Magnesium, EDTA cobalt copper iron manganese zinc complexes

Endpoint summary

Description of key information

Skin: Due to the consistent negative results of different read across substances, the target substance is considered to be non-irritant..
Eye: Only one component of the target substance, EDTA-CuNa2, had irritant properties. However, up to concentrations of 17% of EDTA-CuNa2 the ingredient was not irritant. Due to the fact that the maximal concentration of EDTA-CuNa2 in the target substance is 9% no classification of the target substance is warranted

Key value for chemical safety assessment

Skin irritation / corrosion

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

1.   Hypothesis for the analogue approach

There are no reliable data available for irritation or corrosion ofMagnesium, EDTA cobalt copper iron manganese zinc complexes(CAS 234446-82-3). In order to fulfil the standard information requirements set out in Annex VIII, 8.5, in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006, read-across from structurally related substances, which are ingredients ofMagnesium, EDTA cobalt copper iron manganese zinc complexes(CAS 234446-82-3), was conducted.

In accordance with Article 13 (1) of Regulation (EC) No 1907/2006, "information on intrinsic properties of substances may be generated by means other than tests, provided that the conditions set out in Annex XI are met.” In particular for human toxicity, information shall be generated whenever possible by means other than vertebrate animal tests, which includes the use of information from structurally related substances (grouping or read-across).

Having regard to the general rules for grouping of substances and read-across approach laid down in Annex XI, Item 1.5, of Regulation (EC) No 1907/2006, whereby substances may be predicted as similar provided that their physicochemical, toxicological and ecotoxicological properties are likely to be similar or follow a regular pattern as a result of structural similarity.

 

	Target Substance	Source Substance No 1	Source Substance No 2	Source Substance No 3	Source Substance No 4	Source Substance No 5
Name	Magnesium, EDTA cobalt copper iron manganese zinc complexes	EDTA-MnNa2	EDTA-FeNa	EDTA-ZnNa2	EDTA-CuNa2	EDTA-MgNa2
CAS	234446-82-3	15375-84-5	15708-41-5	14025-21-9	14025-15-1	14402-88-1
% in CAS 234446-82-3	n.a.	30	25	9	9	Excess Mg2+ as counterion
In vivo skin	--	--	Not irritant	--	Not irritant	--
In vitro skin	--	Not irritant	--	Not irritant	--	Not irritant
In vivo eye	--	--	Not irritant	--	Irritant	--
In vitro eye	--	Not irritant	--	Not irritant	Not irritant*	Not irritant

 

n.a.: not applicable; Lack of data for a given endpoint is indicated by “--“;*: up to concentrations of 17%.

 

2.   Analogue approach justification

The above mentioned substances are considered to be similar on the basis of structural similarity resulting in similar properties and/or activities. The available endpoint information of the source substances are used to predict the respective endpoints forMagnesium, EDTA cobalt copper iron manganese zinc complexes(CAS 234446-82-3). A detailed analogue approach justification is provided in the technical dossier (see IUCLID Section 13).

Sincenoskin and eye irritation studies are available forMagnesium, EDTA cobalt copper iron manganese zinc complexes(CAS234446-82-3), a read-across from the structurally related source substancesEDTA-MgNa2 (CAS 15375-84-5), EDTA-FeNa (CAS 15708-41-5), EDTA-ZnNa2 (CAS 14025-21-9), EDTA-CuNa2 (CAS 14025-15-1) and EDTA-MgNa2 (CAS 14402-88-1)were usedin accordance to Regulation (EC) No. 1907/2006 Annex XI, 1.5.

3.   Results

a)   Skin irritation

a.   Target Substance:EDTA cobalt copper iron manganese zinc complexes(CAS 234446-82-3)

There were no studies available with the target substance for skin irritation/corrosion.

 

b.   Source Substances

-      Source Substance No1: EDTA-MnNa2 (CAS 15375-84-5)

The skin corrosion/irritation potential of EDTA-MnNa2 was tested according to draft OECD guideline (Draft OECD guideline - in vitro skin irritation: Reconstructed human Epidermis (RhE) test method) in human reconstructed skin (EPISKIN; Verbaan et al, 2010). At least 10 mg EDTA-MnNa2 powder was applied for 15 min on top of moistened skin tissue. After a 42 hour incubation period, determination of the cytotoxic (irritancy) effect was performed. EDTA-MnNa2 was non-irritant in thein vitroskin irritation test under the experimental conditions.

-      Source Substance No2: EDTA-FeNa (CAS 15708-41-5)

In a skin irritation study according to OECD guideline 404 (Leuschner et al, 2007) 500 mg FeNaEDTA was applied onto the shaved skin of three male rabbits for 4 hours under semiocclusive conditions. Very slight erythema was noted for one animal at 1 hour after patch removal. No other skin effects or systemic effects were noted during the observation period of 72h.

-      Source Substance No3: EDTA-ZnNa2 (CAS 14025-21-9)

The skin corrosion/irritation potential of EDTA-ZnNa2 was tested according to OECD guideline No 439 in human reconstructed skin (EPISKIN; Verbaan et al, 2013). At least 10 mg EDTA-ZnNa2 powder was applied for 15 min on top of moistened skin tissue. After a 42 hour incubation period, determination of the cytotoxic (irritancy) effect was performed. EDTA-ZnNa2 was non-irritant in the in vitro skin irritation test under the experimental conditions.

 

-      Source Substance No4: EDTA-CuNa2 (CAS 14025-15-1)

In a non-GLP study according to OECD guideline No 404, skin irritation potential of the test substance was tested (BASF, 1975). The applicant site of 2.5 x 2.5cm of 3 white Vienna rabbits was exposed for 4h to a 50% dilution of test item using semiocclusive conditions and subsequently was washed with Lutrol and Lutrol/water (1:1) followed by a 72 hours observation period with readings after 30 min and 60 min after removal of the test patches and 24 h, 48 h and 72 h after the beginning of application. Untreated skin sites of the same animal were used as negative control.  No signs of irritation or corrosion were observed at any time.

-      Source Substance No5: EDTA-MgNa2 (CAS 14402-88-1)

The skin corrosion/irritation potential of EDTA-MgNa2 was tested according to OECD guideline No 439 in human reconstructed skin (EPISKIN; Verbaan et al, 2013). About 12 mg EDTA-MgNa2 powder was applied for 15 min on top of moistened skin tissue. After a 42 hour incubation period, determination of the cytotoxic (irritancy) effect was performed. EDTA-MgNa2 was non-irritant in the in vitro skin irritation test under the experimental conditions.

 

b)   Eye irritation

a.   Target Substance:EDTA cobalt copper iron manganese zinc complexes(CAS 234446-82-3)

There were no studies available with the target substance for skin irritation/corrosion.

 

b.   Source Substances

-      Source Substance No1: EDTA-MnNa2 (CAS 15375-84-5)

The eye corrosion/irritation potential of EDTA-MnNa2 was tested according to OECD guideline No 437 in isolated bovine cornea (BCOP; Verspeek et al, 2010). 20% EDTA-MnNa2 solution was applied for 240 min on top of the cornea. After the incubation period, determination of the opacity and permeability values was performed and the read-out was compared to negative controls and the laboratory historical control range. The mean in vitro irritation score (IVIS) was 2.4, thus, EDTA-MnNa2 was considered to be non-irritant.

-      Source Substance No2: EDTA-FeNa (CAS 15708-41-5)

An eye irritation study with three male Himalyan rabbits according to OECD guideline 405 and GLP was performed. A single administration of 100 mg FeNaEDTA per animal into the conjunctival sac of the right eye of three rabbits caused the following effects: Corneal opacity (grade 1) was observed in animal no. three 24 hours to 6 days after instillation. The fluorescein test performed 24 hours after instillation revealed corneal staining in animal no. 3 (up to 1/4 of the surface). Conjunctival redness (grade 1) was observed in animal no. one 60 minutes, in animal no. two 60 minutes to 48 hours and in animal no. three 24 hours to 4 days after instillation. In addition, secretion was observed in all 3 animals 1 hour after instillation. Irises were not affected by instillation of the test item. There were no systemic intolerance reactions.

 

-      Source Substance No3: EDTA-ZnNa2 (CAS 14025-21-9)

The eye corrosion/irritation potential of EDTA-ZnNa2 was tested according to OECD guideline No 437 in isolated bovine cornea (BCOP; Verspeek et al, 2012). Since no workable suspension in physiological saline could be obtained, the test substance was used as delivered (powder) and added pure on top of the moistened corneas (305 to 319 mg). After a 6 hour incubation period, determination of the opacity and permeability values was performed and the read-out was compared to negative controls and the laboratory historical control range. EDTA-ZnNa2 was considered to be not a severe irritant or corrosive in the Bovine Corneal Opacity and Permeability test under the experimental conditions.

