Octadecene, reaction products with hexadecene, hydrogenated

Toxicity to soil macroorganisms except arthropods

Link to relevant study record(s)

Reference

Endpoint:

toxicity to soil macroorganisms except arthropods: short-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

November 7, 2019 to November 22, 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 207 (Earthworm, Acute Toxicity Tests)

Version / remarks:

OECD Guidelines for Testing of Chemicals, No. 207 Earthworm, Acute Toxicity Tests. Paris: OECD & OCED, Adopted 4th April, 1984.

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: National Guidelines

Version / remarks:

Chemical Registration Center of MEP. The Guidelines for the Testing of Chemicals, Effects on Biotic Systems, 207 Earthworm, Acute Toxicity Test [M]. Second edition. Beijing: China Environmental Science Press. 2013: 58-62.

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: National Guideline

Version / remarks:

China National Technical Committee for Standardization of Dangerous Chemical Management. GB/T 21809-2008 Chemical Earthworm, Acute Toxicity Tests [S]. Beijing: China Standardization Press, 2008.

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

No further details specified in the study report.

Analytical monitoring:

not specified

Details on sampling:

Not specified

Vehicle:

yes

Remarks:

quartz sand

Details on preparation and application of test substrate:

Based on the predicted moisture content of artificial soil, approximately 30%, respectively 577 mg of test substance (purity was 100%) was weighed and mixed well with quartz sand to a total weight of about 10 g by grinding. Each mixture was then mixed with about 740 g wet artificial soil to obtain test substance concentrations of 1000 mg/kg (dry weight of artificial soil). Four replicates were prepared. Blank control was prepared by mixing about 10 g of quartz sand and about 740 g wet artificial soil, with four replicates.
The above test media (750 g wet weight) was transferred into 1 L glass beakers. Ten healthy earthworms (which had been washed and dried with filter paper before exposure) were placed randomly on the surface of artificial soil in each container. These earthworms were placed on the surface and allowed to burrow into the test medium by themselves. The containers were covered with perforated plastic film to prevent the artificial soil from drying.

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

Species: Eisenia foetida with obvious clitellum
Batch No.: EEf20190312-1
Age: Older than 2 months
Weight: 395 mg - 486 mg mg per earthworm
Amount: 80 earthworms
Source: Purchased from Fogang Wotu Agricultural Technology Co. Ltd. and raised in the test facility.
Selected reasons: Eisenia foetida (Michaelsen) exist in soil with rich organic matter. Its susceptibility to chemicals resembles that of true soil-inhabiting species. They have a short life cycle and are very prolific. They are available commercially and can be bred readily in a wide range of organic waste materials. Cocoons can be purchased commercially or distributed from a central source to ensure the same strain is used. “The Guidelines for the Testing of Chemicals” recommend the use of this species.
Domesticating: The earthworms which were used in the test were purchased on March 12, 2019, and all earthworms have been raised in the laboratory for 240 d before used. Before test, earthworms were acclimated to the study conditions for about 24 h in the artificial soil.

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

yes

Total exposure duration:

14 d

Post exposure observation period:

No post exposure observation period specified in the study report.

Test temperature:

environment temperature was 19.1 °C – 20.1 °C

pH:

At the start of test, pH value of the artificial soil was determined as 6.20 and not be adjusted. At the end of test, pH values of the artificial soil were determined as 6.30 - 6.37

Moisture:

Moisture content of artificial soil was 30.3% dry weight at the beginning of the test, and was 28.5% ~ 28.9% dry weight at the end of the test

Details on test conditions:

Preparation of artificial soil
The dry constituents were blended in the correct proportions, i.e., 10% sphagnum peat (no visible plant remains), 20% kaolin clay (analytical reagent), and 70% quartz sand (particle size between 50 - 200 µm). Deionized water of about 30% of dry weight was added and the medium was thoroughly mixed in a small electric cement mixer. The complete mixture was moist but not so wet that water appears when the artificial soil was compressed. After the artificial soil had been prepared, the moisture content was then determined by drying a small sample at 105°C ± 5°C and measuring the dry weighing. The pH value of the artificial soil was determined as 6.20.

Test method
Grouping and concentration design
Results of the range-finding test revealed that, after 7 days of exposure, the earthworm mortalities in the test groups of 101 mg/kg, 300 mg/kg, 501 mg/kg and 1000 mg/kg (dry weight of artificial soil) were 0%, 0%, 0% and 0%. Based on the outcome of this dose range finding test, a limit test design was adopted.
The limit test was designed with one blank control and one test group of 1000 mg/kg dry weight of artificial soil. Four replicates in each treatment were adopted.

