Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Administrative data

Description of key information

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

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:
Testing was conducted between the 2nd April 2015 and 16th January 2015.
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)
Deviations:
yes
Remarks:
See principles of method if other tahn guideline part.
Qualifier:
according to guideline
Guideline:
EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))
Deviations:
no
Principles of method if other than guideline:
Deviation No.1

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 three separate days of the study with the maximum humidity achieved being 78.27% 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. QA statement)
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
A sufficient number of male and female Wistar Han™:RccHan™:WIST strain rats were obtained from Harlan Laboratories U.K. Ltd., Oxon, UK. On receipt the animals were examined for signs of ill-health or injury. The animals were acclimatized for seven 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 261g, the females weighed 162 to 184g, 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. A certificate of analysis of the batch of diet used is given in Appendix 24. 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; see deviations from Study Plan.

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.
Route of administration:
oral: gavage
Vehicle:
arachis oil
Details on oral exposure:
Method of administration:
Gavage:
PREPARATION OF DOSING SOLUTIONS:
For the purpose of this study the test item was prepared at the appropriate concentrations as a suspension in Arachis oil BP.


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
- Justification for use and choice of vehicle(if other than water):
Not applicable (N/A)

- Concentration in vehicle:
3, 30 and 200 mg/ml.

- Amount of vehicle (if gavage):
6 ml/kg bodyweight

- Lot/batch no. (if required):
Not stated in report

- Purity:
Not stated in report
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The concentration of test material formulations was determined by high performance liquid chromatography. The test material formulations were sampled and analysed for up to fourteen days. The analytical method has been satisfactorily validated in terms of linearity, specificity and accuracy for the purposes of the study.
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 bw/day
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:
CAGE SIDE 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.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Prior to the start of treatment and on Days 7, 14, 21 and 27, all animals were observed for signs of functional/behavioral toxicity.
- Detailed individual clinical observations were performed for each animal using a purpose built arena. The following parameters were observed:
Gait Hyper/Hypothermia
Tremors Skin color
Twitches Respiration
Convulsions Palpebral closure
Urination Bizarre/Abnormal/Stereotypic behavior
Salivation Defecation
Pilo-erection Transfer arousal
Exophthalmia Tail elevation
Lachrymation

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

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

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations:
Water intake was observed daily, for each cage group, by visual inspection of the water bottles for any overt changes except during Week 3 where water intake was measured gravimetrically.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Hematological and blood chemical investigations were performed on all animals from each test and control group at the end of the study (Day 28).
- Anaesthetic used for blood collection: No data
- Animals fasted:
No
- How many animals:
all animals from each testa nd control group.
- The following parameters were examined:
Hemoglobin (Hb)
Erythrocyte count (RBC)
Hematocrit (Hct)
Erythrocyte indices - mean corpuscular hemoglobin (MCH)
- mean corpuscular volume (MCV)
- mean corpuscular hemoglobin concentration (MCHC)
Total leukocyte count (WBC)
Differential leukocyte count - neutrophils (Neut)
- lymphocytes (Lymph)
- monocytes (Mono)
- eosinophils (Eos)
- basophils (Bas)
Platelet count (PLT)
Reticulocyte count (Retic)
Prothrombin time (CT) was assessed by ‘Innovin’ and Activated partial thromboplastin time (APTT) was assessed by ‘Actin FS’ using samples collected into sodium citrate solution (0.11 mol/L).

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Hematological and blood chemical investigations were performed on all animals from each test and control group at the end of the study (Day 28).

- Animals fasted: Yes

- How many animals: all animals from each test and control group.

- The following parameters were measured on plasma from blood collected into tubes containing lithium heparin anti-coagulant:
Urea Inorganic phosphorus (P)
Glucose Aspartate aminotransferase (ASAT)
Total protein (Tot.Prot.)
Albumin Alkaline phosphatase (AP)
Creatinine (Creat)
Sodium (Na+) Total cholesterol (Chol)
Potassium (K+) Total bilirubin (Bili)
Chloride (Cl-) Bile acids
Calcium (Ca++) Alanine aminotransferase (ALAT)
Albumin/Globulin (A/G) ratio (by calculation)

URINALYSIS: Not tested

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: Prior to the start of treatment and on Days 7, 14, 21 and 27, all animals were observed for signs of functional/behavioral toxicity. Functional performance tests were also performed on all animals during Week 4, together with an assessment of sensory reactivity to different stimuli.
- Dose groups that were examined: All dose groups were examined.
- Battery of functions tested: The following functions were tested sensory activity, grip strength and motor activity.

Prior to the start of treatment and on Days 7, 14, 21 and 27, all animals were observed for signs of functional/behavioral toxicity. Functional performance tests were also performed on all animals during Week 4, together with an assessment of sensory reactivity to different stimuli.

