N-methoxy-1-(2,4,6-trichlorophenyl)propan-2-amine

Toxicity to terrestrial plants

Link to relevant study record(s)

Reference

Endpoint:

toxicity to terrestrial plants: short-term

Remarks:

The seed germination / root elongation toxicity study 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

Study period:

20 Arpil 2015 to 14 October 2015

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: HJ!T 153-2004, The guidelines for the testing of chemicals

Version / remarks:

Beijing: SEPA, 2004

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: CRC-MEP. The Guidelines for the Testing of Chemicals, Degradation and Accumulation

Version / remarks:

2nd edition. Beijing: China Environment Press. 2013: 459-463

Deviations:

no

GLP compliance:

no

Remarks:

The study was conducted under a National quality assurance scheme (HJ/T 155-2004) similar to OECD GLP.

Analytical monitoring:

yes

Details on sampling:

The initial concentration of CA5204A in an additional treatment at the start of the test was determined by UPLC. Samples at different concentrations were prepared as follows: Same volume of stock solution was added to 10 g quartz sand. After evaporated the solvent at room temperature, 50 mL acetonitrile was added and then ultrasonic extraction.
Samples of blank control, solvent control and 100 mg/L: 25.0 mL acetonitrile extract was filtrated by 0.22 µm membrane (discarding the initial 10.0 mL filtrates) and analysed by UPLC.
Samples of 500 mg/L: 5.00 mL acetonitrile extract were diluted to 50.0 mL (diluted 10 fold). Then filtrated by 0.22 µm membrane (discarding the initial 10.0 mL filtrates) and analysed by UPLC.
Samples of 1000 mg/L: 2.50 mL acetonitrile extract were diluted to 50.0 mL (diluted 20 fold). Then filtrated by 0.22 µm membrane (discarding the initial 10.0 mL filtrates) and analysed by UPLC.
The initial concentration of CA5204A was calculated by total amount of test substance in the 50.0 mL acetonitrile extract divide by 30.0 mL (volume of the deionized water).

Vehicle:

yes

Remarks:

acetone

Details on preparation and application of test substrate:

Stock solution (10071 mg/L, adjusted by the purity of 93.2%): directly dissolving 0.5403 g test substance into 50.0 mL acetone.
The test medium was prepared by adding appropriate test solution to 10 g quartz sand. The solvent was evaporated to dry at room temperature. Then 90 g quartz sand was thoroughly mixed and 30 mL deionized water was then added.
Fresh test solutions were added to glass dishes that have been filled with 100g clean quartz sand. The seed was then positioned on the substrate allowing adequate room for anticipated growth. The radicle end of the seed was aligned in the direction of this growth. The dishes were sealed with preservative film with some holes. The dishes were then placed in a climatic chamber.

Species:

Raphanus sativus

Details on test organisms:

The seeds were held in refrigeration facilities in cold storage (5°C) in moisture-proof containers at seed moisture content of less than 10 percent.
The seed was brought in from Nanjing Research Institute of Vegetables and Flowers. The information on seed lot, the seed year or growing season collected and germination percentage was provided by the supplier of the seed. 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 Sizing:
Seeds were separated into appropriate size classes, and that size class containing the most seed was used exclusively for the test. Damaged seed was discarded.

Species:

Glycine max (G. soja)

Details on test organisms:

The seeds were held in refrigeration facilities in cold storage (5°C) in moisture-proof containers at seed moisture content of less than 10 percent.
The seed was brought in from Nanjing Research Institute of Vegetables and Flowers. The information on seed lot, the seed year or growing season collected and germination percentage was provided by the supplier of the seed. 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 Sizing:
Seeds were separated into appropriate size classes, and that size class containing the most seed was used exclusively for the test. Damaged seed was discarded.

Species:

Lycopersicon esculentum

Details on test organisms:

The seeds were held in refrigeration facilities in cold storage (5°C) in moisture-proof containers at seed moisture content of less than 10 percent.
The seed was brought in from Nanjing Research Institute of Vegetables and Flowers. The information on seed lot, the seed year or growing season collected and germination percentage was provided by the supplier of the seed. 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 Sizing:
Seeds were separated into appropriate size classes, and that size class containing the most seed was used exclusively for the test. Damaged seed was discarded.

