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Ecotoxicological information

Toxicity to terrestrial plants

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Reference
Endpoint:
toxicity to terrestrial plants: short-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
This study was conducted between 01 August 2016 and 01 December 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
test procedure in accordance with generally accepted scientific standards and described in sufficient detail
Qualifier:
according to guideline
Guideline:
other: 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]. The second edition. Beijing: China Environmental Science Press. 2013: 459-463.
Version / remarks:
2013
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 850.4200 (Seed Germination/Root Elongation Toxicity Test)
Deviations:
no
GLP compliance:
yes
Remarks:
The guidelines of chemical testing good laboratory practices (HJ/T 155) ,China Metrology Accreditation, China National Accreditation Service for Conformity Assessment, “Principles of GLP (GB/T 22278-2008)” and “OECD Principles on GLP” ENV/MC/CHEM(98)17.
Specific details on test material used for the study:

Chemical name: 1,4-Benzenedicarboxylic acid, mixed Bu and 2-ethylhexyl diester
CAS: 1571954-81-8（A: 1962-75-0; B: 1429441-82-6: C: 6422-86-2）
Molecular formula: A: C16H22O4; B: C20H30O4; C: C24H38O4
Molecular weight: A: 278; B: 334; C: 390
Batch No.: GLFG160607
Purity: 99.8%
Exp. Date: 2017-06-01
Water solubility: 2.19 mg/L (20°C)
Appearance: Colorless liquid
Storage conditions: Room temperature
Analytical monitoring:
yes
Details on sampling:
The initial concentration of GL500 in an additional treatment at the start of the test was determined by UPLC. 300 mL acetonitrile was added to the test substrates for concentration analysis.
Samples of blank control, solvent control and 100 mg/L: 10.0 mL acetonitrile extract was filtrated by 0.22 µm membrane (discarding the initial 5.00 mL filtrates) and analysed by UPLC.
Samples of 500 and 1000 mg/L: 10.0 mL acetonitrile extract were diluted to 50.0 mL, and filtrated by 0.22 µm membrane (discarding the initial 5.00 mL filtrates) and analysed by UPLC.
Vehicle:
no
Details on preparation and application of test substrate:
Dilution Medium
Quartz sand was used as the medium. CAS: 14808-60-7; Lot number: 20160309; Sinopharm Chemical Reagent Co. Ltd.

Preparation of the Test Substrates
Stock solution (10000 mg/L): directly dissolving 1.0036 g test substance into 100 mL acetone.
The test medium was prepared by adding appropriate test solution and to 100 g quartz sand. Then 30 mL deionized water was thoroughly mixed.
The test substrates for concentration analysis were prepared at the same time, while no deionized water added. Details of the media preparation are showed as follows:
No. Concentration (mg/L) Concentration of the Stock solution (mg/L) Volume of the Stock solution added (mL) Deionized water (mL) Total amount of the quartz sand (g)
1 Blank Control - 0 30.0 100
2 Solvent Control - 3.00 mL acetone 30.0 100
3 100 10000 0.30 30.0 100
4 500 10000 1.50 30.0 100
5 1000 10000 3.00 30.0 100

Test Performance
Fresh test solutions were added to glass dishes that were 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. During the test, the following conditions were maintained:
Light: complete darkness.
Temperature: (25±1) °C.
Humidity: (80±5) %.
Seeding densities: 15 seeds /dish for range-finding test.
Replicates: none for range-finding test.
The test period was ended when at least 65 percent of the control seed had germinated and developed roots at least 20 mm long. When both conditions were satisfied, the mean number of seed germinating and mean root length per treatment (and control) were determined. EC50s for germination and root elongation were determined for each species tested and reported along with their 95 percent confidence limits.
Details on test organisms:
SEE TABLE 1 BELOW ANY OTHER INFORMATION ON MATERIALS AND METHODS INCL. TABLES)
Test type:
seed germination/root elongation toxicity test
Study type:
laboratory study
Substrate type:
other: Quartz sand was used as the medium. CAS: 14808-60-7; Lot number: 20160309; Sinopharm Chemical Reagent Co. Ltd.
Limit test:
no
Total exposure duration:
7 d
Remarks:
The seeds were treated with prepared test medium in the dark until at least 65 percent of the control seed have germinated and developed roots that are at least 20 mm long.
Test temperature:
25 ± 1”C
pH:
6.35 - 7.56
Moisture:
Humidity: (80±5) %.
Details on test conditions:
Test Performance
Fresh test solutions were added to glass dishes that were 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. During the test, the following conditions were maintained:
Light: complete darkness.
Temperature: (25±1) °C.
Humidity: (80±5) %.
Seeding densities: 15 seeds /dish for range-finding test.
Replicates: none for range-finding test.
The test period was ended when at least 65 percent of the control seed had germinated and developed roots at least 20 mm long. When both conditions were satisfied, the mean number of seed germinating and mean root length per treatment (and control) were determined. EC50s for germination and root elongation were determined for each species tested and reported along with their 95 percent confidence limits.