Functional Performance Tests:
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.

The following parameters were observed:

Grasp response Touch escape
Vocalization Pupil reflex
Toe pinch Blink reflex
Tail pinch Startle reflex
Finger approach



Sacrifice and pathology:
GROSS PATHOLOGY: Yes
All animals were subjected to a full external and internal examination, and any macroscopic abnormalities were recorded.

On completion of the dosing period all animals were killed by intravenous overdose of a suitable barbiturate agent followed by exsanguination.

Thyroid Hormone Assessment:
At termination, blood samples were taken from the exsanguination procedure and the serum from each animal was stored frozen at approximately -20 °C. No treatment-related effects on the pituitary-thyroid axis were identified, therefore these samples were discarded.

HISTOPATHOLOGY: Yes

All control and high dose animals were subjected to a full histological examination and low and intermediate group animals were routinely subjected to examination of liver and spleen.

All tissues were dispatched to the histology processing Test Site (Propath UK Ltd) for processing (Principal Investigator: N Fower).

Since there were indications of treatment-related kidney, liver and spleen changes, examination was subsequently extended to include similarly prepared sections of kidney, liver and spleen from animals in the low and intermediate groups.

Microscopic examination was conducted by the Study Pathologist (P Millar at Peter Millar Associates Ltd., 3 Queen Charlotte Lane, Edinburgh, EH6 6AY).


Other examinations:
MORTALITY DATA:
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.

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)
Prostate and Seminal Vesicles Uterus with Cervix (with coagulating glands and fluids)
Thyroid/Parathyroid (post fixation)


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 ProvantisTM 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):
There were no unscheduled deaths throughout the study. Neither the type, incidence or distribution of clinical signs indicated an effect of treatment at 30, 300 or 1000 mg/kg bw/day.
Mortality:
no mortality observed
Description (incidence):
There were no unscheduled deaths throughout the study. Neither the type, incidence or distribution of clinical signs indicated an effect of treatment at 30, 300 or 1000 mg/kg bw/day.
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 effect on food consumption or food efficiency was 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 on treatment of food conversion efficiency. Intergroup differences in food conversion efficiency were consistent with differences in body weight gains 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:
effects observed, treatment-related
Description (incidence and severity):
Males treated with 300 or 1000 mg/kg bw/day had a significant reduction in mean corpuscular hemoglobin concentration and a significant increase in platelet count. Females treated with 1000 mg/kg bw/day had a reduction in haemoglobin.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
Males treated with 1000 mg/kg bw/day had a statistically significant increase in cholesterol levels. No such effects were detected in females treated with 1000 mg/kg bw/day or in animals of either sex treated with 300 or 30 mg/kg bw/day.
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. There were no toxicologically significant changes in functional performance. There were no toxicologically significant changes in sensory reactivity.
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
No treatment-related effects were detected on the organ weights measured.
Gross pathological findings:
no effects observed
Description (incidence and severity):
Neither the type, incidence or distribution of macroscopic findings indicated an adverse effect of treatment at 30, 300 or 1000 mg/kg bw/day.
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Microscopic changes were detected in the kidneys, liver and spleen. Please see the any other information on results section for more details.
Histopathological findings: neoplastic:
not examined
Details on results:
Discussion:
The oral administration of FAT 41044/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 was well tolerated, however, treatment related microscopic abnormalities were apparent in animals of either sex treated with 1000 and 300 mg/kg bw/day.

There were no adverse effects of treatment detected in clinical signs, body weight development or food consumption in treated animals. Microscopic examination of the livers revealed hypertrophy or centrilobular hypertrophy in four males and one female treated with 1000 mg/kg bw/day and one male treated with 300 mg/kg bw/day. These findings are suggestive of a response to mixed function oxidase induction, and in the absence of other pathological changes are considered to be adaptive and of limited toxicological significance. These adaptive changes were considered to be responsible for the increased cholesterol levels apparent in males treated with 1000 mg/kg bw/day.

Microscopic assessment of the kidneys revealed an increase in incidence and severity of hyaline droplet accumulation in the tubules of males treated with 1000 mg/kg bw/day. Hyaline droplets can be directly linked to accumulation of alpha 2u-globulin, which is unique to the male rat. This finding is commonly observed in male rats following treatment with some hydrocarbons and is not predictive of any adverse effects in humans.

Increased incidence and severity of extramedullary hematopoiesis was apparent in the spleen of animals of either sex treated with 300 or 1000 mg/kg bw/day. No differences between the controls and treated animals were observed in the bone marrow. Males at 300 and 1000 mg/kg bw/day also showed a statistically significant reduction in mean cell hemoglobin concentration and a statistically significant increase in platelet count. Females treated with 1000 mg/kg bw/day showed a statistically significant reduction in hemoglobin. The majority of individual values for these hematology parameters were within normal range. In the absence of any associated changes in the bone marrow or an obvious indication of red cell depletion, it is likely that the findings apparent in the spleen result from a secondary stress related response to treatment and may be considered adaptive in nature and therefore not adverse.
Dose descriptor:
NOEL
Effect level:
30 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: see 'Remark'
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:
other: see 'Remark'
Critical effects observed:
not specified

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 toxicologically significant 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 effect on food consumption or food efficiency was 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:

Males treated with 300 or 1000 mg/kg bw/day had a statistically significant reduction in mean corpuscular hemoglobin concentration and a statistically significant increase in platelet count. Females treated with 1000 mg/kg bw/day had a statistically significant reduction in hemoglobin.

 

No such effects were detected in females treated with 300 mg/kg bw/day or animals of either sex treated with 30 mg/kg bw/day.

 

Blood Chemistry:

Males treated with 1000 mg/kg bw/day had a statistically significant increase in cholesterol levels.

 

No such effects were detected in females treated with 1000 mg/kg bw/day or in animals of either sex treated with 300 or 30 mg/kg bw/day.

 

Necropsy:

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

 

Organ Weights:

No treatment-related effects were detected on the organ weights measured.

 Histopathology:

Kidneys: In comparison to controls the severity and incidence of intra-epithelial hyaline droplets was marginally greater in males given 1000 mg/kg bw/day. The level of hyaline droplets in the kidneys of males given 30 or 300 mg/kg bw/day were comparable to controls.

 

Incidence and severity of intra-epithelial hyaline droplets in the kidneys of males

Dosage (mg/kg bw/day)

0

30

300

1000

number of animals

5

5

5

5

number examined

5

5

5

5

Hyaline droplets, intra-epithelial, increased

 

 

 

 

- minimal

1

0

2

3

- mild

1

1

1

2

Total

2

1

3

5

 

Liver: Minimal centrilobular hepatocellular hypertrophy was observed in 1/5 males given 300 mg/kg bw/day and 1/5 females and 4/5 the males given 1000 mg/kg bw/day. It was not seen in females given 30 or 300 mg/kg bw/day or in males given 30 mg/kg bw/day. No other morphological changes were found in the liver.

 

Spleen: The incidence and severity of minimal or mild extramedullary hematopoiesis was greater in animals given 300 or 1000 mg/kg bw/day than in controls (see below).

 

Incidence and severity of extramedullary haematopoiesis in the spleen

 

Male

Female

Dosage (mg/kg bw/day)

0

30

300

1000

0

30

300

1000

number of animals

5

5

5

5

5

5

5

5

number examined

5

5

5

5

5

5

5

5

Extramedullary haematopoiesis

 

 

 

 

 

 

 

 

- minimal

1

1

1

2

1

0

2

2

- mild

0

0

1

2

0

0

0

1

Total

1

1

2

4

1

0

2

3

Conclusions:
The oral administration of FAT 41044/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 microscopic abnormalities in animals of either sex treated with 1000 and 300 mg/kg bw/day. The No Observed Effect Level (NOEL) was considered to be 30 mg/kg bw/day for animals of either sex.

The microscopic liver changes apparent in animals of either sex treated with 1000 mg/kg bw/day and males treated with 300 mg/kg bw/day and the associated blood chemistry changes identified in males treated with 1000 mg/kg bw/day were likely to represent an adaptive response to treatment and therefore considered not to represent a serious risk to health.

The microscopic changes apparent in the spleen in the absence of obvious red cell depletion or an associated change in the bone marrow were considered likely to represent a secondary stress related response and therefore were considered not to be adverse. In terms of extrapolation to man, and risk assessment calculations, the effects relating to renal changes in the male rat are species and sex specific and therefore are not relevant.

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).

·        OECD Guideline 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 (Arachis oil BP).

 

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 toxicologically significant 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 effect on food consumption or food efficiency was 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:

Males treated with 300 or 1000 mg/kg bw/day had a statistically significant reduction in mean corpuscular hemoglobin concentration and a statistically significant increase in platelet count. Females treated with 1000 mg/kg bw/day had a statistically significant reduction in hemoglobin.

 

No such effects were detected in females treated with 300 mg/kg bw/day or animals of either sex treated with 30 mg/kg bw/day.

 

Blood Chemistry:

Males treated with 1000 mg/kg bw/day had a statistically significant increase in cholesterol levels.

 

No such effects were detected in females treated with 1000 mg/kg bw/day or in animals of either sex treated with 300 or 30 mg/kg bw/day.

 

Necropsy:

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

 

Organ Weights:

No treatment-related effects were detected on the organ weights measured.

