Magnesium glycerophosphate

Endpoint summary

Description of key information

Magnesium glycerophosphate was administered once daily to group of Wistar rats for a period of 28 days at doses of 100, 500 and 1000 mg/kg bodyweight. In the treated animals, no motality and clinical signs of toxicity were observed. There were no treatment related and toxicologically relevant changes in feed consumption, bodyweight gain, haematology, biochemistry, urinary parameters and organ weights. The gross and histopathology did not attribute to the test item Magnesium glycerophosphate at and up to 1000 mg/kg bw dose level in both sexes. Hence under the conditions of the experiment, the highest dose tested i.e., 1000 mg/kg is considered as NOEL (No Oberved Effect Level).  

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

August 03, 2018 to March 22, 2019

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Clariant India Limited/ INVG003917
- Expiration date of the lot/batch:June 10, 2021
- Purity test date:

RADIOLABELLING INFORMATION (if applicable)
- Radiochemical purity:
- Specific activity:
- Locations of the label:
- Expiration date of radiochemical substance:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room Temperature (20 to 30°C)
- Stability under test conditions:Stable under normal conditions
- Solubility and stability of the test substance in the solvent/vehicle: Stable at 0 and 6 hours
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: Non reactive

TREATMENT OF TEST MATERIAL PRIOR TO TESTING - Not Applicable
- Treatment of test material prior to testing:
- Preliminary purification step (if any):
- Final dilution of a dissolved solid, stock liquid or gel:
- Final preparation of a solid:

FORM AS APPLIED IN THE TEST (if different from that of starting material)

OTHER SPECIFICS:

Species:

rat

Strain:

Wistar

Details on species / strain selection:

Rat is chosen as the test system because this species is commonly used for repeated dose Oral toxicity testing and it meets the regulatory requirement of most of the regulatory agencies

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: RCC Laboratories India Private Limited
- Females (if applicable) nulliparous and non-pregnant: [yes/no]
Age on first Treatment 6 - 7 Weeks
Body weight when treated Males - 120.4 to 168.3 g
Females - 119.4 to 151.8 g
- Fasting period before study: Not Applicable
- Housing: In groups of three and two animals of same sex per cage in Polycarbonate cages (Approximate internal dimensions of 365 mm x 202 mm x 180 mm height) with paddy husk bedding.
- Diet (e.g. ad libitum): Teklad Certified Global 14% Protein Rodent Maintanence Diet. (Lot No. – 2014C-051418MA) was provided ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: Under laboratory conditions 7 days for Step I (Allocation A & B) and
8 days for Step II (Allocation A & B) animals

DETAILS OF FOOD AND WATER QUALITY:
Food and water is checked for proximate and contaminant analysis. the reports were archived at RCC India

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.8°C to 23.7°C
- Humidity (%): 57 to 66%
- Air changes (per hr): >10
- Photoperiod (hrs dark / hrs light):12 hours light :12hours dark

IN-LIFE DATES: From: August 16, 2018 To: November 22, 2018

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:

DIET PREPARATION
- Rate of preparation of diet (frequency):
- Mixing appropriate amounts with (Type of food):
- Storage temperature of food:

VEHICLE
- Justification for use and choice of vehicle (if other than water): As test item soluble in Corn oil
- Concentration in vehicle: 20 mg/mL (500 mg/25 mL) Low dose, 100 mg/mL (2500 mg/25 mL) Intermediate dose, 200 mg/mL (5000 mg/ 25mL) High dose
- Amount of vehicle (if gavage): 5ml/Kg bodyweight
- Lot/batch no. (if required):
- Purity:

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The dose concentration verification was carried out for test item fortified in corn oil pertaining to low dose (20mg/mL), mid dose (100mg/mL) and high dose (200mg/mL). The recovery of test item in the prepared dose formulation was within the guideline specification (i.e. ranged between 95-105%).
The test item was found to be homogeneously dispersed in corn oil, which was evinced from the recovery of test item from different layers (Top, middle and bottom layers of corn oil) was within 95% to 109% for the three different doses (20 mg/mL, 100 mg/mL and 200 mg/mL).

Duration of treatment / exposure:

28 days

Frequency of treatment:

Once daily

Dose / conc.:

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

Remarks:

LOW DOSE

Dose / conc.:

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

Remarks:

INTERMEDIATE DOSE

Dose / conc.:

1 000 mg/kg bw/day (actual dose received)

Remarks:

HIGH DOSE

No. of animals per sex per dose:

5 Aanimals/Sex/ Dose

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Dose Range Finding study (RCC Study No. 138) was carried out before the 28 days repeated dose study to confirm the dose level. Based on the results of the dose range finding study, the dose levels of 100, 500 and 1000 mg/kg body weight for Low, intermediate and High dose respectively were selected

- Rationale for animal assignment (if not random): Animals were selected and grouped based on stratified randomization by using body weights. Computerized statistical analysis was used for randomization

