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Available data for fish, invertebrates, algae and microorganisms indicate a low potential for acute and chronic aquatic toxicity of diiron trioxide in powder and nano-form. Regarding differences in the particle size, the available data for the acute toxicity of diiron trioxide in powder and nano-form to fish further indicate that the particle size does not affect the toxicity potential to any detectable extent since effect concentrations of the powder and nano-form are 100 to 500-fold above the OECD test limit. Based on read-across (category approach) to iron hydroxide oxide yellow and triiron tetraoxide (see attachment "Iron oxides category justification” in IUCLID section 13), available data for fish, invertebrates, algae and microorganisms further indicate a low potential for acute and chronic aquatic toxicity of iron (hydr)oxides. For all reliable studies (RL1 or RL2) with iron (hydr)oxides, unbounded effect concentrations above the OECD test limit were determined (see table below).

Table: Aquatic toxicity of iron (hydr)oxides

Endpoint

Test species

Test results

Test material/form

Reliability: Reference

Short-term toxicity to fish

Danio rerio

LC50 (96h): > 50 g/L nominal

LC0 (96h): >= 50 g/L nominal

diiron trioxide / powder

RL2: Bruns, 1989

.

Danio rerio

LC50 (96h): > 10 g/L nominal

LC0 (96h): >= 10 g/L nominal

triiron tetraoxide / powder

RL2: Caspers, 2000

.

Danio rerio

LC50 (96h): > 10 g/L nominal

LC0 (96h): >= 10 g/L nominal

triiron tetraoxide /powder

RL2: Bayer AG, 1989

.

Danio rerio

LC50 (96h): > 100 g/L nominal

LC0 (96h): >= 100 g/L nominal

zinc ferrite brown spinel / powder

RL2: Weyers, 1989

.

Danio rerio

LC50 (96h): > 100 g/L nominal

LC0 (96h): >= 100 g/L nominal

manganese ferrite black spinel / powder

RL2: Weyers, 1989

.

Danio rerio

LC50 (96h): > 10 g/L nominal

LC0 (96h): >= 10 g/L nominal

diiron trioxide / nano, alpha

RL2: Weyers, 1985

.

Danio rerio

LC50 (96h): > 100 g/L nominal

LC0 (96h): >= 100 g/L nominal

iron hydroxide oxide yellow / nano, alpha

RL2: Weyers & Caspers, 1989

Long-term toxicity to fish

Danio rerio

NOEC (168 h): 10 mg/L nominal

diiron trioxide / nano, alpha

RL3: Zhu et al., 2012

Short-term toxicity to aquatic invertebrates

Daphnia magna

EC50 (48h): > 100 mg/L nominal

diiron trioxide / powder

RL2: Neuhahn, 2008

.

Daphnia magna

EC0 (48 h): >= 10 g/L nominal

triiron tetraoxide /powder

RL2: Caspers, 2000

.

Daphnia magna

EC0 (48 h): >= 10 g/L nominal

zinc ferrite brown spinel / powder

RL2: Caspers, 1999

.

Daphnia magna

EC0 (48 h): >= 10 g/L nominal

manganese ferrite black spinel / powder

RL2: Caspers, 2000

.

Daphnia magna

EC50 (48h): > 100 mg/L nominal

iron hydroxide oxide yellow / nano, alpha

RL2: Neuhahn, 2008

Long-term toxicity to aquatic invertebrates

Daphnia magna

NOEC (21d): >= 20 mg/L nominal

diiron trioxide / nano

RL2: Nogueira et al., 2015

Toxicity to aquatic algae and cyanobacteria

Pseudokirchneriella subcapitata

NOEC (72 h): >= 20 mg/L nominal

EC50 (72 h): > 20 mg/L nominal

diiron trioxide / nano

RL2: Nogueira et al., 2015

Toxicity to microorganisms

Activated sludge

EC50 (3 h): > 10 g/L nominal

triiron tetraoxide / powder

RL2: Bruns, 1989

.

Activated sludge

EC50 (3 h): >= 10 g/L nominal

manganese ferrite black spinel / powder

RL2: Bruns, 1989

.

Activated sludge

EC50 (3 h): > 10 g/L nominal

iron hydroxide oxide/diiron trioxide (75:25) / nano, alpha

RL2: Bruns, 2004

Additional information

Iron oxide category

Based on read-across to micro- and nano-sized iron (hydr)oxides (see attachment "Read-across justification-environ assessment-iron oxides" in section 13), available data for fish, invertebrates, algae and microorganisms indicate a low potential for acute and chronic aquatic toxicity of micro-sized iron (hydr)oxides.

Data available on the short-term toxicity of different iron (hydr)oxides, including micro- and nano-sized iron hydroxide oxide yellow, diiron trioxide, triiron tetraoxide, zinc ferrite brown spinel and manganese ferrite black spinel, to fish (Danio rerio), invertebrates (Daphnia magna) and algae (Pseudokirchneriella subcapitata) indicate a low potential for acute toxicity. Respective unbounded EC/LC50 values amount for fish up to > 100 g/L, a concentration that is 1000-fold above the corresponding OECD test limit for acute toxicity, for invertebrates up to > 10 g/L, a concentration that is 100-fold above the corresponding OECD test limit for acute toxicity and > 20 mg/L for algae.

Iron (hydr)oxides are also not toxic to aquatic micro-organisms as 3-h EC50 values for the respiration inhibition of different iron (hydr)oxides, including micro- and nano-sized iron hydroxide oxide/diiron trioxide (75:25), manganese ferrite black spinel and triiron tetraoxide are > 10 g/L, a concentration that is 10-fold above the corresponding OECD test limit.

Furthermore, chronic toxicity data of diiron trioxide in nanoform indicate a low potential for aquatic toxicity since NOECs available for three trophic levels (algae, invertebrates, fish) are ≥ 10 mg/L, and thus above the corresponding OECD test limit of 10 mg/L for chronic toxicity.

Regarding differences in the particle size, the available data for the acute toxicity of diiron trioxide in powder and nano-form to fish further indicate that the particle size does not affect the toxicity potential to any detectable extent since effect concentrations of the powder and nano-form are up to 500-fold above the OECD test limit. In all acute and chronic toxicity studies with iron (hydr)oxides, effect concentrations were unbounded and/or above the OECD test limit (see Table).

Soluble iron salts are not considered toxic to the aquatic life, i.e. do not meet CLP classification criteria for Acute (short-term) aquatic hazard and Long-term aquatic hazard. The acute and chronic ecotoxicity reference values (ERVs) for iron ions are above the respective CLP classification criteria of 1 mg/L. “Where the acute ERV for the metal ions of concern is greater than 1 mg/l the metals need not be considered further in the classification scheme for acute hazard(ECHA, 2017, Section IV.5.2.1 Classification strategy for determining acute aquatic hazard for metals, Guidance on the application of CLP criteria V. 5.0)”. In accordance with Figure IV.4 “Classification strategy for determining acute aquatic hazard for metal compounds” of ECHA Guidance on the Application of the CLP Criteria (Version 5.0, July 2017) and section 4.1.2.10.2. of Regulation (EC) No 1272/2008, the substance diiron trioxide in powder- and nano-form is poorly soluble and does not meet classification criteria for acute (short-term) aquatic hazard.

Where the chronic ERV for the metal ions of concern is greater than 1 mg/l, the metals need not be considered further in the classification scheme(ECHA, 2017, Section IV.5.2.2.1 Approach based on available chronic toxicity reference data, Guidance on the application of CLP criteria V. 5.0).” In accordance with Figure IV.5 „Classification strategy for determining long-term aquatic hazard for metal compounds “of ECHA Guidance on the Application of the CLP Criteria (V. 5.0, July 2017) and section 4.1.2.10.2. of Regulation (EC) No 1272/2008, the substance diiron trioxide in powder- and nano-form is poorly soluble and does not meet classification criteria for chronic (long-term) aquatic hazard.

The conclusion on a lack of an aquatic hazard potential of diiron trioxide is in accordance with a Tier-1 assessment performed with MECLAS version 5.2 (http://www.meclas.eu/ accessed on 06.02.2020). The Metals Classification Tool (MECLAS) is a web enabled classification tool always updated to the latest classification guidance, legal ruling, information on toxicity references and available self-classifications. In sum, it can safely be assumed that diiron trioxide in powder and nanoform does not need meet classification criteria of an acute or long-term aquatic hazard of Regulation (EC) No 1272/2008.

 “If there are ecotoxicity data showing effects in aquatic organisms, but the substance is not classified as dangerous for the aquatic environment, an aquatic PNEC can nevertheless be derived thus indicating a hazard to the aquatic environment(ECHA guidance on IR & CSA, Part B: Hazard Assessment (V. 2.1, December 2011).” For soluble iron salts and poorly soluble diiron trioxide, there are not any reliable ecotoxicity data showing effects in aquatic organisms, soluble iron salts and diiron trioxide are not classified as dangerous for the aquatic environment, an aquatic PNEC cannot be derived thus not indicating a hazard to the aquatic environment.

Diiron trioxide in powder and nano-form is not classified as harmful, toxic or very toxic to aquatic life or may cause long lasting harmful effects to aquatic life. Diiron trioxide in powder and nano-form is also not an unclassified hazard to the aquatic environment. Based on the poor solubility, bioavailability, lack of a potential for bioaccumulation and toxicity to aquatic organisms and considering ubiquitousness of iron (hydr)oxides in the aquatic compartment and essentiality of iron (as described in section "Nano-Environmental fate and pathways"), diiron trioxide in powder and nano-form is also not considered an unclassified hazard to the aquatic compartment.