Manganate(1-), glycinato-N,O)[sulfato(2-)-O]-, hydrogen

Endpoint summary

Description of key information

- LD50 value for Manganese monolysinate sulfate , is > 2000 mg/kg following single-dose intragastric administration to male and female rats in a study conducted according to OECD guideline 425.

- Manganese sulfate was administered to male and female Wistar rats by oral gavage in several doses. The LD50 was determined by the method of Horn. The determined LD50 is 2150 mg/kg bw.

- Manganese acetate was administered to rats in a dose range of 2.68 to 5.21 g/ kg bw. The LD50 value was determined to be 3730 mg/kg bw. The study was conducted prior to implementation of GLP and OECD guidelines.

- In a study conducted within the NTP rats were administered manganese sulfate in the feed in daily doses of 1,300 mg manganese/kg/day for 14 days and this dose did not affect survival (NTP 1993).

- Glycine was ministered in doses of 500, 1000 and 2000 mg/kg bw to male Sprague Dawley rats for 28 days, results: no mortality, not clinical signs, no differences in body weight, food consumption and water consumption, no changes in urinalysis at necropsy, LD50 > 2000 mg/kg bw, LD0 = 2000 mg/kg bw

- The TOXNET database revealed a LD50 value for glycine of 7930 mg/kg bw in female rats. The study was conducted with either rats, mice or guinea pigs. (Yakuri, 1977).

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:

1993

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

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

Qualifier:

no guideline followed

Principles of method if other than guideline:

- Principle of test: Review of several studies of toxicity in mice and rats. The durations were 14-days, 13 weeks and 2-years. The animlas received manganese (II) sulfate via feed at concentrations of 0, 3130, 6250, 12500, 25000 and 50000 ppm.
- Short description of test conditions: animals were observed for clinical signs once daily day 1-8 and twice daily day 9-14. Feed consumption was recorded weekly by cage. The animals were weighed at study initiation, on day 7, and at the end of the studies.
- Parameters analysed / observed: body weight, food consumption, clinical signs, hematology, necropsy

GLP compliance:

yes

Test type:

other: 14-days repeated dose study

Limit test:

no

Species:

rat

Strain:

Fischer 344

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories
- Age at study initiation: approx. 31 days
- Housing: in groups of five animals in Polycarbonate (Lab Products, Inc., Garfield, NJ), changed twice weekly. Bedding consists of Heat-treated hardwood chips (PWI, Inc., Loweville, NY), changed twice weekly
- Diet (e.g. ad libitum): ad libitum, NIH-07 open formula meal rat and mouse diet (Zeigler Brothers, Inc., Gardners, PA)
- Water (e.g. ad libitum): Automatic watering system (Edstrom Industries, Waterford, WI), available ad libitum
- Acclimation period: 19 days
- Microbiological status when known : Before the beginning of the studies, five male and five female rats and mice were randomly selected for parasite evaluation and gross observation for evidence of disease.
- Method of randomisation in assigning animals to test and control groups

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23.3 ± 2
- Humidity (%): 40-80
- Air changes (per hr): 12
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: feed

Vehicle:

other: in feed

Doses:

0, 3,130, 6,250, 12,500, 25,000, or 50,000 ppm in feed, available ad libitum

No. of animals per sex per dose:

5

Control animals:

yes

Remarks:

control animals were exposed to 92 ppm manganese in feed (background)

Details on study design:

- Duration of observation period following administration: Observed and observations recorded once daily on days 1-8, twice daily on days 9-14 (rats), Animals weighed initially, at the end of 1 week,and at end of the studies. Feed consumption recorded weekly by cage.
- Necropsy of survivors performed: yes
- Clinical signs including body weight
- Other examinations performed: clinical signs, body weight,organ weights, histopathology

Statistics:

The probability of survival was estimated by the product-limit procedure of Kaplan and Meier (1958) and is presented in the form of graphs. Animals were censored from the survival analyses if they were found dead of other than natural causes, were found to be missing, or were missexed; animals dying from natural causes were not censored. Statistical analyses for possible dose-related effects on survival used Cox's (1972) method for testing two groups for equality and Tarone's (1975) life table test to identify dose-related trends. All reported P values for the survival analyses are two sided.
Two approaches were employed to assess the significance of pairwise comparisons between exposed and control groups in the analysis of continuous variables. Organ and body weight data were analyzed using the parametric multiple comparison procedures of Dunnett and Williams. Clinical chemistiy and hematology data and tissue metal concentrations were analyzed using the nonparametric multiple comparison methods of Shirley and Dunn. Average severity values were analyzed for significance using the Mann-Whitney U test.

Key result

Sex:

male/female

Dose descriptor:

other: LOEL

Effect level:

50 000 other: ppm

Based on:

test mat.

Mortality:

No mortality occurred

Clinical signs:

other: Not reported

Gross pathology:

No effects reported

Interpretation of results:

study cannot be used for classification

Conclusions:

In the present study conducted with male and female rats, the animals were exposed to manganese (II) sulfate for 14 days via feed at concentrations of 0, 3130, 6250, 12500, 25000, and 50000 ppm. No Mortality occurred, at the highest concentration decreases in body weight gain was observed in male and female rats. At necropsy the absolute and relative liver weights in male rats of the 50000 ppm group were decreased as compared to the control group.Manganese concentrations in the livers of 50000 ppm males and females were more than twice those of controls. Furthermore, In the hematology evaluation, the total leukocyte and neutrophil counts were significantly increased in 50000 ppm groups, particularly males, other changes in hematology were not considered to be treatment-related.

Based on the presented results manganese (II) sulfate is not considered to be actutely toxic via the oral route according to Regulation (EC) No.1272/2008 (CLP).

Executive summary:

In a 14-days repeated dose toxicity study , fasted, 31 days old Fischer 344 strain rats were fed oral doses of MnSO₄at 0, 3,130, 6,250, 12,500, 25,000, or 50,000 ppm in feed, 5 animals per group 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. The mean body weight gain of the male 50,000 ppm group at the end of the 14-day study was 57% less than that of the control group, and the final mean body weight of this group was 13% lower than that of the controls. The mean body weight gain of 50,000 ppm females was 20% less than that of the controls and the final mean body weight was 7% lower than that of the controls. 

Oral LOEL (rat, females/males) 50000 ppm

Reference 2

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2013

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

other: OECD Guideline 407

Version / remarks:

adopted 03 October, 2008; study was principally conducted according to Partial Revision of Guidelines for Repeated-dose Toxicity
Studies. Notification No. 655 of the Pharmaceutical and Medical Safety Bureau. Ministry of Health and Welfare, Japan. 1999.

GLP compliance:

no

Remarks:

compliance to GLP guidelines was not reported

Test type:

other: 28-days repaeted dose oral toxicity study

Limit test:

no

Species:

rat

Strain:

Crj: CD(SD)

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories Japan Inc. (Kanagawa, Japan)
- Age at study initiation: Five-week-old
- Weight at study initiation: 193.9 to 215.1g
- Housing: individually in metallic bracket cages
- Diet (e.g. ad libitum): allowed ad libitum access to a gamma-ray sterilized powder diet (CRF-1; Oriental Yeast Co., Ltd., Tokyo, Japan).
- Water (e.g. ad libitum): allowed ad libitum access to tap water
- Acclimation period: 6 days prior to experimentation

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

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

Water for injection (D.W., lot 5A88; Otsuka Pharmaceutical Factory Inc., Tokushima, Japan)

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 50, 100 and 200 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg bw
- Lot/batch no. (if required): lot 5A88

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw


CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Absence of toxic effects in other studies with the test substance

Doses:

500, 1000 and 2000 mg/kg bw

No. of animals per sex per dose:

6

Control animals:

yes

Details on study design:

- Duration of observation period following administration: based on study design the duration of exposure was 28-days/once daily, after that period the animals were sacrificed and a gross observation as well as histopathological determination was performed.
- Frequency of observations and weighing: All animals were examined twice daily (pre-dose and post-dose) for clinical signs. Individual body weights were recorded for all animals on days 1, 3, 8, 15, 22 and 28 and on the day of necropsy. Food consumption and water consumption on days 2–3, 7–8, 14–15, 21–22 and 27–28 was measured individually for all animals.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, hematology, urinalysis, blood chemistry

Statistics:

Body weight, food consumption, water consumption, urinalysis, hematology, blood chemistry, and organ weight data were recorded using a MiTOX RDT system. Numerical data obtained during the study were used to calculate group mean values and standard deviations. Group variances for the appropriate parameters were compared using Bartlett’s method. When the differences between group variances were not significant, Dunnett’s multiple comparison method was applied to determine the significance of differences be tween the control group and each glycine-treated group. If the Bartlett’s test indicated significant differences between group variances for a given parameter, that parameter was compared among groups using the Steel’s multiple comparison method for mean ranking. The criterion of significance was an alpha level of 0.05 or 0.01.

Key result

Sex:

male

Dose descriptor:

LD0

Effect level:

2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: under the present condictions there were no toxic effects detectable up to the concentrations used in this test

Mortality:

Not observed

Clinical signs:

other: not adverse effects observed

Gross pathology:

No abnormalties detected

Other findings:

- Other observations: Daily urine volume and total amount of Cl excretion were significantly higher (p<0.05) in the 2000 mg/kg dose group. Statistically significant increases in daily urinary volume and total amount of urinary Cl, and declining trends in urine pH and urinary protein in the glycine 2000 mg/kg dose group were considered to be of little toxicological significance, because there were no histopathological correlates in the kidneys and urinary bladder and no changes in kidney weight or other urinary parameters.

Interpretation of results:

GHS criteria not met

Conclusions:

In the present study conducted similarly to OECD guideline 407 and performed according to Partial Revision of Guidelines for Repeated-dose Toxicity Studies. Notification No. 655 of the Pharmaceutical and Medical Safety Bureau. Ministry of Health and Welfare, Japan. 1999, Glycine showed no adverse effects when administered to male rats for 28 days once daily. However, there were significant changes in urine volume and urinary Cl concentrations but since these effects had no hisotpathologic correlate these effects are regarded not toxicological relevant. Based on the results presented here glycine does not need to be classified according to Regulation (EC) No. 1272/2008 (CLP) and the Harmonized System for Classification and Labelling of Chemcials (GHS).

Executive summary:

In a Repeated Dose, 28-day Oral Toxicity Study performed similar to OECD guideline 407 Glycine was administered to groups of 6 male and 6 female Crj: CD(SD) rats by oral gavage at dose levels of 500, 1000 and 2000 mg/kg bw/day for 28 days. The control group animals received Water for injection (D.W., lot 5A88; Otsuka Pharmaceutical Factory Inc., Tokushima, Japan).

All animals survived to study termination. No test substance-related findings were detected or observed in clinical examinations, body weights, food consumption values, clinical biochemistry and clinical pathology evaluations. During the anatomical pathology examinations, no test substance-related findings were observed or detected during macroscopic examinations, organ weight evaluations or microscopic examinations. Daily urine volume and total amount of Cl excretion were significantly higher (p<0.05) in the 2000 mg/kg dose group. Statistically significant increases in daily urinary volume and total amount of urinary Cl, and declining trends in urine pH and urinary protein in the glycine 2000 mg/kg dose group were considered to be of little toxicological significance, because there were no histopathological correlates in the kidneys and urinary bladder and no changes in kidney weight or other urinary parameters.