Test condition
The test systems were put into an incubator to cultivate. During the test, the light intensity was 400 lx - 800 lx with continuous light, the environment temperature was 20°C±2°C, and the humidity was 65% - 100%. The test duration was 14 days.

Observation and measurement
(1) Normal observation: at 7 d and 14 d of exposure, the test matrix inside the container was gently placed onto a flat plate and the earthworms were sorted from the artificial soil. Any abnormal behavior or pathological symptoms were checked and recorded by provoking rump of the earthworms with a probe.
(2) Index determination
(a) pH value: At the beginning and end of the test, 5 g of air-dried artificial soil was sampled for pH determination. At the beginning of the test, the pH value of the prepared artificial soil was measured directly. At the end of the test, for each test group, a certain amount of artificial soil was taken from each replicate and combined, from which a sample of 5 g air-dried soil was taken and used for pH measurement.
(b) Moisture content of artificial soil: At the beginning and the end of the test, appropriate amounts of artificial soils were dried to constant weight at 105±5 °C to measure moisture of the artificial soil. At the beginning of the test, the moisture of the prepared artificial soil was measured directly. At the end of the test, for each test group, a certain amount of artificial soil was taken from each replicate and combined, from which a sample was taken and dried to determine moisture content.
(c) Environmental conditions: at the start and the end of the test, the light intensity of the test environment above the test container location was determined and recorded. A thermo- hygrometer was set to continuous monitor the culture temperature and humidity automatically, and the environment temperature and humidity were recorded manually every workday.
(d) Weight of earthworms: at the beginning and the end of the test, determined and recorded the amounts and total weights of survival earthworms in each container and calculate the mean weight.

Nominal and measured concentrations:

Nominal concentration: 0 mg/kg dry weight & 1000 mg/kg dry weight

Reference substance (positive control):

yes

Remarks:

chloroacetamide

Key result

Duration:

14 d

Dose descriptor:

LC50

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

Acute toxicity effect
After 7 days and 14 days exposure, the mortalities of earthworms in the blank control were 0% and 0%, and were 0% and 0% in the 1000 mg/kg (dry weight of artificial soil) test group. Under the test conditions, the result for the 14 d median lethal concentration (LC50) was greater than 1000 mg/kg (dry weight of artificial soil).
Test conditions
During the test, the environment temperature was 19.1 °C – 20.1 °C, the humidity was 80.0% - 100.0%. Continuous light, light intensity was 722 lx at the start of the test and 658 lx at the end of the test. Moisture content of artificial soil was 30.3% dry weight at the beginning of the test, and was 28.5% ~ 28.9% dry weight at the end of the test. At the start of test, pH value of the artificial soil was determined as 6.20 and not be adjusted. At the end of test, pH values of the artificial soil were determined as 6.30 - 6.37.

Results with reference substance (positive control):

The sensitivity test was carried out with the reference substance chloroacetamide (C2H4ClNO) based on the artificial soil test of the same batch Eisenia foetida.
The 14 d-LC50 of chloroacetamide was 28.6 mg/kg (dry weight of artificial soil).

Reported statistics and error estimates:

Due to the test substance showed no toxicity to the experimental earthworms, Microsoft Excel was used to analyze the mean weight of live earthworms, the mortality of earthworms and the moisture content of artificial soil.

Deaths and Mortality

Groups	Numbers of earthworm	7 d		14 d
		Deaths of earthworm	Mortality (%)	Deaths of earthworm	Mortality (%)
Blank control	40	0	0	0	0
Test substance group (1000 mg/kg dry weight)	40	0	0	0	0

 

Pathological symptoms and abnormal behavior of earthworm

Groups	7 d	14 d
Blank control	All earthworms were healthy	All earthworms were healthy
Test substance group (1000 mg/kg dry weight)	All earthworms were healthy	All earthworms were healthy

 

Survival number and the average weight of the earthworm

Groups	Replicate	The beginning			The end
		Numbers of earthworm	Average weight (mg)	Mean (mg)	Survival number	Average weight (mg)	Mean (mg)
Blank control	1	10	414	413	10	336	356
	2	10	440		10	375
	3	10	402		10	343
	4	10	395		10	369
Test substance group (1000 mg/kg dry weight)	1	10	422	452	10	380	389
	2	10	486		10	400
	3	10	467		10	399
	4	10	434		10	376

