Reaction products of sodium glucoheptonate with iron sulfate and ammonium hydroxide

Acute Toxicity: oral

Administrative data

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:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Reference

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:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

The animal experiments were designed to compare the toxicologic and hematinic properties of ferrous fumarate with those of ferrous sulphate A.R.,ferrous succinate and ferrous gluconate B.P.C. The ferrous iron contents of ferrous gluconate is 11.7 per cent. Sixty-four adult rabbits having free access to food and water were dosed orally with tablets. The dose was 450 mg Fe/Kg for each ferrous salt, the rabbits receiving twelve tablets of gluconate per Kg. The gluconate tablets contained 36 mg. The gluconate tablets were broken to facilitate administration; the other tablets were administered whole.
The tablets were administered at 10 p.m. and the mortalities were recorded at 10 a.m. next morning. The surviving rabbits were then killed, and the stomachs and livers from all the animals were examined macroscopically and those from three of each group histologically. The macroscopic changes in the gastric mucosa were classified according to the scheme in Table 1, which includes the mortality figures.

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Specific details on test material used for the study:

The substances compared in these tests were ferrous fumarate, ferrous sulphate A.R., ferrous succinate and ferrous gluconate B.P.C. The ferrous iron contents of these four salts are 33.0, 20.0, 24.8 and 11.7 per cent, respectively.

Species:

rabbit

Strain:

not specified

Sex:

not specified

Details on test animals or test system and environmental conditions:

Sixty-four adult rabbits having free access to food and water

Route of administration:

oral: unspecified

Vehicle:

unchanged (no vehicle)

Doses:

450 mg Fe/kg (= 12 tablets of ferrous gluconate each containing 36 mg iron)

No. of animals per sex per dose:

not specified

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 12 hours
- Frequency of observations and weighing: not specified
- Necropsy of survivors performed: no data
- Other examinations performed: percentage mortalities was recorded, no further details available

Sex:

not specified

Dose descriptor:

other: concentration inducing the effects specified under 'Remarks'

Effect level:

450 mg/kg bw

Based on:

element

Remarks on result:

other:

Remarks:

The stomach of one revealed iron incrustation of the mucosa, while that of another showed superficial mucosal necrosis and iron in the vessels. In both, the pylorus showed early inflammation. Iron impregnation was apparent in the lamina propria of one, and there was iron in the vessels of the other. The livers showed a “chemical hepatitis,” the essential features of which were iron impregnation of parenchymal cells, their invasion and replacement by polymorphs, and an increase in intravascular polymorphs.

Mortality:

3/9

Clinical signs:

4/9 animals showed inflamation of the gastric mucosa (12 hours after giving a single dose equivalent to 450 mg Fe/kg). One of them slight and 3 of them severe and extensive inflammation

Body weight:

not examined

Gross pathology:

Gluconate: The stomach of one revealed iron incrustation of the mucosa, while that of another showed superficial mucosal necrosis and iron in the vessels. In both, the pylorus showed early inflammation. Iron impregnation was apparent in the lamina propria of one, and there was iron in the vessels of the other. The livers showed a “chemical hepatitis,” the essential features of which were iron impregnation of parenchymal cells, their invasion and replacement by polymorphs, and an increase in intravascular polymorphs.

The macroscopic changes in the gastric mucosa were classified according to the scheme in table 1, which includes the mortality figures.

The histologic findings were as follows:

Gluconate.-The stomach of one revealed iron incrustation of the mucosa, while that of another showed superficial mucosal necrosis and iron in the vessels. In both, the pylorus

showed early inflammation. Iron impregnation was apparent in the lamina propria of one, and there was iron in the vessels of the other. The livers showed a “chemical hepatitis,” the essential features of which were iron impregnation of parenchymal cells, their invasion and replacement by polymorphs, and an increase in intravascular polymorphs.

In a second experiment, four groups of three adult rabbits were dosed orally with tablets of the four iron compounds. As before, the dose employed was 450 mg Fe/Kg.

On this occasion none of the animals died within 12 hours of being dosed. They were all killed at this stage, and their stomachs were removed, washed with saline, and photographed.