There is no evidence for specific target organ toxicity in this study.

The LD0 is 2000 mg/kg bw/day in this study.

Reference 3

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1991

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

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

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Version / remarks:

dose ranges were not reported, LD50 was calculated by the method of Horn.

GLP compliance:

not specified

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: 100-125g
- Housing: Animals of either sex were kept in polycarbonate cages
- Diet (e.g. ad libitum): The animals were fed Gold Mohur Hind Lever diet ad Iibitum.
- Water (e.g. ad libitum): ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 24 ± 0.5
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

water

Doses:

not reported

No. of animals per sex per dose:

5

Control animals:

yes

Statistics:

LD50 with its fiducial limit was calculated using the method of Horn.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

2 150 mg/kg bw

Based on:

test mat.

Mortality:

not reported

Clinical signs:

other: not reported

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

In the present study manganeous sulfate did not exhibit toxic effects in a test conducted similar to OECD guideline 401. 5 male and female Wistar rats were administered manganeous sulfate in water p.o.. The LD50 value was calculated using the method of Horn. The LD50 of manganeous sulfate was determined to be 2150 mg/kg bw in rats. Thus, manganeous sulfate does not need to be classified according to Regulation (EC) No 1272/2008 (CLP) and is classified accoding the Globally Harmonized System for Classification and Labelling of Chemicals (GHS) Category 5 'Maybe harmful when swallowed'.

Executive summary:

In an acute oral toxicity study according to OECD guideline 401, 100-125 g weighing Wistar strain rats were given a single oral dose of MnSO₄in deionized triple glass distilled water at increasing doses.

LD50 with its fiducial limit was calculated using the method of Horn. 

Oral LD50 (rat, females/males) > 2150  mg/kg bw

Reference 4

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Qualifier:

no guideline followed

Principles of method if other than guideline:

Methods not further described. Rats received by oral gavage Manganous acetate at 0.2g/mL in a concentration range of 2.68g to 5.21g. The mortality was determined.

GLP compliance:

no

Remarks:

Testing was conducted prior to implementation of OECD or GLP Guidelines

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

not specified

Sex:

not specified

Details on test animals or test system and environmental conditions:

Not reported

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 0.2g/mL

MAXIMUM DOSE VOLUME APPLIED: 5.21 g/kg bw

Doses:

2.68 - 5.21 g/kg bw

No. of animals per sex per dose:

not reported

Control animals:

not specified

Details on study design:

not reported

Key result

Sex:

not specified

Dose descriptor:

LD50

Effect level:

3 730 mg/kg bw

Based on:

test mat.

Mortality:

exact number of deaths was not reported but the LD50 value was calculated to be 3730 mg/kg bw

Clinical signs:

other: not reported

Interpretation of results:

study cannot be used for classification

Conclusions:

The present publication reveals handbook data from over 200 substances regarding acute oral toxicity and irritancy. The LD50 of Manganous acetate was determined to be 3730 mg/kg bw. Based on the similarity of the breakdown products of manganous acetate and manganese monoglycinate sulfate and under the assumption that Mn2+ is the toxicity determineing constituent the target substance does not need to be classified.

Executive summary:

In an acute oral toxicity study conducted similar to OECD guideline 401, groups of rats were given a single oral dose of Manganese acetate in water at doses between 2.68 and 5.21 g/kg bw and observed for occurring mortality .

 

Oral LD50 Combined = 3730 mg/kg bw

 

Manganous acetate is of LOW Toxicity based on the LD50 in males / females. According to Regulation (EC) No 1272/2008 (CLP) Manganese acetate does not need to be classified as acute orally toxic.

Reference 5

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1977

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Qualifier:

no guideline followed

Principles of method if other than guideline:

The acute oral toxicity of glycine in rats was investigated.

