination
Key result
Species:
other: Sesamum indicum
Duration:
6 d
Dose descriptor:
NOEC
Effect conc.:
1 012 other: mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
germination
Key result
Species:
Lycopersicon esculentum
Duration:
6 d
Dose descriptor:
NOEC
Effect conc.:
1 012 other: mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
germination
Key result
Species:
other: Capsicum annuum
Duration:
6 d
Dose descriptor:
NOEC
Effect conc.:
1 012 other: mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
germination
Key result
Species:
Raphanus sativus
Duration:
6 d
Dose descriptor:
NOEC
Effect conc.:
1 012 other: mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
germination
Key result
Species:
other: Brassica chinensis L.
Duration:
6 d
Dose descriptor:
NOEC
Effect conc.:
1 012 other: mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
other: root elongation
Key result
Species:
Lactuca sativa
Duration:
6 d
Dose descriptor:
NOEC
Effect conc.:
1 012 other: mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
other: root elongation
Key result
Species:
Oryza sativa
Duration:
6 d
Dose descriptor:
NOEC
Effect conc.:
1 012 other: mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
other: root elongation
Key result
Species:
other: Brassica oleracea
Duration:
6 d
Dose descriptor:
NOEC
Effect conc.:
1 012 other: mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
other: root elongation
Key result
Species:
Cucumis sativus
Duration:
6 d
Dose descriptor:
NOEC
Effect conc.:
1 012 other: mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
other: root elongation
Key result
Species:
Zea mays
Duration:
6 d
Dose descriptor:
NOEC
Effect conc.:
1 012 other: mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
other: root elongation
Key result
Species:
other: Sesamum indicum
Duration:
6 d
Dose descriptor:
NOEC
Effect conc.:
1 012 other: mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
other: root elongation
Key result
Species:
Lycopersicon esculentum
Duration:
6 d
Dose descriptor:
NOEC
Effect conc.:
1 012 other: mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
other: root elongation
Key result
Species:
other: Capsicum annuum
Duration:
6 d
Dose descriptor:
NOEC
Effect conc.:
1 012 other: mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
other: root elongation
Key result
Species:
Raphanus sativus
Duration:
6 d
Dose descriptor:
NOEC
Effect conc.:
1 012 other: mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
other: root elongation
Details on results:
SEED GERMINATION
- Percent seed germination: At the end of the test, over 65 % of the control seeds germinated
- Root length: At the end of the test, control roots were longer than 20 mm
- EC50 values were >1012 mg/L (measured, initial) for all species

SEEDLING EMERGENCE
- Percent seedling emergence: The germination rates of the highest concentration group (1000 mg/L) were < 50 % compared to the blank control
- EC50 values were >1012 mg/L (measured, initial) for all species
Reported statistics and error estimates:
The NOEC (no observed effect concentration) and LOEC (lowest observed effect concentration) for root elongation was determined by comparing the mean growth rate for each concenration with the control mean using a multiple comparison method of Dunnett's tests. The NOEC and LOEC for seed germination was determined using Chi-square Test. The EC50 was not determined as no toxicity was observed.

Table 1. Germination rates for ten seeds

 Initial measured concentration (mg/L) Control     100    519   1012
   Germination rate (%)  Inhibition rate (%)  Germination rate (%)  Inhibition rate (%)  Geramination rate (%)  Inhibition rate (%)  Germination rate (%)  Inhibition rate (%)
 Brassica chinensis L.  100  -  93.3  6.67  93.3  6.67  100 0.00
 Lactuca sativa  100  - 93.3  6.67  93.3  6.67 100  0.00
 Oryza sativa L.  100  -  93.3  6.67  93.3 6.67  93.3 6.67
 Brassica oleracea  93.3  -  93.3  0.00  93.3  0.00  93.3  0.00
 Cucumis sativus 93.3  -  100 -7.14  93.3  0.00  100  -7.14
 Zea mays  100  -  93.3  6.67  100  0.00  93.3  6.67
 Sesamum indicum  93.3  -  100 -7.14  100  -7.14  93.3  0.00
 Lycopersicon esculentum  93.3  -  100  -7.14  86.7  7.14  100  -7.14
Capsicum annuum  100  -  93.3  6.67  100.0 0.00  93.3  6.67
 Raphanus sativus  93.3  - 93.3  0.00  100 -7.14  100 -7.14