Table 1 -Effects of iron gluconate tablets on Rabbit Gastric Mucosa (12 hours after giving a single dose equivalent to 450 mg Fe/kg)
Effects observed	Ferrous gluconate
None	2/9
Slight inflammation	1/9
Severe and extensive inflammation	3/9
Death within 12 hours	3/9

Executive summary:

The animal experiments were designed to compare the toxicologic and hematinic properties of ferrous fumarate with those of ferrous sulphate A.R.,ferrous succinate and ferrous gluconate B.P.C (Berenbaum, 1960). The ferrous iron contents of these four salts are 33.0, 20.0, 24.8 and 11.7 %, respectively. 64 adult rabbits having free access to food and water were dosed orally with tablets of ferrous fumarate, ferrous sulphate compound, ferrous succinate or ferrous gluconate. The dose was 450 mg Fe/Kg, the rabbits receiving seven tablets of fumarate or sulphate compound, or twelve tablets of succinate or gluconate per Kg. The fumarate and sulphate tablets contained 65 mg Fe, the succinate and gluconate tablets 36 mg. The gluconate tablets were broken to facilitate administration; the other tablets were administered whole.

The tablets were administered at 10 p.m. and the mortalities were recorded at 10 a.m. next morning. The surviving rabbits were then killed, and the stomachs and livers from all the animals were examined macroscopically and those from three of each group histologically. The macroscopic changes in the gastric mucosa were classified according to the scheme in Table 1, which includes the mortality figures. The macroscopic changes in the gastric mucosa were classified according to the scheme in table 1, which includes the mortality figures.

The histologic findings were as follows:

Gluconate.-The stomach of one revealed iron incrustation of the mucosa, while that of another showed superficial mucosal necrosis and iron in the vessels. In both, the pylorus showed early inflammation. Iron impregnation was apparent in the lamina propria of one, and there was iron in the vessels of the other. The livers showed a “chemical hepatitis,” the essential features of which were iron impregnation of parenchymal cells, their invasion and replacement by polymorphs, and an increase in intravascular polymorphs.

In a second experiment, four groups of three adult rabbits were dosed orally with tablets of the four iron compounds. As before, the dose employed was 450 mg Fe/Kg. On this occasion none of the animals died within 12 hours of being dosed. They were all killed at this stage, and their stomachs were removed, washed with saline, and photographed.

Cross-reference 2

Reason / purpose for cross-reference:

reference to same study

Reference

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1960

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

The animal experiments were designed to compare the toxicologic and hematinic properties of ferrous fumarate with those of ferrous sulphate A.R.,ferrous succinate and ferrous gluconate B.P.C. The ferrous iron contents of these four salts are 33.0, 20.0, 24.8 and 11.7 per cent, respectively. The sulphate was administered as an aqueous solution and the other three compounds as aqueous suspensions containing 0.1 per cent w/v of tragacanth.
The subchronic oral toxicities of the four ferrous salts were compared in albino rats of the WAG strain. Ferrous sulphate solution and suspensions of the fumarate, gluconate and succinate containing 20 mg Fe/ml were employed in this experiment. 45 male and 45 female rats (40 to 100 g bodyweight) were randomly distributed into nine groups of five males and five females. One group was not dosed and served as controls, while the other eight groups received oral doses of one or other of the iron compounds at a level of 50 or 100 mg Fe/Kg. The animals were individually weighed at intervals and dosed daily, excluding weekends. After 12 weeks’ dosing, the red and total white cell counts and hemoglobin concentrations were determined on two males and two females from each group. All the rats were then killed, and the major organs (liver, spleen, heart, lungs, thymus, kidneys, adrenals, thyroid, testes, prostate, seminal vesicles, ovaries and uterus) were excised, blotted dry, and weighed. The organs from two males and two females from each group dosed at 50 mg Fe/Kg were examined histologically.

GLP compliance:

no

Limit test:

no

Specific details on test material used for the study:

The substances compared in these tests were ferrous fumarate, ferrous sulphate A.R., ferrous succinate and ferrous gluconate B.P.C. The ferrous iron contents of these four salts are 33.0, 20.0, 24.8 and 11.7 per cent, respectively.

Species:

rat

Strain:

other: albino rats of the WAG strain

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: not specified
- Females (if applicable) nulliparous and non-pregnant: not specified
- Age at study initiation: not specified
- Weight at study initiation: 40 to 100 g bodyweight
- Fasting period before study: not specified
- Housing: 5 per group
- Diet (e.g. ad libitum): not specified
- Water (e.g. ad libitum): not specified
- Acclimation period: not specified

DETAILS OF FOOD AND WATER QUALITY: not specified

ENVIRONMENTAL CONDITIONS
- Temperature (°C): not specified
- Humidity (%): not specified
- Air changes (per hr): not specified
- Photoperiod (hrs dark / hrs light): not specified

IN-LIFE DATES: From: To:

Route of administration:

oral: unspecified

Vehicle:

other: suspensions of the gluconate containing 20 mg Fe/mL were employed in this experiment

Details on oral exposure:

not specified

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

12 weeks

Frequency of treatment:

daily

Dose / conc.:

0 other: mg Fe/kg

Dose / conc.:

50 other: mg Fe/kg

Remarks:

corresponding to 427.35 mg ferrous gluconate/kg

Dose / conc.:

100 other: mg Fe/kg

Remarks:

corresponding to 854.70 mg ferrous gluconate/kg

No. of animals per sex per dose:

5

Control animals:

yes

Details on study design:

- Dose selection rationale: not specified
- Rationale for animal assignment (if not random): random
- Rationale for selecting satellite groups: not applicable
- Post-exposure recovery period in satellite groups: not applicable
- Section schedule rationale (if not random): ramdom

Observations and examinations performed and frequency:

observed for clinical signs daily, weighted in intervals, blood examinations (red and total white cell count and hemoglobin concentraitions on 2 males and 2 females from each group)

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes
All the rats were then killed, and the major organs (liver, spleen, heart, lungs, thymus, kidneys, adrenals, thyroid, testes, prostate, seminal vesicles, ovaries and uterus) were excised, blotted dry, and weighed. The organs from two males and two females from each group dosed at 50 mg Fe/Kg were examined histologically.

Clinical signs:

not specified

Mortality:

not specified

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Data showed that at the higher dose level ferrous gluconate significantly depressed growth rate in the male rats, but not in the females.
At the lower dose level the depressions produced in the males by the gluconate were not significant (P = 0.05).

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

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

not specified

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Description (incidence and severity):

No abnormalities were found in the red or total white cell counts or hemoglobin concentrations.

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

None of the organ weights (expressed in mg/100 g bodyweight) in the dosed groups differed significantly (P = 0.05) from those of the controls

Gross pathological findings:

not specified

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Apart from a slight and variable increase in iron deposition in the tissue phagocytes (e.g., Kupffer cells, pulmonary macrophages and adrenal cortical littoral cells), histologic examination of the organs listed above revealed no abnormalities that could be attributed to the drugs.

Histopathological findings: neoplastic:

not specified

Other effects:

not examined

Details on results:

The group mean increases in bodyweight after 12 weeks are given in table 1. Analysis of the combined data showed that at the higher dose level all four compounds significantly depressed growth rate in the male rats, but not in the females. At the lower dose level the depressions produced in the males by the fumarate and the gluconate were not significant (P = 0.05). None of the organ weights (expressed in mg/100 g bodyweight) in the dosed groups differed significantly (P = 0.05) from those of the controls. Likewise no abnormalities were found in the red or total white cell counts or hemoglobin concentrations.
Apart from a slight and variable increase in iron deposition in the tissue phagocytes (e.g., Kupffer cells, pulmonary macrophages and adrenal cortical littoral cells), histologic examination of the organs listed above revealed no abnormalities that could be attributed to the drugs.

Dose descriptor:

NOAEL

Effect level:

427.35 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

haematology

histopathology: non-neoplastic

Remarks on result:

other: calculated based on the ferrous iron content of 11.7 % given for ferrous gluconate

Dose descriptor:

NOAEL

Effect level:

50 other: mg Fe/kg

Based on:

element

Sex:

male/female

Basis for effect level:

body weight and weight gain

haematology

histopathology: non-neoplastic

Critical effects observed:

no

The group mean increases in bodyweight after 12 weeks are given in Table 1. Analysis of the combined data showed that at the higher dose level all four compounds significantly depressed growth rate in the male rats, but not in the females. At the lower dose level the depressions produced in the males by the fumarate and the gluconate were not significant (P = 0.05). None of the organ weights (expressed in mg/100 g bodyweight) in the dosed groups differed significantly (P = 0.05) from those of the controls. Likewise no abnormalities were found in the red or total white cell counts or hemoglobin concentrations.

Apart from a slight and variable increase in iron deposition in the tissue phagocytes (e.g., Kupfer cells, pulmonary macrophages and adrenal cortical littoral cells), histologic examination of the organs listed above revealed no abnormalities that could be attributed to the drugs.

Table 1 -The Effects of 4 Iron Compounds on Growth Rate in Rats
Daily oral dose (mg Fe/kg)	Group mean increases in body weight ± S.E. (Gm.) after 12 weeks' dosing
	Controls		Fumarate		Sulphate		Gluconate		Succinate
	male	female	male	female	male	female	male	female	male	female
0	181 ± 13.6	92 ± 6.6
50			156 ± 11.7	101 ± 6.8	129 ± 10.7	73 ± 11.7	172 ± 10.2	106 ± 11.1	145 ± 6.8	95 ± 8.3
100			136 ± 12.8	87 ± 10.7	113 ± 10.8	84 ± 8.8	136 ± 13.3	85 ± 7.5	135 ± 11.1	96 ± 12.4

Executive summary:

The animal experiments were designed to compare the toxicologic and hematinic properties of ferrous fumarate with those of ferrous sulphate A.R., ferrous succinate and ferrous gluconate B.P.C (Berenbaum, 1960). The ferrous iron contents of these four salts are 33.0, 20.0, 24.8 and 11.7 per cent, respectively. The sulphate was administered as an aqueous solution and the other three compounds as aqueous suspensions containing 0.1 per cent w/v of tragacanth.

The subchronic oral toxicities of the four ferrous salts were compared in albino rats of the WAG strain. Ferrous sulphate solution and suspensions of the fumarate, gluconate and succinate containing 20 mg Fe/mL were employed in this experiment. 45 male and 45 female rats (40 to 100 g bodyweight) were randomly distributed into nine groups of five males and five females. One group was not dosed and served as controls, while the other eight groups received oral doses of one or other of the iron compounds at a level of 50 or 100 mg Fe/Kg. The animals were individually weighed at intervals and dosed daily, excluding weekends. After 12 weeks’ dosing, the  red and total white cell counts and hemoglobin concentrations were determined on two males and two females from each group. All the rats were then killed, and the major organs (liver, spleen, heart, lungs, thymus, kidneys, adrenals, thyroid, testes, prostate, seminal vesicles, ovaries and uterus) were excised, blotted dry, and weighed. The organs from two males and two females from each group dosed at 50 mg Fe/Kg were examined histologically.

Analysis of the combined data showed that at the higher dose level all four compounds significantly depressed growth rate in the male rats, but not in the females. At the lower dose level the depressions produced in the males by the fumarate and the gluconate were not significant (P = 0.05). None of the organ weights (expressed in mg/100 g bodyweight) in the dosed groups differed significantly (P = 0.05) from those of the controls. Likewise no abnormalities were found in the red or total white cell counts or hemoglobin concentrations.

Apart from a slight and variable increase in iron deposition in the tissue phagocytes (e.g., Kupffer cells, pulmonary macrophages and adrenal cortical littoral cells), histologic examination of the organs listed above revealed no abnormalities that could be attributed to the drugs.

Taking into account all given data, a NOAEL of 50 mg Fe/kg bw, corresponding to a value of 427.35 mg Ferrous gluconate/kg bw can be established.

Data source

Reference

Reference Type:

publication

Title:

Animal and Human Studies on Ferrous Fumarate, an Oral Hematinic

Author:

Berenbaum, M.C., Child, K.J., Davis, B., Sharpe, H.M. and Tomich, E.G.

