Product of Semi-Dry Absorption method of Flue Gas Desulphurization (SDA Product)

Toxicity to soil macroorganisms except arthropods

Administrative data

Endpoint:

toxicity to soil macroorganisms except arthropods: short-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

14.03.2022 - 09.08.2022

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2022

Report date:

2022

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 207 (Earthworm, Acute Toxicity Tests)

Version / remarks:

OECD (1984)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

1

Reference substance name:

SDA Product (desulphurization of exhaust gases by semi-dry absorption method from the coal fired power plants)

EC Number:

931-259-6

Molecular formula:

Not available

IUPAC Name:

SDA Product (desulphurization of exhaust gases by semi-dry absorption method from the coal fired power plants)

Test material form:

solid: bulk

Details on test material:

Boundary composition of the test material to cover all member of JS. The analysis of the test material will be performed prior the testing and the exact composition of the test material will be included in study reports:
Calcium sulfate (CAS 7778-18-9) 1-79%
Calcium sulfite hemihydrate (CAS 29501-28-8) 1-75%
Calcium carbonate (CAS 471-34-1) 1-45%
Calcium dihydroxide (CAS 1305-62-0) 0-35%
Calcium dichloride dihydrate (CAS 10035-04-8) 0-35%
Calcium difluoride (CAS 7789-75-5) 0-5%

Test substrate

Vehicle:

yes

Remarks:

deionised water

Details on preparation and application of test substrate:

- Method of application to filter paper (if used):
For the application on the filter paper, the test item was diluted in deionised water and thoroughly agitated to ensure homogeneity.

One ml of test solution was pipetted into each vial and evaporated to dryness in a laboratory drier at 25°C. The control vials were treated with 1 ml of deionised water. After drying, 1 ml of deionised water was added to each vial to moisten the filter paper. Each earthworm was washed, dried, weighed, and put into the vial.



- Method of mixing into soil (if used):

DRF: For the soil application, the test item was thoroughly mixed with the sand. 10 g of the mixture was added in each container.

Artificial soil test: 10 g of treated sand was mixed with the artificial soil in each container. The control containers were treated with 10 g of mere sand. Earthworms were washed, dried, weighed, and placed on the test soil surface.

- Controls: deionised water as negative (vehicle) control; Untreated sand was a negative control in the Artificial soil test.; positive control chloroacetamide as toxic reference item

Test organisms

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

TEST ORGANISM
- Common name: Eisenia fetida
- Source: obtained from a specified commercial supplier (Jiří Tomeček; proRYBY.cz); All control and treatment worms used in the test will be from the same source.
- Age at test initiation (mean and range, SD): adult individuals at least two months old of relatively homogeneous age structure
- Weight at test initiation (mean and range, SD): 300 to 600 mg

ACCLIMATION
- Acclimation period: acclimatised for at least one day
- Acclimation conditions (same as test or not): with the type of artificial soil substrate to be used for the test
- Health during acclimation (any mortality observed): Worms used in the test will be in apparent good health and they will be kept in conditions conforming to proper cultural practices. During this period the worms will be fed by air-dried, finely ground and pasteurised self-collected cow or horse manure.

Study design

Study type:

laboratory study

Substrate type:

artificial soil

Remarks:

10% finely ground sphagnum peat, 20% kaolin clay, approx. 0.3-1% pulverized calcium carbonate (CaCO3) to obtain an initial pH of 6.0 ± 0.5 and 70% fine quartz sand

Limit test:

no

Total exposure duration:

14 d

Test conditions

Test temperature:

Earthworms were kept under suitable conditions (mean temperature 20.2°C, min 19.6 – max 20.6°C; mean humidity 94%, min 71 – max 100%).

pH:

pH of 6.0 ± 0.5

Moisture:

Earthworms were kept under suitable conditions (mean temperature 20.2°C, min 19.6 – max 20.6°C; mean humidity 94%, min 71 – max 100%).

Details on test conditions:

Test conditions
Filter paper test
During the test, vials laid on their sides on trays in dark. Temperature: 20 ± 2°C.

