Octadecene, reaction products with hexadecene, hydrogenated

Toxicity to terrestrial plants

Link to relevant study record(s)

Reference

Endpoint:

toxicity to terrestrial plants: short-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

November 21, 2019 to December 18, 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.4200 (Seed Germination/Root Elongation Toxicity Test)

Version / remarks:

OPPTS. Ecological Effects Test Guidelines, OPPTS 850.4200 Seed Germination/Root Elongation Toxicity Test [S]. US EPA, 1996.

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: National Guidelines: The Guidelines for the Testing of Chemicals Effects on Biotic Systems, 299 Seed Germination/Root Elongation Toxicity Test [M].

Version / remarks:

Chemical Registration Center of MEP. The Guidelines for the Testing of Chemicals Effects on Biotic Systems, 299 Seed Germination/Root Elongation Toxicity Test [M]. Second Edition. Beijing: China Environmental Science Press. 2013: 459-463.

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: National Guidleines: State Environmental Protection Administration of China. HJ/T 153-2004 The Guidelines for the test of chemical [S].

Version / remarks:

State Environmental Protection Administration of China. HJ/T 153-2004 The Guidelines for the test of chemical [S]. Beijing: China Environmental Science Press. 2004.

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: National Guidelines: National Technical Committee for crop seed standardization. GB/T3543.6-1995 Inspection of crop seeds, Determination of moisture content [S].

Version / remarks:

National Technical Committee for crop seed standardization. GB/T3543.6-1995 Inspection of crop seeds, Determination of moisture content [S]. Beijing: China Standards Press. 1996.

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

No further details specified in the study report.

Analytical monitoring:

yes

Details on sampling:

Prior to the beginning of the study, the actual concentration of the above test substance stock solutions were determined after through 0.45μm filter membrane, and that was used for application.

Vehicle:

yes

Remarks:

N-hexane

Details on preparation and application of test substrate:

2000 mg of the test substance was weighed into a beaker and dissolved with appropriate of N-hexane, then this was transferred to 2 L volumetric flask and filled up to the mark with N-hexane, and a stock solution with the concentration of 1000 mg/L was obtained. The purity of the test substance is 100%.
50 mL, 100 mL and 500 mL of the test substance stock solution were respectively taken into three individual 1 L volumetric flasks and filled up to the mark with N-hexane, then test substance stock solutions with the concentration of 50 mg/L, 100 mg/L and 500 mg/L were respectively obtained.
Prior to the beginning of the study, the actual concentration of the above test substance stock solutions were determined after through 0.45μm filter membrane, and that was used for application.

Species:

Lycopersicon esculentum

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

Name: Tomato (Lycopersicon esculentum)
Source:Zhejiang Wuwangnong Seeds Shareholding Co., Ltd.
Batch of laboratory: TLe20191119-2
Moisture content: ≤7.0%

Species:

Cucumis sativus

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

Name: Cucumber (Cucumis sativus)
Source: Beijing Flower-goddess Agriculture Co., Ltd.
Batch of laboratory: TCs20190527-1
Moisture content: ≤8.0%

Species:

Lactuca sativa

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

Name: Lettuce (Lactuca sativa)
Source: Qingxian Xingyun Vegetable Breeding Center
Batch of laboratory: TLs20190527-1
Moisture content: ≤7.0%

Species:

other: Phaseolus radiatus L

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

Name: Mung bean (Phaseolus radiatus L)
Source: Zhejiang Wuwangnong Seeds Shareholding Co., Ltd.
Batch of laboratory: TPr20191119-1
Moisture content: ≤8.0%

Species:

Brassica oleracea var. capitata

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

Name: Cabbage (Brassica oleracea)
Source: Hebei Xingtai Xing Vegetable Seed Co., Ltd.
Batch of laboratory: TBo20191119-1
Moisture content: ≤7%

