isliFe

Endpoint summary

Description of key information

The information on repeated dose toxicity of Acetic acid, oxo-, sodium salt, reaction products with 2-aminoethanol and phenol, sodium hydroxide and iron trichloride (MEAHA-Fe), is based on read-across from the information available for Acetic acid, oxo-, sodium salt, reaction products with ethylenediamine and phenol, iron sodium salts (FeNa-EDDHA, CAS 84539-55-9, EC 283-044-5). The substances are structural analogues and have similar physicochemical properties and are therefore assumed to have similar (eco)toxicity profiles.

In a 90-day oral (gavage) key repeated dose toxicity study in rats (Novartis Crop Protection AG, 1998), the NOAEL for FeNa-EDDHA was established at 10 mg/kg bw/day based on a transient normochromic anaemia present at higher dose levels (estimated from LOAEL). Changes in clinical laboratory parameters noted at higher dose levels and indicative of effects on kidneys and/or liver were without microscopic correlate under the conditions of this study. A supporting and preceding dose range finding subacute oral (gavage) toxicity study (CIBA-GEIGY Limited, 1996a) in rats provided further indication that the haematopoietic system and, at higher dose levels, the kidney represented target organs following repeated oral exposure.

In a key subacute 28-day dermal toxicity study (CIBA-GEIGY Limited, 1996b), the NOEL for FeNa-EDDHA was established at 100 mg/kg bw/day based on slight effects on the liver and skin and due to increased adrenal weight noted at the high dose level of 1000 mg/kg bw/day. No data on repeated inhalation exposure are required. Exposure by the inhalation route is considered to be negligible.      

Similar toxicity is expected for read-across target MEAHA-Fe.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

chronic toxicity: oral

Type of information:

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

Adequacy of study:

key study

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across: supporting information

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

no effects observed

Ophthalmological findings:

no effects observed

Haematological findings:

effects observed, treatment-related

Clinical biochemistry findings:

effects observed, treatment-related

Urinalysis findings:

effects observed, treatment-related

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

not specified

Details on results:

CLINICAL SIGNS AND MORTALITY
There were no test item related clinical findings. No deaths occurred.

BODY WEIGHT AND WEIGHT GAIN
Decreased mean body weights were noted from weeks 5 and 8 onwards at 200 mg/kg bw/day in male and female rats, respectively. The mean body weight gains were decreased by the end of the treatment period in animals treated with 200 mg/kg bw/day. During the recovery period, body weight gain in animals previously treated with 200 mg/kg bw/day was higher as compared to controls.

FOOD CONSUMPTION
Food consumption and food consumption ratios were decreased at 200 mg/kg bw/day in both sexes. During the recovery period food intake improved.

WATER CONSUMPTION
The mean water consumption was not influenced by treatment.

HAEMATOLOGY
At the end of the treatment period, a normochromic anaemia with lower erythrocyte count, haemoglobin concentration and haematocrit was observed in males and females at 200 mg/kg bw/day and males at 50 mg/kg bw/day. A higher reticulocyte count associated with higher MCV and MCH values and reduced white blood cell, basophil, lymphocyte and monocyte counts were confined to males at 200 mg/kg bw/day. A higher platelet count was recorded for males at 50 and 200 mg/kg bw/day and a higher prothrombin activity was recorded for males and females at 200 mg/kg bw/day. Evidence of reversibility for all the above parameters was apparent after the recovery period.

CLINICAL CHEMISTRY
Several clinical chemistry parameters including creatinine, urea, protein, globulin, cholesterol and sodium concentration were increased at 50 and/or 200 mg/kg bw/day. Lower potassium levels were noted in males at 200 mg/kg bw/day. All values were similar to the control group values after the 4-week recovery period.

URINALYSIS
Excretion of more acidic red-brown (males) or yellow-brown (females) discoloured urine was observed at 200 mg/kg bw/day. More acidic urine was also excreted by males at 50 mg/kg bw/day. By the end of the recovery period, the colour and pH of the urine excreted by male and females previously treated at 200 mg/kg bw/day was similar to that of control group animals.

ORGAN WEIGHTS
The mean carcass weights were decreased at the end of the treatment period at 200 mg/kg bw/day. Males at 200 mg/kg bw/day showed an elevated mean heart to body weight ratio which still was higher than the concurrent control value at the end of the recovery period.

GROSS PATHOLOGY and HISTOPATHOLOGY
There were no test item related findings.

Key result

Dose descriptor:

NOAEL

Effect level:

10 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: haematology parameters (anaemia) The NOAEL was estimated based on the LOAEL of 50 mg/kg bw/day applying an assessment factor of 5.

Critical effects observed:

not specified

Conclusions:

Under the conditions of this study, the NOAEL for FeNa-EDDHA when administered by daily oral gavage for three months was 10 mg/kg bw/day (estimated from the LOAEL).
Based on the structural similarity, the similar physico-chemical properties and the similar acute oral toxicity between FeNa-EDDHA and MEAHA-Fe, identical conclusions are drawn for the read-across target MEAHA-Fe.

Executive summary:

In a subchronic toxicity study (Novartis Crop Protection AG, 1998), FeNa-EDDHA in 0.5 % CMC and 0.1 % Tween 80 in distilled water was administered to 10 Sprague-Dawley derived rats/sex/dose level by oral gavage (10 mL/kg bw) at dose levels of 5, 50 or 200 mg/kg bw/day for a period of 90 days. Male and female animals of the concurrent control group were treated with the vehicle only. Additional 10 animals/sex of the high dose and control group were kept on control diet for a 4 -week recovery period before sacrifice. Treatment with the test item resulted in lower food intake and impaired body weight development of rats treated at 200 mg/kg bw/day. Reversible effects on the red blood cell (normochromic anaemia) and white blood cell parameters, and higher values of platelets and prothrombin activity were noted at 50 and/or 200 mg/kg bw/day. In addition, there were changes of blood chemistry and urine parameters concerning the liver and kidneys. The body weight relative heart weight was increased in males at 200 mg/kg bw/day. Under the conditions of this study, the NOAEL for FeNa-EDDHA when administered by daily oral gavage for three months was 10 mg/kg bw/day (estimated from the LOAEL).

This subchronic toxicity study in the rat is acceptable and satisfies the guideline requirement for a subchronic oral study (OECD 408).

Estimation of NOAEL:

A more realistic NOAEL was estimated from the LOAEL of 50 mg/kg bw/day applying an assessment factor of 5. This method is applicable and scientifically justified for this test, as only slight adverse effects were observed at 50 mg/kg bw/day (haematology parameters - anaemia). To take the relatively large concentration gap between 5 (clear NOEL) and 50 mg/kg bw/day into account, 5 mg/kg bw/day was not taken as NOAEL, but extrapolated form the LOAEL. The guidance on information requirements and CSA, R.8 (ECHA, 2008 -2010) recommends a factor between 3 and 10 for extrapolation from LOAEL to NOAEL. An assessment factor of 5 seems to be appropriate and conservative enough for the current study as only slight effects were observed at the LOAEL of 50 mg/kg bw/day. Consequently a NOAEL of 10 mg/kg bw/day is calculated for this study and used for DNEL and PNEC(oral) derivation.

Based on the structural similarity, the similar physico-chemical properties, similar predicted toxicokinetic behavior and the similar acute oral and local toxicity profiles between FeNa-EDDHA and MEAHA-Fe, identical conclusions are drawn for the read-across target MEAHA-Fe.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

10 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

100 mg/kg bw/day

Study duration:

subacute

Species:

rat

Additional information

The information on repeated dose toxicity of Acetic acid, oxo-, sodium salt, reaction products with 2-aminoethanol and phenol, sodium hydroxide and iron trichloride (MEAHA-Fe), is based on read-across from the information available for Acetic acid, oxo-, sodium salt, reaction products with ethylenediamine and phenol, iron sodium salts (FeNa-EDDHA, CAS 84539-55-9, EC 283-044-5). The substances are structural analogues and have similar physicochemical properties and are therefore assumed to have similar (eco)toxicity profiles.

Oral route

The subchronic toxicity by the oral route was investigated in rats (Novartis Crop Protection AG, 1998). The test item FeNa-EDDHA was administered to 10 Sprague-Dawley derived rats/sex/dose level by oral gavage at 5, 50 or 200 mg/kg bw/day for 90 days. A concurrent control group was treated with the vehicle only. Additional 10 animals/sex of the high dose and control group were kept on control diet for a 4 -week recovery period before sacrifice. Treatment with the test item resulted in lower food intake and impaired body weight development at 200 mg/kg bw/day. Reversible effects on red blood cell (normochromic anaemia) and white blood cell parameters, and higher values of platelets and prothrombin activity were noted at 50 and/or 200 mg/kg bw/day. In addition, changes of blood chemistry and urine parameters concerning the liver and kidneys were noted. The body weight relative heart weight was increased in males at 200 mg/kg bw/day. Under the conditions of this study, the NOAEL for FeNa-EDDHA when administered by daily oral gavage for three months was 10 mg/kg bw/day (estimated from the LOAEL).