Observations & Measurements
As different seed germination and root elongation varies dependent on species, the exposure duration for each test species were shown in followed table. When the exposure ended, numbers of seeds that germinated were counted and root lengths measured. All root elongation measurements for a given species were made sequentially before proceeding to the next species. Root length was measured from the transition point between the hypocotyl and root to the tip of the root.
Temperature in the germination facility was reported at the start and the end of the test. The pH of the test solutions was recorded at the initiation of the test.

Validity of Test
(1) Control group
One blank control and one solvent control series were also run in addition to the treatment series.
(2) Validity of test result
During the whole test period, more than 65 percent of the blank control and solvent control seed germinated at the end of test. Therefore the test was considered valid.

Data Processing
For the estimation of the Lowest Observed Effect Concentration (LOEC), and hence the No Observed Effect Concentration (NOEC), for effects of the chemical on root elongation, the mean growth rate for each concentration were compared with the control mean using a multiple comparison method of Dunnett’s tests (ANOVA, SPSS 17.0).
For the estimation of the Lowest Observed Effect Concentration (LOEC), and hence the No Observed Effect Concentration (NOEC), for effects of the chemical on germination, Chi-square Test was used (SPSS 17.0).
Probit analysis (SPSS 17.0) was used to calculate the EC50 and EC50 confidence limits. This method was not carried out because of the low toxicity of the test substance.
Nominal and measured concentrations:
Measured
Reference substance (positive control):
no
Key result
Species:
other: All Species tested
Duration:
7 d
Dose descriptor:
LOEC
Effect conc.:
> 1 000 other: mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Key result
Species:
other: All species tested
Duration:
7 d
Dose descriptor:
NOEC
Effect conc.:
> 1 000 other: mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Key result
Species:
other: All species tested
Duration:
7 d
Dose descriptor:
EC50
Effect conc.:
> 1 000 other: mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Key result
Species:
other: All species tested
Duration:
7 d
Dose descriptor:
LOEC
Effect conc.:
> 1 000 other: mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Root elongation
Key result
Species:
other: All species tested
Duration:
7 d
Dose descriptor:
NOEC
Effect conc.:
> 1 000 other: mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: root elongation
Key result
Species:
other: All species tested
Duration:
7 d
Dose descriptor:
EC50
Effect conc.:
> 1 000 other: mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: root elongation
Details on results:
Analysis Results of the Test Concentration
(1) Specificity
Under the above UPLC condition, the retention times of test substance was about 2 min, 3.9 min and 9.3 min (see Figure 2), while there was no chromatographic peak at the retention time for the control sample (see Figure 3). The results showed that the chromatographic peak was the characteristic peak of the test substance. And the analytical method had specificity.
(2) Calibration curve
A series of standard solutions with concentrations of 0, 0.50, 1.00, 2.00, 5.00, 10.0 and 20.0 mg/L were measured under the UPLC conditions mentioned above. Based on the test result, a linear regression equation was obtained between the concentration and the sum of 3 peak area: A=102565c-5988.5, with good linearity of r2 = 0.9999, where A represents peak area (AU); and c is the concentration of the test substance (mg/L) (see Figure 1). The results showed that linearity for concentration range of 0 to 20.0 mg/L was good.
(3) Detection limit and quantitation limit
The detection limit was 0.06 mg/L (S/N ≥ 3), and the quantitation limit was 0.20 mg/L (S/N ≥ 10).
(4) Analysed results of the test concentration
According to the test result, the analysed concentration for nominal concentration of 100 mg/L, 500 mg/L and 1000 mg/L were 99.1 mg/L, 497 mg/L and 1015 mg/L respectively (Table 1). The measured concentrations were more than 90 percent of the nominal concentrations, so the final test results were based on nominal concentrations.

