Zeolite, cuboidal, crystalline, synthetic, non-fibrous

Toxicity to soil macroorganisms except arthropods

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

toxicity to soil macroorganisms except arthropods: long-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Mar. 8-12, 2021

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

other: ISO 10872

Version / remarks:

2020

Deviations:

no

Remarks:

All validity criteria were met in the controls of all tests.

GLP compliance:

no

Remarks:

As these data are not used to assess extrapolation to humans, a study according to GLP was not regarded as necessary.

Specific details on test material used for the study:

• Chemical name: Aluminium Magnesium Vanadium Oxide
• Trade name: Na Y (Y Zeolite)
• Acronym in Report: NaY
• Batch number: RZ200094/1
• Composition: Aluminosilicate (Al/Si ~ 0.3)
• CAS No.: 1318-02-1
• Purity: 100 %
• Specific Surface Area (BET): not stated
• Grain size (diameter): 5.723

Vehicle:

yes

Remarks:

M9 medium

Details on preparation and application of test substrate:

0.073 ml M9-medium (ISO 10872; 6 g Na2HPO4/L, 3 g KH2PO4/L, 5 g NaCl, 0.25 g MgSO4 7H2O /L) were added to 0.5 g dry soil. Together with 0.1 ml of bacterial suspension (0.073 + 0.1 = 0.173 ml) soil moisture was adjusted to 80% of the WHCmax (corresponding to 74.3 % dry weight).

Test organisms (species):

Caenorhabditis elegans

Animal group:

nematods

Details on test organisms:

Strain: N2
Genotype: C. elegans wild type
Source: Caenorhabditis Genetic Centre; University of Minnesota; Minneapolis; MN 55455; USA.
Cultivation: C. elegans were cultivated according to ISO 10872.

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Total exposure duration:

96 h

Test temperature:

19.5-20.5 °C

pH:

5.6 (± 0.4)

Details on test conditions:

Soil Properties (according to LUFA; all values refer to dry soil):
- Soil type: loamy sand
- Organic carbon: 1.61 (± 0.44) %
- pH (0.01 M CaCl2): 5.6 (± 0.4)
- maximal Water Holding Capacity (WHCmax): 43.3 (± 5.1) g/100g
- Particle size distribution: <0.002 mm: 8.9 (± 0.8) %; 0.002-0.05 mm: 13.9 (± 1.3) %; 0.05-2.0 mm: 77.2 (± 1.9) %
• Air-dried soil was used for preparing spiked test material
• 4 g of dry soil were thoroughly mixed in 10-ml glas jars with the respective mass of test item

Nominal and measured concentrations:

0, 3125, 6250, 12500, 25000, 50000 mg/kg d.w.

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

6 250 mg/kg soil dw

Nominal / measured:

nominal

Basis for effect:

reproduction

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

25 000 mg/kg soil dw

Nominal / measured:

nominal

Basis for effect:

growth

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

3 125 mg/kg soil dw

Nominal / measured:

nominal

Basis for effect:

reproduction

Details on results:

Compared to the control (Lufa St. 2.2 soil without test item), first inhibitory effects of NaY on growth could be observed at the highest tested nominal concentration of 50,000 mg/kg dry soil (p = 0.002; one-way ANOVA [df = 5; F = 5.553]; posthoc Dunnett: p = 0.002. Thus, a NOEC of 25,000 mg/kg dry soil could be calculated for growth. No significant dose-response curve could be fitted to the toxicity data.
Reproduction was first significantly inhibited at 6,250 mg/kg soil (p < 0.001; one-way ANOVA [df = 5; F = 13.667], posthoc Dunnett: p = 0.002). Thus, a NOEC of 3,125 mg/kg dry soil could be calculated. For reproduction a proper dose-response curve could be established, as effects of > 50% inhibition were observed. Based on the logistic function that significantly fitted the dose-response curve to the observed toxicity data, EC10 and EC50 values were calculated as 1,836 and 8,778 mg/L, respectively. For reproduction, the EC10 was only slightly lower than the NOEC value. However, according to the %MDD for growth and reproduction, significant effects could have been expected already at lower concentrations.

Validity criteria fulfilled:

yes

Conclusions:

NOECs for growth and reproduction were 25,000 and 3,125 mg/kg dry soil, respectively.

