Hydrogen (glycinato-N,O)[sulphato(2-)-O,O']ferrate(1-)

Endpoint summary

Description of key information

- study conducted according to OECD guideline 425, ferrous lysinate sulfate was applied to male and female Sprague Dawley rats by oral gavage at doses of 175, 550, 2000 mg/kg bw, the animals were observed for 14 days, no mortality or any sign of toxicity occurred, LD50 > 2000 mg/kg bw, read-across

- study conducted according to OECD guideline 401, male and female rats were administered increasing doses of ferrous glycinate up to a dose of 2800 mg/kg bw, no mortality or clinical signs of intoxication were reported, LD50 > 2800 mg/kg bw.

- study conducted similar to OECD guideline 401, study was conducted prior to implementation of GLP and OECD guidelines, male and female Harlan-Wistar rats were administered ferrous glycinate sulfate in a dose range of 4454 to 7014 mg, the LD50 was reported 5590 mg/kg bw.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2019-07-26 to 2019-10-31

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 425 (Acute Oral Toxicity: Up-and-Down Procedure)

Version / remarks:

Federal Law No. 61–FZ “On the Circulation of Medicinal Products” of 12 April 2010 (as amended);Federal Law No. 323–FZ “On the Basics of Health Protection of the Citizens in the Russian Federation” of 21 November 2011 (as amended)

GLP compliance:

yes

Test type:

up-and-down procedure

Limit test:

no

Species:

rat

Strain:

not specified

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: NPO House of Pharmacy (inhouse bred)
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 8-12 weeks
- Fasting period before study: 16h
- Housing: individually in standard transparent plastic cages. Wood pellets were used as bedding.
- Diet (e.g. ad libitum): Feed for laboratory animals PK-120-1 prepared in accordance with GOST R50258-92 "Compound Feeds for Laboratory Animals. Specifications” was given ad libitum
- Water (e.g. ad libitum): purified water normalized in respect of organoleptic properties, pH, solids, reducing substances, carbon dioxide, nitrates and nitrites, ammonia, chlorides, sulphates, calcium and heavy metals in accordance with SanPiN 2.1.4.1074-01 "Drinking Water. Hygienic Requirements for the Quality of Water from Centralized Drinking Water Supply Systems. Quality Control". Water in standard drinking bowls with steel nose caps was given ad libitum.
- Acclimation period: 5 days
- Method of randomisation in assigning animals to test and control groups: Randomization was not expected in this study, since dosing occurred in stages and individually. The main criterion for including an animal in the experiment was its body weight.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-26 °C
- Humidity (%): 30-70%
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From:
No. 2.0-31.05/19 of 31 May 2019
No. 2.0-30.06/19 of 01 July 2019

Route of administration:

oral: gavage

Vehicle:

other: 1% starch solution

Details on oral exposure:

VEHICLE
- Concentration in vehicle: Before administration, suspensions of the test article was prepared with concentrations of ≈ 29.2 mg/mL for 175 mg/kg, ≈ 91.7 mg/ml for 550 mg/kg and 333.3 mg/ml for 2000 mg/kg.
- Amount of vehicle (if gavage): 1.5 mL per 250g rat
- Lot/batch no. (if required): М-4.38/19

MAXIMUM DOSE VOLUME APPLIED: 333.3 mg/mL

CLASS METHOD
- Rationale for the selection of the starting dose:since the test article is presumably a low toxicity substance, 175 mg/kg was selected as the starting dose for a single intragastric administration to one male and one female rat.

Doses:

175, 550, 2000 mg/kg bw

No. of animals per sex per dose:

175 mg/kg bw: 1 animal; 550 mg/kg bw: 1 animal; 2000 mg/kg bw 3 animals

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: On day 1, 2, 7 and 15
- Necropsy of survivors performed: yes
- Clinical signs: daily
- Other examinations performed: other:Clinical examination on day 2, 7, 14 and local tolerance evaluation on day 15

Statistics:

Calculation of LD50 with confidence intervals was performed using the AOT 425 StatPgm program (Westat, USA).

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No deaths were observed

Clinical signs:

other: No clinical signs of intoxication were observed.

