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Diss Factsheets

Toxicological information

Repeated dose toxicity: oral

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Administrative data

Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
24 Nov 2021 - 07 Jun 2022
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2022
Report date:
2022

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Version / remarks:
June 2018
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.26 (Sub-Chronic Oral Toxicity Test: Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Version / remarks:
May 2008
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.3100 (90-Day Oral Toxicity in Rodents)
Version / remarks:
August 1998
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Limit test:
no

Test material

Constituent 1
Reference substance name:
Reaction products of 1H-Imidazole-1-ethanol, 4,5-dihydro-, 2-(C11-C13 odd-numbered alkyl) derivs. and sodium hydroxide and chloroacetic acid
EC Number:
938-645-3
Molecular formula:
Not applicable (a generic molecular formula cannot be provided for this specific UVCB substance)
IUPAC Name:
Reaction products of 1H-Imidazole-1-ethanol, 4,5-dihydro-, 2-(C11-C13 odd-numbered alkyl) derivs. and sodium hydroxide and chloroacetic acid
Test material form:
liquid
Details on test material:
- Physical appearance: clear yellow liquid
- Storage conditions: At room temperature
Specific details on test material used for the study:
- Physical appearance: clear yellow liquid
- Storage conditions: At room temperature
- Test item handling: No specific handling conditions required

Test animals

Species:
rat
Strain:
Wistar
Remarks:
Crl: WI(Han)
Details on species / strain selection:
At this time, studies in laboratory animals provide the best available basis for extrapolation to humans and are required to support regulatory submissions. Acceptable models that do not use live animals currently do not exist. The Wistar Han rat was chosen as the animal model for this study as it is an accepted rodent species for nonclinical toxicity test by regulatory agencies. The total number of animals to be used in this study is considered to be the minimum required to properly characterize the effects of the test item. This study has been designed such that it does not require an unnecessary number of animals to accomplish its objectives.
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 6-7 weeks old
- Weight at study initiation: 169 – 218 g (males); 112 – 155 g (females)
- Fasting period before study: no
- Housing: Polycarbonate cages containing sterilized wooden fibers as bedding material equipped with water bottles. Up to 5 animals of the same sex and same dosing group together. Animals were socially housed for psychological/environmental enrichment and were provided with items such as devices for hiding in, paper and/or objects for chewing, except when interrupted by study procedures/ activities
- Diet: ad libitum, SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany
- Water: ad libitum, municipal tap water
- Acclimation period: 15 days

DETAILS OF FOOD AND WATER QUALITY: It is considered that there are no known contaminants in the feed and water that could interfere with the outcome of the study.

ENVIRONMENTAL CONDITIONS
- Temperature: 21 to 23°C (actual daily mean)
- Humidity: 37 to 52% (actual daily mean)
- Air changes (per hr): ten or more
- Photoperiod: 12 hours light and 12 hours dark

IN-LIFE DATES: From: 09 Dec 2021 To: 08 Apr 2022

Administration / exposure

Route of administration:
oral: gavage
Details on route of administration:
The oral route of exposure was selected because this is a possible route of human exposure during manufacture, handling or use of the test item and is specifically requested by ECHA.
Vehicle:
water
Remarks:
Elix
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
Test item dosing formulations (w/w) were homogenized to visually acceptable levels at appropriate concentrations to meet dose level requirements. Test item dosing formulation were stored in the refrigerator (4°C) until dosing. Adjustment were made for specific gravity of the test item. No correction was made for the purity/composition of the vehicle. A factor of 2 was used to correct for the purity/composition of the test item. High dose group (group 4) was dosed using pure test item.