Species:

other: Citrullus lanatus

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

Name: Watermelon (Citrullus lanatus)
Source: Qingxian Chunfeng Vegetable Breeding Ground
Batch of laboratory: TCl20190527-1
Moisture content: ≤8.0%

Species:

Daucus carota

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

Name: Carota (Daucus carota)
Source: Hebei Aisen Vegetable Promotion Center
Batch of laboratory: TDc20191016-1
Moisture content: ≤8.0%

Species:

other: Brassicaceae Brassica

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

Name: Mustard (Brassicaceae Brassica)
Source: Beijing Nanwo Science & Trade Co., Ltd.
Batch of laboratory: TBb20190527-1
Moisture content: ≤8.0%

Species:

Oryza sativa

Plant group:

Monocotyledonae (monocots)

Details on test organisms:

Name: Paddy (Oryza sativa)
Source: Zhongken Jinxiu Huanong Wuhan Technology Co., Ltd
Batch of laboratory: TOs20191114-1
Moisture content: 7.5

Species:

Zea mays

Plant group:

Monocotyledonae (monocots)

Details on test organisms:

Name: Corn (Zea mays)
Source: Qingxian Lilifeng Seeds Shareholding
Batch of laboratory: TZm20190527-1
Moisture content: ≤8.0%

Test type:

seed germination/root elongation toxicity test

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Test temperature:

24.2°C - 25.7°C

pH:

5.91 (Deionized water)

Moisture:

Not specified

Details on test conditions:

The selection and preparation of artificial substrate
Quartz sand was used for artificial substrate. The quartz sand was washed by tap water for three times and rinsed by deionized water for one time, and then was dry-heat sterilized at 160°C - 170°C for 2 h or 180°C - 200°C for 1 h after filtrated. The quartz sand was determined non-biotoxicity before used. The pH of the quartz sand was 6.76. Before the test, the saturation moisture content of quartz sand was measured as 65 mL per 300 g of quartz sand, and the test solution were applied on the basic with 80% of the saturation moisture content.

Test design
For each species of seeds, one blank control, one solvent control and four test groups (50 mg/L, 100 mg/L, 500 mg/L, 1000 mg/L) were performed, with one replicate in each group and 15 seeds per replicate.

Test procedure
(1) On the day before the test, each test substance solution was mixed with quartz sand evenly. The solvent control replaced the test substance solution of N-hexane, and then evaporated the N-hexane overnight.
(2) After the N-hexane was completely evaporated, a certain volume of deionized water were respectively added into the quartz sand of blank control, solvent control and each test group and mixed uniformly, and then sub-packed to Petri dishes evenly.
(3) The seeds of each species were positioned on each Petri dish, and then completely filled with quartz sand. All tested seed of per species were from the same batch and same size class, and the radical end of the seed was aligned in the direction of growth.
(4) The Petri dishes were sealed with PE cling wrap.
(5) All the Petri dishes were placed into the incubator at a slight angle to facilitate linear root growth.
(6) The test period was ended when the seeds in control had germinated at least 65 percent and the length of roots were at least 20 mm.

Test conditions
All Petri dishes were incubated in the darkness at 25°C±1°C.

Nominal and measured concentrations:

Detailed in table form - see "Any other information"

Reference substance (positive control):

not specified

Key result

Species:

Lycopersicon esculentum

Dose descriptor:

EC50

Effect conc.:

> 1 000 other: mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: germination and root elongation

Remarks on result:

other: Duration not specified

Key result

Species:

Cucumis sativus

Dose descriptor:

EC50

Effect conc.:

> 1 000 other: mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: germination and root elongation

Remarks on result:

other: Duration not specified

Key result

Species:

Lactuca sativa

Dose descriptor:

EC50

Effect conc.:

> 1 000 other: mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: germination and root elongation

Remarks on result:

other: Duration not specified

Key result

Species:

other: Phaseolus radiatus L

Dose descriptor:

EC50

Effect conc.:

> 1 000 other: mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: germination and root elongation

Remarks on result:

other: Duration not specified

Key result

Species:

Brassica oleracea var. capitata

Dose descriptor:

EC50

Effect conc.:

> 1 000 other: mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: germination and root elongation

Remarks on result:

other: Duration not specified

Key result

Species:

other: Citrullus lanatus

Dose descriptor:

EC50

Effect conc.:

> 1 000 other: mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: germination and root elongation

Remarks on result:

other: Duration not specified

Key result

Species:

Daucus carota

Dose descriptor:

EC50

Effect conc.:

> 1 000 other: mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: germination and root elongation

Remarks on result:

other: Duration not specified

Key result

Species:

other: Brassicaceae Brassica

Dose descriptor:

EC50

Effect conc.:

> 1 000 other: mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: germination and root elongation

Remarks on result:

other: Duration not specified

Key result

Species:

Oryza sativa

Dose descriptor:

EC50

Effect conc.:

> 1 000 other: mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: germination and root elongation

Remarks on result:

other: Duration not specified

Key result

Species:

Zea mays

Dose descriptor:

EC50

Effect conc.:

> 1 000 other: mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: germination and root elongation

Remarks on result:

other: Duration not specified

Details on results:

(1) At the end of the test, above 65% of test seeds (86.7% - 100%, 73.3% - 100%) in blank controls and solvent controls had germinated. The means of root length were in the range of 27.18 mm - 96.01 mm and 28.63 mm - 85.11 mm, which above 20 mm long. The above were satisfied the validity of the test. The significant probability of the root length in the solvent control (based on the blank control) was within 0.07 - 0.89, which were greater than 0.05, indicating that N-hexane had been completely volatilized and had no significant effect on seed germination and root elongation.
(2) Due to the poor water solubility of the test substance, N-hexane was used to dissolve the test substance and then volatilized completely at the time of test. Recovery test with 50 mg/L and 1003 mg/L quartz sand systems containing test substance were conducted to assess the loss of test substance during the process of N-hexane volatile. When the N-hexane volatilized completely, each of three replicates were sampled for extracting to detect the recovery and uniformity of the test substance. The results showed that, recoveries of test substance were 117%, 113%, 113% and 102%, 102%, 114%, which revealed that there were no obvious losses of the test substance during the process of N-hexane volatilization, and the distribution of the test substance in quartz sand was homogeneous.
(3) At the beginning of the test, the test substance concentrations in each test group had been measured, and the measured concentrations were 92.6% - 108% of the theoretical concentrations.
(4) Under the present conditions of the study, the test results revealed that EC50 of germination and root elongation inhibition rates of the 10 seeds were all greater than 1000 mg/L.

Results with reference substance (positive control):

Not specified

Reported statistics and error estimates:

Detailed in table for see "Any other information" for details.

 Design of Range-Finding test

Test groups		Test substance stock solution (mL)	Quartz sand (g)	Deionized water (mL)	Number of dishes	Number of seeds per dish
Blank control		--	3000	520	10	15
Solvent control		520 (N-hexane)	3000	520	10	15
Test group (mg/L)	50	652	3000	520	10	15
	100	663	3000	520	10	15
	500	813	3000	520	10	15
	1000	643.6	3000	520	10	15

 

