Registration Dossier

Diss Factsheets

Administrative data

Endpoint:
repeated dose toxicity: oral, other
Remarks:
Repeated Dose 90-day Oral Toxicity Study in Rodents
Type of information:
experimental study
Adequacy of study:
key study
Study period:
04 May 2016 - 25 November 2016
Reliability:
1 (reliable without restriction)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report date:
2016

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Version / remarks:
Revised version, adopted 21st September 1998.
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
Sulphuric acid, compound with graphite
EC Number:
235-819-4
EC Name:
Sulphuric acid, compound with graphite
Cas Number:
12777-87-6
Molecular formula:
Cm+ HSO4-* n H2SO4
IUPAC Name:
methane; sulfuric acid
Details on test material:
ECGA97914xH(21T9-2)
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL- batch No.of test material: 8T5-2 - Expiration date of the lot/batch: 01.01.2021 - Purity test date: 16.04.2015 STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL- Storage condition of test material: Room Temperature (20 ± 5°C) - Solubility and stability of the test substance in the solvent/vehicle: H2O: 0,1-1 g/l (in water)TREATMENT OF TEST MATERIAL PRIOR TO TESTING- Treatment of test material prior to testing: The test item was suspended in water. The suspensions were prepared freshly before each application. To avoid sedimentation, the suspension was stirred or shaken until application. - Preliminary purification step (if any): noFORM AS APPLIED IN THE TEST (if different from that of starting material): The test item consists of non-soluble grey/black crystals ..and was therefore applied as a suspension of homogeneous/uniform flakes.OTHER SPECIFICS: The test item is known to be stable for more than 5 years after production and its stability was not affected by being suspended in water. Therefore the daily documentation of weights and volumes of each suspension as well as the volume applied to each individual animal is sufficient to meet the requirements of guideline 21 CFR 58.31(d). An additional testing of the suspensions for identity, strength, purity, stability, and uniformity is not applicable for graphite crystals.

Test animals

Species:
rat
Strain:
Wistar
Details on species / strain selection:
Species: Rattus norvegicus Strain: Wistar Han IGS (Crl:WI(Han))
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS- Source: Charles River, Sandhofer Weg 7, 97633 Sulzfeld, Germany- Females (if applicable) nulliparous and non-pregnant: yes- Age at study initiation: 5-6 weeks - Housing: Air conditioning with approx. 15 air changes per hour, temperature at 22 ± 3°C, relative humidity at 30-70% (aimed limits - due to meteorological circumstances, relative humidity was above 70% for short term), artificial lighting, 12 h light/12 h dark- Diet (e.g. ad libitum): Maintenance diet rat/mouse, pellets, No. 1324 TPF (Altromin GmbH & Co. KG, 32791 Lage), ad lib. - Water (e.g. ad libitum): Sterilized community tap water ad lib- Acclimation period: 7-10 days ENVIRONMENTAL CONDITIONS- Temperature (°C): 22 ± 3°C- Humidity (%): relative humidity at 30-70% - Air changes (per hr): 15- Photoperiod (hrs dark / hrs light): artificial lighting, 12 h light/12 h dark IN-LIFE DATES: From: To: The first day of application was indicated as day 1. The in-life phase was terminated on day 91.

