Creosote oil, acenaphthene fraction

Toxicity to terrestrial arthropods

Administrative data

Endpoint:

toxicity to terrestrial arthropods: long-term

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

ISO 11267 (Inhibition of Reproduction of Collembola by Soil Pollutants)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Application method:

soil

Test material

Test material information

Constituent 1

Reference substance name:

8001-58-9

Cas Number:

8001-58-9

IUPAC Name:

8001-58-9

Constituent 2

Reference substance name:

Creosote

EC Number:

232-287-5

EC Name:

Creosote

IUPAC Name:

232-287-5

Constituent 3

Reference substance name:

creosote

IUPAC Name:

creosote

Details on test material:

- Name of test material (as cited in study report): Creosote Grade B
- Physical state: brown liquid
- Analytical purity: 100% creosote (Creosote Grade B according to EU standard EN 13991, August 2003)
- Impurities (identity and concentrations): not applicable
- Composition of test material, percentage of components: complex mixture of numerous aromatic hydrocarbons, phenolics,
nitrogen-containing heterocycles and others
- Expiration date of the lot/batch: 2047-03-09
- Storage condition of test material: at room temperature, protected from moisture and light

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
--

Sampling and analysis

Analytical monitoring:

no

Details on sampling:

--

Test substrate

Vehicle:

yes

Details on preparation and application of test substrate:

- Method of mixing into soil: a stock solution of test substance in acetone (5,575 g in 25 mL) was prepared. 1 mL of this stock solution was applied onto the surface of 10 g quartz. After evaporation of the solvent, the quartz sand was mixed with 557.5 g soil (dw) to get a concentration of 400 mg/kg dw. For the other test concentrations, 0.625, 1.25, 2.50, and 5.00 mL stock solution were diluted to 10 mL with aceton. 1 mL ot each of this dilutions was applied onto 10 g quartz sand and after evaporation mixed with 557.5 g soil (dw) to result the respective test concentrations.
- Controls: artificial soil containing 10 g quartz sand treated with 1 mL acetone moistened with deionized water without test substance.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone
- Concentration of vehicle in test medium (stock solution and final test solution): 1 mL of test item solution in acetone was added to 10 g quartz sand. After evaporation of the vehicle, the quartz sand coverd with test substance was mixed with artificial soil (557.5 g)
- Evaporation of vehicle before use: yes
- Volume of test solution applied: 1 mL

Test organisms

Test organisms (species):

Folsomia candida

Animal group:

Collembola (soil-dwelling springtail)

Details on test organisms:

TEST ORGANISM
- Common name: springtail
- Source: own breeding in testing facility
- Age at test initiation (mean and range, SD): 10 - 11 days
- Kept according to standard practices: yes

Study design

Study type:

laboratory study

Limit test:

no

Total exposure duration:

28 d

Post exposure observation period:

no

Test conditions

Test temperature:

19 - 22°C

pH (if soil or dung study):

Day 0: 6.04 - 5.97; day 28: 5.93 - 5.89

Humidity:

Soil moisture: day 0: 33.5 - 33.9%; day 15: 32.7 - 33.3%; day 28: 32.2 - 33.0%

Photoperiod and lighting:

16/8 h light/dark; 427 - 667 lux

Details on test conditions:

TEST SYSTEM
- Test container (material, size): glass beakers, volume 100 mL, covered with PARAFILM
- Amount of soil or substrate: 30 g test medium (artifical soil including quartz sand) wet weight
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 5
- No. of replicates per control: 8

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Soil classification system: artificial soil
- Composition (if artificial substrate): 10% peat, air dried and finely ground (2 mm)
20% kaoline clay, kaolinite content > 30%
approx. 69% air dried quartz sand (sand with > 50% particles size of 0.05 to 0.20 mm)
approx. 0.5% CaCO3
pH was determined to be 5.69
- Organic carbon (%): no data (10 % peat)
- Maximum water holding capacity (in % dry weigth): 68.6%
- Pretreatment of soil: Three days before starting the test, the artificial soil was pre-moistened by adding enough deionised water
to get 25% of the maximal water holding capacity of the artificial soil.
- Storage (condition, duration): in firmly closed containers
- Stability and homogeneity of test material in the medium: no data

OTHER TEST CONDITIONS
- Photoperiod: 16/8 h, light/dark
- Light intensity: 427 - 667 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): number of surviving adults, number of instars (at day 28)

TEST CONCENTRATIONS
- Spacing factor for test concentrations:2
- Range finding study: yes
- Test concentrations: 0.1, 1.0, 10, 100, 1000 mg/kg dw
- Results used to determine the conditions for the definitive study: mortality increased from 13 to 100 % for the 100 and 1000 mg/kg dw group respectively.