Results of Seed Germination/Root Elongation

No.	Seeds	Groups (mg/L)	Root Length (mm)
			1	2	3	4	5	6	7	8	9	10	11	12	13	14	15
1	Tomato	Blank control	22.52	67.62	31.06	43.07	67.86	69.07	51.74	54.01	46.85	25.77	30.89	84.20	75.83	26.12	11.02
		Solvent control	30.46	22.92	81.60	39.09	31.49	71.69	52.53	61.90	89.16	54.03	33.46	95.22	47.31	62.97	0
		50	51.88	26.10	38.47	75.19	24.85	46.72	58.79	74.97	79.47	38.60	90.10	78.32	89.57	0	0
		100	54.41	29.57	38.28	66.21	70.67	29.03	61.62	31.47	39.39	52.29	85.31	78.41	57.50	0	0
		500	36.66	32.37	69.53	34.86	50.65	51.82	75.23	105.22	115.25	93.24	24.57	66.60	81.79	31.93	122.54
		1000	26.68	24.68	36.38	55.35	82.81	27.92	85.06	63.25	38.14	45.16	40.65	61.30	68.17	0	0
2	Cucumber	Blank control	66.26	60.00	48.99	76.97	65.34	69.66	76.86	63.38	56.55	65.18	53.33	65.49	69.22	62.44	71.98
		Solvent control	60.64	55.01	54.70	56.77	61.38	68.98	63.43	61.40	51.50	65.88	67.81	67.63	52.12	49.38	0
		50	53.86	50.28	62.03	59.34	50.59	54.24	52.71	54.77	67.15	61.19	56.27	55.16	52.98	65.53	65.29
		100	67.61	60.27	46.46	58.57	70.92	71.75	58.92	48.08	52.19	31.11	79.30	73.28	58.59	48.21	59.11
		500	64.01	55.89	28.13	56.50	68.26	60.93	81.14	52.45	50.80	53.69	59.63	53.04	52.13	0	0
		1000	74.19	53.39	63.49	54.15	67.71	53.63	58.93	51.80	56.97	57.19	57.68	70.72	68.99	0	0
3	Mung bean	Blank control	47.47	79.36	68.84	75.77	77.08	50.91	53.34	50.50	72.97	45.50	78.20	75.28	54.61	69.98	0
		Solvent control	65.07	62.26	48.00	58.59	57.06	67.49	65.75	30.08	40.22	41.57	62.65	54.64	58.97	63.04	0
		50	60.91	54.05	65.63	60.30	58.04	65.13	76.47	70.52	61.89	47.01	70.02	50.62	67.60	52.47	44.96
		100	66.88	51.26	70.49	38.00	65.10	55.70	37.46	42.91	46.57	57.66	79.80	78.81	69.04	69.26	50.87
		500	57.13	49.13	78.28	64.60	45.93	69.27	81.05	54.65	74.88	34.46	69.96	52.45	47.06	66.55	48.77
		1000	39.88	74.51	24.88	15.32	24.42	67.93	52.75	57.49	76.71	71.39	72.01	83.60	72.32	56.41	0
4	Watermelon	Blank control	94.10	87.52	112.94	110.73	70.37	121.05	102.01	92.54	73.89	89.40	107.62	132.48	76.49	99.99	68.98
		Solvent control	114.44	59.67	92.16	60.09	100.55	91.25	76.24	82.99	84.59	98.80	82.23	74.18	77.38	96.43	85.61
		50	43.91	65.92	109.75	100.69	83.36	44.80	66.19	61.67	113.73	96.27	81.51	61.86	83.84	91.64	0
		100	65.86	21.20	80.91	107.26	81.50	85.70	99.08	99.88	43.49	96.14	76.48	99.60	79.38	94.73	83.18
		500	69.91	84.49	68.84	112.95	88.92	115.57	109.92	108.38	49.04	91.87	84.56	99.43	90.30	64.80	0
		1000	82.42	95.01	34.17	69.52	98.00	68.43	89.06	56.88	103.18	85.52	81.33	43.58	50.08	96.30	0
5	Paddy	Blank control	84.99	59.73	22.65	11.86	3.09	80.69	76.01	82.61	86.92	43.20	63.23	61.10	78.43	18.31	84.74
		Solvent control	31.44	80.39	63.76	90.23	67.31	27.04	90.23	47.24	50.86	80.10	78.03	16.46	64.46	77.37	48.94
		50	57.10	57.78	49.56	61.88	25.94	13.23	17.46	80.47	47.06	5.09	16.08	25.96	66.99	32.27	0
		100	3.07	48.97	82.59	61.97	70.63	20.14	63.61	8.15	17.27	96.46	59.46	53.51	79.30	40.43	22.91
		500	41.16	48.46	28.53	29.12	15.70	55.58	53.58	27.60	63.63	64.96	29.61	30.07	11.88	22.03	49.93