 

Histopathology:

The following microscopic changes were detected:

 

Kidneys: In comparison to controls the severity and incidence of intra-epithelial hyaline droplets was marginally greater in males given 1000 mg/kg bw/day. The level of hyaline droplets in the kidneys of males given 30 or 300 mg/kg bw/day were comparable to controls.

 

Liver: Minimal centrilobular hepatocellular hypertrophy was observed in 1/5 males given 300 mg/kg bw/day and 1/5 females and 4/5 the males given 1000 mg/kg bw/day. It was not seen in females given 30 or 300 mg/kg bw/day or in males given 30 mg/kg bw/day. No other morphological changes were found in the liver.

 

Spleen: The incidence and severity of minimal or mild extramedullary haematopoiesis was greater in animals given 300 or 1000 mg/kg bw/day than in controls.

 

Conclusion:

The oral administration of FAT 41044/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 microscopic abnormalities in animals of either sex treated with 1000 and 300 mg/kg bw/day. The No Observed Effect Level (NOEL) was considered to be 30 mg/kg bw/day for animals of either sex.

 

The microscopic liver changes apparent in animals of either sex treated with 1000 mg/kg bw/day and males treated with 300 mg/kg bw/day and the associated blood chemistry changes identified in males treated with 1000 mg/kg bw/day were likely to represent an adaptive response to treatment and therefore considered not to represent a serious risk to health.

 

The microscopic changes apparent in the spleen in the absence of obvious red cell depletion or an associated change in the bone marrow were considered likely to represent a secondary stress related response and therefore were considered not to be adverse. In terms of extrapolation to man, and risk assessment calculations, the effects relating to renal changes in the male rat are species and sex specific and therefore are not relevant.

 

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
Quality of whole database:
The available information comprises adequate, reliable (Klimisch score 1) and consistent studies, and is thus sufficient to fulfil the standard information requirements set out in Annex VIII, 8.6, of Regulation (EC) No 1907/2006.

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 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 (Arachis oil BP). 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 toxicologically significant 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 effect on food consumption or food efficiency was 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: Males treated with 300 or 1000 mg/kg bw/day had a statistically significant reduction in mean corpuscular hemoglobin concentration and a statistically significant increase in platelet count. Females treated with 1000 mg/kg bw/day had a statistically significant reduction in hemoglobin. No such effects were detected in females treated with 300 mg/kg bw/day or animals of either sex treated with 30 mg/kg bw/day.

Blood Chemistry: Males treated with 1000 mg/kg bw/day had a statistically significant increase in cholesterol levels. No such effects were detected in females treated with 1000 mg/kg bw/day or in animals of either sex treated with 300 or 30 mg/kg bw/day.

Necropsy: Neither the type, incidence or distribution of macroscopic findings indicated an adverse effect of treatment at 30, 300 or 1000 mg/kg bw/day.

Organ Weights: No treatment-related effects were detected on the organ weights measured.

Histopathology: The following microscopic changes were detected:

Kidneys:In comparison to controls the severity and incidence of intra-epithelial hyaline droplets was marginally greater in males given 1000 mg/kg bw/day. The level of hyaline droplets in the kidneys of males given 30 or 300 mg/kg bw/day were comparable to controls.

Liver:Minimal centrilobular hepatocellular hypertrophy was observed in 1/5 males given 300 mg/kg bw/day and 1/5 females and 4/5 the males given 1000 mg/kg bw/day. It was not seen in females given 30 or 300 mg/kg bw/day or in males given 30 mg/kg bw/day. No other morphological changes were found in the liver.

Spleen:The incidence and severity of minimal or mild extramedullary haematopoiesis was greater in animals given 300 or 1000 mg/kg bw/day than in controls.

Conclusion: The oral administration of FAT 41044/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 microscopic abnormalities in animals of either sex treated with 1000 and 300 mg/kg bw/day. The No Observed Effect Level (NOEL) was considered to be 30 mg/kg bw/day for animals of either sex.

The microscopic liver changes apparent in animals of either sex treated with 1000 mg/kg bw/day and males treated with 300 mg/kg bw/day and the associated blood chemistry changes identified in males treated with 1000 mg/kg bw/day were likely to represent an adaptive response to treatment and therefore considered not to represent a serious risk to health.

The microscopic changes apparent in the spleen in the absence of obvious red cell depletion or an associated change in the bone marrow were considered likely to represent a secondary stress related response and therefore were considered not to be adverse. In terms of extrapolation to man, and risk assessment calculations, the effects relating to renal changes in the male rat are species and sex specific and therefore are not relevant.

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:
The selected study is the most adequate and reliable study based on overall assessment of quality, duration and dose descriptor level (refer to the endpoint discussion for further details).

Justification for classification or non-classification