Glass vials will be used. Their sides will be lined with filter paper cut to a suitable size so it does not overlap appreciably. Each vial will be sealed with a cap with a small ventilation hole. Five replicates, each consisting of one worm per vial, will be used for each treatment in the Range-finding test. For the Definitive test, ten replicates per treatment will be conducted.


Artificial soil test
Test containers will be stored at 20 ± 2°C and continuous light conditions (mean illuminance of 481 lux (Range Finding test) and 623 lux (Definitive test)).
Untreated soil pH at the beginning of the trial was 6.1 (Range Finding test) and 6.5 (Definitive test). Initial soil moisture was 50% of maximal water holding capacity.

Test containers made of glass of one-litre capacity were filled with 750 g of artificial soil. The containers were covered by a perforated transparent cover permitting gaseous exchange between the substrate and the atmosphere and access to light whilst preventing the worms from escaping. Two replicates, each consisting of ten worms per test container, will be used for each treatment in the Range-finding test. For the Definitive test, four replicates per treatment will be conducted.


The test temperature and humidity were monitored using a bluematic datalogger (Appendix II).
Range Finding test
- temperature average 20.3 °C (range 19.9 – 21.6 °C)
- relative humidity average 72 % (range 63 – 83 %)

Definitive test
- temperature average 20.4 °C (range 19.9 – 23.5 °C)*
- relative humidity average 85 % (range 66 – 95 %)


TEST SYSTEM
Definitive Filter test
- Test container (material, size): glass vial
- Amount of soil or substrate: test item was diluted in deionised water and thoroughly agitated to ensure homogeneity. 1 mL of the solution was applied in each vial and evaporated to dryness in a laboratory drier at 25°C.
- No. of organisms per container (treatment): 1
- No. of replicates per treatment group: 10
- No. of replicates per control: 10
- No. of replicates per vehicle control: 10

Definitive Artificial soil test
- Test container (material, size): glass of one-litre capacity
- Amount of soil or substrate: 750 g of artificial soil
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4
- No. of replicates per control: 4
- No. of replicates per vehicle control: 4

- Composition (if artificial substrate): 10% finely ground sphagnum peat, 20% kaolin clay, and 70% fine quartz sand and 0.1% of pulverized calcium carbonate (CaCO3) to obtain optimal initial pH
- Maximum water holding capacity (in % dry weigth): Artificial soil was premoistened to 50% of its maximal water holding capacity.

OTHER TEST CONDITIONS
- Photoperiod and Light intensity: continuous dark (filter test) or continuous light conditions with mean illuminance of 481 lux (Range Finding test) and 623 lux (Definitive test)

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality at 24 and 48 h and mean weight (filter test), mortality at 7 and 14 d after treatment application and mean weight (artificial test)

VEHICLE CONTROL PERFORMED: yes, deionised water

TEST CONCENTRATIONS
- Range finding study
The range-Finding test was conducted at five rates with the highest rate of 5 t of product/ha for the Filter paper test and 10 g of product/kg dry weight for the Artificial soil test and dilution step of 10.
A Range-finding test will be carried out with 5 rates in geometric series with dilution step of 10, one application rate of the toxic reference and untreated (water only) control. The highest tested rate will be 5 t of product/ha for filter paper test and 10 g of product/kg dry weight for soil test.

- Test concentrations:
The Definitive test was conducted at five rates with the highest rate of 0.5 t of product/ha for the Filter paper test and 10 g of product/kg dry weight for the Artificial soil test and dilution step of 1.8.

- Results used to determine the conditions for the definitive study:
Filter paper test
Average mortality of earthworms after exposure to the rate of 0.5 t of test item/ha was 100%, this rate was used as the highest tested rate in the Definitive test.
Artificial soil test
Average mortality of earthworms after exposure to the highest rate of 10 g of test item/ kg dry weight was 0% at 7 days. Mean mortality at 14 days was 5%, this rate was used as the highest tested rate in the Definitive test. No unusual behaviour or morphology was observed.