		1000	22.06	49.88	61.08	22.69	16.38	59.92	56.31	37.14	77.82	65.22	15.22	15.09	26.92	0	0
6	Corn	Blank control	74.69	60.98	58.43	73.58	53.42	71.69	33.10	67.50	62.02	77.45	68.91	89.20	63.52	58.69	0
		Solvent control	54.29	41.55	48.29	45.24	59.33	66.45	67.51	72.67	56.29	49.65	55.08	45.49	39.84	0	0
		50	40.05	51.56	85.22	66.48	43.50	41.78	67.90	37.01	50.82	42.70	49.16	44.29	47.28	0	0
		100	55.41	43.36	64.30	67.64	77.73	53.49	49.40	53.97	14.61	45.78	40.51	63.13	38.53	57.95	42.01
		500	51.33	50.03	53.78	48.52	52.11	68.93	61.27	33.13	49.99	57.58	48.30	38.82	63.98	43.67	75.17
		1000	61.68	50.66	43.63	41.21	50.14	41.59	66.17	92.30	46.49	49.47	48.17	50.49	35.60	46.73	0
7	Lettuce	Blank control	22.65	26.19	38.60	52.37	48.23	52.73	58.20	40.58	13.85	65.19	20.63	52.44	27.59	68.62	57.12
		Solvent control	48.14	17.71	16.64	55.68	59.61	31.10	23.71	43.41	24.49	46.16	21.03	30.12	48.09	29.80	0
		50	36.77	39.42	25.13	41.74	16.29	58.74	51.97	46.39	46.81	24.49	57.25	26.69	13.77	33.40	13.89
		100	16.57	25.66	20.17	18.18	24.17	26.57	51.70	27.08	52.78	21.74	27.32	36.65	36.98	49.02	0
		500	57.58	17.06	31.85	22.53	20.04	19.27	47.62	17.42	44.82	22.35	57.11	24.47	63.32	49.53	0
		1000	55.37	16.66	14.03	55.23	20.98	55.32	29.37	68.27	20.05	39.68	30.60	20.94	38.07	11.98	16.99
8	Cabbage	Blank control	66.95	47.72	82.99	55.29	76.90	73.18	65.15	58.74	53.44	58.14	79.66	36.23	41.22	0	0
		Solvent control	38.52	75.90	63.01	29.64	57.06	40.38	34.34	44.83	67.87	50.83	48.91	0	0	0	0
		50	33.61	42.62	21.49	65.50	31.37	43.66	26.22	57.25	36.46	43.45	48.06	0	0	0	0
		100	39.93	17.31	21.24	49.11	41.58	15.27	56.67	39.60	69.99	26.50	66.68	21.73	0	0	0
		500	63.37	79.73	60.68	69.81	75.82	23.45	41.96	57.25	49.34	23.94	63.55	37.56	87.73	26.95	0
		1000	42.64	69.26	36.49	51.08	47.79	29.07	53.97	77.54	18.33	43.23	32.89	0	0	0	0
9	Mustard	Blank control	24.69	28.87	21.17	27.01	23.40	13.05	42.52	24.05	27.93	39.27	29.12	23.26	28.97	0	0
		Solvent control	21.65	22.37	54.24	26.40	31.42	22.10	18.64	26.33	29.73	26.81	33.25	30.65	0	0	0
		50	34.56	18.87	27.36	26.60	44.80	9.08	10.38	20.71	32.11	22.06	25.64	28.17	22.72	0	0
		100	25.77	13.98	23.45	37.92	16.01	21.46	21.37	25.39	17.62	30.59	24.94	0	0	0	0
		500	13.38	17.08	3.71	20.93	20.86	27.14	24.47	35.39	17.60	8.02	23.41	25.22	20.73	0	0
		1000	28.72	26.34	29.77	18.98	30.90	19.25	24.66	24.86	27.13	29.64	23.98	20.28	0	0	0
10	Carrot	Blank control	16.74	21.19	41.10	38.98	37.44	43.66	41.99	20.43	36.04	26.34	38.03	27.07	49.03	21.15	0
		Solvent control	37.08	25.82	23.48	19.37	16.65	46.81	32.67	60.32	37.76	40.85	35.10	34.01	25.91	0	0
		50	40.92	48.47	26.96	40.97	33.98	25.75	18.98	26.30	42.01	42.86	20.60	30.04	33.74	42.15	0
		100	48.26	30.80	41.62	59.00	32.57	22.05	30.81	23.48	14.09	17.16	49.92	42.31	48.85	34.34	0
		500	21.10	42.71	36.88	49.11	45.94	33.74	56.07	61.53	44.94	67.15	44.48	28.16	15.60	0	0
		1000	44.23	41.27	47.83	53.42	28.86	41.87	35.74	39.44	52.47	37.84	59.42	35.05	43.47	29.74	0

