1,1'-(ethane-1,2-diyl)bis[pentabromobenzene]

Toxicity to soil macroorganisms except arthropods

Administrative data

Endpoint:

toxicity to soil macroorganisms except arthropods: long-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

May 6 2003 to July 1, 2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study performed according to international guidelines and GLPs using commercial product as test article and published in the literature.

Data source

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

2003

Reference 2

Reference Type:

publication

Title:

Terrestrial toxicity evaluation of decabromodiphenyl ethane on organisms from three trophic levels

Author:

Hardy et al.

Year:

2011

Bibliographic source:

Ecotoxicol Envirohn Saf 74(4):703-10.

Reference 3

Reference Type:

publication

Title:

Results of terrestrial and aquatic studies on the brominated flame retardant, decabromodipehnyl ethane, a.k.a. Ethane 1,2 bis(pentabromophenyl)

Author:

Hardy et al.

Year:

2009

Bibliographic source:

SETAC Annual Meeting, Nov 2009, New Orleans, LA

Reference 4

Reference Type:

publication

Title:

Environmental Risk Evaluation Report: 1,1'-(Ethane-1,2-diyl)bis[pentabromobenzene] (CAS no. 84852-53-9)

Author:

Dungey SM, Akintoye L

Year:

2007

Bibliographic source:

http://publications.environment-agency.gov.uk/epages/eapublications.storefront/48771dd8022bcfe8273fc0a8029606a9/Search/Run

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.6200 (Earthworm Subchronic Toxicity Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 207 (Earthworm, Acute Toxicity Tests)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

OECD Guideline 222 (Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei))

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

1,1'-(ethane-1,2-diyl)bis[pentabromobenzene]

EC Number:

284-366-9

EC Name:

1,1'-(ethane-1,2-diyl)bis[pentabromobenzene]

Cas Number:

84852-53-9

Molecular formula:

C14H4Br10

IUPAC Name:

1,2,3,4,5-pentabromo-6-[2-(2,3,4,5,6-pentabromophenyl)ethyl]benzene

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

98.34 % Decabromodiphenyl ethane
1.66 % Nonabromodiphenyl ethane

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Soil samples were collected on days 0, 28 and 56 for analytical confirmation of DBDPEthane test concentrations. The day 28 and 56 samples were composite samples of each treatment level. Homogeneity was determined for the low and high dose replicates (top, middle, bottom of container) just prior to test initiation.

Test substrate

Vehicle:

no

Details on preparation and application of test substrate:

Soils were mixed in a rotary mixer for at least 24 hours after addition of the test article, hydrated to 60% of their water holding capacity (WHC) with deionized water, and re-mixed to a uniform consistency. Approximately 615 g of soil (i.e., 500 g dry soil) was added to each replicate.

Test organisms

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

Eisenia fetida (Vicker’s Farms, Orlando, FL) were from a synchronous population and no greater than four weeks apart in age.

Study design

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Total exposure duration:

56 d

Remarks:

28 d for survival; 56 d for reproduction

Post exposure observation period:

no

Test conditions

Test temperature:

18.8 to 20.3 degrees C

pH:

Day 0: 6.6 to 6.7; Day 56: 6.0 to 6.6

Moisture:

Day O: 21.9 to 22.7%; Day 28: 33.0 to 34.6%; Day 56: 33.5 to 37.5%

Details on test conditions:

Photoperiod: 16 hr light: 8hr dark
Light Intensity: 463 to 492 lux

Nominal and measured concentrations:

Nominal: 0, 313, 625, 1250, 2500, 5000 mg/kg dry soil
Measured:

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

3 720 mg/kg soil dw

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

28 d

Effect concentrations 2

Key result

Duration:

56 d

Dose descriptor:

NOEC

Effect conc.:

1 970 mg/kg soil dw

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

DBDPEthane was not detected in the control soils from the earthworm study (
On day 0 and at test termination, the pH of soils ranged from 6.6 to 6.7 and 6.0 to 6.6, respectively. At initiation, soil moisture ranged from 21.9 to 22.7% of the dry weight; the percent moisture at 60% of the soil’s water holding capacity was 23%. After 28 days, soil moisture ranged from 33.0 to 34.6%. At test termination, moisture ranged from 33.5 to 37.5% of the dry weight. Temperatures recorded with the electronic data logger ranged from 18.8 to 20.3°C throughout the test. The 56-d average temperature of the water bath was 19.8 ± 0.1°C. The light intensity ranged from 463 to 692 lux.
Exposure to soils containing DBDPEthane for 28 d had no adverse effect on mortality (e.g. <10% effect), burrowing or avoidance behaviors. The EC10 and EC50 for survival were estimated greater than the highest concentration tested. No abnormal burrowing or avoidance behaviors were observed during the first 60 minutes of testing. After 28 d of exposure, the percent mortality in the adult earthworms was 0, 0, 2, 0, 0, and 0 in the control, 231, 440, 761, 1,910 and 3,720 mg/kg dry soil groups, respectively (Table 1). One out of 10 worms in one of the four replicates at 440 mg/kg soil was not found, and considered dead. All live earthworms were normal in appearance and behavior. The control worms gained an average of 1.92 g/replicate or 42% in replicate mass during the initial 28 d. The mean replicate weight gain in the treatment groups ranged from 1.77 to 2.27 g or 39 to 48% in replicate mass over the initial 28 d. No concentration dependent response in weight gain was observed.

Reproductive output in the controls ranged from 31 to 76 juvenile worms/replicate with an average of 44 juveniles/replicate, which was within acceptable limits. The percent coefficient of variation (%CV) for the control data was 37% with the reproductive data from the E replicate a statistical outlier. When these data were removed, the calculated %CV was 26%. However, the inclusion or exclusion of these data did not alter the NOEC. Therefore, the data reported and the statistical parameters are inclusive of this control replicate. The average reproduction for the treatment groups were 62, 45, 62, 48 and 18 juveniles/replicate for the 231, 440, 761, 1,910 and 3,720 mg/kg groups, respectively. Reproduction at the highest dose level was statistically lower from the control group (p<0.05).

Exposure to DBDEthane did not adversely affect (i.e., >10% effect) earthworm survival, and the 28-d survival NOEC was 3,720 mg/kg dry soil (measured) or 5,000 mg/kg dry soil (nominal), the highest dose tested, with the EC10 and EC50 > 3,720 mg/kg dry soil. The 56-d reproduction NOEC and LOEC were 1,970 and 3,720 mg kg dry soil, respectively. The estimated 28-d reproduction EC10 was 1,860 mg/kg dry soil. The estimated 28-reproduction EC50 was 3,180 mg/kg dry soil, the highest dose tested.

Reported statistics and error estimates:

Earthworm data was analyzed using SAS for Windows, Relase 6.12 and ToxCalcTM version 5.0 (p<0.05). The NOEC for earthworm survival data was determined using Fisher’s exact test. A Hochberg adjustment was used to control the experiment-wise error rate for the Fisher’s test. The reproduction NOEC value was estimated using a one-way analysis of variance and a one-tailed Dunnet’s test. A Shapiro-Wilk’s and Levene’s tests were performed to determine data normality and homogeneity of the treatment variances. The reproduction data exhibited normality and homogeneity of variance, and a parametric analysis was performed on the untransformed data. DBDPEthane exposure did not adversely effect (i.e., >10% effect) survival, and the EC10 and EC50 for survival were estimated greater than the highest concentration tested. The reproduction data did not display a strong concentration response curve, and non-linear interpolation was utilized to estimate the EC10 and EC50 concentrations for reproduction. The alpha level, p<=0.05, was used for all tests.

Any other information on results incl. tables

Table 1. Measured Soil Concentrations of Saytex 8010 During an Exposure of the Earthworm (Eisenia fetida).