Nominal and measured concentrations:

Filter paper test: 0.048, 0,086, 0.154, 0.278, 0.5 t of product/ha and
Artificial soil test: 0.95, 1.71, 3.09, 5.56, 10 g of product/kg dry soil weight

Reference substance (positive control):

yes

Remarks:

Chloroacetamide Application rate: 2.7 μg a.i. /cm2 for the Filter paper test 38.5 mg a.i./kg dry weight of soil for he Artificial soil test (Edwards and Bater 1992)

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

48 h

Dose descriptor:

LR50

Effect conc.:

>= 0.09 other: t/ha

95% CI:

> 0.063 - < 1.122

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

mortality

Remarks on result:

other: Filter paper test

Effect concentrations 2

Duration:

48 h

Dose descriptor:

other: NOER

Effect conc.:

0.048 other: t/ha

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

mortality

Remarks on result:

other: Filter paper test

Effect concentrations 3

Duration:

48 h

Dose descriptor:

other: LOER

Effect conc.:

0.086 other: t/ha

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

mortality

Remarks on result:

other: Filter paper test

Effect concentrations 4

Key result

Duration:

14 d

Dose descriptor:

other: NOER

Effect conc.:

> 10 other: g/kg dry soil weight

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

mortality

Remarks on result:

other: Artificial soil test: no significant effect on mortality within the tested rates

Results with reference substance (positive control):

- Results with reference substance valid? Yes
Application rate:
2.7 μg a.i. /cm2 for filter paper test
38.5 mg a.i./kg dry weight for soil test (Edwards and Bater 1992)

Reported statistics and error estimates:

Statistical analyses were performed in ToxRat 3.3. (ToxRat Solutions GmbH 2018) in accordance with OECD Environment Health and Safety Publications Series on Testing and Assessment (2006).
The lethal rates (LRx) for the Filter paper test were calculated using Probit analysis using maximum likelihood regression. NOER was determined by the Step-down Cochran-Armitage test procedure. This method was selected based on the results of the Qualitative trend analysis by contrasts (test of monotonicity of rate/response) and Tarone test procedure (test for extra-binomial variance).
The lethal rates (LRx) for the Artificial soil test were calculated using Probit analysis using maximum likelihood regression. NOER was determined by a 2x2 Table test with Bonferroni correction. This method was selected based on the results of the Qualitative trend analysis by contrasts (test of monotonicity of rate/response).

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

The mortality in the controls should not exceed 10 per cent at the end of either test.

Conclusions:

Application of SDA PRODUKT ODSÍŘENÍ T700 in Filter paper test significantly affected the survival of Eisenia fetida. The value for LR50 was established to be 0.09 t of product/ha. The values for NOER and LOER were established to be 0.048 t/ha and 0.086 t/ha, respectively.
Application of the product in artificial soil had no significant effect on mortality within the tested rates. NOER appears to be higher than 10 g of product/ kg dry soil weight.

Executive summary:

A GLP laboratory study was conducted to evaluate the lethal effects of the test item SDA PRODUKT ODSÍŘENÍ T700 on the earthworm Eisenia fetida. The study followed OECD guideline, Test no. 207. The study was conducted at i2L Research Europe s.r.o. in České Budějovice, Czech Republic.

Mortality of Eisenia fetida was determined after their exposure to fresh residues of the test item on filter paper and a mixture of test item with artificial soil. The test was performed with five application rates, an untreated control (water only) and a toxic reference (2-Chloroacetamide). Range-Finding test was conducted at five rates with the highest rate of 5 t of product/ha for the Filter paper test and 10 g of product/kg dry soil weight for the Artificial soil test and dilution step of 10.

Definitive test was conducted at five rates with the highest rate of 0.5 t of product/ha for the Filter paper test and 10 g of product/kg dry soil weight for the Artificial soil test and dilution step of 1.8. The test has met all validity criteria.

Application of SDA PRODUKT ODSÍŘENÍ T700 in the Filter paper test significantly affected the survival of Eisenia fetida. The value for LR50 was established to be 0.09 t of product/ha. The values for NOER and LOER were established to be 0.048 t/ha and 0.086 t/ha, respectively.

Application of the product in artificial soil had no significant effect on mortality within the tested rates. NOER appears to be higher than or equal to 10 g of product/ kg dry soil weight.