GLP compliance:

no

Remarks:

Study was conducted prior to implementation of GLP Guidelines

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:

Age at study initiation: at 7 weeks of age
Weight at study initiation: No data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23±2 °C
- Humidity (%): 55 ± 5 °C
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): no data

Route of administration:

oral: gavage

Vehicle:

physiological saline

Details on oral exposure:

Manufacture, supplier, source of supply: Iwai Chemicals Co.
Lot/batch No.: no data

Doses:

more than 4 doses (unspecified)

No. of animals per sex per dose:

5-10 animals per dose

Control animals:

not specified

Details on study design:

Duration of observation period following administration: 7 days
Frequency of observations and weighing:no data
Necropsy of survivors performed: yes
Other examinations performed: clinical signs

Statistics:

According to Litchfield and Wilcoxon

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

7 930 mg/kg bw

Based on:

test mat.

95% CL:

> 7 329 - < 8 580

Key result

Sex:

male

Dose descriptor:

LD50

Effect level:

9 550 mg/kg bw

Based on:

test mat.

95% CL:

> 8 917 - < 10 228

Mortality:

Not reported

Clinical signs:

other: Decrease in in locomotor activity, slightly respiratory depression, piloerection and ptosis were observed in female and male rats at 15 to 35 min after dosing. Most of the animals stopped breathing after muscle weakness, cyanosis, circular movement, tonic

Gross pathology:

Congestion in lungs and dot hemorrhage in glandular stomach were observed in some animals. No abnormal gross findings in the other organs were observed.

Interpretation of results:

GHS criteria not met

Conclusions:

The information gathered during research of literature revealed the present results from the TOXNET database. In the study of Yakuri et al., 1977 mice and rats were treated with several doses of glycine either intraperitoneal, subcutaneous or orally. The LD50 value was calculated according to the method of Litchfield and Wilcoxon. The LD50 value for female rats was determined to be 7930 mg/kg bw and for male rats 9550 mg/kg bw.
Based on these results glycine does not need to be classified according to Regulation (EC) No 1272/2008 (CLP) and the Globally Harmonized System for Classification and Labelling of Chemicals (GHS).

Executive summary:

In an acute oral toxicity study , Wistar strain rats were administered single oral doses of glycine at at least four increasing doses in physiological saline, 5 animals per group, and were observed for 7 days.

The LD50 value was calculated according to Litchfield and Wilcoxon.

The number of animals that had died was not reported, only the calculated LD50 Values were provided. Clinical signs reported were decrease in in locomotor activity, slightly respiratory depression, piloerection and ptosis were observed in female and male rats at 15 to 35 min after dosing. Most of the animals stopped breathing after muscle weakness, cyanosis, circular movement, tonic convulsion. Some animals stopped breathing after coma without tonic convulsion. But some rats became a coma and caused breathing to stop without tonic convulsion. Necropsy findings were: Congestion in lungs and dot hemorrhage in glandular stomach were observed in some animals. No abnormal gross findings in the other organs were observed.

Oral LD50 (rat, females) 7930 mg/kg bw

Oral LD50 (rat, males) 9550 mg/kg bw

Reference 6

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2019-08-02 to 2019-09-10

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, manganese lysinate sulfate, at 2000 mg/kg following single-dose intragastric administration had a moderate local irritant effect in 33% of the animals, 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 the oral route

Conclusions:

The LD50 value for the test article, manganese lysinate sulfate, is > 2000 mg/kg following single-dose intragastric administration to male and female rats. The test article, manganese lysinate sulfate can be classified as toxicity category 5 according to the international GHS classification: 2000 < LD50 < 5000 mg/kg.

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 manganese 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, manganese lysinate sulfate, at 2000 mg/kg following single-dose intragastric administration had a moderate local irritant effect in 33% of the animals, which was manifested as catarrhal gastritis detected by a histological examination of the stomach tissues.