Gross pathology:

No gross abnormalities in the internal organs were identified at the scheduled necropsy in experimental animals treated with a single intragastric dose

Other findings:

The test article, ferrous monolysinate sulfate, at 175 mg/kg (in 1 male) and at 2000 mg/kg (in 1 male and 1 female) following single-dose intragastric administration had a moderate local irritant effect, which was manifested as catarrhal gastritis detected by a histological examination of the stomach tissues.

Interpretation of results:

other: According to Regulation (EU) No. 1272/2008 (CLP) the substance does not need to be classified as acutely toxic by oral route

Conclusions:

LD50 for ferrous monolysinate sulfate is > 2000 mg/kg following single-dose intragastric administration to male and female rats. Ferrous monolysinate sulfate can be classified as toxicity category 5 according to the international GHS classification.

Executive summary:

In an acute oral toxicity study according to OECD guideline 425, fasted, 8-12 weeks old Sprague Dawley strain rats were given a single oral dose of ferrous monolysinate sulfate in 1% starch solution by gavage at a dose of 175 mg/kg bw: 1 animal; 550 mg/kg bw: 1 animal; 2000 mg/kg bw 3 animals and were observed for 14 days.

None of the animals died. No clinical signs were observed in the treated animals. No gross abnormalities in the internal organs were identified at the scheduled necropsy in experimental animals treated with a single intragastric dose. The test article, at 175 mg/kg (in 1 male) and at 2000 mg/kg (in 1 male and 1 female) following single-dose intragastric administration had a moderate local irritant effect, which was manifested as catarrhal gastritis detected by a histological examination of the stomach tissues.

 

Oral LD50 (rat, females/males) > 2000  mg/kg bw

Reference 2

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Version / remarks:

study performed similar to OECD Guideline 401 adopted 24th February 1987

Deviations:

no

Remarks:

number of dose groups were not reported

Principles of method if other than guideline:

- Principle of test: This study compares the effects of Iron-supplementation using ferrous sulfate, iron amino acid chelate and iron polymaltose complex.
- Short description of test conditions: Sprague-Dawley rats were administered orally different doses of these compounds either once or repeated for 4 weeks or 4 months. At the end of the repeated dose studies additionally biochemistry parameters were determined as well as immunohistopathology and gross necropsy were performed. For the determination of the LD50 only mortality was reported.

GLP compliance:

not specified

Test type:

acute toxic class method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: not specified
- Weight at study initiation: 200-220 g
- Fasting period before study: 18h prior to dosing
- Diet (e.g. ad libitum): ad libitum, standard rat chow (Cooperación, Argentina)
- Water (e.g. ad libitum): ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C

Route of administration:

oral: gavage

Vehicle:

not specified

Doses:

increasing doses up to 2800 mg/kg bw

No. of animals per sex per dose:

6

Control animals:

yes

Remarks:

only vehicle was administered

Details on study design:

- Duration of observation period following administration: Clinical signs of the animals were observed 4 times on the dose day and thereafter once a day. The number of deaths was counted at 24 h after treatment.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 800 mg/kg bw

Based on:

test mat.

Remarks on result:

other: no deaths occurred up to the highest dose administered no clinical signs were reported.

Mortality:

none

Clinical signs:

other: none reported

Gross pathology:

no

Other findings:

not reported

Interpretation of results:

study cannot be used for classification

Conclusions:

In the present publication of Toblli et al. (2008) female and male Sprague Dawley rats were administered various doses of a ferrous amino acid chelate by stomach tube. Clinical signs were recorded 4 times at the dosing day and once thereafter. The number of deaths were counted after 24h after dosing. The mean LD50 value reported was > 2800 mg Fe/kg bw. Thus, ferrous amino acid chelates should be considered to be classified as Category 5 'may be harmful if swallowed' according to the Globally Harmonized System for Classification and Labelling of Chemicals (GHS). According to Regulation (EU) No. 1272/2008 (CLP) no classification is necessary.

Executive summary:

In an acute oral toxicity study similar to OECD guideline 401, fasted, Sprague Dawley strain rats weighing 200-220g were given a single oral dose of a not further specified range of ferrous amino acid chelates by gavage and were observed for 24 h.

The exact number of animals that died was not reported. No clinical signs were not reported. The LD50 value was reported to be > 2800 mg Fe/kg bw..