- Rationale for selecting satellite groups: To establish recovery of treatm,ent related effects if any
- Post-exposure recovery period in satellite groups: 14 days
- Section schedule rationale (if not random):

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule:Twice daily on first 3 days of treatment and Once daily thereafter
- Cage side observations checked in table [No.?] were included. Yes

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Week 4 (towards end of treatment)

BODY WEIGHT: Yes
- Time schedule for examinations:
Body Weights
Treatment Period : Once Weekly
Recovery Period : Once Weekly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes / No / Not specified
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / Not specified

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / Not specified

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes / No / Not specified
- Time schedule for examinations:

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: Once during acclimatization ; during week 4 in all animals
- Dose groups that were examined: Groups 1 and 4

HAEMATOLOGY: Yes
- Time schedule for collection of blood:
Blood sampling schedule:
Week 4 Group 1 to 4 (Allocation A)
Week 6 Group 1R and 4R (Allocation B)
- Anaesthetic used for blood collection: Yes (identity)
Isoflurane
- Animals fasted: Yes
- How many animals: All animals
- Parameters checked in table [No.?] were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood:
Blood sampling schedule:
Week 4 Group 1 to 4 (Allocation A)
Week 6 Group 1R and 4R (Allocation B)
- Animals fasted: Yes
Isoflurane
- How many animals: All animals
- Parameters checked in table [No.?] were examined.
URINALYSIS: Yes
- Time schedule for collection of urine:
sampling schedule:
Week 4 Group 1 to 4 (Allocation A)
Week 6 Group 1R and 4R (Allocation B)
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- Parameters checked in table [No.?] were examined.

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations:
Week 4
- Dose groups that were examined: Group 1 to 4
- Battery of functions tested: grip strength / motor activity

IMMUNOLOGY: No
- Time schedule for examinations:
- How many animals:
- Dose groups that were examined:
- Parameters checked in table [No.?] were examined.

OTHER:

Sacrifice and pathology:

GROSS PATHOLOGY: Yes

HISTOPATHOLOGY: Yes

Statistics:

• Data are summarized in tabular form. Statistical analysis was performed using “Statplus” program.
• All the data was checked for normality with Shapiro-Wilk W test
• All the data was checked for homogeneity with Bartlett’s Chi-square test
• For comparing more than 2 groups, If data passes Normality & Homogeneity then ANOVA was done, Otherwise Kruskal Wallis Test be done followed by appropriate Post-hoc tests if p < 0.05
• For comparing 2 groups, if data passes Normality & Homogeneity then Student T-test was done, Otherwise Mann-Whitney U Test was done. If p < 0.05 the compared groups are said to be significantly different
• Values was summarized as mean ± standard deviation (SD)

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

No adverse effect on body weight was observed for treated groups when compared with control group. There was significant decrease in the bodyweight of female animals on day 1 was observed in high dose group when compared with low dose and intermediate dose group. This was not considered treatment related due to the early incidence.

Food consumption and compound intake (if feeding study):

no effects observed

Ophthalmological findings:

no effects observed

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

In males, MCH decreased in intermediate dose group (G3) when compared with vehicle control (G1) and low dose (G2) group. MCH and MCHC values were increased in high dose group (G4) when compared with intermediate group (G3). Monocyte count increased in high dose recovery (G4R) when comapred with vehicle control recovery (G1R). The chnges inHaematology parameters observed as a single incidence with no dose-response hence it is incidental and without toxicological relevance

Clinical biochemistry findings:

effects observed, non-treatment-related

Description (incidence and severity):

In females, glucose levels were decreased in intermediate dose group (G3) and triglyceride levels increased in high dose group (G4) when comapred with vehicle control group (G1). Chloride levels increased in high dose dose recovery (G4R) when compared with vehicle control recovery group(G1R). Thease changes in the biochemical parameters observed as a single incidence with no dose-response hence it is incidental and without toxicological relevance

Urinalysis findings:

effects observed, non-treatment-related

Description (incidence and severity):

In males, urine analysis revealed increased pH in low (G2), intermediate (G3) and high dose (G4) when compared with the vehicle control group (G1). Urinary crystals were increased in low (G2) and high dose group (G4) when compared with control group (G1). Urinary crystals decreased in intermedate group(G3) when compared with low dose (G2).
In females, leukocytes (LEU) was increased when compared with low (G2) and intermediate dose (G3) group. Proteins were increased in high dose group (G4) when compared with intermediate group (G3) and Crystal is also increased in low dose (G2) when compared with vehicle control group (G1).
Changes in the urinary parameters did not show dose response and toxicological relevance.

