Registration Dossier

Administrative data

Description of key information

A 28-day oral toxicity study performed according to OECD/EC guideline and GLP principles is available with the NOAEL found to be 1000 mg/kg bw/d based on no adverse effects observed.

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:
Study conducted between 9th April 2014 and 12th December 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study
Qualifier:
according to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
Deviations:
yes
Remarks:
See principles of method if other than guideline part
Qualifier:
according to
Guideline:
EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))
Deviations:
yes
Remarks:
See principles of method if other than guideline part.
Principles of method if other than guideline:
Deviation No.1:
Clinical Observations:
On Day 2, the one-hour post dose clinical observations were performed approximately 25 minutes late in error.
All animals showed no clinical signs of toxicity, therefore, the late observations were considered not to affect the purpose or integrity of the study.

Necropsy:
The pancreas of Female No. 29 was misplaced at necropsy; therefore this tissue was not retained as specified in the Study Plan.

There were no macroscopic abnormalities detected in the pancreas for any treated animal and this tissue was not required for processing or examination. Therefore the absence of this tissue in one animal was considered not to affect the purpose or integrity of the study.

Animal Husbandry - Environment:
The Study Plan target values for temperature and relative humidity controls were 22 ± 3ºC and 50 ± 20% respectively. There was no deviation from the target range for temperature but the target range for relative humidity were exceeded for limited periods of four separate days of the study with the maximum humidity achieved being 75.54% RH. In general, the deviations from target ranges observed were minimal. While it is accepted that these deviations from the target range for relative humidity were less than ideal, overall it is considered that these deviations had no adverse impact on the scientific purpose of the study.
GLP compliance:
yes (incl. certificate)
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
A sufficient number of male and female Wistar Han™ RccHan™ WIST strain rats were obtained from Harlan Laboratories UK. Ltd., Oxon, UK. On receipt the animals were examined for signs of ill-health or injury. The animals were acclimatized for at least five days during which time their health status was assessed. A total of forty animals (twenty males and twenty females) were accepted into the study. At the start of treatment the males weighed 214 to 240g, the females weighed 161 to 188g, and were approximately six to eight weeks old.

The animals were housed in groups of five by sex in solid floor polypropylene cages with stainless steel mesh lids and softwood flake bedding (Datesand Ltd., Cheshire, UK). The animals were allowed free access to food and water. A pelleted diet (Rodent 2014C Teklad Global Certified Diet, Harlan Laboratories U.K. Ltd., Oxon, UK) was used. Mains drinking water was supplied from polycarbonate bottles attached to the cage. Environmental enrichment was provided in the form of wooden chew blocks and cardboard fun tunnels (Datesand Ltd., Cheshire, UK). The diet, drinking water, bedding and environmental enrichment were considered not to contain any contaminant at a level that might have affected the purpose or integrity of the study.

The animals were housed in a single air-conditioned room within the Harlan Laboratories Ltd., Shardlow, UK Barrier Maintained Rodent Facility. The rate of air exchange was at least fifteen air changes per hour and the low intensity fluorescent lighting was controlled to give twelve hours continuous light and twelve hours darkness. Environmental conditions were continuously monitored by a computerized system, and print-outs of hourly temperatures and humidities are included in the study records. The Study Plan target ranges for temperature and relative humidity were 22 ± 3 °C and 50 ± 20% respectively. Short term deviations from these targets were considered not to have affected the purpose or integrity of the study.

The animals were randomly allocated to treatment groups using a stratified body weight randomization procedure and the group mean body weights were then determined to ensure similarity between the treatment groups. The cage distribution within the holding rack was also randomized. The animals were uniquely identified within the study by an ear punching system routinely used in these laboratories.

Justification:
The rat was selected for this study as it is a readily available rodent species historically used in safety evaluation studies and is acceptable to appropriate regulatory authorities.

The study was performed between 09 April 2014 and 12 December 2014 (date of final histopathology report). The in-life phase of the study was conducted between 29 May 2014 (first day of treatment) and 26 June 2014 (final day of necropsy).

Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
Method of administration: The test item was administered daily, for up to twenty-eight consecutive days, by gavage using a stainless steel cannula attached to a disposable plastic syringe.

PREPARATION OF DOSING SOLUTIONS:
For the purpose of this study the test item was prepared at the appropriate concentrations as a solution in Distilled water. The stability and homogeneity of the test item formulations were determined by Harlan Laboratories Ltd., Shardlow, UK, Analytical Services. Results show the formulations to be stable for at least 22 days. Formulations were therefore prepared twice during the treatment period and stored at approximately 4 ºC in the dark.

The test item was administered daily, for up to twenty-eight consecutive days, by gavage using a stainless steel cannula attached to a disposable plastic syringe. Control animals were treated in an identical manner with 5 mL/kg of Distilled water.

DIET PREPARATION
- Rate of preparation of diet (frequency):
Not applicable

- Mixing appropriate amounts with (Type of food):
Not applicable.

- Storage temperature of food:
Not applicable.

VEHICLE:
A control group of five males and five females was dosed with vehicle alone (Distilled water).
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Samples of each test item formulation were taken and analyzed for concentration of FAT 40863/A TE at Harlan Laboratories Ltd., Shardlow, UK, Analytical Services.

The test item concentration in the test samples was determined by high performance liquid chromatography with UV detection (HPLC/UV). The results indicate that the prepared formulations were within acceptable ranges for the purpose of this study.

The following instrumental settings were used:
HPLC: Agilent Technologies 1100, incorporating autosampler and workstation
Column: Gemini 5µ C18 (100 x 4.6 mm id)
Mobil Phase: Eluent A: Water
Eluent B: Acetonitrile
Eluent C: 10g tetrabutylammonium bromide in 500 mL of acetonitrile

Time % A %B %C
0 65 30 5
5 0 95 5
7 0 95 5
7.1 65 30 5
12 65 30 5

Flow Rate: 1 mL/min
UV detector wavelength: 254 nm
Injection Volume: 100µL
Retention time: 4 minutes

Results:
Linearity: The data was found to have a linear correlation within the calibration range. The R2 fit of the calibration curve to the data was 0.999 and considered to be acceptable.

Test item formulations:
The formulations investigated during the study were found to comprise the test item in the range of 101% to 111%.

Conclusion:
In conclusion the results indicate the accurate use of the test itema nd distilled water as vehicle during this study. The formulations were found to be homogeneously prepared and sufficient formulation stability under storage conditions was approved.
Duration of treatment / exposure:
Test duration: 28 days
Frequency of treatment:
Dosing regime: 7 days/week
Remarks:
Doses / Concentrations:
dose levels of 0, 30, 300 and 1000 mg/kg/day EXAMPLE:
Basis:
actual ingested
No. of animals per sex per dose:
Male and Female: 5 animals per sex at 0 mg/kg/day
Male and Female: 5 animals per sex at 30 mg/kg/day
Male and Female: 5 animals per sex at 300 mg/kg/day
Male and Female: 5 animals per sex at 1000 mg/kg/day
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale:
Based on available toxicity information and compliance with regulatory hazard classification.

- 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):
Not applicable
Positive control:
None
Observations and examinations performed and frequency:
CLINICAL OBSERVATIONS: Yes
- Time schedule:
All animals were examined for overt signs of toxicity, ill-health or behavioral change immediately before dosing, up to thirty minutes post dosing and one hour after dosing. All observations were recorded.

BODY WEIGHT: Yes
- Time schedule for examinations:
Individual bodyweights were recorded on Day 1 (prior to start of treatment) and at weekly intervals thereafter. Body weights were also performed prior to terminal kill.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
Food consumption was recorded for each cage group at weekly intervals throughout the study.

FOOD EFFICIENCY: Yes
- Weekly food efficiency (bodyweight gain/food intake) was calculated.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations:
Water intake was visually observed daily except for Week 3 when gravimetric measurement was employed.

OPHTHALMOSCOPIC EXAMINATION: Not recorded

HAEMATOLOGY: Yes
- Time schedule for collection of blood: At the end of the treatment period (Day 28).
- Anaesthetic used for blood collection: No data: Not applicable
- Animals fasted: No
- How many animals: Five males and five females

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: At the end of the treatment period (day 28)

- Animals fasted: No

- How many animals: Hematological and blood chemical investigations were performed on all animals from each test and control group at the end of the study (Day 28). Blood samples were obtained from the lateral tail vein.

URINALYSIS: Not performed

NEUROBEHAVIOURAL EXAMINATION: Yes
Behavioural Assessments were undertaken for all animals at weekly intervals throughout the study. Motor Activity, Forelimb/Hindlimb Grip Strength and Sensory Reactivity were undertaken for all animals during the final week of treatment.

Motor Activity. Twenty purpose built 44 infra-red beam automated activity monitors were used to assess motor activity. Animals of one sex were tested at each occasion and were randomly allocated to the activity monitors. The tests were performed at approximately the same time each occasion (at least two hours after dosing), under similar laboratory conditions. The evaluation period was one hour for each animal. The time in seconds each animal was active and mobile was recorded for the overall one hour period and also during the final 20% of the period (considered to be the asymptotic period, Reiter and Macphail 1979).

Forelimb/Hindlimb Grip Strength. An automated grip strength meter was used. Each animal was allowed to grip the proximal metal bar of the meter with its forepaws. The animal was pulled by the base of the tail until its grip was broken. The animal was drawn along the trough of the meter by the tail until its hind paws gripped the distal metal bar. A record of the force required to break the grip for each animal was made. Three consecutive trials were performed for each animal. The assessment was developed from the method employed by Meyer et al (1979).

Sensory Reactivity:
Each animal was individually assessed for sensory reactivity to auditory, visual and proprioceptive stimuli. This assessment was developed from the methods employed by Irwin (1968) and Moser et al (1988). The scoring system used is outlined in The Key to Scoring System and Explanation for Behavioral Assessments and Sensory Reactivity Tests.

Sacrifice and pathology:
GROSS PATHOLOGY: Yes

On completion of the dosing period all animals were killed by intravenous overdose of sodium pentobarbitone followed by exsanguination.
All animals were subjected to a full external and internal examination, and any macroscopic abnormalities were recorded.

HISTOPATHOLOGY: Yes

All control and high dose animals were subjected to a full histological examination.

Since there were indications of treatment-related stomach, kidney and thyroid changes, examination was subsequently extended to include similarly prepared sections of the stomach and kidney for animals of either sex from animals in the low and intermediate groups and the sections of thyroid glands from males in the low and intermediate groups.

Other examinations:
OTHER INVESTIGATIONS:
MORTALITY DATA: All animals were examined for signs of ill-health immediately before dosing, up to thirty minutes post dosing and one hour after dosing. All
observations were recorded.

ORGAN WEIGHTS:
The following organs, removed from animals that were killed either at the end of the dosing period or at the end of the treatment-free period, were dissected free from fat and weighed before fixation:
Adrenals, Liver, Brain, Ovaries, Epididymides, Spleen, Heart, Testes, Kidneys, Thymus, Pituitary (post-fixation), Thyroid/Parathyroid (post fixation),
Prostate and Seminal Vesicles (with coagulating glands and fluids) and the Uterus with Cervix.

Statistics:
Where considered appropriate, quantitative data was subjected to statistical analysis to detect the significance of intergroup differences from control; statistical significance was achieved at a level of p < 0.05. Statistical analysis was performed on the following parameters:

Grip Strength, Motor Activity, Body Weight Change, Hematology, Blood Chemistry, Absolute Organ Weights, Body Weight-Relative Organ Weights.

Data were analyzed using the decision tree from the Provantis TM Tables and Statistics Module as detailed as follows:

Where appropriate, data transformations were performed using the most suitable method. The homogeneity of variance from mean values was analyzed using Bartlett’s test. Intergroup variance were assessed using suitable ANOVA, or if required, ANCOVA with appropriate covarities. Any transformed data were analyzed to find the lowest treatment level that showed a significant effect using the Williams Test for parametric data or the Shirley Test for non-parametric data. If no dose response was found but the data shows non-homogeneity of means, the data were analyzed by a stepwise Dunnett’s (parametric) or Steel (non-parametric) test to determine significant difference from the control group. Where the data were unsuitable for these analyses, pair-wise tests was performed using the Student t-test (parametric) or the Mann-Whitney U test (non-parametric).

Probability values (p) are presented as follows:

p<0.01 **
p<0.05 *
p>0.05 (not significant)
Clinical signs:
no effects observed
Description (incidence and severity):
One male and two females treated with 1000 mg/kg bw/day had an isolated incidence of orange staining of the fur. This observation was considered to reflect the coloration of the test item and was considered not to be toxicologically significant. No mortalities were observed either.
Mortality:
no mortality observed
Description (incidence):
One male and two females treated with 1000 mg/kg bw/day had an isolated incidence of orange staining of the fur. This observation was considered to reflect the coloration of the test item and was considered not to be toxicologically significant. No mortalities were observed either.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No toxicologically significant effects on body weight development were detected at 30, 300 or 1000 mg/kg bw/day.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No treatment-related effects on food consumption or food efficiency were detected at 30, 300 or 1000 mg/kg bw/day.
Food efficiency:
no effects observed
Description (incidence and severity):
No consistent pattern was apparent to indicate an effect of treatment on food efficiency. Intergroup differences in food efficiency were consistent with differences in body weight gain observed.
Water consumption and compound intake (if drinking water study):
no effects observed
Description (incidence and severity):
No obvious effect on water consumption was detected for any treated animal.
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Description (incidence and severity):
There were no toxicologically significant changes in any of the hematological parameters measured.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
All males had a statistically significant increase in bilirubin. All females had lower triglyceride levels compared to control. Females treated with 1000 mg/kg bw/day also had a statistically significant increase in albumin/globulin ratio.
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
There were no treatment-related changes in the behavioral parameters measured.
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
There were no toxicologically significant effects detected in the organ weights measured.
Gross pathological findings:
no effects observed
Description (incidence and severity):
No abnormalities were detected.
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Abnormalities were detected in the kidneys, stomach and thyroid glands
Histopathological findings: neoplastic:
not examined
Details on results:
Discussion:
The oral administration of FAT 40863/A TE to rats by gavage for a period of twenty-eight consecutive days at dose levels of 30, 300 and 1000 mg/kg bw/day showed treatment related microscopic abnormalities in animals of either sex treated with 1000 or 300 mg/kg bw/day and in females treated with 30 mg/kg bw/day.

No clinical signs of toxicity or any adverse effects on body weight development or food consumption was evident in treated animals. Although there were some statistically significant differences in treated animals from controls for the hematological parameters measured, these differences were considered not to be of toxicological significance.

Microscopic examination of the thyroids revealed minimal to mild diffuse hypertrophy of the follicular epithelium in all males given 1000 mg/kg bw/day and minimal hypertrophy in a control male, two males treated with 30 mg/kg bw/day and two males treated with 300 mg/kg bw/day. The findings in the low and intermediate dosage groups were considered to have arisen spontaneously, as a result of individual animal variation given the labile nature of the follicular epithelium in male rats. The incidence of diffuse hypertrophy of the thyroid follicular epithelium in females treated with 1000 mg/kg bw/day was comparable to controls. Although histopathological examination of the liver did not reveal any treatment-related microscopic changes, signs of altered metabolism were evident following blood chemical evaluations. All treated males had a statistically significant increase in bilirubin and all treated females had lower triglyceride levels in comparison to control. The reduction was dose dependent and differences in females treated with 300 or 1000 mg/kg bw/day achieved statistical significance. Females treated with 1000 mg/kg bw/day also had a statistically significant increase in albumin/globulin ratio. All individual values for these parameters were within background control ranges and the changes were considered not to represent an adverse effect of treatment, however, the intergroup differences in these parameters may still be indicative of a degree of hepatocellular induction. The appearance of follicular cell hypertrophy in the thyroid in this instance may, therefore, be considered likely to result from an initial increase in hepatic metabolism in response to treatment. Increases in hepatic metabolism lead to increased turnover of T3 and T4 resulting in increased stimulation of the thyroid gland due to inadequate negative feedback of TSH production. As the changes in the thyroid were considered likely to be a consequence of an adaptive response to treatment, the microscopic changes in the thyroid of males treated with 1000 mg/kg bw/day were considered to be adaptive in nature.

Minimal, mild or moderate hypertrophy of mucous neck cells was present in the glandular stomach of one female treated with 30 mg/kg bw/day, two males and one female treated with 300 mg/kg bw/day and four males and one female treated with 1000 mg/kg bw/day. The finding was not present in controls or males treated with 30 mg/kg bw/day. The single incidences of this finding in the treated female groups was considered most likely to have arisen spontaneously since there was no dosage relationship and although the findings in the intermediate and high dose males were greater, no hypertrophy was observed in low dose males. This finding, in males, was attributed to mild irritation of the stomach mucosa rather than a consequence of true systemic toxicity.

Minimal or mild increased intra-epithelial brown pigment was present in the cortical epithelial cells of the kidneys of females given 30 mg/kg bw/day and both sexes given 300 or 1000 mg/kg bw/day. This pigment was considered to be the test item or a metabolite of the test item and in the absence of any associated changes in kidney weights or any microscopic degenerative changes, the pigment accumulation was considered not to represent an adverse effect of treatment.
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: see 'Remark'
Dose descriptor:
NOEL
Effect level:
30 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male
Basis for effect level:
other: see 'Remark'
Critical effects observed:
not specified

Results:

Mortality: There were no unscheduled deaths.

Clinical Observations: Neither the type, incidence or distribution of clinical signs indicated an effect of treatment at 30,

300 or 1000 mg/kg bw/day.

Behavioral Assessment: There were no treatment-related changes in the behavioral parameters measured.

Functional Performance Tests - There were no toxicologically significant changes in functional performance.

Sensory Reactivity Assessments - There were no treatment-related changes in sensory reactivity.

Body Weight: No toxicologically significant effects on body weight development were detected at 30, 300 or 1000 mg/kg bw/day.

Food Consumption: No treatment-related effects on food consumption or food efficiency were detected at 30, 300 or 1000 mg/kg bw/day.

Water Consumption: No obvious effect on water consumption was detected for any treated animal.

Hematology: There were no toxicologically significant changes in any of the hematological parameters measured.

Blood Chemistry: All treated males had a statistically significant increase in bilirubin. All treated females had lower triglyceride levels in comparison to control; the reduction was dose dependent and differences in females treated with 300 or 1000 mg/kg bw/day achieved statistical significance. Females treated with 1000 mg/kg bw/day also had a statistically significant increase in albumin/globulin ratio.

Necropsy: No macroscopic abnormalities were detected.

Organ Weights:There were no toxicologically significant effects detected in the organ weights measured.

Histopathology:

Kidneys: In comparison to controls, minimal or mild increased intra-epithelial brown pigment was observed in the renal cortical epithelium of 2/5 females given 30 mg/kg bw/day, 3/5 males and 5/5 females given 300 mg/kg bw/day and all the animals given 1000 mg/kg bw/day.

Stomach, Glandular: Minimal to moderate hypertrophy of mucous neck cells was present in the glandular stomach of 1/5 females given 30 mg/kg bw/day, 2/5 males and 1/5 females given 300 mg/kg bw/day and 4/5 males and 1/5 females given 1000 mg/kg bw/day. The finding was not seen in controls or males given 30 mg/kg bw/day.

Thyroid Glands: The incidence and severity of minimal or mild diffuse hypertrophy of the follicular epithelium was greater in males given 1000 mg/kg bw/day than in controls. The incidence of this finding in females given the highest dosage and males given 30 or 300 mg/kg bw/day was comparable to controls.

Conclusions:
The oral administration of FAT 40863/A TE to rats by gavage for a period of twenty-eight consecutive days at dose levels of 30, 300 and 1000 mg/kg bw/day resulted in treatment-related effects in animals of either sex treated with 300 or 1000 mg/kg bw/day and females treated with 30 mg/kg bw/day. Therefore, a No Observed Effect Level (NOEL) for females cannot be established. The No observed Effect Level (NOEL) for males is 30 mg/kg bw/day.

Pigment deposits in the kidneys of animals of either sex treated with 1000 or 300 mg/kg bw/day and females treated with 30 mg/kg bw/day were considered to be the test item or a metabolite of the test item and the presence of this pigment was not associated with any pathological processes. Therefore the pigment accumulation was considered not to represent an adverse effect of treatment. The microscopic changes in the thyroid of males treated with 1000 mg/kg bw/day and the increased thyroid weights in all treated females were considered to result from an adaptive response to treatment and were therefore not adverse. Hypertrophy of the mucous neck cells of the glandular stomach in males treated with 300 or 1000 mg/kg bw/day was considered to result from local irritation and is not indicative of systemic toxicity.

For these reasons, 1000 mg/kg bw/day may be regarded as a 'No Observed Adverse Effect Level' (NOAEL) for animals of either sex.
Executive summary:
Introduction:

The study was designed to investigate the systemic toxicity of the test item and is compatible with the following regulatory guidelines:

 

·        Commission Directive 96/54/EC (Method B7).

·        The OECD Guidelines for Testing of Chemicals No. 407 "Repeated Dose 28 Day Oral Toxicity Study in Rodents" (adopted 03 October 2008).

·        Commission Regulation (EC) No 440/2008 of 30 May 2008, test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH).

 

Methods:.

The test item was administered by gavage to three groups, each of five male and five female Wistar Han™:RccHan™:WIST strain rats, for twenty-eight consecutive days, at dose levels of 30, 300 and 1000 mg/kg bw/day. A control group of five males and five females was dosed with vehicle alone (Distilled water).

 

Clinical signs,functional observations, body weight change, dietary intake and water consumption were monitored during the study. Hematology and blood chemistry were evaluated for all animals at the end of the study.

 

All animals were subjected to gross necropsy examination and histopathological evaluation of selected tissues was performed.

 

Results:

Mortality

There were no unscheduled deaths.

 

Clinical Observations

Neither the type, incidence or distribution of clinical signs indicated an effect of treatment at 30, 300 or 1000 mg/kg bw/day.

 

Behavioral Assessment

There were no treatment-related changes in the behavioral parameters measured.

 

Functional Performance Tests

There were no toxicologically significant changes in functional performance.

 

Sensory Reactivity Assessments

There were no treatment-related changes in sensory reactivity.

 

Body Weight

No toxicologically significant effects on body weight development were detected at 30, 300 or 1000 mg/kg bw/day.

 

Food Consumption

No treatment-related effects on food consumption or food efficiency were detected at 30, 300 or 1000 mg/kg bw/day.

 

Water Consumption

No obvious effect on water consumption was detected for any treated animal.

 

Hematology

There were no toxicologically significant changes in any of the hematological parameters measured.

 

Blood Chemistry

All treated males had a statistically significant increase in bilirubin. All treated females had lower triglyceride levels in comparison to control; the reduction was dose dependent and differences in females treated with 300 or 1000 mg/kg bw/day achieved statistical significance. Females treated with 1000 mg/kg bw/day also had a statistically significant increase in albumin/globulin ratio.

 

Necropsy

No macroscopic abnormalities were detected.

 

Organ Weights

There were no toxicologically significant effects detected in the organ weights measured.

 

Histopathology:

Kidneys: In comparison to controls, minimal or mild increased intra-epithelial brown pigment was observed in the renal cortical epithelium of 2/5 females given 30 mg/kg bw/day, 3/5 males and 5/5 females given 300 mg/kg bw/day and all the animals given 1000 mg/kg bw/day.

 

Stomach, Glandular: Minimal to moderate hypertrophy of mucous neck cells was present in the glandular stomach of 1/5 females given 30 mg/kg bw/day, 2/5 males and 1/5 females given 300 mg/kg bw/day and 4/5 males and 1/5 females given 1000 mg/kg bw/day. The finding was not seen in controls or males given 30 mg/kg bw/day.

 

Thyroid Glands: The incidence and severity of minimal or mild diffuse hypertrophy of the follicular epithelium was greater in males given 1000 mg/kg bw/day than in controls. The incidence of this finding in females given the highest dosage and males given 30 or 300 mg/kg bw/day was comparable to controls.

 

Conclusion:

The oral administration of FAT 40863/A TE to rats by gavage for a period of twenty-eight consecutive days at dose levels of 30, 300 and 1000 mg/kg bw/day resulted in treatment-related effects in animals of either sex treated with 300 or 1000 mg/kg bw/day and females treated with 30 mg/kg bw/day. Therefore, a No Observed Effect Level (NOEL) for females cannot be established. The No observed Effect Level (NOEL) for males is 30 mg/kg bw/day.

 

Pigment deposits in the kidneys of animals of either sex treated with 1000 or 300 mg/kg bw/day and females treated with 30 mg/kg bw/day were considered to be the test item or a metabolite of the test item and the presence of this pigment was not associated with any pathological processes. Therefore the pigment accumulation was considered not to represent an adverse effect of treatment. The microscopic changes in the thyroid of males treated with 1000 mg/kg bw/day and the increased thyroid weights in all treated females were considered to result from an adaptive response to treatment and were therefore not adverse. Hypertrophy of the mucous neck cells of the glandular stomach in males treated with 300 or 1000 mg/kg bw/day was considered to result from local irritation and is not indicative of systemic toxicity.

For these reasons, 1000 mg/kg bw/day may be regarded as a 'No Observed Adverse Effect Level' (NOAEL) for animals of either sex.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
Study Klimisch 1.

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

The oral administration of FAT 40863/A TE to rats by gavage for a period of twenty-eight consecutive days at dose levels of 30, 300 and 1000 mg/kg bw/day resulted in treatment-related effects in animals of either sex treated with 300 or 1000 mg/kg bw/day and females treated with 30 mg/kg bw/day. Therefore, a No Observed Effect Level (NOEL) for females cannot be established. The No observed Effect Level (NOEL) for males is 30 mg/kg bw/day.

Pigment deposits in the kidneys of animals of either sex treated with 1000 or 300 mg/kg bw/day and females treated with 30 mg/kg bw/day were considered to be the test item or a metabolite of the test item and the presence of this pigment was not associated with any pathological processes. Therefore the pigment accumulation was considered not to represent an adverse effect of treatment. The microscopic changes in the thyroid of males treated with 1000 mg/kg bw/day and the increased thyroid weights in all treated females were considered to result from an adaptive response to treatment and were therefore not adverse. Hypertrophy of the mucous neck cells of the glandular stomach in males treated with 300 or 1000 mg/kg bw/day was considered to result from local irritation and is not indicative of systemic toxicity.

For these reasons, 1000 mg/kg bw/day may be regarded as a 'No Observed Adverse Effect Level' (NOAEL) for animals of either sex.


Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
Only this study is available

Justification for classification or non-classification

The available data on repeated dose toxicity of the test substance do not meet the criteria for classification according to Regulation (EC) 1272/2008 or Directive 67/548/EEC, and are therefore conclusive but not sufficient for classification.