Effects on Germination Rates and Root elongations
Table 3 showed germination rates and germination inhibition rates observed for the seeds of different species. Table 4 to Table 13 showed root lengths and root lengths inhibition rates observed for the seeds of different species
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%.
The mean root length for each concentration was compared with the solvent control mean using an multiple comparison method of Dunnett’s tests (ANOVA PSS 17.0). The results showed that no significant difference was observed for 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), Glycine max (Soybean) and Raphanus sativus (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).
Under valid test conditions, the LOEC, NOEC and EC50 for 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), Glycine max (Soybean), Raphanus sativus (Radish) tested were shown in the table below.
Seed Germination Root Elongation
LOEC NOEC EC50 LOEC NOEC EC50
Cabbage > 1000 1000 > 1000 > 1000 1000 > 1000
Lettuce > 1000 1000 > 1000 > 1000 1000 > 1000
Rice > 1000 1000 > 1000 > 1000 1000 > 1000
Kohlrabi > 1000 1000 > 1000 > 1000 1000 > 1000
Cucumber > 1000 1000 > 1000 > 1000 1000 > 1000
Corn > 1000 1000 > 1000 > 1000 1000 > 1000
Sesame > 1000 1000 > 1000 > 1000 1000 > 1000
Tomato > 1000 1000 > 1000 > 1000 1000 > 1000
Soybean > 1000 1000 > 1000 > 1000 1000 > 1000
Radish > 1000 1000 > 1000 > 1000 1000 > 1000
Reported statistics and error estimates:
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).

Table 3 Germination Rates for Ten Seeds

Nominal Concentration

mg/L

Analysed concentration

mg/L

Cabbage

Lettuce

Rice

Kohlrabi

Cucumber

germination rate(%)

inhibition rate(%)

germination rate(%)

inhibition rate(%)

germination rate(%)

inhibition rate(%)

germination rate(%)

inhibition rate(%)

germination rate(%)

inhibition rate(%)

1CK

2ND

100

-

93.3

-

93.3

-

93.3

-

100

-

1SK

2ND

93.3

-

93.3

-

93.3

-

100

-

100

-

100

99.1

93.3

0.00

86.7

7.1

93.3

0.00

93.3

6.67

93.3

6.67

500

497

93.3

0.00

93.3

0.0

100

-7.14

100

0.00

93.3

6.67

1000

1015

86.7

7.1

93.3

0.0

93.3

0.00

93.3

6.67

100

0.00

EC50

> 1000 mg/L (measured concentration 1015 mg/L)

LOEC

> 1000 mg/L (measured concentration 1015 mg/L)

NOEC

= 1000 mg/L (measured concentration 1015 mg/L)

Nominalconcentration

mg/L

Analysed concentration

mg/L

Corn

Sesame

Tomato

Soybean

Radish

germination rate(%)

inhibition rate(%)

germination rate(%)

inhibition rate(%)

germination rate(%)

inhibition rate(%)

germination rate(%)

inhibition rate(%)

germination rate(%)

inhibition rate(%)

1CK

2ND

100

-

93.3

-

93.3

-

100

-

100

-

1SK

2ND

100

-

100

-

93.3

-

100

-

93.3

-

100

99.1

100

0.00

93.3

6.67

86.7

7.14

100

0

93.3

0.00

500

497

100

0.00

100

0.00

93.3

0.00

100

0

100

-7.14

1000

1015

100

0.00

93.3

6.67

100

-7.14

93.3

6.67

93.3

0.00

EC50

> 1000 mg/L (measured concentration 1015 mg/L)

LOEC

> 1000 mg/L (measured concentration 1015 mg/L)

NOEC

= 1000 mg/L (measured concentration 1015 mg/L)

Table 4 Root Lengths Observed forBrassica chinensisL. (Cabbage)

Nominalcon.

Analysed con.

Root Length(mm)

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

Mean

RSD%

4I.rate %

1CK

2ND

30.8

24.6

23.5

27.1

24.4

31.4

25.9

34.8

28.5

28.1

21.2

32.9

36

40.5

28.7

29.2

17.9

-

1SK

2ND

29.9

24.6

24.5

28.4

25.5

26.7

24.3

30.4

29.7

29.1

25.4

26.7

30.4

29.7

3NA

27.5

8.61

-

100

99.1

30.1

28.1

37.7

25.2

30.2

25.9

25.9

27.4

33.3

29.8

32.1

29.4

34.7

24.9

3NA

29.6

12.8

-7.63

500

497

32.0

26.2

23.3

27.1

29.8

31.2

28.8

25.4

23.5

22.4

36.2

32.1

27.8

37.2

3NA

28.8

16.0

-4.59

1000

1015

26.4

28.8

23.9

23.6

26.9

24.0

27.3

30.9

30.7

26.2

24.8

26.0

27.1

3NA

3NA

26.7

8.91

3.12

EC50

> 1000 mg/L (measured concentration 1015 mg/L)

LOEC

> 1000 mg/L (measured concentration 1015 mg/L)

NOEC

= 1000 mg/L (measured concentration 1015 mg/L)

1: CK was abbreviation of blank control, and SK was abbreviation of solvent control.2:not detectable because it was lower than the limit of detection.3NA: not available because the seed did not germinated.4:I.ratewas abbreviation of inhibition rate.

Table 5 Root Lengths Observed forLactuca sativa(lettuce)

Nominalcon.

Analysed con.

Root Length(mm)

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

Mean

RSD%

5I.rate %

1CK

2ND

28.1

22.9

29

20.7

24.6

28.3

21.6

26.2

26.9

26.5

25.6

23.5

24.5

27.4

3NA

25.4

10.0

-

1SK

2ND

28.3

21.5

27.4

21.5

23.1

28.4

22.2

24.3

25.7

25.9

26.4

22.9

26.1

27.4

3NA

25.1

9.84

-

100

99.1

27.0

23.0

19.6

24.7

27.4

18.0

24.9

26.3

22.1

23.6

21.8

24.7

24.3

3NA

3NA

23.6

11.7

5.71

500

497

25.7

21.0

22.4

18.6

27.3

19.3

31.3

22.2

29.1

25.9

22.6

17.4

21.5

20.7

3NA

23.2

17.6

7.43

1000

1015

24.3

19.9

21.3

25.7

23.1

22.7

30.9

19.3

23.5

22.3

22.3

18.3

31.3

23.3

3NA

23.4

16.2

6.52

EC50

> 1000 mg/L (measured concentration 1015 mg/L)

LOEC

> 1000 mg/L (measured concentration 1015 mg/L)

NOEC

= 1000 mg/L (measured concentration 1015 mg/L)

1: CK was abbreviation of blank control, and SK was abbreviation of solvent control.2:not detectable because it was lower than the limit of detection.3NA: not available because the seed did not germinated.4:I.ratewas abbreviation of inhibition rate.

Table 6 Root Lengths Observed forOryza sativa L.(Rice)

Nominalcon.

Analysed con.

Root Length(mm)

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

Mean

RSD%

4I.rate %

1CK

2ND

56.8

48.6

53.3

46.5

59.0

46.6

46.5

51.5

54.0

46.8

52.4

44.6

48.6

49.7

3NA

50.4

8.53

-

1SK

2ND

55.8

50.2

55.1

48.9

53.2

46.5

45.9

52.3

53.9

45.1

52.3

44.8

48.5

49.6

3NA

50.2

7.40

-

100

99.1

47.5

50.4

49.1

56.8

53.3

47.2

47.3

54.1

50.0

43.9

51.0

49.1

46.0

54.6

3NA

50.0

7.28

0.26

500

497

57.7

50.5

49.1

51.5

58.9

48.8

48.6

43.2

47.9

53.9

42.9

45.7

47.5

48.3

51.4

49.7

9.15

0.84

1000

1015

52.4

53.1

45.9

44.6

46.4

43.1

45.4

48.3

53.7

52.7

43.2

49.2

50.6

45.7

3NA

48.2

7.84

3.96

EC50

> 1000 mg/L (measured concentration 1015 mg/L)

LOEC

> 1000 mg/L (measured concentration 1015 mg/L)

NOEC

= 1000 mg/L (measured concentration 1015 mg/L)

1: CK was abbreviation of blank control.2:not detectable because it was lower than the limit of detection.3NA: not available because the seed did not germinated.