Species:

other: Sesamum indicum

Details on test organisms:

The seeds were held in refrigeration facilities in cold storage (5°C) in moisture-proof containers at seed moisture content of less than 10 percent.
The seed was brought in from Nanjing Research Institute of Vegetables and Flowers. The information on seed lot, the seed year or growing season collected and germination percentage was provided by the supplier of the seed. 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 Sizing:
Seeds were separated into appropriate size classes, and that size class containing the most seed was used exclusively for the test. Damaged seed was discarded.

Species:

Zea mays

Details on test organisms:

The seeds were held in refrigeration facilities in cold storage (5°C) in moisture-proof containers at seed moisture content of less than 10 percent.
The seed was brought in from Nanjing Research Institute of Vegetables and Flowers. The information on seed lot, the seed year or growing season collected and germination percentage was provided by the supplier of the seed. 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 Sizing:
Seeds were separated into appropriate size classes, and that size class containing the most seed was used exclusively for the test. Damaged seed was discarded.

Species:

Cucumis sativus

Details on test organisms:

The seeds were held in refrigeration facilities in cold storage (5°C) in moisture-proof containers at seed moisture content of less than 10 percent.
The seed was brought in from Nanjing Research Institute of Vegetables and Flowers. The information on seed lot, the seed year or growing season collected and germination percentage was provided by the supplier of the seed. 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 Sizing:
Seeds were separated into appropriate size classes, and that size class containing the most seed was used exclusively for the test. Damaged seed was discarded.

Species:

Brassica oleracea var. capitata

Details on test organisms:

The seeds were held in refrigeration facilities in cold storage (5°C) in moisture-proof containers at seed moisture content of less than 10 percent.
The seed was brought in from Nanjing Research Institute of Vegetables and Flowers. The information on seed lot, the seed year or growing season collected and germination percentage was provided by the supplier of the seed. 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 Sizing:
Seeds were separated into appropriate size classes, and that size class containing the most seed was used exclusively for the test. Damaged seed was discarded.

Species:

Oryza sativa

Details on test organisms:

The seeds were held in refrigeration facilities in cold storage (5°C) in moisture-proof containers at seed moisture content of less than 10 percent.
The seed was brought in from Nanjing Research Institute of Vegetables and Flowers. The information on seed lot, the seed year or growing season collected and germination percentage was provided by the supplier of the seed. 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 Sizing:
Seeds were separated into appropriate size classes, and that size class containing the most seed was used exclusively for the test. Damaged seed was discarded.

Species:

Lactuca sativa

Details on test organisms:

The seeds were held in refrigeration facilities in cold storage (5°C) in moisture-proof containers at seed moisture content of less than 10 percent.
The seed was brought in from Nanjing Research Institute of Vegetables and Flowers. The information on seed lot, the seed year or growing season collected and germination percentage was provided by the supplier of the seed. 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 Sizing:
Seeds were separated into appropriate size classes, and that size class containing the most seed was used exclusively for the test. Damaged seed was discarded.

Species:

Brassica campestris var. chinensis

Details on test organisms:

The seeds were held in refrigeration facilities in cold storage (5°C) in moisture-proof containers at seed moisture content of less than 10 percent.
The seed was brought in from Nanjing Research Institute of Vegetables and Flowers. The information on seed lot, the seed year or growing season collected and germination percentage was provided by the supplier of the seed. 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 Sizing:
Seeds were separated into appropriate size classes, and that size class containing the most seed was used exclusively for the test. Damaged seed was discarded.

Test type:

seed germination/root elongation toxicity test

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Remarks:

The exposure period ended when at least 65 percent of the control seed had germinated and developed roots that are 20 mm long.

Test temperature:

24°C to 25°C

pH:

between 6.75 and 7.04

Moisture:

Humidity: (80±5) %.

Details on test conditions:

Seeding densities: 15 seeds /dish for range-finding test.
Fresh test solutions were added to glass dishes that have been filled with 1 OOg clean quartz sand. The seed was then positioned on the substrate allowing adequate room for anticipated growth. The radicle end of the seed was aligned in the direction of this growth. The dishes were sealed with preservative film with some holes. The dishes were then placed in a climatic chamber.