Table 2. Root lengths for ten seeds

 Initial measured concentration (mg/L)  Control        100     519        1012      
   Mean (mm)  RSD (%)  Inhibition rate (%)  Mean (mm)  RSD (%)  Inhibition rate (%)  Mean (mm)  RSD (%)  Inhibition rate (%)  Mean (mm)  RSD (%)  Inhibition rate (%)
 Lactuca sativa  25.2  10.6  - 23.6 12.0 6.08  22.8  19.2  9.23  22.6  18.0  10.2
 Oryza sativa L.  50.4  6.84  -  50.7  7.10  -0.56  50.0  10.27  0.80  49.0  7.02  2.70
 Brassica oleracea  34.1  12.7  -  34.1  11.7  -0.13  31.3 17.4  8.16  30.0 11.9  11.8
 Cucumis sativus  50.7  8.33  -  52.2  8.19 -3.02  49.8  8.03  1.75  49.5 5.46  2.31
 Zea mays 56.2  13.3  -  57.5  12.7  -2.22  58.8  9.97 -4.49  55.9  11.6  0.57
 Sesamum indicum  26.5  11.6  -  26.8  18.0  -1.27  25.2  15.9  4.78  24.0  14.2  9.26
 Lycopersicon esculentum  29.5  13.2  -  29.5  15.7  0.11  29.3  13.8  0.50 26.2  9.0  11.1
 Capsicum annuum  25.2  6.89  -  23.6  12.0 6.65  23.2 15.7  8.11  22.7  16.4  9.96
 Raphanus sativus  30.1  15.5  -

 28.9

 12.8

 3.92

 28.9

 12.6

 3.99

 28.2

 9.90

 6.27

  Brassica chinensis L.

 32.9

14.2 

 -

31.3 

 14.2

 4.91

 30.1

15.2 

8.71 

29.9

13.2 

9.11 

 

Table 3. 6-day LOEC and EC50 values for ten plant species

 Seed

 Germination   

 Root elongation   

 

 LOEC (mg/L)

 EC50 (mg/L)

 LOEC (mg/L)

 EC50 (mg/L)

 Brassica chinensis L.

 > 1012

 > 1012

 > 1012

 > 1012

 Lactuca sativa

 > 1012

 > 1012

 > 1012

 > 1012

 Oryza sativa L.

 > 1012

 > 1012

 > 1012

 > 1012

 Brassica oleracea

 > 10120

 > 1012

 > 1012

 > 1012

 Cucumis sativus

 > 1012

 > 1012

 > 1012

 > 1012

 Zea mays

 > 1012

 > 1012

 > 1012

 > 1012

 Sesamum indicum

 > 1012

 > 1012

 > 1012

 > 1012

 Lycopersicon esculentum

 > 1012

 > 1012

> 1012

> 1012 

 Capsicum annuum

 > 1012

 > 1012

 > 1012

 > 1012

 Raphanus sativus

 > 1012

 > 1012

 > 1012

 > 1012

Initial measured concentrations are reported, based on test material

Validity criteria fulfilled:
yes
Remarks:
> 65 % of the control seed germinated at the end of the test
Conclusions:
Based on initial measured concentration, the NOEC for seed germination and root elongation of the test material to Brassica chinensis L. (cabbage), Lactuca sativa (lettuce), Oryza sativa L. (rice), Brassica oleracea (kohlrabi), Cucumis sativus (cucumber), Zea mays (corn), Sesamum indicum (sesame), Lycopersicon esculentum (tomato), Capsicum annuum (pepper) and Raphanus sativus (radish) was 1012 mg/L (nominal concentration: 1000 mg/L), the highest concentration tested.
Executive summary:

The effects of the test material on seed germination and root elongation of cabbage (Brassica chinensis L.), lettuce (Lactuca sativa), rice (Oryza sativa L), kohlrabi (Brassica oleracea), cucumber (Cucumis sativus), corn (Zea mays), sesame (Sesamum indicum), tomato (Lycopersicon esculentum), pepper (Capsicum annuum) and radish (Raphanus sativus) was determined in a 6-day range-finding test according to SEPA HJ/T 153-2004 and under GLP. Seedlings were exposed to initial measured concentrations of 100, 519 and 1012 mg/L, alongside a blank control. Each replicate contained 15 seeds and only one replicate was tested for each treatment. Based on initial measured concentration, the NOEC for seed germination and root elongation of the test material was 1012 mg/L for all plant species tested (nominal concetration: 1000 mg/L). Based on initial measured concentration, the LOEC and EC50 were >1012 mg/L (nominal concentration: 1000 mg/L).

Description of key information

6-day NOEC = 1012 mg/L, seed germination and root elongation, 10 plant species, SEPA HJ/T 153 -2004, Zhen 2016

Key value for chemical safety assessment

Additional information

The effects of the test material on seed germination and root elongation of cabbage (Brassica chinensis L.), lettuce (Lactuca sativa), rice (Oryza sativa L), kohlrabi (Brassica oleracea), cucumber (Cucumis sativus), corn (Zea mays), sesame (Sesamum indicum), tomato (Lycopersicon esculentum), pepper (Capsicum annuum) and radish (Raphanus sativus) was determined in a 6-day range-finding test according to SEPA HJ/T 153-2004 and under GLP. Seedlings were exposed to initial measured concentrations of 100, 519 and 1012 mg/L, alongside a blank control. Each replicate contained 15 seeds and only one replicate was tested for each treatment. Based on initial measured concentration, the NOEC for seed germination and root elongation of the test material was 1012 mg/L for all plant species tested (nominal concentration: 1000 mg/L). Based on initial measured concentration, the LOEC and EC50 were >1012 mg/L (nominal concentration: 1000 mg/L).