4:I.ratewas abbreviation of inhibition rate.

Table 7 Root Lengths Observed forBrassica oleracea(Kohlrabi)

Nominalcon.

Analysed con.

Root Length(mm)

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

Mean

RSD%

4I.rate %

1CK

2ND

31.6

27.9

30.5

31.3

34.1

30.9

23.1

35.1

38.6

31.7

36.3

35.1

35.6

34.6

3NA

32.6

12.0

-

1SK

2ND

30.5

28.6

28.9

30.5

29.4

34.5

31.1

32.5

35.8

33.4

35.6

37.1

34.6

35.1

34.1

32.8

8.42

-

100

99.1

27.8

30.4

36.3

35.5

32.7

35.4

39.2

32.5

36.7

29.2

32.1

28.7

33.7

38.4

3NA

33.5

10.8

-2.11

500

497

28.3

30.5

26.4

28.1

29.8

30.5

30.7

27.6

40.9

24.2

35.6

32.2

28.9

35.2

27.9

30.5

13.7

7.10

1000

1015

33.2

32.7

28.7

24.7

33.8

33.6

28.8

28.7

32.6

28.2

30.6

26.6

32.9

29.5

3NA

30.3

9.46

7.48

EC50

> 1000 mg/L (measured concentration 1015 mg/L)

LOEC

> 1000 mg/L (measured concentration 1015 mg/L)

NOEC

= 1000 mg/L (measured concentration 1015 mg/L)

1: CK was abbreviation of blank control, and SK was abbreviation of solvent control.2:not detectable because it was lower than the limit of detection.3NA: not available because the seed did not germinated.4:I.ratewas abbreviation of inhibition rate.

Table 8 Root Lengths Observed forCucumis sativus(cucumber)

Nominalcon.

Analysed con.

Root Length(mm)

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

Mean

RSD%

4I.rate %

1CK

2ND

51.1

47.4

55.5

40.5

56.6

50.7

46.1

48.2

53.5

45.3

55.8

51.3

46.1

53.8

47.5

50.0

9.21

-

1SK

2ND

48.9

49.6

56.1

45.2

49.6

53.8

49.5

50.1

52.6

46.8

53.6

50.8

48.7

51.9

49.2

50.4

5.57

-

100

99.1

49.1

56.7

47.4

43.3

47.9

57.5

52.6

56.8

53.1

56.8

58.7

53.3

49.0

48.8

3NA

52.2

9.00

-3.54

500

497

54.7

56.8

47.5

47.6

50.0

44.0

48.5

41.7

52.2

43.8

52.8

49.8

55.1

52.4

3NA

49.8

9.15

1.29

1000

1015

53.7

53.2

50.0

44.5

45.2

48.7

52.0

46.2

47.7

52.8

53.7

47.6

50.8

48.6

47.1

49.5

6.31

1.93

EC50

> 1000 mg/L (measured concentration 1015 mg/L)

LOEC

> 1000 mg/L (measured concentration 1015 mg/L)

NOEC

= 1000 mg/L (measured concentration 1015 mg/L)

1: CK was abbreviation of blank control, and SK was abbreviation of solvent control.2:not detectable because it was lower than the limit of detection.3NA: not available because the seed did not germinated.4:I.ratewas abbreviation of inhibition rate.

Table 9 Root Lengths Observed forZea mays(corn)

Nominalcon.

Analysed con.