Behaviour (functional findings):

effects observed, non-treatment-related

Description (incidence and severity):

Fore limb and hind limb gripstrength decreased in high dose male and female when compared with vehicle control (G1), low (G2) and intermediate (G3) dose groups. Whereas hind limbs grip strength increased in male and female animals of intermediate dose (G3) when compared with vehicle control (G1).
In open field activity there was increased foot splay noted in high dose group female animals when compared with low dose group (G2). Eventhough there was significance observed in grip strength, this signficance might be due increase or decrease in individual values and not to represent any significance. No significat and toxicological relevant changes between treated and control group observed

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Description (incidence and severity):

In recovery animals, significant decrease in absolute and relative weight of prostate with seminal vesicle and coagulating glands and brain in male and significant decrease in absolute weight of liver in female was observed in high dose group when compared to control group. This can be considered incidental in the absence of weight changes in these organs of main group anim

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

There were no treatment related macroscopic findings observed in the study. Bilateral small sized testes and epididymides was observed in one male animal (Animal No. 23) of intermediate (G3) dose group. Microscopically, degenerative changes of seminiferous tubules in testes and oligospermia in epididymides of moderate severity were observed. Single incidence of these lesions were considered incidental

Histopathological findings: non-neoplastic:

effects observed, non-treatment-related

Description (incidence and severity):

There were no treatment related histopathological findings attributable to the test item magnesium glycerophosphate.
Single incidence of microscopic findings observed in heart, testes and epididymides were considered incidental

Details on results:

Magnesium glycerophosphate was administered once daily to group of Wistar rats for a period of 28 days at doses of 100, 500 and 1000 mg/kg bodyweight. In the treated animals, no motality and clinical signs of toxicity were observed. There were no treatment related and toxicologically relevant changes in feed consumption, bodyweight gain, haematology, biochemistry, urinary parameters and organ weights. The gross and histopathology did not attribute to the test item Magnesium glycerophosphate at and up to 1000 mg/kg bw dose level in both sexes. Hence under the conditions of the experiment, the highest dose tested i.e., 1000 mg/kg is considered as NOEL (No Observed Effect Level).
No Observed Effect Level (NOEL): 1000 mg/kg body weight

Key result

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

clinical biochemistry

clinical signs

dermal irritation

food consumption and compound intake

gross pathology

haematology

mortality

organ weights and organ / body weight ratios

Key result

Critical effects observed:

no

Conclusions:

Magnesium glycerophosphate was administered once daily to group of Wistar rats for a period of 28 days at doses of 100, 500 and 1000 mg/kg bodyweight. In the treated animals, no motality and clinical signs of toxicity were observed. There were no treatment related and toxicologically relevant changes in feed consumption, bodyweight gain, haematology, biochemistry, urinary parameters and organ weights. The gross and histopathology did not attribute to the test item Magnesium glycerophosphate at and up to 1000 mg/kg bw dose level in both sexes. Hence under the conditions of the experiment, the highest dose tested i.e., 1000 mg/kg is considered as NOEL (No Oberved Effect Level).

Executive summary:

Repeated Dose 28 Days Oral Toxicity Study with Magnesium glycerophosphate in Rats was conducted based on the OECD Guidelines for the Testing of Chemicals, Section 4, Health Effects, Number 407, of 3 October 2008.

The purpose of this study was to assess the toxicity ofMagnesium glycerophosphate when administered once daily to Wistar rats by oral gavage for a period of 28 days. Dose levels for main study were selected based on dose range finding study results. The doses selected for main study was 100, 500 and 1000 mg/kg body weight. In addition, recovery groups for vehicle control and high dose were included to know the reversibility of treatment related effects.

The test item was formulated in corn oil and administered once daily for 28 days to Wistar rats at dose levels of 100 (low dose, G2), 500 (intermediate dose, G3) and 1000 (High dose, G4) mg/kg body weight. The rats of the control (G1) received the vehicle, corn oil. In this study, recovery groups for vehicle control (G1R, vehicle control recovery) and high dose (G4R, high dose recovery) were included to know the reversibility of treatment related effects.Each group consists of five males and five female animals. The control, test item were administered at a volume of 5 ml/kg body weight. The observation and examinations performed in the study were mortality/viability, clinical signs, detailed clinical signs once during acclimatiozation, functional observation battery, ophthalmoscopy, body weights, food consumption, clinical pathology, macroscopy at termination, organ weights and histopathology on the preserved organs and tissues of all animals in the control and high dose groups.

All animals survived till the end of their scheduled study period. No clinical signs of illness were observed in any of the animals during treatment and recovery period.No treatment-related neuromuscular and physiological abnormolities were observed in any of the animalsduring treatment and recovery period.No abnormalities were observed in the ophthalmological examination.

Test item did not had any effect on body weight and body weight gain (%). No toxicological relevant findings were observed in the haematology , biochemistry and urine anlysis parameters.

There were no treatment related gross and histopathological findings attributable to the test item magnesium glycerophosphat. Under the conditions of the experiment, the highest dose tested i.e., 1000 mg/kg is considered as NOEL (No Oberved Effect Level).  

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat