Sodium 1H-benzotriazolide

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:

biological phase: 20.10.-15.12.2020

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:

2016-07-29

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Product name: 1,2,3 Benzotriazole Fine Granular
- Source (i.e. manufacturer or supplier) and lot/batch number of test material:
Connect Chemicals, Ratingen, Germany, Batch 190720
- Purity, including information on contaminants, isomers, etc.: 99.9 %

RADIOLABELLING INFORMATION (if applicable)
no

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Store cool (SDS)
- Stability and homogeneity of the test material in the vehicle/solvent under test conditions (e.g. in the exposure medium) and during storage: stable
- Stability in the medium, i.e. sensitivity of the test material to hydrolysis and/or photolysis: assumed stable
- Solubility and stability of the test material in the solvent/vehicle and the exposure medium: assumed stable, Water solubility (25 °C): 20 g/L
- Reactivity of the test material with the incubation material used (e.g. plastic ware): assumed no reactivity

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
please refer to following sections

FORM AS APPLIED IN THE TEST (if different from that of starting material)
same as starting material

INFORMATION ON NANOMATERIALS
no nanomaterial

TYPE OF BIOCIDE/PESTICIDE FORMULATION (if applicable)
no biocide/pesticide

OTHER SPECIFICS
n/a

Analytical monitoring:

no

Details on sampling:

- Sampling method: At the end, the adults were removed. For each container the total number and mass of living adult earthworms was recorded. After allowing the offspring to develop, the number of offspring per test container hatched from the cocoons was counted by hand selection
- Sample storage conditions before analysis: not indicated

Vehicle:

no

Details on preparation and application of test substrate:

- Method of application to filter paper (if used): -
- Method of mixing into soil (if used): -
- Method of application to soil surface (if used): direct application as aqueous solution
- Controls: adjusted to 60 % WHCmax
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): -
- Concentration of vehicle in test medium (stock solution and final test solution): -
- Evaporation of vehicle before use: -

Test organisms (species):

Eisenia andrei

Animal group:

annelids

Details on test organisms:

TEST ORGANISM
- Common name: Earthworm
- Source: in-house bred IME (origin: Regenwurmfarm Tacke, Klosterdiek 61, 46325 Borken)
- Age at test initiation (mean and range, SD): two months - 1 year
- Weight at test initiation (mean and range, SD): wet mass 250 - 600 mg

ACCLIMATION
- Acclimation period: 4 days
- Acclimation conditions (same as test or not): in artificial soil, same food as in test in sufficient amount
- Health during acclimation (any mortality observed): not indicated

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Total exposure duration:

4 wk

Post exposure observation period:

4 weeks

Test temperature:

18 – 21 °C

pH:

6.0 ± 0.5

Moisture:

40-60% of max. water-holding capacity

Details on test conditions:

TEST SYSTEM
- Test container (material, size): plastic, area of 17x12cm, height 6cm, covered with perforated transparent plastic caps to prevent escaping
- Amount of soil or substrate: 500g dry mass, substrate depth of about 5-6 cm
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4
- No. of replicates per control: 8

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Composition (if artificial substrate): Sphagnum peat, air-dried, finely ground (10%) Kaolinite, air-dried (20%) Industrial quartz sand, air-dried (70%)
- Maximum water holding capacity (in % dry weigth): 49.7 – 58.4
- Pretreatment of soil: pH, water-holding capacity adjusted
- pH: 5.6 – 5.8 (start), 5.9 – 6.0 (end)
- Storage (condition, duration): not indicated

OTHER TEST CONDITIONS
- Photoperiod: 16/8 light/dark
- Light intensity: 602 – 752 lx

FOOD
- Description: air-dried, finely ground horse manure
- Source: Elisabeth Hardebusch, 57392 Schmallenberg, Germany
- Known treatments: Horses from which manure was obtained by self-collecting were not subject to medication or treatment with substances, such as growth promoters, nematicides or similar veterinary products that could adversely affect the worms during the test (self-declaration by owner of the horses)
- Feeding: day 1 after introduction (25g remoistened finely ground horse manure, corresponding to 5g dw)/container, further feeding once weekly

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): total number and mass of living adult earthworms, number of offspring per test container hatched from cocoons

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Range finding study: yes
- Test concentrations: not indicated
- Results used to determine the conditions for the definitive study: The concentrations were chosen based on a non-GLP range finding test suggesting a start of reproduction impairment at 10 mg test item/kg dry mass soil and an EC50 in the range of 100 – 1000 mg dry mass/kg dry mass soil.

Nominal and measured concentrations:

2.5, 5, 10, 20, 40, 80 and 160 mg test item/kg soil dw (nominal)

Reference substance (positive control):

yes

Remarks:

Carbendazim

Key result

Duration:

56 d

Dose descriptor:

EC10

Effect conc.:

19.1 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Remarks:

95% CL: 2.2 – 162.8 mg TI/kg DM

Duration:

28 d

Dose descriptor:

EC50

Effect conc.:

> 160 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth

Remarks:

weight change

Duration:

28 d

Dose descriptor:

LC50

Effect conc.:

> 160 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

56 d

Dose descriptor:

NOEC

Effect conc.:

40 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

>= 160 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth

Remarks:

weight change

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

>= 160 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Results with reference substance (positive control):

- Results with reference substance valid? yes, not longer than 12 months before
- Relevant effect levels: NOEC for survival was found to be ≥ 5 mg/kg, no concentration dependence occurred. Reproduction was significantly affected by carbendazim. The NOEC was suggested to be < 0.56 mg/kg. The EC10 and EC50 were 0.31 mg/kg and 4.47 mg/kg, respectively.

Reported statistics and error estimates:

ANOVA followed by Williams` Multiple Sequential t-Test or Dunnett’s Multiple t-Test for metric response, ToxRat Professional by ToxRat® Solutions GmbH

Validity criteria fulfilled:

yes

Conclusions:

In this valid study, no decreased survival rate and no impaired weight change were detected up to the highest nominal concentration tested (160 mg test item/kg soil dw) compared to the control. No concentration response could be computed.
There was a concentration relating inhibition of reproduction, therefore Benzotriazole showed chronic effects on the Earthworm.

Executive summary:

The effect of Benzotriazole on survival rate, biomass, and reproduction of adult earthworms of the species Eisenia andrei was examined in this study. Survival rate and biomass were determined after 28 days, reproduction after 56 days. 
The used concentrations were 2.5, 5, 10, 20, 40, 80 and 160 mg test item/kg soil dw, based on a non-GLP range finding test.
The EC₁₀ for survival rate and weight change were >160 mg test item/kg soil dw (NOEC= >160 mg test item/kg soil dw) , EC₁₀ for reproduction was 19.1 mg test item/kg soil dw (95% CL: 2.2 – 162.8 mg TI/kg DM) (NOEC =20 mg test item/kg soil dw)

Benzotriazole showed chronic effects on Eisenia andrei under the test conditions.

Reference 2

Endpoint:

toxicity to soil macroorganisms except arthropods: long-term

Type of information:

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

Adequacy of study:

key study

Justification for type of information:

See read-across justification in Section 13.2 titled "140704_NaBZT_rationale_RA 8.4.16".

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

56 d

Dose descriptor:

EC10

Effect conc.:

22.6 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Remarks:

95% CL: 2.6 – 192.9 mg TI/kg DM

Remarks on result:

other: Results are the source substance results corrected for molecular weight of the target substance.

Duration:

28 d

Dose descriptor:

EC50

Effect conc.:

> 189.6 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth

Remarks:

weight change

Remarks on result:

other: Results are the source substance results corrected for molecular weight of the target substance.

Duration:

28 d

Dose descriptor:

LC50

Effect conc.:

> 189.6 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

56 d

Dose descriptor:

NOEC

Effect conc.:

47.4 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

>= 189.6 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth

Remarks:

weight change

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

>= 189.6 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Results with reference substance (positive control):

- Results with reference substance valid? yes, not longer than 12 months before
- Relevant effect levels: NOEC for survival was found to be ≥ 5 mg/kg, no concentration dependence occurred. Reproduction was significantly affected by carbendazim. The NOEC was suggested to be < 0.56 mg/kg. The EC10 and EC50 were 0.31 mg/kg and 4.47 mg/kg, respectively.

Reported statistics and error estimates:

ANOVA followed by Williams` Multiple Sequential t-Test or Dunnett’s Multiple t-Test for metric response, ToxRat Professional by ToxRat® Solutions GmbH

Validity criteria fulfilled:

yes

Conclusions:

In this valid study on the source substance, no decreased survival rate and no impaired weight change were detected up to the highest nominal concentration tested of the source substance of 160 mg test item/kg soil dw. A one-to-one read-across approach with correction for molecular weight has been used to derive endpoints for the target substance.

As read-across has been shown to be applicable (IUCLID Section 13.2), but there are differences in molecular weight between the source and target substance the endpoint has been corrected for this difference (target endpoint: 189.6 mg/kg soil dw). No concentration response could be computed.

The source substance reduced reproduction in earthworms and the NOEC was 40 mg/kg soil dw. Corrected for molecular weight this gives a NOEC of 47.4 mg/kg soil dw for the target substance. The EC10 for reporductions was 19. mg/kg soil dw for the source substance resulting in an EC10 of 22.6 mg/kg soil dw for the target substance. The source and target substance show chronic effects on the Earthworm.

Executive summary:

A one-to-one read-across approach with correction for molecular weight has been used to derive endpoints for the target substance. The read-across was from benzotriazole (EC No.: 202-394-1) to sodium 1H-benzotriazolide (EC No. 239-269-6).  In line with ECHA guidance the reported endpoint values are those relevant to the target substance and have been corrected for molecular weight (ECHA How to prepare registration and PPORD dossiers, v10.0, 2021). The only differences between the substances is the addition of a sodium salt in the target substance.  However, at relevant environmental pH ca. 6-9, protonation will occur yielding the neutral form of the target substance (See Section 13.2 for more information). 

The effect of Benzotriazole (source substance) on survival rate, biomass, and reproduction of adult earthworms of the species Eisenia andrei was examined in this study. Survival rate and biomass were determined after 28 days, reproduction after 56 days. 
The used concentrations were 2.5, 5, 10, 20, 40, 80 and 160 mg test item/kg soil dw, based on a non-GLP range finding test.
The EC₁₀ for survival rate and weight change were >160 mg test item/kg soil dw (NOEC= >160 mg test item/kg soil dw) , EC₁₀ for reproduction was 19.1 mg test item/kg soil dw (95% CL: 2.2 – 162.8 mg TI/kg DM).

This translates to expected effects of the target substance after molecular weight correction of: EC₁₀ for survival rate and weight change were >189.6 mg test item/kg soil dw (NOEC= >189.6 mg test item/kg soil dw) , EC₁₀ for reproduction was 22.6 mg test item/kg soil dw.

The test item showed chronic effects on Eisenia andrei under the test conditions.

Description of key information

Eisenia andrei reproduction EC10: 22.6 mg/kg soil dw, OECD 222. Simon, 2021.

Eisenia andrei growth, mortality EC50: ≥ 189.6 mg/kg soil dw, OECD 222. Simon, 2021.

Key value for chemical safety assessment

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

22.6 mg/kg soil dw

Additional information

A one-to-one read-across approach with correction for molecular weight has been used to derive endpoints for the target substance. The read-across was from benzotriazole (EC No.: 202-394-1) to sodium 1H-benzotriazolide (EC No. 239-269-6).  In line with ECHA guidance the reported endpoint values are those relevant to the target substance and have been corrected for molecular weight. The only differences between the substances is the addition of a sodium salt in the target substance.  However, at relevant environmental pH ca. 6-9, protonation will occur yielding the neutral form of the target substance (See Section 13.2 for more information). 

The effect of Benzotriazole (source substance) on survival rate, biomass, and reproduction of adult earthworms of the species Eisenia andrei was examined in this study. Survival rate and biomass were determined after 28 days, reproduction after 56 days. The used concentrations were 2.5, 5, 10, 20, 40, 80 and 160 mg test item/kg soil dw, based on a non-GLP range finding test.
The EC₁₀ for survival rate and weight change were >160 mg test item/kg soil dw (NOEC= >160 mg test item/kg soil dw) , EC₁₀ for reproduction was 19.1 mg test item/kg soil dw (95% CL: 2.2 – 162.8 mg TI/kg DM).

This translates to expected effects of the target substance after molecular weight correction of: EC₁₀ for survival rate and weight change were >189.6 mg test item/kg soil dw (NOEC= >189.6 mg test item/kg soil dw) , EC₁₀ for reproduction was 22.6 mg test item/kg soil dw.

The test item showed chronic effects on Eisenia andrei under the test conditions.