VEHICLE
- Concentration in vehicle: 59, 177, 590 mg a.i./mL
- Adjusted concentration in vehicle (with a correction factor 2): 118, 354, 1180 mg/mL
- Amount of vehicle (if gavage): 1.695 mL/kg bw
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Accuracy and homogeneity were determined for formulations prepared for use on Week 1 and in Week 6 and Week 12.
Concentrations of groups 1, 2 and 3 were analysed and homogeneity of groups 2 and 3 were determined.
Analyses were performed using a validated analytical procedure (Test Facility Study No. 20297952).
Acceptance criteria for concentration analysis: mean sample concentration results within or equal to ± 10% of theoretical concentration.
Acceptance criteria for homogeneity: relative standard deviation (RSD) of concentrations of ⩽ 10% for each group.
Duration of treatment / exposure:
90 days
Frequency of treatment:
Once daily, 7 days a week
Doses / concentrationsopen allclose all
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Remarks:
group 2 - low dose group
Dose / conc.:
300 mg/kg bw/day (actual dose received)
Remarks:
group 3 - mid dose group
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
Remarks:
group 4 - high dose group
dosed as pure test item
No. of animals per sex per dose:
- control and high dose group: 15 animals per sex per dose (10 in main group and 5 in satellite group)
- low and mid dose group: 10 animals per sex per dose
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The dose levels were selected based on information provided by the results of a (14-day repeated dose toxicity study with oral exposure of PC-2020-926 in rats, Test Facility Study No. 20299085), and in an attempt to produce graded responses to the test item. There were no test item-related microscopic alterations following the administration of the test substance by Oral Gavage for 14 Days to Wistar Han Rats. A few microscopic findings of note were present in the kidney, including mixed cell inflammation in the renal pelvis of a single female dosed at 500 mg/kg bw/day) and a focus of nephroblastomatosis in another female dosed at 1000 mg/kg bw/day). These findings were within the range of background pathology encountered in rats of this age and strain. The lesion seen in the female dosed at 500 mg/kg bw/day (inflammation in the renal pelvis) is most likely the result of an ascending infection from the lower urinary tract. Regarding the focus of nephroblastematosis noted in the second female dosed at 1000 mg/kg bw/day, this is a spontaneous lesion that is encountered sporadically in specific colonies of rats, which suggest a predisposing genetic basis. For both cases, these observations are interpreted to be a spontaneous change and not related to the test item, given the single incidence in this study and the fact that we can see these lesions in untreated female rats of this strain. The high-dose level should produce some toxic effects, but not excessive lethality that would prevent meaningful evaluation. The mid-dose level is expected to produce minimal to moderate toxic effects. The low-dose level should produce no observable indications of toxicity.
- Fasting period before blood sampling for clinical biochemistry: yes (overnight with a maximum of 24 hours)
- Rationale for selecting satellite groups: satellite recovery groups scheduled for follow-up observations were kept for an appropriate period without treatment (28 days) to detect persistence of, or recovery from toxic effects
- Post-exposure recovery period in satellite groups: 28 days
- Section schedule rationale: random
- Dose range finding studies: yes (see information under "dose selection rationale")
- Other: In groups 1 (control) and 4 (high dose) 5 animals per sex per group were assigned to the recovery satellite groups.

Examinations

Observations and examinations performed and frequency:
MORTALITY: Yes
- Time schedule: at least twice daily, except on days of receipt and necropsy where frequency was at least once daily

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: At least once daily; from Day 1 at 0 to 1 hours postdose during dosing

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Weekly; from Week 1 and throughout the study, and on the day of necropsy.

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly; from at least Day 1 and throughout the study.

FOOD CONSUMPTION: Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Time schedule for examinations: Weekly; from at least Day 1 and throughout the study.

WATER CONSUMPTION: Yes
- Time schedule for examinations: on regular basis throughout the study by visual inspection of the water bottles.