Administration / exposure

Route of administration:
oral: gavage
Details on route of administration:
The test item SAT graphite was administered suspended to sterile community tap water.
Vehicle:
water
Remarks:
Community tap water, sterile filtered
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS: The test item was suspended in water. The suspensions were prepared freshly before each application. To avoid sedimentation, the suspension was stirred or shaken until application. For each individual animal, the syringes were inverted upside down immediately before oral application to ensure a homologues suspension. The preparation was intended for an application volume of 5 mL per kg body weight per gavage. VEHICLE - Justification for use and choice of vehicle (if other than water): - - Concentration in vehicle: 1000, 250 and 62,5 mg/kg BW/day - Amount of vehicle (if gavage): 5 mL/kg BW
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
The test item is known to be stable for more than 5 years after production and its stability was not affected by being suspended in water. Therefore the daily documentation of weights and volumes of each suspension as well as the volume applied to each individual animal is sufficient to meet the requirements of guideline 21 CFR 58.31(d). An additional testing of the suspensions for identity, strength, purity, stability, and uniformity is not applicable for graphite crystals.
Duration of treatment / exposure:
90 days
Frequency of treatment:
Administration was performed daily
Doses / concentrationsopen allclose all
Dose / conc.:
62.5 mg/kg bw/day (actual dose received)
Dose / conc.:
250 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: In preparation for this study a dose range finding (DRF) study with dose escalation was performed. Hence, the test item was administered daily by oral gavage at escalating dose levels over a time period of 18 days. Doses were increased every three day; the highest dose (1000 mg/kg BW) was applied for six days. No acute toxicological effects were observed in the study. In accordance with the Sponsor, and based on the regulatory requirements, the maximum dose for the 90-day toxicity study was 1000 mg/kg body weight. For the endorsed two lower dose-levels a factor of 4 was chosen, to ensure an appropriate NOAEL for the test item.- Stratification and randomization: This study was designed as a randomized controlled trial (RCT). Upon arrival at the test facility, rats were weighed individually and grouped into three weight clusters per sex. Individual animals from the central clusters were placed consecutively into prepared cages until the cluster was depleted. Animals from the remaining clusters were caged alternately in the same manner. - Rationale for randomization: To avoid a potential bias during animal distribution it is in accordance with current scientific knowledge that a pseudo-randomization as described above is sufficient to generate the modest amount of unpredictability required for preclinical and clinical studies.
Positive control:
no

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes - Time schedule: performed daily throughout the study up to the day of necropsy- Cage side observations checked in table were included.DETAILED CLINICAL OBSERVATIONS: Yes - Time schedule: General clinical signs were monitored daily during the in-life phase. Detailed clinical signs were recorded once weekly. BODY WEIGHT: Yes - Time schedule for examinations: weeklyFOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): yes - once weekly- 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 specifiedFOOD 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 specifiedWATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes / No / Not specified- Time schedule for examinations:OPHTHALMOSCOPIC EXAMINATION: Yes / No / Not specified- Time schedule for examinations:- Dose groups that were examined:HAEMATOLOGY: Yes / No / Not specified- Time schedule for collection of blood:- Anaesthetic used for blood collection: Yes (identity) / No / Not specified- Animals fasted: Yes / No / Not specified- How many animals:- Parameters checked in table [No.?] were examined.CLINICAL CHEMISTRY: Yes / No / Not specified- Time schedule for collection of blood:- Animals fasted: Yes / No / Not specified- How many animals:- Parameters checked in table [No.?] were examined.URINALYSIS: Yes / No / Not specified- Time schedule for collection of urine:- Metabolism cages used for collection of urine: Yes / No / Not specified- Animals fasted: Yes / No / Not specified- Parameters checked in table [No.?] were examined.NEUROBEHAVIOURAL EXAMINATION: Yes / No / Not specified- Time schedule for examinations:- Dose groups that were examined:- Battery of functions tested: sensory activity / grip strength / motor activity / other:IMMUNOLOGY: Yes / No / Not specified- 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 (see table) HISTOPATHOLOGY: Yes (see table)
Statistics:
Descriptive statistics: The arithmetic mean, standard deviation and median were calculated for all grouped numerical data originating from the monitoring of grip strength, gross pathology (organ weights) and parts of the hemogram (for details see appendix). Where appropriate, detailed column statistics were applied (minimum / maximum data, 25% quartiles, standard error, upper and lower confidence interval 95%). Data of the IRWIN- and the beam walking test were analyzed using a scoring system as described in previous chapters of the report.Inductive statistics:For basic analysis the respective test item groups were compared to the vehicle group by assessing of statistical significance using a two-tailed unpaired Student´s t-test. For all calculations, the significance level was set to 0,05. In case the basic analysis returns a statistical significance, additional inductive statistical analysis was applied as outlined in the main decision tree (see attachment). Except for individual blood parameters (monocytes, eosinophils, basophils; only descriptive statistics) it was assumed that collected blood data study were metrically scaled and normally distributed (Gaussian). Most statistical hypotheses in this study were best characterized as “many to one”– comparing a vehicle control vs. the treatment groups, respectively. Therefore the adequate analysis methods was one-way ANOVA (analysis of variance) followed by a post- hoc Dunnett´s t-test (for hemograms and organ weights), or two-way ANOVA followed by a post-hoc Bonferroni test (for body weight analysis). For all calculations, the significance level was set to 0,05. This further inductive statistics were calculated using Graph Pad Prism for Mac, Version 5. All statistical data were documented within the appendix. Whenever the term “significant” is used in the study report, it stands for “statistically significant”.

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Description (incidence and severity):
General clinical signs were monitored daily during the in-life phase. Detailed clinical signs were recorded once weekly. Throughout the course of the study no test item related effect was observed in any experimental animal group.
Mortality:
no mortality observed
Description (incidence):
There were no test item related fatalities.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Males: The mean body weight and body weight gain observed in male experimental animals were within normal range for male rats of this strain and age. Using Bonferroni post hoc t-test (after Two-way ANOVA), no differences between the test item treated animals of the high- medium- or low dose group to the vehicle control group were found. No test item related effects on the body weight or body weight gain were found in any dose group. Females:The mean body weight and body weight gain observed in female experimental animals, was within normal range for female rats of this strain and age. Using Bonferroni post hoc t-test (after ANOVA), no differences between the test item treated animals of the high- medium- or low dose group to the vehicle control group were found. No test item related effects on the body weight or body weight gain were found in any dose group.In summary, no indication for toxicological effects on body weight or body weight gain was observed in any animal (male and female) of the experimental dose groups during the in-life phase.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Males: The food consumption observed in male experimental animals was within the normal range for male rats of this strain and age. The differences in food consumption were low during the entire in life phase. Normalized to the body weight, differences in food consumption of 2% ± 1% (high dose), 3% ± 2% (medium dose) or 4% ±2 % (low dose group) compared to the male vehicle group were observed, respectively. Females:The food consumption observed in experimental female animals was within normal range for female rats of this strain and age. The differences in food consumption were low during the entire in life phase. Normalized to the body weight, differences in food consumption of 3% ± 2% (high dose), 5% ± 4% (medium dose) or 5% ±2 % (low dose group) compared to the female vehicle group were observed, respectively. In summary, few and low differences in food consumption were observed between the experimental dose groups over the study course. The differences were within the natural occurring variability not considered test item related.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
no effects observed
Description (incidence and severity):
Males:The water consumption observed in experimental male animals, was within normal range for male rats of this strain and age. The differences in water consumption were low during the entire in life phase. Normalized to the body weight, differences in food consumption of 6% ± 4% (high dose), 5% ± 5% (medium dose) or 5% ±4 % (low dose group) compared to the male vehicle group were observed, respectively. Females:The majority of water consumption observed in experimental female animals, was within normal range for female rats of this strain and age. The differences in water consumption were low during the entire in life phase. Normalized to the body weight, differences in food consumption of 4% ± 3% (high dose), 4% ± 3% (medium dose) or 7% ± 4 % (low dose group) compared to the male vehicle group were observed, respectively.In summary, few and low differences in water consumption were observed between the experimental dose groups over the study course. The differences were within the natural occurring variability not considered test item related.
Ophthalmological findings:
no effects observed
Description (incidence and severity):
Ophthalmological examination of the high dose animals and the vehicle control revealed no treatment related effects on the eyes. In accordance with the study plan, the medium and low dose groups were not examined.
Haematological findings:
no effects observed
Description (incidence and severity):
The statistical analysis of the male high dose group revealed a higher level of relative neutrophil granulocytes [NEU] (20,9% ±2,4%) compared to respective control (16,3% ±3,01%). However, when compared to the respective control, for the absolute numbers of neutrophil granulocytes [NEUA], no significant change could be observed. Furthermore, this increase was within the 5%-95% percentile of the laboratory´s historical data (10% -23%). In the male medium dose group, hemoglobin [HB] and hematocrit [HKT] values were slightly below those of the control group, but within the 5% -95% percentile of the laboratory´s historical data. The statistical analysis of the male low dose group revealed a lower level of relative Lymphocytes [LYM] (74% ±5,8%) compared to respective control (79,1% ±3,31%) However, when compared to the respective control, for the absolute numbers of lymphocytesl [LYMA], no significant change could be observed. Furthermore, this increase was within the 5%-95% percentile of the laboratory´s historical data (71% -87%) In the female low dose group, hemoglobin [HB] values were slightly below those of the control group, but within the 5% -95% percentile of the laboratory´s historical data. Therefore a biological relevance could be excluded with high probability.
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
The statistical analysis of all parameters of clinical biochemistry yielded incidental (and partly contradictory) significances. E.g. lower concentrations of the electrolytes calcium [CA] and chloride [Cl] compared to the control group were found in the male high dose group. In contrast, the male low dose group showed a lower calcium [CA] and chloride [Cl] concentrations, compared to the control group. None of the findings was supported by any positive correlate after histopathological analysis of the relevant organs. Summarized, all significant discrepancies were within the 5%-95% percentiles of the laboratory´s historical data Therefore, it can be summarized, that no relevant or adverse test item induced effects on any hematological and clinical biochemistry parameter could be detected in any treatment group. Therefore, all blood analysis findings were toxicologically considered as not adverse.
Urinalysis findings:
no effects observed
Description (incidence and severity):
No abnormalities detected
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
Overall, the IRWIN tests returned no abnormal clinical signs or test item related changes in any of the animal’s behavior during the course of this study.
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
Analysis of organ weight data (Dunnett’s post-hoc t-test after One-way ANOVA) returned no significant changes in the absolute organ weights of the male or female animals, when comparing test item treated groups to the respective control groups. The relative heart weight of the male low dose and the female medium dose group were slightly reduced compared with their respective vehicle controls. Moreover, the relative weight of the right ovary was slightly reduced in the female high dose animals. Since neither the absolute weights of these organs nor the absolute body weights differ significantly, this is considered to be an incidental finding and not a test item related effect.
Gross pathological findings:
no effects observed
Description (incidence and severity):
Based on gross and microscopic pathology, no dose-limiting toxicity was detected.
Neuropathological findings:
no effects observed
Description (incidence and severity):
No abnormalities detected.
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
There were no microscopic observations that were attributed to the test item, that were consistent with the spectrum of background pathology in Wistar Han rats of this age.
Histopathological findings: neoplastic:
no effects observed
Description (incidence and severity):
There were no microscopic observations that were attributed to the test item, that were consistent with the spectrum of background pathology in Wistar Han rats of this age.
Details on results:
Viability and general clinical signs: During the whole in-life phase no test item related abnormalities regarding general clinical signs and behavior were observed in any of the remaining experimental animals. Detailed clinical signs (IRWIN test): On a weekly basis, the health status and overall physiological condition of all animals was assessed based on 30 endpoints according to appearance, motor activity, excitation, stereotypies, and abnormal behavior. In week 7, an increased exploration behavior was observed for one animal of the female high dose group (E240/2). This observation was rated as a natural variability of individual activities. There were no pathologic findings. Overall, the IRWIN tests returned no abnormal clinical signs or test item related changes in any of the animal’s behavior during the course of this study. Beam walk test and grip strength: The beam-walking scores of all animals were evenly distributed over all dose groups and the vehicle control group, both male and female, showing only slight differences in their respective latency periods. The noted differences were within the normal range of variability found among healthy animals. No test item related effects were found. All animals of the male vehicle group returned grip strength values from 592 g to 1307 g (977 ± 237 g), whereas the animals from the high dose group returned significantly lower grip strength values from 297 g to 867 g (622 ± 210 g). The values of the medium- (593 g to 1302 g (899 ± 235 g)) and low- dose group (278 g to 1036 g (715 ± 276 g)) did not differ significantly when compared to the vehicle control group. The grip strength values of the female high- (214 g to 985 g (766 ± 218 g)), medium (277 g to 1277 g (775 ± 293 g)) and low- dose group (431 g to 1095 g (721 ± 241 g)) did not differ significantly differ when compared to the vehicle group (306 g to 1034 g (593 ± 236 g)). Given the facts that male and female high- medium- and low- dose groups are similar to those of the respective vehicle groups, these findings were considered to be accidental. Moreover, few animals refused to stick to the grid properly, resulting in a high variability of the results. Small and inconsistent variation in grip strength between the dose groups are regarded as an accidental finding and was assessed to be not test item related. Ophthalmological examination: Ophthalmological examination of the high dose animals and the vehicle control revealed no treatment related effects on the eyes. In accordance with the study plan, the medium and low dose groups were not examineBody weight: In summary, no indication for toxicological effects on body weight or body weight gain was observed in any animal (male and female) of the experimental dose groups during the in-life phase.Food consumption: In summary, few and low differences in food consumption were observed between the experimental dose groups over the study course. The differences were within the natural occurring variability not considered test item related. Water consumption: The water consumption of male and female animals was calculated as a mean of each experimental group per animal and day, as well as in relation to the animal body weight. In summary, few and low differences in food consumption were observed between the experimental dose groups over the study course. The differences were within the natural occurring variability not considered test item related.Hematology and clinical biochemistry: It can be summarized, that no relevant or adverse test item induced effects on any hematological and clinical biochemistry parameter could be detected in any treatment group. Therefore, all blood analysis findings were toxicologically considered as not adverse. Necropsy - Macroscopic observations : There were no macroscopic observations that were attributed to the test item Analysis of organ weights : Analysis of organ weight data (Dunnett’s post-hoc t-test after One-way ANOVA) returned no significant changes in the absolute organ weights of the male or female animals, when comparing test item treated groups to the respective control groups.Histopathology findings: There were no microscopical findings attributed to the test item.

Effect levels

Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
clinical biochemistry
clinical signs
food consumption and compound intake
haematology
histopathology: neoplastic
histopathology: non-neoplastic
mortality
water consumption and compound intake

Target system / organ toxicity

Critical effects observed:
no

Any other information on results incl. tables

Summary of results:

 

Dose Level

High dose

Medium dose

Low dose

Vehicle

 

Sex

M

F

M

F

M

F

M

F

Parameter

 

 

 

 

 

 

 

 

 

Viability / general clinical signs

 

NAD

NAD

NAD

NAD

NAD

NAD

NAD

NAD

Clinical signs / behavior

 

NAD

NAD

NAD

NAD

NAD

NAD

NAD

NAD

Body weight

 

NAD

NAD

NAD

NAD

NAD

NAD

NAD

NAD

Food consumption

 

NAD

NAD

NAD

NAD

NAD

NAD

NAD

NAD

Water consumption

 

NAD

NAD

NAD

NAD

NAD

NAD

NAD

NAD

Hematology

 

[NEU]↑

NAD

[HB]↓

[HKT] ↓

NAD

[LYM] ↓

[HB] ↓

NAD

NAD

Clinical biochemistry

 

[CA] ↓

[CL] ↓

[ALB] ↓

[GLOBU] ↓

[PROT↑

[GLUC] ↑

NAD

[NA] ↑

[GLUC] ↑

[CA] ↓

[CL] ↓

[ALB] ↓

 

[HST] ↑

[NA] ↑

[CA] ↑

[CL] ↑

[GLOBU] ↑

[PROT]↑

NAD

NAD

Necropsy*

 

NAD

NAD

Focal bone alteration

(1 of 10 animals)

Adhesion of a liver lobe to the pylorus (1 of 10 animals)

NAD

NAD

NAD

NAD

Organ weight

 

NAD

NAD

NAD

NAD

NAD

NAD

NAD

NAD

Histopathology*

 

NAD

NAD

Exostosis in the femur

Hemorrhage, inflammation necrosis (liver)

n.e.

n.e.

NAD

NAD

* only toxicological relevant findings are listed

M = Male animals

F = Female animals

NAD = No abnormality detected

n.e. = not examined

↑= higher compared to control

↓= lower compared to control

Applicant's summary and conclusion

Conclusions:
A continuous oral administration of the test item SAT Graphite to Wistar rats at dose levels of 1000, 250 and 62,5 mg/kg bodyweight over a period of 91 days resulted in some minor changes in clinical biochemistry blood parameters and lymphoid organs. However, in conclusion with the histopathological result these particular observations were toxicologically considered as not adverse. For this reason, the NOAEL of SAT Graphite after administration over 91 days is determined at 1000 mg/kg BW per day.
Executive summary:

The aim of the study was to assess data on the potential subacute toxicity of SAT graphite after oral administration in Wistar Han rats in a 90-day oral toxicity test. The test item SAT graphite was administered suspended to sterile community tap water over a period of 90-days. Administration was performed daily by oral gavage at dosages of 62,5, 250 or 1000 mg/kg bodyweight per animal and day, respectively. A control group of 10 male and 10 female rats received vehicle (sterile community tab water) as control; all other parameters were identical.

General clinical signs, viability, reactions to treatment and conspicuous behavioral traits of all experimental animals were monitored daily during the in-life phase. Detailed clinical signs, individual body weight as well as group food and water consumption was recorded once weekly.

At the end of the treatment period, blood samples from all animals were drawn to provide data on hematology, clinical biochemistry and blood clotting time. All animals were sacrificed after bleeding and examined by gross necropsy. According to the study plan, weights of selected organs were recorded, and tissues and organs were preserved. Samples from the high dose groups and the vehicle groups were processed for histopathological examination.

Results:

Fatalities: There were no test item related fatalities.

General and detailed clinical signs: Throughout the course of the study no test item related effect was observed in any experimental animal group.

Body weight, food and water consumption: No abnormal differences regarding body weight and body weight gain were observed between all test item treated animals (neither male nor female) and the respective vehicle control animals. No test item related changes in food and water consumption of all test item treated animals (males and females) were observed throughout the whole in-life phase when compared to their respective control animals.

Hematology and clinical biochemistry: In all test animals no relevant or adverse test item induced effects on any hematological parameter could be detected. Most prominently in the high dose groups (males and females) some changes in clinical biochemistry parameters were observed. The results from the histopathological examination of selected organs revealed no positive correlates. Therefore all observed clinical parameters were regarded as not adverse.

Necropsy: In summary, all macroscopic findings were regarded as not treatment related. Except the relative (not absolute) heart weight of the female low dose group, no significant changes in organ weights were noted.

Histopathological examination: There were no microscopic observations that were attributed to the test item, that were consistent with the spectrum of background pathology in Wistar Han rats of this age.

A continuous oral administration of the test item SAT Graphite to Wistar rats at dose levels of 1000, 250 and 62,5 mg/kg bodyweight over a period of 91 days resulted in some minor changes in clinical biochemistry blood parameters and lymphoid organs. However, in conclusion with the histopathological result these particular observations were toxicologically considered as not adverse. For this reason, the NOAEL of SAT Graphite after administration over 91 days is determined at 1000 mg/kg BW per day.