 

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

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

One study according to OECD guideline 425 with the structurally similar Manganese monolysinate sulfate is available. Several studies investigated the effects of Manganese sulfate and glycine. Most of the data was gathered from studies that were performed prior to implementation of GLP and/or OECD guidelines. Only studies with sufficient documentation have been considered for a weight of evidence approach. Thus, the quality of the database is considered to be sufficient for classification.

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

Data for the target substance manganese monoglycinate sulfate are not available. Thus, the acute oral toxicity of manganese monoglycinate was determined by a read-acorss approach.

This read-across hypothesis is based on transformation of the target and source substances to common compounds (scenario1 of the RAAF). For a more detailed information please refer to the generl justification for read-across attached to section 13 of this IUCLID file.

The target substance manganese monoglycinate sulfate and the source substances manganese sulfate, manganese Chloride, and other manganese compounds are ionic and consist of the Mn²⁺cation and the respective anion (glycine).

Acute toxicity data for the source substances manganese sulfate and glycine are listed below.

LD50 value for manganese monolysinate sulfate , is > 2000 mg/kg following single-dose intragastric administration to male and female rats in a study conducted according to OECD guideline 425.

Female and male Sprague Dawley rats were administered manganese monolysinate sulfate in the following doses: 175, 550, 2000 mg/kg bw. After the single oral administration the animals were observed for further 14 days. No Mortality was reported and no clinical signs of intoxication were observed.

Manganese sulfate was administered to male and female Wistar rats by oral gavage in several doses. The LD50 was determined by the method of Horn. The determined LD50 is 2150 mg/kg bw.

Manganese acetate was administered to rats in a dose range of 2.68 to 5.21 g/ kg bw. The LD50 value was determined to be 3730 mg/kg bw. The study was conducted prior to implementation of GLP and OECD guidelines.

In a study conducted within the NTP rats were administered manganese sulfate in the feed in daily doses of 1,300 mg manganese/kg/day for 14 days and this dose did not affect survival (NTP 1993).

Williams et al. summarized the effects of various manganese salts in humans and animals, in their report further studies were listed which were performed using manganese chloride as manganese source. The measured LD50 values for 1-21 days of exposure range from 225 to 1,082 mg manganese/kg/day in mice and rats (Holbrook et al. 1975; Kostial et al. 1978, 1989; Rehnberg et al. 1980; Singh and Junnarkar 1991; Smyth et al. 1969). The authors discussed that “these results suggest that gavage dosing with a bolus of a concentrated soluble manganese compound in water may not be a good model for determining the toxic effects of manganese ingested by humans from environmental sources.It should be noted that the concentrations used in the bolus dosing studies are much higher than even excess levels to which certain humans are typically exposed.”( Williams, M., Todd, G. D., Roney, N., Crawford, J., Coles, C., McClure, P. R., & Citra, M. (2012). Toxicological profile for manganese.)

For the other source substance glycine two studies are available.

Shibui et al. (2012) investigated the effects of glycine in doses of 500, 1000, and 2000 mg/kg bw administered by oral gavage in a 28 days repeated dose toxicity study in male Sprague Dawley rats. Parameters observed were clinical signs, body weight changes, food and water consumption, urinalysis, gross pathology. No changes in the observed parameters was detected, no mortality occurred up to the highest dose tested.

A research of database information revealed results of a study in which the LD50 of glycine was determined in rats, mice and guinea pigs. The LD50 was determined at 7930 mg/kg bw for rats.

Based on the available information and on the hypothesis of the read-across approach the acute oral toxicity of manganese monoglycinate sulfate is expected to be determined by manganese and the lowest determined LD50 value for manganese of 2150 mg/kg bw and for the structurally similar manganese monolysinate complex of ≥ 2000 mg/kg bw, the lowest LD50 detectable is ≥ 2000 mg/kg bw.

Justification for classification or non-classification

Based on the available information Manganese monoglycinate sulfate is not classified respect to acute oral toxicity as described in Regulation (EU) No. 1272/2008 (CLP).