Nominal Concentration (mg/kg dry soil)	Measured Concentration as mg/kg dry soil (% of Nominal)			Mean Measured Concentration As mg/kg dry soil (% of Nominal)
	Day 0	Day 28	Day 56
0 313 625 1250 2500 5000	<MQL^a 289 (92) 546 (87) 1020 (82) 2210 (88) 4280 (86)	<MQL^a 227 (73) 448 (72) 516^c(41) 1760 (70) 3330 (67)	<MQL^b 178 (57) 326 (52) 746 (60) 1750 (70) 3560 (71)	<MQL 231 (74) 440 (70) 761 (61) 1910 (76) 3720 (74)
QC Fortifications Low – 299 mg/kg High – 5410 mg/kg High 5430 – mg/kg High – 5940 mg/kg	282 (94) 4740 (88) --^d --^d	235 (79) --^d 4620 (85) --^d	217 (73) --^d --^d 5420 (91)

Table 2. Measured Soil Concentrations of Saytex 8010 within the Homogeneity Samples.

Nominal Concentration (mg/kg dry soil)	Measured Concentration as mg/kg dry soil (% of Nominal)			Mean Measured Concentration As mg/kg dry soil (% of Nominal)
	Top	Middle	Bottom
313 5000	281 (90) 4220 (84)	293 (94) 4030 (81)	297 (95) 4300 (86)	290 (8.33); CV = 3% 4180 (139); CV = 3%
QC Fortifications Low – 299 mg/kg High – 5220 mg/kg	279 (93) 4210 (81)

Table 3. Mortality and Behavioral Observations During the Adult Earthworm Exposure to Saytex 8010.

Mean Measured Concentration (mg/kg dry soil)	Repetition	Day 28 Mortality^a (% Mortality)	Day 28 Observations	Treatment % Mortality
O	A B C D E F G H	0 0 0 0 0 0 0 0	10N 10N 10N 10N 10N 10N 10N 10N	0
231	A B C D	0 0 0 0	10N 10N 11N 10N	0
440	A B C D	0 0 0 1(10)	10N 10N 10N 9N, 1NF	2
761	A B C D	0 0 0 0	10N 10N 10N 10N	0
1910	A B C D	0 0 0 0	10N 10N 10N 10N	0
3720	A B C D	0 0 0 0	10N 10N 10N 10N	0

Table 4. Worm Weights Measured During the Adult Earthworm Exposure to Saytex 8010.

Mean Measured Concentration (mg/kg dry soil)	Repetition	Initial Replicate Weight (g)	Final Replicate Weight (g)a	Weight Change per Replicate (g)	Mean Weight Change per Treatment (g)	% Weight Change per Treatment
O	A B C D E F G H	4114 4571 4567 4852 5253 4410 4629 3735	5585 6650 6598 6891 7311 6642 5554 6242	1.471 2.079 2.031 2.039 2.058 2.232 0.925 2.507	1918	42
231	A B C D	4999 4080 5093 4783	7286 5702 6001 7184	2.287 1.622 0.908 2.401	1805	38
440	A B C D	5041 4164 5199 4521	7389 6340 6303 6899	2.348 2.176 1.104 2.378	2002	42
761	A B C D	4827 5127 4876 4072	7647 6898 7032 6426	2.820 1.771 2.156 2.354	2275	48
1910	A B C D	4866 4336 4445 4749	7574 6303 5183 6418	2.708 1.967 0.738 1.669	1771	39
3720	A B C D	4278 4292 4130 4027	6170 6564 7123 4666	1.892 2.272 2.993 0.639	1949	47

Table 5. Reproduction During the Earthworm Exposure to Saytex 8010.

Mean Measured Concentration (mg/kg dry soil)	Repetition	Number of Surviving Adult Worms	Number of Juvenile Worms	Average Reproduction^a
O	A B C D E F G H	10 10 10 10 10 10 10 10	46 32 31 36 76 31 59 39	44 (CV = 37%)
231	A B C D	10 10 10 10	33 56 68 92	62
440	A B C D	10 10 10 10	63 42 15^b 29	45
761	A B C D	10 10 10 10	58 90 49 49	62
1910	A B C D	10 10 10 10	55 63 39 33	48
3720	A B C D	10 10 10 10	21 27 9 13	18*

^aThe mean number of juveniles produced is calculated as the summation of the number of juveniles per container divided by the number of replicate containers. These values were rounded to two significant figure.

^bThis sample was spilled during the juvenile enumeration and was not used in the calculation of the mean.

*Statistically significant (p<0.05) reduction in treatment reproductive output as compared to the control value.

Table 6. Effect Concentrations (EC10, EC50). No-Observed-Effect (NOEC), and Lowest-Observed-Effect (LOEC) Concentrations During the Earthworm Exposure to Saytex 8010.

Biological Parameter	Statistical Endpoints
	LC10 or EC10 (95% CI^a)	LC50 or EC50 (95% CI^a)	NOEC	LOEC
28-day Survival (mg/kg dry soil) Reproduction (mg/kg dry soil)	>3720 (N/C^b)   1860 (0 – 2390)	>3720 (N/C)   3180 (N/C)	3720   1910	>3720   3720

^aCI = Confidence Interval

^bN/C = Could not be calculated

Table 7. Soil pH and Moisture Measurements Taken During the Earthworm Exposure to Saytex 8010.

Mean Measured Soil Concentration (mg/kg dry soil)	Day 0^a pH	Day 56^b pH	Day 0 % Moisture^c	Day 28 % Moisture^c	Day 56 % Moisture^c
0 231 440 761 1910 3720	6.7 6.7 6.7 6.6 6.7 6.6	6.6 6.1 6.0 6.0 6.2 6.3	22.1 21.9 22.3 22.2 22.7 22.4	33.3 33.0 33.4 34.6 33.5 33.5	37.5 34.2 34.8 34.4 33.8 33.5

^apH recorded from parent soil sample

^bpH recorded from replicate A soil sample

^cSoil moisture of the parent soil at initiation and from a composite sample on days 28 and 56. The soil moisture was measured as a percentage of the dry soil weight. At 60% of the maximum water holding capacity the percent moisture was 23%.

Table 8. Light Measurements Taken During the Earthworm Exposure to Saytex 8010.

Study Day	Experimental Position	Treatment Replicate	Light Intensity (lux)
0 0 0 0 0 0 0 0 0	1 7 12 18 19 24 22 25 27	Level 4D Level 1A Control F Control D Level 5C Level 1D Control C Level 2D Level 3B	692 563 628 463 558 527 472 506 470

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Exposure to DBDEthane did not adversely affect (i.e., >10% effect) earthworm survival, and the 28-d survival NOEC was 3,720 mg/kg dry soil (measured) or 5,000 mg/kg dry soil (nominal), the highest dose tested, with the EC10 and EC50 > 3,720 mg/kg dry soil. The 56-d reproduction NOEC and LOEC were 1,970 and 3,720 mg kg dry soil, respectively. The estimated 28-d reproduction EC10 was 1,860 mg/kg dry soil. The estimated 28-reproduction EC50 was 3,180 mg/kg dry soil, the highest dose tested.

Executive summary:

Exposure to DBDEthane did not adversely affect (i.e., >10% effect) earthworm survival, and the 28-d survival NOEC was 3,720 mg/kg dry soil (measured) or 5,000 mg/kg dry soil (nominal), the highest dose tested, with the EC10 and EC50 > 3,720 mg/kg dry soil. The 56-d reproduction NOEC and LOEC were 1,970 and 3,720 mg kg dry soil, respectively. The estimated 28-d reproduction EC10 was 1,860 mg/kg dry soil. The estimated 28-reproduction EC50 was 3,180 mg/kg dry soil, the highest dose tested.