Year:

1960

Bibliographic source:

Blood 1960 15:540-550

Materials and methods

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

The animal experiments were designed to compare the toxicologic and hematinic properties of ferrous fumarate with those of ferrous sulphate A.R.,ferrous succinate and ferrous gluconate B.P.C. The ferrous iron contents of ferrous gluconate was 11.7 per cent. The ferrous gluconate was administered as aqueous suspension containing 0.1 per cent w/v of tragacanth. Groups of 10 male fawn mice ( GFF strain, bodyweights 17 to 22 gr ) were dosed orally and then observed for seven days, when the percentage mortalities were recorded. The LD50 values, which were calculated according to do Beer (1945) and expressed in mg Fe/Kg.

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Iron digluconate

EC Number:

206-076-3

EC Name:

Iron digluconate

Cas Number:

299-29-6

Molecular formula:

C12H22FeO14

IUPAC Name:

Iron; (2R,3R,4S,5S)-2,3,4,5,6-pentahydroxyhexanoic acid

Details on test material:

not available

Specific details on test material used for the study:

The substances compared in these tests were ferrous fumarate, ferrous sulphate A.R., ferrous succinate and ferrous gluconate B.P.C. The ferrous iron contents of these four salts are 33.0, 20.0, 24.8 and 11.7 per cent, respectively.

Test animals

Species:

mouse

Strain:

other: GFF

Sex:

male

Details on test animals or test system and environmental conditions:

Groups of 10 male fawn mice (GFF strain, bodyweights 17 to 22 gr)

Administration / exposure

Route of administration:

oral: unspecified

Vehicle:

other: aqueous suspensions containing 0.1 per cent w/v of tragacanth.

Details on oral exposure:

VEHICLE
- Concentration in vehicle: not specified
- Amount of vehicle (if gavage): not specified
- Justification for choice of vehicle: not stated
- Lot/batch no. (if required): not stated
- Purity: not stated

MAXIMUM DOSE VOLUME APPLIED: not stated

DOSAGE PREPARATION (if unusual): aqueous suspensions containing 0.1 per cent w/v of tragacanth.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: not specified

Doses:

not specifed only LD50 values are given

No. of animals per sex per dose:

presumably 10 males

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 7 days
- Frequency of observations and weighing: not specified
- Necropsy of survivors performed: no data
- Other examinations performed: percentage martalities was recorded, no further details available

Statistics:

LD50 values were calculated according to do Beer (1945) and are expressed in mg Fe/Kg.

Results and discussion

Effect levels

Effect levels 1

Sex:

male

Dose descriptor:

LD50

Effect level:

320 mg/kg bw

Based on:

element

Effect levels 2

Sex:

male

Dose descriptor:

LD50

Effect level:

2 735 mg/kg bw

Based on:

test mat.

Remarks on result:

other: calculated according to the ferrous iron content (11.7%) given for ferrous gluconate

Mortality:

no details available, only LD50 value is given

Clinical signs:

no details available, only LD50 value is given

Body weight:

no details available, only LD50 value is given

Gross pathology:

no details available, only LD50 value is given

Any other information on results incl. tables

The LD50 value for ferrous gluconate was calculated taking into account the ferrous iron content given for this compound (11.7 %) and is 2735 mg/kg bw.

Applicant's summary and conclusion

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

The LD50 value for ferrous gluconate was calculated taking into account the ferrous iron content given for this compound (11.7 %) and is 2735 mg/kg bw.

Executive summary:

The substances compared in these acute oral toxicity tests in mice were ferrous fumarate, ferrous sulphate A.R., ferrous succinate and ferrous gluconate B.P.C (Berenbaum, 1960). The ferrous iron contents of ferrous gluconate was 11.7 per cent. It was administered as aqueous suspensions containing 0.1 per cent w/v of tragacanth.

Groups of 10 male fawn mice (GFF strain, bodyweights 17 to 22 gr) were dosed orally and then observed for seven days, when the percentage mortalities were recorded. The LD50 values, which were calculated according to do Beer (1945) and expressed in mg Fe/Kg, were fumarate 630, succinate 560, gluconate 320 and sulphate 230. Thus, the relative toxicities were fumarate 1 , succinate 1 .1 , gluconate 2.0 and sulphate 2.7.

The LD50 value for ferrous gluconate was calculated taking into account the ferrous iron content given for this compound (11.7 %) and is 2735 mg/kg bw.