Root Length(mm)

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

Mean

RSD%

3I.rate %

1CK

2ND

61.3

50.5

49.1

54.1

51.5

56.8

51.4

62.0

68.9

45.6

71.2

62.0

57.0

53.1

48.3

56.2

13.4

-

1SK

2ND

62.5

55.5

50.1

53.2

51.1

55.6

53.2

59.8

64.2

55.1

53.6

57.8

56.7

54.5

50.1

55.5

7.50

-

100

99.1

64.1

65

67.1

53

49.3

56.1

50.5

66.2

51.9

53.6

60.4

58.2

60.2

60.5

53.4

58.0

10.2

-4.38

500

497

52.5

65.1

57.2

55.1

60.2

61.2

54.6

49.3

51.2

64

67.1

53.2

61.8

54.7

55.3

57.5

9.40

-3.54

1000

1015

55.8

59.2

64.1

58.4

49.8

53.9

54.8

64.9

51.2

48.9

54.7

67.4

50.9

48.2

61.5

56.2

10.9

-1.28

EC50

> 1000 mg/L (measured concentration 1015 mg/L)

LOEC

> 1000 mg/L (measured concentration 1015 mg/L)

NOEC

= 1000 mg/L (measured concentration 1015 mg/L)

1: CK was abbreviation of blank control, and SK was abbreviation of solvent control.2:not detectable because it was lower than the limit of detection.3:I.ratewas abbreviation of inhibition rate.

Table 10 Root Lengths Observed forSesamum indicum(sesame)

Nominalcon.

Analysed con.

Root Length(mm)

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

Mean

RSD%

4I.rate %

1CK

2ND

29.7

30.7

26.3

27.5

23.4

28.6

18.6

32.3

21.8

28.5

25

28.9

24.2

26

3NA

26.5

14.0

-

1SK

2ND

30.1

30.1

28.9

25.6

23.4

24.8

20.5

30.4

26.7

24.6

26.8

27.9

23.7

25.7

24.3

26.2

10.9

-

100

99.1

31.2

30.8

32.2

17.8

28.7

22.1

22.0

23.4

31.9

25.4

31.8

28.5

29.4

30.8

3NA

27.6

16.7

-5.10

500

497

28.2

22.5

18.9

28.7

22.7

17.4

23.3

21.4

29.7

20.8

30.0

28.6

29.6

26.2

29.2

25.1

17.3

4.14

1000

1015

21.0

18.9

20.6

31.0

26.2

28.8

22.0

22.8

28.6

28.2

22.6

26.3

27.4

24.6

3NA

24.9

14.7

4.97

EC50

> 1000 mg/L (measured concentration 1015 mg/L)

LOEC

> 1000 mg/L (measured concentration 1015 mg/L)

NOEC

= 1000 mg/L (measured concentration 1015 mg/L)

1: CK was abbreviation of blank control, and SK was abbreviation of solvent control.2:not detectable because it was lower than the limit of detection.3NA: not available because the seed did not germinated.4:I.ratewas abbreviation of inhibition rate.

Table 11 Root Lengths Observed forLycopersicon esculentum(tomato)

Nominalcon.

Analysed con.

Root Length(mm)

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

Mean

RSD%

4I.rate %

1CK

2ND

30.5

32.1

22.8

30.7

30.7

33.3

21.6

31.3

27.1

27.5

31.2

24.6

29.9

32.7

3NA

29.0

12.8

-

1SK

2ND

28.6

29.3

24.5

28.4

30.5

32.4

30.5

29.8

29.9

28.6

30.5

26.7

30.2

31.1

3NA

29.4

6.69

-

100

99.1

30.4

32.2

24.5

27.0

31.6

28.4

27.8

31.7

30.1

22.7

27.8

21.0

26.7

3NA

3NA

27.8

12.5

5.17

500

497

26.3

28.4

30.0

23.0

34.1

27.5

29.0

32.3

34.9

25.8

26.0

23.2

30.8

25.3

3NA

28.3

13.2

3.50

1000

1015

31.8

24.4

30.7

22.7

31.0

25.8

27.2

30.5

24.7

23.2

28.8

28.4

26.1

24.8

29.1

27.3

11.0

7.12

EC50

> 1000 mg/L (measured concentration 1015 mg/L)

LOEC

> 1000 mg/L (measured concentration 1015 mg/L)

NOEC

= 1000 mg/L (measured concentration 1015 mg/L)

1: CK was abbreviation of blank control, and SK was abbreviation of solvent control.2:not detectable because it was lower than the limit of detection.3NA: not available because the seed did not germinated.4:I.ratewas abbreviation of inhibition rate.

Table 12 Root Lengths Observed forGlycine max(Soybean)

Nominalcon.

Analysed con.

Root Length(mm)

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

Mean

RSD%

4I.rate %

1CK

2ND

69.2

62.9

68.3

66.8

65.5

64.1

69.4

57.6

69.2

53.3

52.7

57.6

62.9

68.0

53.4

62.7

9.92

-

1SK

2ND

67.8

60.6

61.1

51.6

62.1

52.2

66.5

50.1

69.5

69.1

52.3

67.3

69.1

52.5

57.2

60.6

12.2

-

100

99.1

68.5

68.0

61.1

53.6

57.5

51.7

58.8

58.4

52.5

51.4

65.5

53.6

64.2

67.0

54.6

59.1

10.6

2.49

500

497

54.1

53.4

56.2

57.3

66.4

66.3

59.3

67.6

66.3

61.7

53.0

59.8

61.5

54.9

56.1

59.6

8.63

1.64

1000

1015

58.9

50.8

51.8

56.6

62.4

50.1

63.7

53.5

68.9

59.4

54.1

55.2

57.7

62.4

3NA

57.5

9.51

5.07

EC50

> 1000 mg/L (measured concentration 1015 mg/L)

LOEC

> 1000 mg/L (measured concentration 1015 mg/L)

NOEC

= 1000 mg/L (measured concentration 1015 mg/L)

1: CK was abbreviation of blank control, and SK was abbreviation of solvent control.2:not detectable because it was lower than the limit of detection.3NA: not available because the seed did not germinated.4:I.ratewas abbreviation of inhibition rate.

Table 13 Root Lengths Observed forRaphanus sativus(Radish)

Nominalcon.

Analysed con.

Root Length(mm)

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

Mean

RSD%

4I.rate %

1CK

2ND

32.7

32.3

29.5

37.1

32.9

28.3

24.5

41.9

28.7

26.9

23.9

25.8

29.5

28.6

30.1

30.2

15.7

-

1SK

2ND

31.8

33.3

29.8

35.9

31.7

29.6

25.6

36.9

33.5

29.8

24.1

22.8

23.6

28.1

3NA

29.8

15.1

-

100

99.1

33.2

32.4

24.5

28.6

34.6

24.9

29.0

34.5

30.6

23.5

27.3

23.8

25.2

32.8

3NA

28.9

14.2

2.79

500

497

26.8

28.7

22.2

26.3

23.9

28.0

29.2

27.6

36.2

25.9

28.3

25.3

34.7

29.5

27.8

28.0

12.9

5.79

1000

1015

31.1

29.0

28.4

31.1

30.0

27.1

25.9

25.6

28.5

23.2

36.0

25.5

28.3

26.9

3NA

28.3

11.1

4.78

EC50

> 1000 mg/L (measured concentration 1015 mg/L)

LOEC

> 1000 mg/L (measured concentration 1015 mg/L)

NOEC

= 1000 mg/L (measured concentration 1015 mg/L)

1: CK was abbreviation of blank control, and SK was abbreviation of solvent control.2:not detectable because it was lower than the limit of detection.3NA: not available because the seed did not germinated.4:I.ratewas abbreviation of inhibition rate

Validity criteria fulfilled:
yes
Conclusions:
Under valid test conditions, the LOEC, NOEC and EC50 for 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), Glycine max (Soybean), Raphanus sativus (Radish) tested were shown in the table below.
Seed Germination Root Elongation
LOEC NOEC EC50 LOEC NOEC EC50
Cabbage > 1000 1000 > 1000 > 1000 1000 > 1000
Lettuce > 1000 1000 > 1000 > 1000 1000 > 1000
Rice > 1000 1000 > 1000 > 1000 1000 > 1000
Kohlrabi > 1000 1000 > 1000 > 1000 1000 > 1000
Cucumber > 1000 1000 > 1000 > 1000 1000 > 1000
Corn > 1000 1000 > 1000 > 1000 1000 > 1000
Sesame > 1000 1000 > 1000 > 1000 1000 > 1000
Tomato > 1000 1000 > 1000 > 1000 1000 > 1000
Soybean > 1000 1000 > 1000 > 1000 1000 > 1000
Radish > 1000 1000 > 1000 > 1000 1000 > 1000
Executive summary:

Under valid conditions, the phytotoxicity of the test substance (GL500) to 10 terrestrial plant species was conducted according to “The guidelines for the testing of chemicals” (HJ/T 153-2004).

In the test, 10 plant species including Brassica chinensisL. (Cabbage), Lactuca sativa (Lettuce), Oryza sativaL.(Rice), Brassica oleracea (Kohlrabi), Cucumis sativus (Cucumber) , Zea mays (Corn),Sesamum indicum (Sesame), Lycopersicon esculentum (Tomato), Glycine max (Soybean), Raphanus sativus (Radish) (15 seeds/group/species) were exposed to 100, 500 and 1000 mg/Lof the test substance. A blank control and a solvent control without the test substance were run at the same time. At the beginning of the test, the pH values of the test solution were between 6.35and7.56. During the test, the temperature in the germination facility was maintained in the range of 24.5°C to 25.2°C. At the end of the test, more than 65 percent of the control seeds germinated and the roots developed were longer than. So the study met the acceptability criteria prescribed by the protocol. Therefore the test was considered valid.

The initial concentration of GL500 in the test solution at the start of the test was determined by UPLC. According to the test result, the measured concentration of 100mg/L, 500mg/L and 1000 mg/L were 99.1 mg/L, 497 mg/L and 1015 mg/L respectively.The measured concentrations were more than 90 percent of the nominal concentrations, so the final test results were based on nominal concentrations.

The results showed that there was no significant phytotoxicity observed at rates of100mg/L, 500 mg/L and 1000 mg/L in the range-finding test for Brassica chinensisL.(Cabbage), Lactuca sativa (Lettuce),Oryza sativaL. (Rice), Brassica oleracea (Kohlrabi), Cucumis sativus (Cucumber), Zea mays (Corn), Sesamum indicum (Sesame), Lycopersicon esculentum (Tomato), Glycine max (Soybean),Raphanus sativus (Radish) (p> 0.05).

Under valid test conditions,the LOEC, NOEC and EC50of the test substance (GL500) for all10 plant species tested are shown in the table below.

Seed

1Germination

1Root Elongation

LOEC

(mg/L)

NOEC 

(mg/L)

EC50 

(mg/L)

LOEC 

(mg/L)

NOEC 

(mg/L)

EC50 

(mg/L)

Cabbage

> 1000

1000

> 1000

> 1000

1000

> 1000

Lettuce

> 1000

1000

> 1000

> 1000

1000

> 1000

Rice

> 1000

1000

> 1000

> 1000

1000

> 1000

Kohlrabi

> 1000

1000

> 1000

> 1000

1000

> 1000

Cucumber

> 1000

1000

> 1000

> 1000

1000

> 1000

Corn

> 1000

1000

> 1000

> 1000

1000

> 1000

Sesame

> 1000

1000

> 1000

> 1000

1000

> 1000

Tomato

> 1000

1000

> 1000

> 1000

1000

> 1000

Soybean

> 1000

1000

> 1000

> 1000

1000

> 1000

Radish

> 1000

1000

> 1000

> 1000

1000

> 1000

1: The measured concentration for1000mg/Lwas 1015 mg/L.


Description of key information

Under validtestconditions, theLOEC, NOEC and EC50forBrassica chinensisL.(Cabbage),Lactuca sativa(Lettuce),Oryza sativaL.(Rice),Brassica oleracea(Kohlrabi),Cucumis sativus(Cucumber) ,Zea mays(Corn),Sesamum indicum(Sesame),Lycopersicon esculentum(Tomato),Glycine max(Soybean),Raphanus sativus(Radish)testedwere >1000 mg/L

Key value for chemical safety assessment

Additional information