 

Statistical results

No.	Seeds	Groups (mg/L)	Germination percentage (%)	Germination Inhibition rate (%)	Mean of root length (mm)	Reduction inhibition in growth (%)	S.D.	Significance probability	EC50 (mg/L)
1	Tomato	Blank control	100	--	47.18	--	22.3	--	>1000
		Solvent control	93.3	--	55.99	--	23.0	0.57
		50	86.7	7.1	59.46	-6.2	23.3	0.94
		100	86.7	7.1	53.40	4.6	18.8	0.54
		500	100	-7.1	66.15	-18.2	32.2	0.21
		1000	86.7	7.1	50.43	9.9	20.6	0.38
2	Cucumber	Blank control	100	--	64.78	--	7.9	--	>1000
		Solvent control	93.3	--	59.76	--	6.6	0.07
		50	100	-7.1	57.43	3.9	5.6	0.72
		100	100	-7.1	58.96	1.3	12.6	0.56
		500	86.7	7.1	56.66	5.2	12.0	0.37
		1000	86.7	7.1	60.68	-1.5	7.5	0.66
3	Mung bean	Blank control	93.3	--	64.27	--	13.0	--	>1000
		Solvent control	93.3	--	55.39	--	11.3	0.25
		50	100	-7.1	61.04	-10.2	8.9	0.08
		100	100	-7.1	58.65	-5.9	13.9	0.24
		500	100	-7.1	59.61	-7.6	13.6	0.18
		1000	80.0	14.3	56.40	-1.8	22.1	0.91
4	Watermelon	Blank control	100	--	96.01	--	19.0	--	>1000
		Solvent control	100	--	85.11	--	14.8	0.09
		50	93.3	6.7	78.94	7.3	22.3	0.19
		100	100	0	80.96	4.9	23.0	0.56
		500	93.3	6.7	88.50	-4.0	20.0	0.77
		1000	93.3	6.7	75.25	11.6	21.9	0.08
5	Paddy	Blank control	93.3	--	61.03	--	26.6	--	>1000
		Solvent control	100	--	60.92	--	23.2	0.70
		50	93.3	6.7	39.78	34.7	23.3	0.01
		100	93.3	6.7	51.81	15.0	26.9	0.19
		500	100	0	38.12	37.4	17.0	0
		1000	86.7	13.3	40.44	33.6	22.1	0.01
6	Corn	Blank control	93.3	--	65.23	--	13.1	--	>1000
		Solvent control	86.7	--	53.98	--	10.3	0.08
		50	86.7	0	51.37	4.8	13.8	0.78
		100	100	-15.4	51.19	5.2	15.0	0.52
		500	100	-15.4	53.11	1.6	11.0	0.32
		1000	93.3	-7.7	51.74	4.2	14.1	0.84
7	Lettuce	Blank control	100	--	43.00	--	17.3	--	>1000
		Solvent control	93.3	--	35.41	--	14.4	0.12
		50	100	-7.1	35.52	-0.3	15.1	0.67
		100	93.3	0	31.04	12.3	12.4	0.48
		500	93.3	0	35.36	0.1	17.1	1.00
		1000	100	-7.1	32.90	7.1	18.1	0.98
8	Cabbage	Blank control	86.7	--	61.20	--	14.6	--	>1000
		Solvent control	73.3	--	50.10	--	14.5	0.09
		50	73.3	0	40.88	18.4	13.0	0.45
		100	80.0	-9.1	38.80	22.6	19.0	0.53
		500	93.3	-27.3	54.37	-8.5	21.1	0.14
		1000	73.3	0	45.66	8.9	17.2	0.73
9	Mustard	Blank control	86.7	--	27.18	--	7.5	--	>1000
		Solvent control	80.0	--	28.63	--	9.2	0.89
		50	86.7	-8.3	24.85	13.2	9.5	0.78
		100	73.3	8.3	23.50	17.9	6.8	0.25
		500	80.0	0	21.19	26.0	8.2	0.22
		1000	80.0	0	25.38	11.4	4.2	0.58
10	Carrot	Blank control	93.3	--	32.80	--	10.3	--	>1000
		Solvent control	86.7	--	33.53	--	11.8	0.77
		50	93.3	-7.7	33.84	-0.9	9.2	0.64
		100	93.3	-7.7	35.38	-5.5	13.5	0.50
		500	86.7	0	42.11	-25.6	15.0	0.28
		1000	93.3	-7.7	42.19	-25.8	8.8	0.07

Note: The seeds with root length less than 5mm were not used to calculate the mean root length and standard deviation (S.D.)

 

Results for test substance concentration at the start of the test

Groups (mg/L)	Quartz sand containing test substance (g)	N-hexane (mL)	Theoretical concentration of the test substance (mg/L)	Measured concentration of the test substance (mg/L)	Recovery rate (%)
50	30	15	17.3	17.2	99.4
100	20	20	17.3	18.0	104
500	20	50	34.7	37.4	108
1000	10	50	34.7	32.1	92.6

 

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of the test, the EC50 for both germination and root elongation of Tomato, Cucumber, Lettuce, Mung bean, Cabbage, Watermelon, Carrot, Mustard, Paddy and Corn were all greater than 1000 mg/L.

Executive summary:

Purpose of the study

To evaluate the inhibition effect if the test substance on seed germination and root elongation toxicity of the test species after the seeds were exposed to a certain chemical concentration series over the test period.

 

Guidelines

[1] Chemical Registration Center of MEP. The Guidelines for the Testing of Chemicals Effects on Biotic Systems, 299 Seed Germination/Root Elongation Toxicity Test [M]. Second Edition. Beijing: China Environmental Science Press. 2013: 459-463.

[2] OPPTS. Ecological Effects Test Guidelines, OPPTS 850.4200 Seed Germination/Root Elongation Toxicity Test [S]. US EPA, 1996.

[3] State Environmental Protection Administration of China. HJ/T 153-2004. The Guidelines for the test chemical [S]. Beijing: China Environmental Science Press. 2004.

[4] National Technical Committee for crop seed standardization. GB/T3543.6-1995 Inspection of crop seeds, Determination of moisture content [S]. Beijing: China Standards Press. 1996.

 

Principle

Test seeds were exposed to a certain chemical concentration series. The test period was ended when at least 65 percent of the control seeds have germination and developed roots that were at least 20 mm long. When both conditions were satisfied, the menta number of seed germinating and mean root length per treatment (and control) were determined, and the inhibition effect of the test substance was evaluated.