 

-      Source Substance No4: EDTA-CuNa2 (CAS 14025-15-1)

o  in-vivo study

The eye irritancy potential of EDTA-CuNa2 was determined according to OECD guideline No 405 (BASF, 1985). 0.1ml bulk volume (about 61 mg) of the comminuted test substance was applied to the conjunctival sac of one eye of three white Vienna rabbits, respectively. The saline-treated respective adjacent eye served as control. The eyes were not washed out after 24 hours as specified in OECD guideline 405. Irritancy parameters were recorded after 1, 24, 48, 72 hours and on day 8. Average scores after 24/48/72 hours were 1.3/1.6 and 2.0 for conjunctival redness, 1.0/0.7/1.0 for corneal opacity. Because 2/3 animals showed corneal opacity grade 1 (mean 24, 48 and 72 h) which was reversible in one animal after 8 days and is expected to be fully reversible within an observation period of 21 days, the test material was classified as eye irritant. 

Due to the fact that EDTA-CuNa2 is ingredient of the target substance in concentrations of only up to 9% and in order to recheck this borderline result, an in vitro study with EDTA-CuNa2 (CAS 14025-15-1) was performed (see below).

o  in vitro study

The potential of Librel RMX 8 (17% EDTA-CuNa2) to cause ocular irritation was assessed by a single topical application of 50μL bulk volume (about 35 mg) of the undiluted test substance in a GLP-compliant non-guideline study to a reconstructed three dimensional human cornea model (EpiOcular™; BASF 2013).

Tissue samples were incubated with the test substance for 90 minutes followed by an 18-hours post-incubation period. Cytotoxicity was determined using the MTT-method, i.e. by measuring the metabolic activity of the tissue after exposure/post-incubation using a colorimetric test. The test substance was able to reduce MTT directly. However, this ability of direct MTT reduction did not impair the study result as demonstrated by the concurrently performed exposure of control tissues inactivated by freezing. The mean viability of the test-substance treated tissues was 89 %.

Based on the observed results and applying the evaluation criteria it was concluded, that the test substance does not show an eye irritation potential in the EpiOcular™ eye irritation test under the test conditions chosen.

-      Source Substance No5: EDTA-MgNa2 (CAS 14402-88-1)

The eye corrosion/irritation potential of EDTA-MgNa2 was tested according to OECD guideline No 437 in isolated bovine cornea (BCOP; Verspeek et al, 2010). 20% EDTA-MgNa2 solution was applied for 240 min on top of the cornea. After the incubation period, determination of the opacity and permeability values was performed and the read-out was compared to negative controls and the laboratory historical control range. The mean in vitro irritation score (IVIS) was 2.4, thus, EDTA-MgNa2 was considered to be non-irritant.

 

4.   Key study assignment

There are no information according skin irritation available for the target substance. In a weight of evidence approach in vivo or in vitro studies with five different source substances, which are ingredients of the target substance, where used to assess the skin irritancy potential of the target substance. All studies are well documented, reliable and reveal consistent negative results for skin irritation.

 

There are no information according eye irritation available for the target substance. In a weight of evidence approach in vivo or in vitro studies with five different source substances, which are ingredients of the target substance where used to assess the eye irritancy potential of the target substance. All studies are well documented, reliable and most reveal consistent negative results for eye irritation. EDTA-CuNa2 was the only ingredient with irritant properties, however, up to a concentration of 17% EDTA-CuNa2 no irritation potential was determined in vitro.

 

5.   Conclusion

Several reliable studies with different read-across substances (CAS15375-84-5, CAS 15708-41-5, CAS 14025-21-9, CAS 14025-15-1, CAS 14402-88-1), which are also ingredients of the target substance,areavailable to assess the skin irritancy potential of the target substance.Due to the consistent negative results the target substance is considered to be non-irritant..

Several reliable studies with different read-across substances ((CAS15375-84-5, CAS 15708-41-5, CAS 14025-21-9, CAS 14025-15-1, CAS 14402-88-1), which are also ingredients of the target substance,areavailable to assess the eye irritancy potential of the target substance. Only one component of the target substance, EDTA-CuNa2, had irritant properties. However, up to concentrations of 17% of EDTA-CuNa2 the ingredient was not irritant. Due to the fact that the maximal concentration of EDTA-CuNa2 in the target substance is 9% no classification of the target substance is warranted.

 

In conclusion, it is not warranted to classify the target substance as skin or eye irritant.

Justification for selection of skin irritation / corrosion endpoint:
Weight of evidence approach with five different source substances, which are ingredients of the target substance.

Justification for selection of eye irritation endpoint:
Weight of evidence approach with five different source substances, which are ingredients of the target substance.

Justification for classification or non-classification

Justification

Based on the above information received for structurally similar source substances, which are ingredients of the target substance, a classification of the target substance as skin or eye irritant according to Regulation (EC) 1272/2008 or Directive 67/548/EEC is not justified.

- GHS: no classification

- DSD: no classificatio