Magnesium, EDTA cobalt copper iron manganese zinc complexes

Endpoint summary

Description of key information

Structurally related source substances (EDTA-MnNa, EDTA-CuNa and EDTA-FeNa) ingredients of the target substance, showed no adverse effects relevant for classification after repeated oral administration. Therefore it is not justified to classify the target substance Magnesium, EDTA cobalt copper iron manganese zinc complexes (CAS 234446-82-3).

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LOAEL

150 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Additional information

1.   Hypothesis for the analogue approach

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). ). In this case the related substances mentioned below are ingredients of Magnesium, EDTA cobalt copper iron manganese zinc complexes(CAS 234446-82-3).

 

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
Name	Magnesium, EDTA cobalt copper iron manganese zinc complexes	EDTA-MnNA2	EDTA-FeNa	EDTA-CuNa2
CAS	234446-82-3	15375-84-5	15708-41-5	14025-15-1
% in CAS 234446-82-3	n.a.	30	25	9
Subchronic oral toxicity study (90d)	--	NOAEL: 500 mg/kg/d	NOAEL: >=84 mg/kg/d (highest tested concentration)	NOEL: ~150 mg/kg/d

 

Lack of data for a given endpoint is indicated by “--“.

 

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 is used to predict the respective endpoints forMagnesium, EDTA cobalt copper iron manganese zinc complexes(CAS 234446-82-3).

 

Since no subchronic studies investigating the repeated dose toxicity are available forMagnesium, EDTA cobalt copper iron manganese zinc complexes(CAS234446-82-3), a read-across from the structurally related source substancesEDTA-MnNA2 (CAS 15375-84-5), EDTA-FeNa (CAS 15708-41-5) and EDTA-CuNa2 (CAS 14025-15-1)were usedin accordance to Regulation (EC) No. 1907/2006 Annex XI, 1.5.

 

3.   Results

 

3.2 Oral

a.   Target Substance:

EDTA cobalt copper iron manganese zinc complexes(CAS 234446-82-3)

There were no studies available with the target substance for repeated dose toxicity via inhalation.

 

b.   Source substances

-      Source Substance No1: EDTA-MnNA2 (CAS 15375-84-5)

Repeated doses of 150, 500 or 1500 mg/kg bw/d of EDTA-MnNa2 (gavage) was determined in an extended repro-screening study equivalent or similar to OECD guideline No 408 (Wolterbeek, 2010).

No relevant treatment-related effects were observed in clinical observations, for neurobehavior, body weights and body weight changes,food consumption , haematology and clinical chemistry or necropsy.

Water consumption was measured during 2 consecutive days of two weeks during the premating period. During all these 4 days, water consumption of particularly male and also of female animals treated with the highest concentration of the test item was increased. Most probably, this effect was due to the high sodium exposure of these animals via the test item.

Urinary volume was increased in the male animals of the mid- and highest dose groups and in the female animals of the high dose group which resulted in an increased concentration of creatinine. The absolute amount of creatinine excreted was not affected. The sodium concentration and the sodium/creatinine ratio was statistically significantly increased in both male and female animals of the two groups treated with the highest concentration of the test item (irrespectively of dietary zinc supplementation).

Absolute and relative weights of the kidneys of the male and females of the two groups treated with the highest concentration of the test item (irrespectively of dietary zinc supplementation) were statistically significantly increased. Furthermore, the absolute and relative weights of the spleen of the female animals of the highest dose group with supplementary zinc was statistically significantly decreased.

In the two groups treated with the highest concentration of the test item (irrespectively of dietary zinc supplementation) an increase in the incidence of rats showing very slight diffuse subcortical tubular dilatation was observed in the kidneys, reaching the level of statistically significance in the female animals only.

Based on the results of this repeated dose study (specifically water consumption, urinary sodium concentration, weight of and histopathological effects in kidneys as observed in the animals treated with the highest concentration of the test item), the No Observed Adverse Effect Level (NOAEL) was 500 mg/kg body weight/day.

 

-      Source substance No 2: EDTA-FeNa (CAS 15708-41-5)

Repeated dose toxicity of EDTA-FeNa was determined according to OECD guideline No 407 (Appel, 2001). A study was performed to provide data on the disposition, accumulation and toxicity of sodium iron EDTA in comparison with iron (II) sulfate in rats on administration via the diet for 31 and 61 days. Clinical signs, body weights, food consumption, food conversion efficiency, hematology, clinical chemistry and pathology of selected organs were used as criteria for disclosing possible harmful effects. Determination of iron and total iron binding capacity in blood plasma and non-heme iron analysis in liver, spleen and kidneys were used to assess the disposition and accumulation of iron originating from sodium iron EDTA or iron (II) sulfate.

It was concluded that, under the conditions of the present study, iron is accumulated from the diet in liver, spleen and kidneys in a dose-dependent manner, and iron derived from FeEDTA is taken up and/or accumulated less efficiently in liver and spleen than iron from FeSO4.

Moreover, feeding iron up to 11.5 and 11.2 mg/kg body weight/day, derived from FeSO4 and FeEDTA, respectively, did not result in tissue iron excess nor in any other toxicologically significant effects. The latter value corresponds to 84 mg EDTA-FeNa.3H2O per kg bw/day.

-      Source substance No 3: EDTA-CuNa2 (CAS 14025-15-1)

Repeated dose toxicity of 150, 500 or 1500/1050 mg/kg bw/d of EDTA-CuNa2 (gavage) was determined in an extended repro-screening study equivalent or similar to OECD guideline No 408 (Lina, 2013). Due to mortality, the high-dose level was reduced to 1050 mg/kg bw/day from day 9 of the study.

No treatment-related effects were found for neurobehavior and motor activity, ophthalmoscopy.

Clinical signs observed in rats of the high-dose group and, to a lesser extent, in the mid-dose group included thin appearance, hunched posture, piloerection, blepharospasm, swollen abdomen, soft faeces and green watery discharge around perineum.

Body weights were decreased in males of the high-dose group, and, from the end of the premating period, in males of the mid-dose group. During lactation, female body weights were increased in the remaining treatment groups. Feed intake was reduced in males of the high-dose group.

Haematology and clinical chemistry was conductedin females on day 65 (at the end of the premating period), and in males on day 85 (a few days prior to necropsy). At these time points, all males and most females of the high-dose group had died or had been killed.

The following changes in haematology were noted in the mid-dose group (now representing the highest dose level):

-       Prothrombin time was decreased in both sexes.

-       Red blood cell count was increased and MCV and MCH were decreased in males.

-       Total white blood cell counts and absolute neutrophils and monocytes counts were increased in males. 

The following changes in clinical chemistry were noted:

-       ALP, ASAT, ALAT and GGT activity were increased in males of the mid-dose group. In females ALP activity (mid-dose group) and ALAT activity (low- and mid-dose group) were decreased.

-       Bilirubin was increased in males of the mid-dose group

-       Creatinine was increased in the mid-dose group in both sexes

-       Urea, inorganic phosphate and calcium were increased in mid-dose males

-       Sodium was increased in males of the low- and mid-dose group. Potassium was increased in females of the mid-dose group.

The surviving rats (control, low- and mid-dose group) were killed on day 90 (males) or on day 4 of lactation (females).

-       Terminal body weights were decreased in males and increased in females of the mid-dose group.

-       The absolute and the relative weights of the heart were decreased in the mid-dose group in both sexes.

-       The relative weight of the kidneys was increased in males of the mid-dose group. In females of this group, the absolute kidney weight was increased.

-       The absolute and the relative weights of the spleen were increased in the mid-dose females.

-       The absolute and the relative weights of the ovaries were decreased in females of the mid-dose group.

-       The absolute weight of the testis was decreased in mid-dose males. 

The main gross finding in animals of the mid-dose group (and in intercurrently killed animals of the high-dose group) were enlarged intestines with green/watery contents, a pale and/or green appearance of the liver and kidneys, small epididymides and seminal vesicles, enlarged dark spleen, small thymus and a variety of changes in the stomach.

 

Microscopic examination revealed histopathological changes in the kidneys, the liver and the spleen.

-       The histopathological changes in the kidneys were characterised by tubular necrosis and degeneration, tubular epithelial cell karyomegaly and accumulation of brown pigment. These changes were mainly present in the mid-dose animals. However, tubular epithelial brown pigment was also noted in the kidneys of 6/10 low-dose males.

-       The changes in the liver were accumulation of periportal macrophages, especially in the mid-dose males, hepatocellular karyomegaly, brown pigment accumulation, bile duct hyperplasia and (multi)focal infiltration of mononuclear inflammatory cells. These changes were mainly present in the mid-dose animals. However, mononuclear cell infiltrate was also noted in the liver of 6/10 low-dose males and 3/10 low-dose females.

-       The changes in the spleen were accumulation of brown pigment and accumulation of macrophages in the white pulp in animals of the mid-dose group.

The decedent high-dose animals were subjected to microscopical examination only on the basis of macroscopic observations. The microscopic observations in this group confirmed the above histopathological changes.

 

Based on the histopathological effects in liver and kidneys noted in animals of the low-dose group, the no-observed-effect level (NOEL) for parental toxicity was lower than 150 mg/kg bw/day. However, because only limited effects were observed in the low-dose group, it was speculated that the NOEL was close to 150 mg/kg bw/day.

 

Key study assignment:

 

Since no subchronic study investigating the repeated dose oral toxicity are availableMagnesium, EDTA cobalt copper iron manganese zinc complexes(CAS234446-82-3), its repeated dose toxicity was assessed in a weight of evidence approach. Available studies from structurally related analogue substancesEDTA-MnNA2 (CAS 15375-84-5), EDTA-FeNa (CAS 15708-41-5) and EDTA-CuNa2 (CAS 14025-15-1) were usedin accordance to Regulation (EC) No. 1907/2006 Annex XI, 1.5.

 

Conclusion/Disussion

a)   inhalation

Inhalation exposure of rats to EDTA-Na2H2 for 90 days (65 exposures) according to OECD guideline No 413 did not lead to any substance-related clinical signs of toxicity(BASF, 2014) nor were there any effect in clinical chemistry, hematology. Histological examination revealed some effects in larynx at the highest tested concentration of 15 mg/m³ in female animals. No signs of systemic toxicity were observed up to a concentration of 15 mg/m³. Signs of local toxicity were observed only at the high concentration of 15 mg/m³ in female animals. The NOAEC was 3 mg/m3 under the study conditions. However, although a structurally related substance, EDTA-Na2H2, was used in the inhalation study, the results cannot be transferred directly to the target substance. The local effects induced by EDTA-Na2H2 most likely are related to chelation of bivalent physiological ions, such as Ca2+ or Zn2+, and depletion of bivalent cations from the respiratory tract (larynx) membrane. However, if the chelant is already bound to bivalent ions with equal or higher conditional complex building constants or if such bivalent ions are present, as it is the case for the target substance, no effects after repeated exposure are expected. This is supported by the fact that the acute inhalation toxicity of the target substance (see endpoint acute toxicity) is much lower than for the “empty” chelate, EDTA-Na2H2, that is loaded with monovalent ions and that induced acute toxicity by induction of local effects. In conclusion, no adverse effects after repeated inhalation of the target substance which are relevant for classification and labelling are expected.

b)   oral

In conclusion, ingredients of the target substance were tested for repeated oral toxicity. the available data on repeated dose toxicity via the oral route with structurally relatedsource substancesshowed no adverse effects which are relevant for classification. The NOAEL for EDTA-MnNa2, which is included in the target substance to 30%, is 500 mg/kg bw/d and the LOAEL for EDTA-CuNa2, which is included in the target substance to 9%, is 150 mg/kg bw/d.

  

c)   Dermal

No data are available for the target substance on repeated exposure via the dermal route. However, dermal uptake is expected to be very low (<0.001%) based on results obtained with EDTA and because of its structure.

Altogether, structurally relatedsource substances, ingredients of the target substance,showed no adverse effects relevant for classification after repeated oral administration. Therefore it is not justified to classify the target substanceMagnesium, EDTA cobalt copper iron manganese zinc complexes(CAS234446-82-3).

 

References

 

BASF (2014), Trilon BD - 90-day inhalation study in Wistar rats dust exposure, RepNo. 50I0628/09I127

 

Lina BAR (2013) Combined oral repeated dose toxicity study with reproduction/developmental toxicity screening test with EDTA-CuNa2 in rats. TNO

Triskelion, Zeist, the Netherlands. Report no. V20114

 

Wolterbeek APM (2010) Combined oral repeated dose toxicity study with the reproduction/developemental toxicity screening test with EDTA-MnNa2 in rats. TNO

Quality of Life, Zeist, the Netherlands, Report no. V8650

 

 

 

 

 

 

 

 [SB1]Include Reference

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
Most relevant and reliable study

Justification for classification or non-classification

There are currently no data available leading to a classification ofMagnesium, EDTA cobalt copper iron manganese zinc complexes(CAS234446-82-3)for its repeated oral, dermal and inhalation toxicity according to Regulation No 1272/2008/EC (CLP) or Directive 67/548/EEC.

 

Labelling repeated dose toxicity:

GHS: no classification

DSD: no classification