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations and dose groups examined: pretreatment period (all animals); week 13 of dosing period (main group 1 and 4)

HAEMATOLOGY: Yes
- Time schedule for collection of blood: On days of scheduled or unscheduled necropsy Between 7.00 and 10.30 from the retro-orbital sinus at the end of the treatment period and at the end of the recovery period respectively
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes overnight with a maximum of 24 hours)
- How many animals: all main and recovery animals
- Parameters examined: White Blood Cell (WBC), Neutrophils (absolute), Lymphocytes (absolute), Monocytes (absolute), Eosinophils (absolute), Basophils (absolute), Large unstained cells (LUC) (absolute), Red Blood Cell (RBC), Reticulocytes (absolute), Red Blood Cell Distribution Width (RDW), Hemoglobin, Hematocrit, Mean corpuscular volume (MCV), Mean corpuscular hemoglobin (MCH), Mean corpuscular hemoglobin concentration (MCHC), Platelets

COAGULATION: Yes
- Time schedule for collection of blood: see HEMATOLOGY
- Animals fasted: see HEMATOLOGY
- How many animals: see HEMATOLOGY
- Parameters examined: Prothrombin time (PT), Activated partial thromboplastin time (APTT)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: see HEMATOLOGY
- Animals fasted: see HEMATOLOGY
- How many animals: see HEMATOLOGY
- Parameters examined: Alanine aminotransferase (ALT), Triglycerides, Aspartate aminotransferase (AST), HDL and LDL Cholesterol, Alkaline Phosphatase (ALP), Sodium, Total protein, Potassium, Albumin, Chloride, Total Bilirubin, Calcium, Urea, Inorganic Phosphate (Inorg. Phos), Creatinine, Glucose, Cholesterol

SERUM HORMONES: Yes
- Time of blood sample collection: see HEMATOLOGY
- Animals fasted: see HEMATOLOGY
- How many animals: see HEMATOLOGY
- Parameters examined: Triiodothyronine (T3), Thyroxine (T4), Thyroid-Stimulating Hormone (TSH)

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: once during week 13 of the dosing period
- Dose groups that were examined: the first 5 animals per sex per group of all dose groups (1-4)
- Battery of functions tested: sensory activity, grip strength and motor activity

ESTROUS STAGE DETERMINATION: Yes
- Time schedule: end of treatment on the day of necropsy

IMMUNOLOGY: No
Sacrifice and pathology:
SACRIFICE
All main and recovery animals surviving until scheduled euthanasia were weighed, and deeply anesthetized using isoflurane and subsequently exsanguinated and subjected to a full post mortem examination. All animals surviving to scheduled necropsy were fasted overnight with a maximum of 24 hours before necropsy. Water was available.

GROSS NECROPSY
- All animals were subjected to a full post mortem examination, which included evaluation of the carcass and musculoskeletal system; all external surfaces and orifices; cranial cavity and external surfaces of the brain; and thoracic, abdominal, and pelvic cavities with their associated organs and tissues.

-Tissue collection and preservation:
Representative samples of the tissues indicated in Table 1 below were collected from all animals and preserved in 10% neutral buffered formalin: Testes and epididymis were preserved in modified Davidson’s fixative and transferred to formalin after fixation for at least 24 hours. Gross lesions or masses were also collected from all animals if found.

HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in Table 1 below were prepared for microscopic examination and weighed, respectively for the selected and remaining animals.
For all mail males and females of group 1 and 4 the full list of tissues mentioned in table 1 below (see section "Any other information on materials and methods incl. tables") was examined histopathologically. For all remaining animals (main animals of group 2 and 3 and recovery animals of group 1 and 4) only gross lesions and target tissues were examined histopathologically.
Target tissues identified: rectum of males and stomach of females
Statistics:
All statistical tests were conducted at the 5% significance level. All pairwise comparisons were conducted using two sided tests and were reported at the 1% or 5% levels. The following pairwise comparisons were made:
Group 2 vs. Group 1
Group 3 vs. Group 1
Group 4 vs. Group 1

Numerical data collected on scheduled occasions for all variables were analyzed as indicated according to sex and occasion. Descriptive statistics number, mean and standard deviation (or %CV or SE when deemed appropriate) were reported whenever possible.