Executive summary:

The toxicity of the Alumininosilicate (NaY Zeolite; NaY) via spiked water, sediment and soil was tested with the nematode Caenorhabditis elegans, using the ISO guideline 10872. In spiked soil, nematode growth was slightly, but significantly inhibited at the highest tested concentration, so that a NOEC = 25,000 mg/kg dry soil could be calculated. First effects on reproduction occurred at 6,250 mg/kg dry sediment (NOEC: 3,125 mg/kg dry soil).

Reference 2

Endpoint:

toxicity to soil macroorganisms except arthropods: long-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

May 25 - Nov. 21, 2021; experimental phase: Jun. 10 - Aug. 10, 2021

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

Jul. 29, 2016

Deviations:

yes

Remarks:

s. below

Principles of method if other than guideline:

Deviations from the Guideline:
- The temperature was not raised from 40 to 60 °C at first when the glass jars stood in the water bath. As all juveniles were counted, this deviation can be stated as uncritical.
- The temperature in the experiment was in the range 19.0 – 23.1 °C, and therefore not in the desired range (18 - 22 °C). As all validity criteria were met and normal reproduction was observed in the blank control this was stated as uncritical.
- The water content at the end of the test of the control deviated by more than 10 % of the start water content. As normal reproduction in the blank controls was observed and this deviation was only minor, it was stated as uncritical.
- The medium for the animal husbandry contained horse dung, sphagnum peat mineral
mix and urge rock powder instead of only horse dung and sphagnum peat. As the composition has no influence on the outcome of the study, this deviation can be stated as uncritical.

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Name: SYLOSIV A4 (A Zeolite)
Batch no.: 1000361205
CAS no.: 1318-02-1
Composition: Zeolite (crystalline Aluminium silicate) 95-100 %
Storage: room temperature (20 ± 5 °C)
Expiry date: 31. Jan. 2022
Stability: stable under storage conditions
Appearance: off-white powder
Purity: not applicable, UVCB
Homogeneity: homogeneous
Production date: 01. Dec. 2020
EC no.: 215-283-8 (930-915-9)
Molecular formula: not stated
Molecular weight: not stated
Vapour pressure: 0 hPa

Details on preparation and application of test substrate:

For the treatments, two mixtures of 10 ± 1 g of quartz sand once with 2205.0 mg (500 mg/kg nominal) and once with 4410.0 mg (1000 mg/kg) test item were prepared for the test and mixed with the artificial soil.

For the test, the whole artificial soil was divided into 3 portions of an appropriate amount.
The quartz sand was mixed with the respective amount of the test item for each treatment. For the blank control, only quartz sand was used. The quartz sand with test item was mixed with the artificial soil portion for each treatment.
Then, demineralised water was used to adjust the soil to a water content of 40 % to 60 % of the maximum water holding capacity calculated in relation to dry matter. Afterwards, it was mixed with each soil portion, the soil portions are divided into portions à 500 – 600 g dry weight and each portion is given into a 1 L glass preserving jar.

Test organisms (species):

Eisenia andrei

Animal group:

annelids

Details on test organisms:

Species/Strain: Eisenia andrei BOCHÉ
Age: 2 - 12 months with clitellum (adult animals)
Weight: 283 – 600 mg (range in this study)

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

yes

Total exposure duration:

56 d

Post exposure observation period:

none

Test temperature:

19.0-23.1 °C

pH:

6.7-7.0

Moisture:

water content of 40 % to 60 % of the maximum water holding capacity calculated in relation to dry matter

Details on test conditions:

Lightning: 595 lux
Photo period: 16 hours light / 8 hours dark, using LED’s

Replicates
Treatments: 8 vessels, each containing 500 – 600 g artificial soil (dry weight) with quartz sand, test item and 10 worms
Blank Control: 8 vessels, each containing 500 – 600 g artificial soil (dry weight) with quartz sand and 10 worms

Nominal and measured concentrations:

1000 mg/kg

Reference substance (positive control):

yes

Remarks:

The 56d-EC50 of Benomyl (CAS-No. 17804-35-2) was tested in a current reference test.

Duration:

56 d

Dose descriptor:

LOEC

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Basis for effect:

reproduction

Key result

Duration:

56 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

nominal

Basis for effect:

reproduction

Duration:

28 d

Dose descriptor:

LC50

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Basis for effect:

mortality

Details on results:

No mortality of adult animals after 28 days occurred in any replicate of the control and the treatments. As the mean number of juveniles in the test concentration 1000 mg/kg was higher than in the control, no inhibition of reproduction after 56 days occurred at this concentration.