Estimation of NOAEL:

A more realistic NOAEL was estimated from the LOAEL of 50 mg/kg bw/day applying an assessment factor of 5. This method is applicable and scientifically justified for this test, as only slight adverse effects were observed at 50 mg/kg bw/day (haematology parameters - anaemia). To take the relatively large concentration gap between 5 (clear NOEL) and 50 mg/kg bw/day into account, 5 mg/kg bw/day was not taken as NOAEL, but extrapolated form the LOAEL. The guidance on information requirements and CSA, R.8 (ECHA, 2008 -2010) recommends a factor between 3 and 10 for extrapolation from LOAEL to NOAEL. An assessment factor of 5 seems to be appropriate and conservative enough for the current study as only slight effects were observed at the LOAEL of 50 mg/kg bw/day. Consequently a NOAEL of 10 mg/kg bw/day is calculated for this study and used for DNEL and PNEC(oral) derivation.

In a dose range-finding subacute oral toxicity study (CIBA-GEIGY Limited, 1996a), FeNa-EDDHA was administered to 5 Sprague-Dawley derived rats/sex/dose level by oral gavage at 50, 200 or 1000 mg/kg bw/day for 28 days. A concurrent control group was treated with the vehicle only. Treatment with the test item resulted in impaired body weight development at 200 and 1000 mg/kg bw/day and correspondent lower food intake. An anaemia without erythropoietic response was noted at 200 and 1000 mg/kg bw/day. At the same dose levels, the kidney was revealed as target organ by microscopical examination, by blood chemistry data evaluation and by organ weight evaluation. In addition, body weight relative organ weight changes were noted in the heart, adrenals and spleen. However, the relevance of these findings was considered as equivocal. This study was used as scientific basis for dose level selection for the above-mentioned 90 -day repeated dose oral toxicity study in the rat.

Dermal route

In a repeated dose dermal toxicity study (CIBA-GEIGY Limited, 1996b), FeNa-EDDHA was administered to the skin of 5 Sprague-Dawley derived rats/sex/dose level at 10, 100 or 1000 mg/kg bw/day for 28 days (5 days/week). A concurrent control group was treated with the vehicle only. Dermal treatment with the test item resulted in no mortality, no relevant clinical signs, no changes in food consumption, no effects on haematology and clinical chemistry parameters and no gross findings. A transient slight body weight loss was noted in females at 1000 mg/kg bw/day during the first week of treatment. There was an increase in adrenal weight in males at 1000 mg/kg bw/day. Microscopically, the skin application sites of females at 1000 mg/kg bw/day revealed epidermal hyperkeratosis associated with an increased severity of acanthosis. In 2/5 males at 1000 mg/kg bw/day centrilobular hypertrophy of hepatocytes was noted. Based on the slight effects on the liver and skin and due to the increased adrenal weight noted at 1000 mg/kg bw/day, the NOEL for FeNa-EDDHA was established at 100 mg/kg bw/day.

Inhalation route

In accordance with column 2 of REACH Annex IX, the most appropriate route of administration, having regard to the likely route of human exposure must be tested for repeated dose toxicity.  Inhalation exposure is regarded negligible as the particle size distribution for particles below 100 µm were found to approximately 10% and no particles were detected with a diameter below 10 µm. In addition, the substance showed a very low vapour pressure (<0.0015 Pa at 20°C) and only very low toxicity after acute inhalation exposure with an LD50 of greater than 4200 mg/m³ in the rat.

Repeated dose toxicity: via oral route - systemic effects (target organ) cardiovascular / hematological: hematopoiesis

Justification for classification or non-classification

Based on the results of the key repeated dose toxicity studies with special regard to specific target organ toxicity after repeated exposure and considering the NOAEL of 10 mg/kg bw/day established for the oral route, MEAHA-Fe is not subject to classification and labelling according to Regulation No 1272/2008 (CLP) since the slight and reversible changes in haematology parameters noted at higher dose levels were of minimal toxicological relevance.