In-life, clinical chemistry, hematology, coagulation, organ weights data:
- Parametric/Non-parametric: Levene’s test was used to assess the homogeneity of group variances. The groups were compared using an overall one-way ANOVA F-test if Levene’s test was not significant or the Kruskal-Wallis test if it was significant. If the overall F-test or Kruskal-Wallis test was found to be significant, then pairwise comparisons were conducted using Dunnett’s or Dunn’s test, respectively.
- ANCOVA: The data corresponding to a response variable of interest and to a related covariate was submitted to an analysis of covariance, including only groups with at least three non-missing paired values and when found to be significant, then pairwise comparisons was conducted using Dunnett’s test.
-Incidence: A Fisher’s exact test was used to conduct pairwise group comparisons.

Functional tests data:
- Parametric: Datasets with at least 3 groups (the designated control group and 2 other groups) were compared using Dunnett-test (many-to-one-t-test).
- Non-parametric: Datasets with at least 3 groups was compared using a Steel-test (many-to-one rank test).
- Incidence: An overall Fisher’s exact test was used to compare all groups at the 5% significance level. Pairwise comparisons were conducted using Fisher’s exact test whenever the overall test was significant.

Results and discussion

Results of examinations

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Abnormal breathing sounds were noted for animals dosed at all dose levels, with a dose dependent increase in incidence with increasing dose. Abnormal breathing sounds were also noted for one control female. For a few individual animals, additional deep, laboured or shallow breathing was incidentally noted across the test material-treated groups. Moreover, retching was noted for males at 100 and 1000 mg/kg bw/day and females at 100, 300 and 1000 mg/kg bw/day. Finally, hunched posture was noted for one male each at 300 and 1000 mg/kg bw/day and one female at 1000 mg/kg bw/day, and erected fur was noted for two males at 300 mg/kg bw/day, four males at 1000 mg/kg bw/day and one female each at 300 and 1000 mg/kg bw/day.
Salivation, on some occasions accompanied by ploughing, was noted in animals dosed at 300 and 1000 mg/kg bw/day. Taking into account the nature and minor severity of the effect and its time of occurrence (i.e., after dosing), salivation was considered to be a physiological response rather than a sign of systemic toxicity. Ploughing was considered to be an adaptive response to salivation, and not directly related to treatment with the test material.
During the Recovery Period, abnormal breathing sounds and/or laboured breathing were noted on few occasions for one female at 1000 mg/kg bw/day. At the incidence observed, and as this concerned the treatment-free period, this was considered to be unrelated to treatment with the test material. No test material-related clinical signs were noted during weekly arena observations for the Main and Recovery animals.
Any other clinical signs noted during the Treatment or Recovery Period occurred within the range of background findings to be expected for rats of this age and strain which are housed and treated under the conditions in this study and/or did not show any apparent dose-related trend. At the incidence observed, these were considered to be unrelated to treatment with the test material.
(see Table 29-30 under "Any other information on results incl. tables")
Mortality:
no mortality observed
Description (incidence):
No mortality occurred during the study period.
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
Body weights and body weight gain were considered to have been unaffected by treatment with the test material.
At 1000 mg/kg bw/day, body weight gain was significantly lower for males over Days 78-85, and significantly higher for females over Days 1-8 when compatred to controls. For one male at 1000 mg/kg bw/day, a body weight loss of 9% was noted over Days 57-64. In addition, mean body weight gain over the complete Recovery Period was significantly higher for males at 1000 mg/kg bw/day, when compared to control. As they did not result in significant differences in mean body weights and showed no trend regarding duration of dosing, these slight and incidental changes were considered to be unrelated to treatment with the test material.
(see Table 5-12 under "Any other information on results incl. tables")
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Food consumption was considered to have been unaffected by treatment with the test material.
At 300 mg/kg bw/day, food intake was slightly low for females during the Treatment Period. As this did not result in significant differences in mean food consumption, showed no trend regarding duration of dosing and did not result in a dose response, these slight and incidental changes were considered to be unrelated to treatment with the test material.
For females at 1000 mg/kg bw/day, food intake was slightly low during the Recovery Period (7.5% lower when compared to control). As this concerned the Recovery Period only, this was considered to be unrelated to treatment with the test material.
Ophthalmological findings:
no effects observed
Description (incidence and severity):
No ophthalmology findings were noted that were considered to be related to the test material.
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
Hematology parameters were considered not to have been affected by the test material in females up to 1000 mg/kg bw/day and in males up to 300 mg/kg bw/day.
In males at 1000 mg/kg bw/day, red blood cell (RBC) count and red blood cell distribution width (RDWG) were significantly decreased (0.92x and 0.95x of control, respectively), and mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH) were significantly increased (1.03x and 1.05x of control, respectively) at the end of the Treatment Period. All reported changes reached statistical significance but were within historical control for this rat strain at the site.
At the end of the Recovery Period, RBC counts were comparable to control, while RDWG was still decreased, and MCV and MCH were still increased (not statistically significant) in males at 1000 mg/kg bw/day.
Any other differences in hematology parameters, regardless of statistical significance, were considered to be unrelated to the test material as these occurred in the absence of a dose-related trend or only at the end of the Recovery Period.