Nominal and measured concentrations:

Nominal concentrations: 25, 50, 100, 200, and 400 mg/kg dw

Reference substance (positive control):

yes

Remarks:

E 605 Forte, Bayer AG (active ingredient: Parathion 520 g/L) (0.05, 0.1, and 0.2 mg/kg dw)

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

28 d

Dose descriptor:

LC10

Effect conc.:

ca. 3.4 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: original LC10 normalised to standard soil with 3.4 % organic matter

Effect concentrations 2

Duration:

28 d

Dose descriptor:

LC10

Effect conc.:

ca. 10 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: value estimated from test data obtained in soil with 10 % organic matter

Effect concentrations 3

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

< 25 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Effect concentrations 4

Duration:

28 d

Dose descriptor:

LOEC

Effect conc.:

25 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Effect concentrations 5

Duration:

28 d

Dose descriptor:

LC50

Effect conc.:

15.9 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: original LC50 normalised to standard soil with 3.4 % of organic matter

Effect concentrations 6

Duration:

28 d

Dose descriptor:

LC50

Effect conc.:

46.8 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: 95% CL 21.2 - 103 mg/kg soil dw

Effect concentrations 7

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

50 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Effect concentrations 8

Duration:

28 d

Dose descriptor:

LOEC

Effect conc.:

100 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Effect concentrations 9

Duration:

28 d

Dose descriptor:

EC50

Effect conc.:

111 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Remarks on result:

other: 95% CL: 74.6 - 165 mg/kg soil dw

Details on results:

- Mortality at end of exposure period: is the more sensitive parameter (see table below)
- No. of offspring produced: see table below
- Morphological abnormalities: Compared to the control, the juveniles at all test item concentrations were on the average visibly smaller
(not quantitatively evaluated).

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Relevant effect levels: mortality: LOEC 0.05 mg/kg soil dw
reproduction: LOEC 0.40 mg/kg soil dw; NOEC 0.20 mg/kg soil dw

Any other information on results incl. tables

Adult Mortality [%] after 28 Days of Exposure and Statistics

 

Test item [mg/kg dw]	Replicate								Mortality
	1	2	3	4	5	6	7	8	Mean±SD	Corrected
25	50	50	50	60	60	-	-	-	54.0 ± 5.48	43.4*
50	40	20	70	50	60	-	-	-	48.0 ± 19.2	36.0*
100	50	70	10	70	80	-	-	-	56.0 ± 27.9	45.8*
200	90	90	100	80	90	-	-	-	90.0 ± 7.07	87.7*
400	100	100	100	100	100	-	-	-	100 ± 0.0	100**
Control	20	0	10	20	20	30	20	30	18.8 ± 9.91	-

 

SD = Standard deviation

*) = Statistically significant different compared to the control (t-test, p = 0.05)

**) = Statistically significant different compared to the control (Mann-Whitney Rank Sum Test, p = 0.05)

- = No observation

 

Reproduction of the Springtails after 28 Days of Exposure

 

Test item [mg/kg dw]	Number of juveniles/Replicate								Reproduction rate
									Mean+SD	CV [%]	Inhibition relative to control [%]
	1	2	3	4	5	6	7	8
25	562	267	236	494	407	-	-	-	393 ± 141	35.9	18.8
50	10	595	255	700	314	-	-	-	375 ± 276	73.7	22.6
100	194	333	302	286	266	-	-	-	276 ± 52	18.9	43.0*
200	0	216	60	94	0	-	-	-	74.0 ± 89.0	120	84.7*
400	0	0	0	0	0	-	-	-	0.0 ± 0.0	-	100*
Control	397	592	480	420	669	382	628	306	484 ± 131	27.1	-

 

SD = Standard deviation

CV = Coefficient of variation

*) = Statistically significant different compared to the control (t-test, p = 0.05)

 

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

criteria according to TG ISO 11267

Conclusions:

In a valid 28 d mortality and reproduction test according to test guideline ISO 11267 (Inhibition of Reproduction of Collembola by Soil Pollutants), using Folsomia candida (soil-dwelling springtail) as test organism, a NOEC < 25 mg/kg soil dw for mortality (LOEC = 25 mg/kg soil dw) and a NOEC of 50 mg/kg soil dw for reproduction (LOEC = 100 mg/kg soil dw) was determined. LC50 and EC50 were 46.8 and 111 mg/kg soil dw respectively. Reproduction seems to be the less sensitive parameter to creosote exposure compared to mortality.
In a range finding pre-test with a shorter exposure period of 14 days, mortality was only 13% at a test concentration of 100 mg/kg soil dw rising to 100% at a test concentration of 1000 mg/kg soil dw. This indicates that survival was markedly dependent on exposure time.
Test substance concentrations were not monitored during the test. Due to the long exposure time (28 d), test substance concentrations in soil may have declined during the experiment. Thus, nominal concentrations may result in higher effect levels than actually present and underestimate the toxic potential of creosote.