 

Test conditions

Test substance: SynNova Base Oil

Seeds: Tomato, Cucumber, Lettuce, Mung bean, Cabbage, Watermelon, Carrot, Mustard, Paddy and corn.

Test groups: For each species of seeds, four test groups (50 mg/L, 100 mg/L, 500 mg/L, 1000 mg/L) were performed with one replicate. At the meantime, one blank control and one solvent control were also performed.

Seeds number: 15 pieces per replicate

Addition of the test substance: The test substance was dissolved by N-hexane and mixed uniformity with quartz sand, then the solvent was volatized completely and the test substance was adhered to quartz sand uniformly.

Test temperature: 24.2 °C – 25.7 °C

Light: Dark

pH: 5.91 (Deionized water)

 

Results

[1] At the end of the test, above 65% of test seeds (86.7% - 100%, 73.3% - 100%) in blank controls and solvent controls had germinated. The means of root length were in the range of 28.18 mm – 95.01 mm and 28.63 mm – 85.11 mm, which above 20 mm long.

[2] Due to poor water solubility of the test substance, N-hexane was used to dissolve the test substance and then volatilized completely at the time of test. Recovery test with 50 mg/L and 1003 mg/L quartz sand systems containing test substance were conducted to assess the loss of test substance during the process of N-hexane volatile. When the N-hexane volatilized completely, each of three replicates were sampled for extracting to detect the recovery and uniformity of the test substance. The results showed that, recoveries of test substance were 117%, 113%, 113% and 102%, 102%, 114% which revealed that there were no obvious losses of the test substance during the process of N-hexane volatilization, and the distribution of the test substance in quartz sand was homogenous.

[3] At the beginning of the test, the test substance concentrations in each test group had been measured, and the measured concentrations were 92.6% - 108% of the theoretical concentrations.

[4] Under the present conditions of the study, the test results revealed that EC50 of germination and root elongation inhibition rates of the 10 seeds were greater than 1000 mg/L.

 

Conclusion

Under the conditions of the test, the EC50 for both germination and root elongation of Tomato, Cucumber, Lettuce, Mung bean, Cabbage, Watermelon, carrot, Mustard, Paddy and Corn were all greater than 1000 mg/L.

Description of key information

EC50 for both germination and root elongation of Tomato, Cucumber, Lettuce, Mung bean, Cabbage, Watermelon, carrot, Mustard, Paddy and Corn were all greater than 1000 mg/L.

Key value for chemical safety assessment

Short-term EC50 or LC50 for terrestrial plants:

1 000 mg/kg soil dw

Additional information

At the end of the test, above 65% of test seeds (86.7% - 100%, 73.3% - 100%) in blank controls and solvent controls had germinated. The means of root length were in the range of 28.18 mm – 95.01 mm and 28.63 mm – 85.11 mm, which above 20 mm long.

Due to poor water solubility of the test substance, N-hexane was used to dissolve the test substance and then volatilized completely at the time of test. Recovery test with 50 mg/L and 1003 mg/L quartz sand systems containing test substance were conducted to assess the loss of test substance during the process of N-hexane volatile. When the N-hexane volatilized completely, each of three replicates were sampled for extracting to detect the recovery and uniformity of the test substance. The results showed that, recoveries of test substance were 117%, 113%, 113% and 102%, 102%, 114% which revealed that there were no obvious losses of the test substance during the process of N-hexane volatilization, and the distribution of the test substance in quartz sand was homogenous.

At the beginning of the test, the test substance concentrations in each test group had been measured, and the measured concentrations were 92.6% - 108% of the theoretical concentrations.

 

Conclusion

Under the conditions of the test, the EC50 for both germination and root elongation of Tomato, Cucumber, Lettuce, Mung bean, Cabbage, Watermelon, carrot, Mustard, Paddy and Corn were all greater than 1000 mg/L.