As the concentration 1000 mg/kg showed no significant effect on the reproduction, the juveniles of the concentration 500 mg/kg were not counted and therefore not used for the biological evaluation.

After counting the juveniles, two replicates of the blank control clearly showed a much too low number and were therefore not used for the evaluation. If these replicates were taken into account, the scatter and therefore, the coefficient of variation in the blank control replicates would be extraordinarily and implausible high, but quite possible in a biological system. The exclusion of the two replicates can be stated as uncritical, because the test result is unequivocal (non-toxic), the number of juveniles in the replicates of the test concentration 1000 mg/kg is higher than in the control. The scatter and the coefficient of variation in the test replicates is much lower than in the control replicates and lay in the normal range.

Results with reference substance (positive control):

The value was determined as 1.9 mg/kg, lying within the required range of OECD Guideline 222 of 1 – 5 mg/kg.

Validity criteria fulfilled:

yes

Conclusions:

No mortality of adult animals after 28 days occurred in any replicate of the control and the treatments. As the mean number of juveniles in the test concentration 1000 mg/kg was higher than in the control, no inhibition of reproduction after 56 days occurred at this concentration.

Executive summary:

This study was performed as a limit test in order to evaluate the toxicity of the test item SYLOSIV A4 in a chronic reproduction test towards reproduction of Eisenia andrei. The not typical soil species Eisenia andrei Bouché, which belongs to the family of lumbricidae, was chosen because of its susceptibility to chemicals resembles that of soil-inhabiting species. No effects on mortality or reproduction were observed.

Reference 3

Endpoint:

toxicity to soil macroorganisms except arthropods: long-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Apr. 20-24, 2021

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

other: ISO 10872

Version / remarks:

2020

Deviations:

no

Remarks:

All validity criteria were met in the controls of all tests.

GLP compliance:

no

Remarks:

As these data are not used to assess extrapolation to humans, a study according to GLP was not regarded as necessary.

Specific details on test material used for the study:

• Chemical name: Aluminosilicate
• Trade name: Zeocros (P Zeolite)
• Acronym in Report: Zeo
• Batch number: 2020-531
• Composition: Aluminosilicate (Al/Si ~ 0.8)
• CAS No.: 1318-02-1
• Purity: 100 %
• Specific Surface Area (BET): not stated
• Grain size (diameter): 1.115 μm

Vehicle:

yes

Remarks:

M9 medium

Details on preparation and application of test substrate:

0.073 ml M9-medium (ISO 10872; 6 g Na2HPO4/L, 3 g KH2PO4/L, 5 g NaCl, 0.25 g MgSO4 7H2O /L) were added to 0.5 g dry soil. Together with 0.1 ml of bacterial suspension (0.073 + 0.1 = 0.173 ml) soil moisture was adjusted to 80% of the WHCmax (corresponding to 74.3 % dry weight).

Test organisms (species):

Caenorhabditis elegans

Animal group:

nematods

Details on test organisms:

Strain: N2
Genotype: C. elegans wild type
Source: Caenorhabditis Genetic Centre; University of Minnesota; Minneapolis; MN 55455; USA.
Cultivation: C. elegans were cultivated according to ISO 10872.

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Total exposure duration:

96 h

Test temperature:

19.5-20.5 °C

pH:

5.6 (± 0.4)

Details on test conditions:

Soil Properties (according to LUFA; all values refer to dry soil):
- Soil type: loamy sand
- Organic carbon: 1.61 (± 0.44) %
- pH (0.01 M CaCl2): 5.6 (± 0.4)
- maximal Water Holding Capacity (WHCmax): 43.3 (± 5.1) g/100g
- Particle size distribution: <0.002 mm: 8.9 (± 0.8) %; 0.002-0.05 mm: 13.9 (± 1.3) %; 0.05-2.0 mm: 77.2 (± 1.9) %
• Air-dried soil was used for preparing spiked test material
• 4 g of dry soil were thoroughly mixed in 10-ml glas jars with the respective mass of test item

Nominal and measured concentrations:

0, 781, 1563, 3125, 6250, 12500, 25000, 50000 mg/kg d.w.

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

25 000 mg/kg soil dw

Nominal / measured:

nominal

Basis for effect:

growth

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

12 500 mg/kg soil dw

Nominal / measured:

nominal

Basis for effect:

reproduction

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

12 500 mg/kg soil dw

Nominal / measured:

nominal

Basis for effect:

growth

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

6 250 mg/kg soil dw

Nominal / measured:

nominal

Basis for effect:

reproduction

Details on results:

Compared to the control (Lufa St. 2.2 soil without test item), first inhibitory effects of Zeo on growth could be observed at the a nominal concentration of 25,000 mg/kg dry soil (p = 0.002; one-way ANOVA [df = 7; F = 19.264]; posthoc Games-Howell: p = 0.029). Thus, a NOEC of 12,500 mg/kg dry soil could be calculated for growth.
Reproduction was first significantly inhibited at 12,500 mg/kg soil (p < 0.001; one-way ANOVA [df = 7; F = 23.956], posthoc Dunnett: p = 0.002). Thus, a NOEC of 6,250 mg/kg dry soil could be calculated.
For growth a partial dose-response curve could be set up (highest effect < 50%). Based on the logistic function that significantly fitted the dose-response curve to the observed toxicity data, an EC10 of 9,534 mg/kg dry sediment has been determined.For reproduction a proper dose-response curve could be established, as effects of > 50% inhibition were observed. Based on the logistic function that significantly fitted the dose-response curve to the observed toxicity data, EC10 and EC50 values were calculated as 892 and 6735 mg/L, respectively.
For growth, the EC10 was only slightly lower than the NOEC value. %MDD values for growth could properly predict first significant effects. However, for reproduction the EC10 was considerably lower than the NOEC. This was due to the phenomenon that only effects >50% yielded a significant difference, although %MDD values predicted significant effects already at > 21%. The variance of the data showed to be very heterogenous (Levene’s test: p = 0.002), so that a more robust posthoc test (Games-Howell) is recommended. Using a one-sided Dunnett posthoc test would reveal a NOEC of 781 mg/kg dry soil, matching the EC10 of 892 mg/kg dry soil quite well.

Validity criteria fulfilled:

yes

Conclusions:

NOECs for growth and reproduction were 12,500 and 6,250 mg/kg dry soil, respectively.

Executive summary:

The toxicity of the zeolite Zeocros (Zeo) via spiked water, sediment and soil was tested with the nematode Caenorhabditis
elegans, using the ISO guideline 10872. In spiked soil,
nematode growth was first significantly inhibited at a concentration of 25,000 mg/kg dry soil, so that a NOEC = 12,500 mg/kg dry soil could be determined. First effects on reproduction occurred at 12,500 mg/kg dry soil (NOEC: 6,250 mg/kg dry soil).

Reference 4

Endpoint:

toxicity to soil macroorganisms except arthropods: long-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Mar. 8-12 and 25-29, 2021

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

other: ISO 10872

Version / remarks:

2020

Deviations:

no

Remarks:

All validity criteria were met in the controls of all tests.

GLP compliance:

no

Remarks:

As these data are not used to assess extrapolation to humans, a study according to GLP was not regarded as necessary.

Specific details on test material used for the study:

• Chemical name: Aluminosilicate
• Trade name: Sylosiv A4 (A Zeolite)
• Acronym in Report: Syl
• Batch number: 1000361205
• Composition: Aluminosilicate (Al/Si ~ 1)
• CAS No.: 1318-02-1
• Purity: 100 %
• Specific Surface Area (BET): not stated
• Grain size (diameter): 8.687 μm

Vehicle:

yes

Remarks:

M9 medium

Details on preparation and application of test substrate:

0.073 ml M9-medium (ISO 10872; 6 g Na2HPO4/L, 3 g KH2PO4/L, 5 g NaCl, 0.25 g MgSO4 7H2O /L) were added to 0.5 g dry soil. Together with 0.1 ml of bacterial suspension (0.073 + 0.1 = 0.173 ml) soil moisture was adjusted to 80% of the WHCmax (corresponding to 74.3 % dry weight).

Test organisms (species):

Caenorhabditis elegans

Animal group:

nematods

Details on test organisms:

Strain: N2
Genotype: C. elegans wild type
Source: Caenorhabditis Genetic Centre; University of Minnesota; Minneapolis; MN 55455; USA.
Cultivation: C. elegans were cultivated according to ISO 10872.

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Total exposure duration:

96 h

Test temperature:

19.5-20.5 °C

pH:

5.6 (± 0.4)

Details on test conditions:

Soil Properties (according to LUFA; all values refer to dry soil):
- Soil type: loamy sand
- Organic carbon: 1.61 (± 0.44) %
- pH (0.01 M CaCl2): 5.6 (± 0.4)
- maximal Water Holding Capacity (WHCmax): 43.3 (± 5.1) g/100g
- Particle size distribution: <0.002 mm: 8.9 (± 0.8) %; 0.002-0.05 mm: 13.9 (± 1.3) %; 0.05-2.0 mm: 77.2 (± 1.9) %
• Air-dried soil was used for preparing spiked test material
• 4 g of dry soil were thoroughly mixed in 10-ml glas jars with the respective mass of test item

Nominal and measured concentrations:

Test 1: 3125, 6250, 12500, 25000, 50000 mg/kg d.w.
Test 2: 781, 1563, 3125, 50000 mg/kg d.w.

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

25 000 mg/kg soil dw

Nominal / measured:

nominal

Basis for effect:

growth

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

3 125 mg/kg soil dw

Nominal / measured:

nominal

Basis for effect:

reproduction

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

12 500 mg/kg soil dw

Nominal / measured:

nominal

Basis for effect:

growth

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

1 563 mg/kg soil dw

Nominal / measured:

nominal

Basis for effect:

reproduction

Details on results:

Compared to the control (Lufa St. 2.2 soil without test item), first inhibitory effects of Syl on growth could be observed at the a nominal concentration of 25,000 mg/kg dry soil (soil test 1: p < 0.001; one-way ANOVA [df = 5; F = 6.902]; posthoc Dunnett: p = 0.036. Thus, a NOEC of 12,500 mg/kg dry soil could be determined for growth. For growth, no dose-response curves could be set up.
Reproduction was first significantly inhibited at 3,125 mg/kg soil (soil test 1: p < 0.001; oneway ANOVA [df = 5; F = 41.652], posthoc Dunnett: p < 0.001). In the second soil test, no significant inhibition of reproduction occurred till 3250 mg/kg soil (one-way ANOVA, posthoc Dunnett: p > 0.05). Thus, using the data of both tests, a NOEC of 1,563 mg/kg dry
soil could be determined.
For reproduction a proper dose-response curve could be established, as effects of > 50% inhibition were observed. Based on the logistic function that significantly fitted the doseresponse curve to the observed toxicity data, EC10 and EC50 values were calculated as 668 and 6317 mg/L, respectively.
For reproduction the EC10 was considerably lower than the NOEC. This was due to the phenomenon that only effects >50% yielded a significant difference, although %MDD values predicted significant effects already at > 21%. The variance of the data showed to be very heterogenous (Levene’s test: p = 0.002), so that a more robust posthoc test (Games-Howell) is
recommended. Using a one-sided Dunnett posthoc test would reveal a NOEC of 781 mg/kg dry soil, matching the EC10 of 892 mg/kg dry soil quite well.

Validity criteria fulfilled:

yes

Conclusions:

NOECs for growth and reproduction were 12,500 and 1,563 mg/kg dry soil, respectively.

Executive summary:

The toxicity of the zeolite Sylosiv via spiked water, sediment and soil was tested with the nematode Caenorhabditis elegans, using the ISO guideline 10872. In spiked soil, nematode growth was first significantly inhibited at a concentration of 25,000 mg/kg dry soil, so that a NOEC of 12,500 mg/kg dry soil could be determined. First effects on reproduction occurred at 3,125 mg/kg dry soil (NOEC: 1,563 mg/kg dry soil).

Reference 5

Endpoint:

toxicity to soil macroorganisms except arthropods: long-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Mar. 8-12, 2021

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

other: ISO 10872

Version / remarks:

2020

Deviations:

no

Remarks:

All validity criteria were met in the controls of all tests.

GLP compliance:

no

Remarks:

As these data are not used to assess extrapolation to humans, a study according to GLP was not regarded as necessary.

Specific details on test material used for the study:

• Chemical name: Alumininosilicate
• Trade name: Köstrolith (X Zeolite)
• Acronym in Report: Kt
• Batch number: 2006090001
• Composition: Aluminosilicate (Al/Si ~ 0.7)
• CAS No.: 1318-02-1
• Purity: 100 %
• Specific Surface Area (BET): not stated
• Grain size (diameter): 3.432 μm

Vehicle:

yes

Remarks:

M9 medium

Details on preparation and application of test substrate:

0.073 ml M9-medium (ISO 10872; 6 g Na2HPO4/L, 3 g KH2PO4/L, 5 g NaCl, 0.25 g MgSO4 7H2O /L) were added to 0.5 g dry soil. Together with 0.1 ml of bacterial suspension (0.073 + 0.1 = 0.173 ml) soil moisture was adjusted to 80% of the WHCmax (corresponding to 74.3 % dry weight).

Test organisms (species):

Caenorhabditis elegans

Animal group:

nematods

Details on test organisms:

Strain: N2
Genotype: C. elegans wild type
Source: Caenorhabditis Genetic Centre; University of Minnesota; Minneapolis; MN 55455; USA.
Cultivation: C. elegans were cultivated according to ISO 10872.

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Total exposure duration:

96 h

Test temperature:

19.5-20.5 °C

pH:

5.6 (± 0.4)

Details on test conditions:

Soil Properties (according to LUFA; all values refer to dry soil):
- Soil type: loamy sand
- Organic carbon: 1.61 (± 0.44) %
- pH (0.01 M CaCl2): 5.6 (± 0.4)
- maximal Water Holding Capacity (WHCmax): 43.3 (± 5.1) g/100g
- Particle size distribution: <0.002 mm: 8.9 (± 0.8) %; 0.002-0.05 mm: 13.9 (± 1.3) %; 0.05-2.0 mm: 77.2 (± 1.9) %
• Air-dried soil was used for preparing spiked test material
• 4 g of dry soil were thoroughly mixed in 10-ml glas jars with the respective mass of test item

Nominal and measured concentrations:

Test 1: 3125, 6250, 12500, 25000, 50000 mg/kg d.w.
Test 2: 781, 1563, 3125, 50000 mg/kg d.w.

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

25 000 mg/kg soil dw

Nominal / measured:

nominal

Basis for effect:

reproduction

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 50 000 mg/kg soil dw

Nominal / measured:

nominal

Basis for effect:

growth

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

12 500 mg/kg soil dw

Nominal / measured:

nominal

Basis for effect:

reproduction

Details on results:

Compared to the control (Lufa St. 2.2 soil without test item), no inhibitory effects of Kt on growth could be observed unil the highest tested nominal concentration of 50,000 mg/kg dry soil (p > 0.05; one-way ANOVA; df = 5; F = 2.229). Thus, a NOEC of ≥ 50,000 mg/kg dry soil could be calculated for growth. No significant dose-response curve could be fitted to the toxicity data.
Reproduction was first significantly inhibited at 25,000 mg/kg soil (p < 0.001; one-way ANOVA [df = 5; F = 10.663], posthoc Dunnett: p = 0.030). Thus, a NOEC of 12,500 mg/kg dry soil could be calculated. For reproduction a dose-response curve could be established, as effects of > 50% inhibition were observed. However, it has to be noted that the fit to the data using a logistic function was not statistically significant (p = 0.106; r² = 0.78). EC10 and EC50 values were calculated as 12,879 and 33,535 mg/L, respectively. EC10 was only slightly lower than the NOEC value. However, according to the %MDD for growth and reproduction, significant effects could have been expected already at lower concentrations. NOEC and EC10 values matched quite nicely for reproduction.

Validity criteria fulfilled:

yes

Conclusions:

NOECs for growth and reproduction were >= 50,000 and 12,500 mg/kg dry soil, respectively.

Executive summary:

The toxicity of the zeolite Köstrolith (Kt) via spiked water, sediment and soil was tested with the nematode Caenorhabditis elegans, using the ISO guideline 10872. In spiked soil, C. elegans’s growth was not affected
till the highest concentration (NOEC ≥ 50,000 mg/kg dry sediment), while first effects on reproduction occurred at 25,000 mg/kg dry sediment (NOEC: 12,500 mg/kg dry soil).

Description of key information

96-hour experiments in soil were conducted with the nematode Caenorhabditis elegans. The NOECs for growth and reproduction ranged between 12500 to >= 50000 and 1563 to 12500 mg/kg dry soil, respectively. 

Also, a 56-day experiment in Eisenia andrei was conducted with the a type A zeolite. No toxicity to reproduction was detected at the limit dose of 1000 mg/kg dry soil.

Key value for chemical safety assessment

Long-term EC10, LC10 or NOEC for soil macroorganisms:

1 563 mg/kg soil dw