 

Oral LD50 (rat, females/males) > 2000  mg/kg bw

Reference 3

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

yes

Remarks:

Determination of LD50 value in rats after 24h, no further observation

Principles of method if other than guideline:

Male (250 to 350 g) and female (150 to 250 g) Harlan-Wistar rats in mixed groups of 6 received the iron compounds i.g. in attempts to determine 24-hour LD50 in this species.

GLP compliance:

no

Remarks:

The study was conducted prior to implementation of the GLP Guideline

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: male: 250 to 350g, female: 150 to 250g.

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: not reported

DOSAGE PREPARATION (if unusual): Tablets of ferroglycine sulfate (Ferronord®, one tablet conteining 400 mg of the substance corresponding to 75 mg iron) were dissolved in water

Doses:

not reported, dose range 4454 to 7014 mg, dose at which the LD50 was reported 5590 mg/kg bw

No. of animals per sex per dose:

6

Control animals:

no

Details on study design:

- Duration of observation period following administration: 24 h
- Necropsy of survivors performed: no

Statistics:

The animals were observed closely for several hours and the LD50 and 95% confidence limits were determined at the end of 24 hours by the method of Litchfield and Wilcoxon.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

5 590 mg/kg bw

Based on:

test mat.

Mortality:

not reported for the dose range tested 4454 to 7014 mg

Clinical signs:

other: not reported

Gross pathology:

Not performed for the test in rats

None of the tested compounds are very toxic following i.g. administration to rats and evidently there are only slight differences in the lethal effects of ferrous sulfate, ferrous gluconate and ferroglycine sulfate complex. There was no significant difference between the 1- and 7-day LD50 for ferrous sulfate and iron-carbohydrate complex in rats.

Interpretation of results:

GHS criteria not met

Conclusions:

In the present study the LD50 value of ferroglycine sulfate was determined in male and female Harlan Wistar rats. The study was conducted prior to implementation of OECD and GLP guidelines but the performance is similar to the standard acute method. The LD50 value was determined to be 5590 mg/kg bw. Thus, the substance is not classified according to Regulation (EC) No 1272/2008 (CLP) and the Globally Harmonized System for Classification and Labelling of Chemicals (GHS) with respect to acute oral toxicity.

Executive summary:

In an acute oral toxicity study similar to OECD guideline 401 (study was conducted prior to implementation of OECD and GLP guidelines), groups of 6 female and male Harlan Wistar rats were given a single oral dose of ferroglycine sulfate in a dose range of 4454 to 7016 mg/kg bw in water and observed for 1 day.

 

Oral LD50 Female/Male Combined = 5590 mg/kg bw

 

Ferroglycine sulfate is of LOW Toxicity based on the LD50 in males and females and thus not classified according to Regulation (EC) No 1272/2008 (CLP) and the Globally Harmonized System for Classification and Labelling of Chemicals (GHS).                                                                               

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

The study used for classification of the substance was performed according to OECD guideline 425 and under GLP-conditions and is therefore considered adequate.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Two studies are available investigating the effects of ferrous glycinate sulfate in rats after single oral administration. Both studies (Toblli, 2008 and Weaver, 1961) have acceptable limitations as the documentation was not detailed enough (Toblli, 2008) and the study was performed prior to implementation of OECD and GLP guidelines (Weaver, 1961). However, a third study was conducted according to OECD guideline 425 in male and female Sprague Dawley rats with the similar substance ferrous lysinate sulfate. Since the amino acid moiety contained either in ferrous glycinate sulfate or in ferrous lysinate sulfate is not considered to be the constituent responsible for the toxicity of the substance ferrous glycinate sulfate, the results obtained from the study conducted with ferrous lysinate sulfate are considered the most reliable. as described in the justification for read-across in Chapter 13 acute oral toxicity data are available (data matrix) indicating that lysine exhibits even a lower acute oral toxicity than compared to glycine when administered to rats (Breglia et al., 1973). Thus, in both cases the metal cation is the constituent that determines the toxicity of the target substance. Therefore, the maximum dose used in this study was 2000 mg/kg bw. No mortality or clinical signs of intoxication were observed in this study, thus, the LD50 for ferrous glycinate sulfate is set to > 2000 mg/kg bw.

Justification for classification or non-classification

Based on the available data ferrous glycinate sulfate is not classified with regard to acute oral toxicity. According to Regulation (EU) No.1272/2008 (CLP) no classification is required.