 

The pH of the artificial soil

Time	Groups	Dry soil (g)	pH
The beginning of test	Artificial soil without adding test substance	5.017	6.20
The end of test	Blank control	5.012	6.30
	Test substance group (1000 mg/kg dry weight)	5.017	6.37

 

Moisture content of the artificial soil

Time	Groups	Measure (%)	Mean (%)
The beginning of test	Artificial soil without adding test substance	29.95	30.3
		30.98
		29.84
The end of test	Blank control	29.00	28.9
		29.00
		28.72
	Test substance group (1000 mg/kg dry weight)	28.50	28.5
		28.38
		28.47

 

Validity criteria fulfilled:

yes

Conclusions:

Under the test conditions, the result for the 14 d median lethal concentration (LC50) was greater than 1000 mg/kg (dry weight of artificial soil).

Executive summary:

Purpose of the study

Evaluate the acute toxicity of the test substance to earthworm by observing the mortality and pathological symptoms.

 

Guidelines

[1] Chemical Registration Center of MEP. The Guidelines for the Testing of Chemicals, Effects on Biotic Systems, 207 Earthworm, Acute Toxicity Test [M]. Second edition. Beijing: China Environmental Science Press. 2013: 58-62.

[2] China National Technical Committee for Standardization of Dangerous Chemical Management. GB/T 21809-2008 Chemical Earthworm, Acute Toxicity Tests [S]. Beijing: China Standardization Press, 2008.

[3] OECD. OECD Guidelines for Testing of Chemicals, No. 207 Earthworm, Acute Toxicity Tests. Paris: OECD & OCED, Adopted 4th April, 1984.

 

Principle

The artificial soil test involves keeping earthworms in samples of a precisely defined artificial soil which 1000 mg/kg (dry weight of artificial soil) of the test substance had been applied. Mortality was assessed 7 and 14 days after application. Then the acute toxicity of the test substance was determined by the results of the mortality.

 

Test conditions

Test substance: SynNova Base Oil

Type of test: Acute toxicity test in artificial soil test, limit test

Test organism: Eisenia foetida with obvious clitellum

Test duration: 14 d

Test concentration: One test group of 1000 mg/kg (dry weight of artificial soil) was designed

Control: A blank control without test substance was designed

Replicates: 4 replicates per group

Test organism number: 10 earthworms per replicate

Preparation of test system: Based on the predicted moisture content of artificial soil, approximately 30%, respectively 577 mg of test substance (purity was 100%) was weighed and mixed well with quartz sand to a total weight of about 10 g by grinding. Each mixture was then mixed with about 740 g wet artificial soil to obtain test substance concentrations of 1000 mg/kg (dry weight of artificial soil). Four replicates were prepared. Blank control was prepared by mixing about 10 g of quartz sand and about 740 g wet artificial soil, with four replicates. 

Temperature in incubator: 19.1 °C – 20.1 °C

Ambient humidity: 80.0% - 100.0%

Soil moisture content: 30.3% (dry weight, at the beginning of the test); 28.5% - 28.9% (dry weight, at the end of the test)

Soil pH: 6.20(at the beginning of the test); 6.30 - 6.37 (at the end of the test)

Light condition: Continuous light, the light intensity was 722 lx at the beginning of the test and 658 lx at the end of the test.

 

Results

Based on the results of range-finding test, a limit test was conducted to evaluate the acute toxicity of the test substance to earthworm (Eisenia foetida). After 7 days and 14 days exposure, the mortalities of earthworms in the blank control were 0% and 0%, and were 0% and 0% in the 1000 mg/kg (dry weight of artificial soil) test group.

Under the test conditions, the result for the 14 d median lethal concentration (LC50) was greater than 1000 mg/kg (dry weight of artificial soil).

Description of key information

14 d median lethal concentration (LC50) was greater than 1000 mg/kg (dry weight of artificial soil).

Key value for chemical safety assessment

Short-term EC50 or LC50 for soil macroorganisms:

1 000 mg/kg soil dw

Additional information

Based on the results of range-finding test, a limit test was conducted to evaluate the acute toxicity of the test substance to earthworm (Eisenia foetida). After 7 days and 14 days exposure, the mortalities of earthworms in the blank control were 0% and 0%, and were 0% and 0% in the 1000 mg/kg (dry weight of artificial soil) test group.

Under the test conditions, the result for the 14 d median lethal concentration (LC50) was greater than 1000 mg/kg (dry weight of artificial soil).