Nominal and measured concentrations:

The following nominal concentrations of the test substance (CA5204A) were tested in range-finding test: 100, 500 and 1000 mg/L. A blank control and a solvent control without the test substance were run at the same time.
There was no significant phytotoxicity observed in the range-finding test. Therefor, no definitive testing was carried out.
According to the test result, the analysed concentration for nominal concentration of 100 mg/L, 500 mg/Land 1000 mg/L were 107 mg/L, 545 mg/Land 1109 mg/L respectively.

Reference substance (positive control):

no

Species:

other: all tested species

Duration:

15 d

Dose descriptor:

NOEC

Effect conc.:

1 000 other: mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

germination

Species:

other: all tested species

Duration:

15 d

Dose descriptor:

EC50

Effect conc.:

> 1 000 other: mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

germination

Species:

other: all tested species

Duration:

15 d

Dose descriptor:

NOEC

Effect conc.:

1 000 other: mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: Root elongation

Species:

other: all tested species

Duration:

15 d

Dose descriptor:

EC50

Effect conc.:

> 1 000 other: mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: Root elongation

Details on results:

At the end of the test, over 65% of the control seeds germinated and the roots developed were longer than 20 mm. Compared with the germination rates of the blank control and solvent control groups, the germination rates of the highest concentration group (1000 mg/L) were< 50%.

Reported statistics and error estimates:

The mean root length for each concentration were compared with the solvent control mean using an multiple comparison method ofDunnett's tests (ANOVA, SPSS 17.0). The results showed that no significant difference was observed for Brassica chinensis L. (Cabbage), Lacluca saliva (lettuce), Oryza saliva L. (Rice), Brassica oleracea (Kohlrabi), Cucumis salivus (cucumber), Zea mays (com), Sesamum indicum (sesame), Lycopersicon esculenlum (tomato), Glycine max (soybean) and Raphanus salivus (Radish) (p > 0.05).
According to the statistical results of the germination rate for each seed by analysis of Chi-square Test (SPSS 17.0), the germination rates of all treatments for the 10 seeds shown no significant different (p > 0.05).

Validity criteria fulfilled:

yes

Conclusions:

For germination, the LOEC and EC50 of the test substance (CA5204A) for all 10 plant species tested were found to be > 1000 mg/L.
For root elongation, the NOEC of the test substance (CA5204A) for all 10 plant species tested were found to be =1000 mg/L.

Executive summary:

Phytotoxicity of the test substance (CA5204A) to 10 terrestrial plant species was conducted according to "The guidelines for the testing of chemicals" (HJff 153-2004).

In the test, 10 plant species including Brassica chinensis L. (Cabbage), Lacluca saliva (Lettuce), Oryza saliva L. (Rice), Brassica oleracea (Kohlrabi), Cucumis salivus (Cucumber) , Zea mays (Corn), Sesamum indicum (Sesame), Lycopersicon esculenlum (Tomato), Glycine max (Soybean), Raphanus sativus (Radish) (15 seeds/group/species) were exposed to 100, 500 and 1000 mg/L of the test substance. A blank control and a solvent control without the test substance were run at the same time.

The initial concentration of CA5204A in the test solution at the start of the test was determined by UPLC. According to the test result, the measured concentration of 100 mg/L, 500 mg/L and 1000 mg/L were 107 mg/L, 545 mg/L and 1109 mg/L respectively.

During the whole test period, the pH values of the freshly prepared control and test media were between 6.75 and 7.04, and the temperature in the germination facility was maintained in the range of 24°C to 25°C. At the end of the test, more than 65 percent of the control seeds germinated and the roots developed were longer than 20 mm. So the study met the acceptability criteria prescribed by the protocol. Therefore the test was considered valid.

The results showed that under valid test conditions, there was no significant phytotoxicity observed in the range-finding test for Brassica chinensis L. (Cabbage), Lacluca saliva (Lettuce), Oryza saliva L. (Rice), Brassica oleracea (Kohlrabi), Cucumis sativus (Cucumber) , Zea mays (Corn), Sesamum indicum (Sesame), Lycopersicon esculentum (Tomato), Glycine max (Soybean), Raphanus salivus (Radish) (p > 0.05).

Description of key information

In a seed germination and root elongation toxicity test on 10 species of the substance, the germination LOEC and EC50 of the test substance (CA5204A) for all 10 plant species tested were found to be > 1000 mg/L and the root elongation NOEC of the test substance (CA5204A) for all 10 plant species tested was found to be =1000 mg/L.