Coagulation parameters were considered not to have been affected by treatment with the test material.

Actual mean values:
Red Blood Cell Count males: control group mean = 9.302; high dose group mean = 8.558
Red Blood Cell Distribution Width males: control group mean =12.67 ; high dose group mean = 12.04
Mean Corpuscular Volume males: control group mean = 51.75 ; high dose group mean = 53.47
Mean Corpuscular Hemoglobin males: control group mean = 17.30; high dose group mean = 18.19

HCD
HCD Red Blood Cell Count males: mean = 8.802; P5-P1: 7.9–9.55
HCD Red Blood Cell Distribution Width males: mean = 427; P5-P1: 11.3–13.6
HCD Mean Corpuscular Volume males: mean = 51.61; P5-P1: 48.9–54.6
HCD Mean Corpuscular Hemoglobin males: mean = 17.21; P5-P1: 16.3–18.9

(see Table 13-16 under "Any other information on results incl. tables")
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
Clinical chemistry parameters were considered not to have been affected by the test material in females up to 1000 mg/kg bw/day and in males up to 300 mg/kg bw/day.
In males at 1000 mg/kg bw/day, triglyceride concentration was increased (1.40x of control, not statistically significant and within historical control data) at the end of the Treatment Period.
At the end of the Recovery Period, triglyceride concentration in males at 1000 mg/kg bw/day was partly recovered.
Remaining differences in clinical chemistry parameters, regardless of statistical significance, were considered not test material-related based on the absence of a dose response, general overlap of individual values with the range of control values, variation in direction of change, were considered to have arisen as a result of slightly high control values, were seen at end of recovery only and/or were of a magnitude of change commonly observed in rats under similar study conditions.

Actual mean values:
Triglyceride males: control group mean = 0.956; high dose group mean = 1.338

HCD
HCD Triglyceride males: mean = 399; P5-P1: 0.36–1.52

(see Table 17-20 under "Any other information on results incl. tables")
Endocrine findings:
effects observed, treatment-related
Description (incidence and severity):
No effect was noted on T3 and TSH levels in males and females, and on T4 levels in males up to 1000 mg/kg bw/day.
In females at 100 and 1000 mg/kg bw/day, T4 levels were increased at the end of the Treatment Period (1.22x and 1.37x of control, not statistically significant at 100 mg/kg bw/day). Mean values remained within the historical control range .
At the end of the Recovery Period, T4 levels in females at 1000 mg/kg bw/day were comparable to control.

Since no histopathological changes were observed in the thyroid, the observed increase in the study was considered not toxicologically relevant.

Actual mean values:
T4 (ng/mL) – females: control group mean = 24.72; high dose group mean = 33.98

HCD
HCD T4 (ng/mL) – females: mean = 29.87; P2.5-P-97.5: 13.92 - 51.38

(see Table 17-20 under "Any other information on results incl. tables")
Behaviour (functional findings):
effects observed, treatment-related
Description (incidence and severity):
Hearing ability, pupillary reflex and static righting reflex were normal in all examined animals. Grip strength was considered unaffected by treatment with the test material.
Motor activity was considered unaffected by treatment with the test material in males at all dose levels and in females up to 300 mg/kg bw/day.
For females at 1000 mg/kg bw/day, the mean total movements and ambulations were both lower compared to control (0.59x and 0.61x of control, respectively; not statistically significant for ambulations). Mean values remained within the historical control range, albeit close to the lower limit.
At the end of the Recovery Period, mean total movements and ambulations were within the same range for control animals and animals treated at 1000 mg/kg bw/day.
All groups showed a similar motor activity habituation profile with a decreasing trend in activity over the duration of the test period.

Actual mean values:
Total movements – females: control group mean = 4615; high dose group mean = 2739
Total ambulations – females: control group mean = 1095; high dose group mean = 667

HCD
HCD Total movements – females: mean = 4740; P5-P95 = 2393-8076
HCD Total ambulations – females: mean = 1335; P5-P95 = 638-2271

(see Table 21-22 under "Any other information on results incl. tables")
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
Test material-related significantly higher kidney and liver weights (absolute and relative to body weight) were noted in females at 1000 mg/kg bw/day (see Table 2 under "Any other information on results incl. tables"). These differences were within the historical control data of the test site.
Higher kidney weight (absolute and relative to body weight, statistically significant) was present in females at 1000 mg/kg bw/day at the end of the Treatment Period. Following the Recovery Period, there was no significant difference in the kidney weight of test material-treated females compared to the control females, suggesting full recovery.
Higher liver weight (absolute and relative to body weight, statistically significant) was present in females at 1000 mg/kg bw/day at the end of the Treatment Period. Following the Recovery Period, there was no significant difference in the liver weight of test material-treated females compared to the control females, suggesting full recovery.
Other statistically significant changes at the end of the Treatment Period (lower relative pituitary gland weight, females at 100 mg/kg bw/day) or Recovery Period (higher relative brain weight, females at 1000 mg/kg bw/day) were considered to be not test material-related due to the lack of a dose-related pattern (pituitary gland) or was regarded related to the lower final body weight (brain).

Since the higher kidney and liver weights in females at 1000 mg/kg bw/day remained within the historical control data, the changes were no longer observed after recovery and in the absence of any concomitant histopathological changes, these effects were considered non-adverse.

Actual mean values:
Liver weight: control group mean = 5.3753; high dose group mean = 6.4360
Liver relative to body weight: control group mean = 2.43901; high dose group mean = 2.79153
Kidney weight: control group mean = 1.4672; high dose group mean = 1.7272
Kidney relative to body weight: control group mean = 0.66594; high dose group mean = 0.74781

HCD
HCD Liver weight: Mean = 5.56; P5-P95 = 4.634-6.472
HCD Liver relative to body weight: Mean = 2.50; P5-P95 = 2.226-2.876
HCD Kidney weight: Mean = 1.53; P5-P95 = 0.526-1.80
HCD Kidney relative to body weight: Mean = 0.69; P5-P95 = 0.588-0.807

(see also Table 23-26 under "Any other information on results incl. tables")
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
There were no test material-related gross observations.
All the recorded macroscopic findings were within the range of background gross observations encountered in rats of this age and strain.
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Test material-related microscopic findings were noted in the rectum of males at 1000 mg/kg bw/day and the stomach of females at 300 and 1000 mg/kg bw/day (see Table 3 under "Any other information on results incl. tables").
In the rectum, goblet cell hyperplasia was recorded in 3/10 males at 1000 mg/kg bw/day (minimal) at the end of the Treatment Period. Following the Recovery Period this finding was not present, suggesting full recovery.
In the stomach, in the non-glandular stomach, squamous cell hyperplasia often accompanied by hyperkeratosis was recorded in 1/10 females at 300 mg/kg bw/day (minimal) and in 3/10 females at 1000 mg/kg bw/day (up to mild degree) at the end of the Treatment Period. Following the Recovery Period this finding was not present, suggesting full recovery (see Table 4 under "Any other information on results incl. tables").
Focal erosion of the non-glandular and glandular portion of the stomach (minimal) was noted in 1/10 females at 1000 mg/kg/day (Main Group females) and 1/5 females at 1000 mg/kg bw/day (Recovery Group females). At this low incidence and severity, these findings were considered not to represent a test material-related effect but rather represent a procedure-related effect or an incidental finding.
There were no other test material-related histologic changes. The remainder of the recorded microscopic findings were considered to be incidental findings or were within the range of background pathology encountered in rats of this age and strain. There was no test material related alteration in the prevalence, severity, or histologic character of those incidental tissue alterations.

(see also Table 27-28 under "Any other information on results incl. tables")

Effect levels

Key result
Dose descriptor:
NOAEL
Effect level:
>= 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Remarks:
corrected for water content
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects

Target system / organ toxicity

Key result
Critical effects observed:
no

Any other information on results incl. tables

The accuracy and homogeneity of the formulations of Group 4 were not determined because it was pure test material.
Accuracy
The concentrations analyzed in the formulations of Groups 2 and 3 were in agreement with target concentrations (i.e., mean sample concentration results were within or equal to 90-110% of target concentration).
No test material was detected in the Group 1 formulations.
Homogeneity
The formulations of Groups 2 and 3 were homogeneous (i.e., coefficient of variation ≤ 10%).


 


Table 2: Mean Percent kidney and liver Weight Differences from Control Groups




























































 



Main Females



Recovery Females



Dose level (mg/kg bw/day):



100



300



1000



1000



KIDNEYS



 



 



 



 



               Absolute



8



0



18**



-3



               Relative to body weight



5



4



12**



2



LIVER



 



 



 



 



               Absolute



1



-5



20**



-7



               Relative to body weight



-2



-2



14**



-2



**: P≤0.01


 


Table 3: Summary Test Material-Related Microscopic Findings - Males















































 



Main Males



Recovery Males



Dose level (mg/kg bw/day):



0



100



300



1000



0



1000



RECTUM a



10



10



10



10



5



5



     Hyperplasia, goblet cell



 



 



 



 



 



 



       Minimal



-



-



-



3



-



-



a  =  Number of tissues examined from each group.


 


Table 4: Summary Test Material-Related Microscopic Findings - Females
























































 



Main Females



Recovery Females



Dose level (mg/kg bw/day):



0



100



300



1000



0



1000



STOMACH a



10



10



10



10



5



5



       Hyperplasia, squamous cell, 


      non-glandular



 



 



 



 



 



 



        Minimal



-



-



1



2



-



-



        Mild



-



-



-



1



-



-



a  =  Number of tissues examined from each group.


Table 5: Body weights (gram) summary (males) -Main








































































































































































































































































































Sex: Male



Day(s) Relative to Start Date


 


 



1



8



15



22



29



36



43



50



57



64



71



78



85



91



0



Mean



193.5



235.9



270.2



296.3



317.1



338.5



350.3



366.9



377.4



387.4



395.7



401.7



411.6



415.7



mg/kg/day



SD



10.6



14.1



18.5



20.5



23.0



23.3



25.8



25.4



29.4



28.9



28.9



30.9



31.8



32.1



Group 1



N



15



15



15



15



15



15



15



15



15



15



15



15



15



15



100



Mean



199.5



241.6



275.0



299.5



321.2



342.0



356.8



374.6



383.8



393.9



404.5



412.1



420.6



423.6



mg/kg/day



SD



9.3



14.7



17.5



19.0



20.1



22.6



25.0



27.2



27.6



29.7



33.4



34.0



34.4



35.8



Group 2



N



10



10



10



10



10



10



10



10



10



10



10



10



10



10



 



%Diff



3.1



2.4



1.8



1.1



1.3



1.0



1.8



2.1



1.7



1.7



2.2



2.6



2.2



1.9



300



Mean



197.8



237.0



268.0



292.0



312.3



334.5



350.6



365.2



377.2



386.4



397.7



406.6



414.0



418.6



mg/kg/day



SD



14.1



17.1



18.4



24.4



26.1



29.2



28.2



31.8



32.3



34.4



33.3



34.4



35.6



37.6



Group 3



N



10



10



10



10



10



10



10



10



10



10



10



10



10



10



 



%Diff



2.2



0.5



-0.8



-1.4



-1.5



-1.2



0.1



-0.5



-0.1



-0.3



0.5



1.2



0.6



0.7



1000



Mean



196.5



236.8



269.8



298.0



318.3



337.9



350.3



365.3



373.1



381.3



390.4



398.5



401.1



408.1



mg/kg/day



SD



12.1



16.0



19.1



20.8



23.9



26.0



28.3



30.4



31.5



38.2



36.4



34.8



35.1



37.0



Group 4



N



15



15



15



15



15



15



15



15



15



15



15



15



15



15



 



%Diff



1.6



0.4



-0.1



0.6



0.4



-0.2



0.0



-0.5



-1.1



-1.6



-1.3



-0.8



-2.5



-1.8



 


Table 6: Body weigths (gram) summary (Females) -Main








































































































































































































































































































Sex: Female



Day(s) Relative to Start Date



1 [G]



8 [G]



15 [G1]



22 [G]



29 [G]



36 [G]



43 [G1]



50 [G]



57 [G]



64 [G]



71 [G]



78 [G]



85



91



0



Mean



131.6



147.7



169.2



183.3



195.1



206.5



212.5



219.9



225.8



228.9



233.2



236.0



240.5



241.3



mg/kg/day



SD



9.6



9.4



11.2



14.5



13.8



13.5



12.2



15.3



14.7



13.7



14.5



16.4



16.0



15.1



Group 1



N



15



15



15



15



15



15



15



15



15



15



15



15



15



15



100



Mean



130.9



150.5



166.5



184.1



196.0



208.2



213.7



223.1



226.8



231.7



234.0



238.0



240.2



241.1



mg/kg/day



SD



7.9



10.7



11.8



10.7



12.3



14.4



15.3



13.9



15.2



16.5



17.9



16.7



15.6



17.0



Group 2



N



10



10



10



10



10



10



10



10



10



10



10



10



10



10



 



%Diff



-0.5



1.9



-1.6



0.5



0.4



0.8



0.5



1.5



0.4



1.2



0.3



0.8



-0.1



-0.1



300



Mean



123.8



139.5



158.3



174.6



185.8



195.9



202.6



210.5



213.0



217.4



220.8



224.9



229.0



229.6



mg/kg/day



SD



11.0



11.1



15.5



15.7



17.4



18.4



17.3



18.4



18.2



18.6



20.3



18.3



17.1



17.0



Group 3



N



10



10



10



10



10



10



10



10



10



10



10



10



10



10



 



%Diff



-5.9



-5.6



-6.4



-4.7



-4.8



-5.1



-4.7



-4.3



-5.7



-5.0



-5.3



-4.7



-4.8



-4.8



1000



Mean



135.7



156.1



175.5



192.5



205.7



214.1



219.3



231.4



234.9



236.6



239.5



243.7



245.0



244.8



mg/kg/day



SD



8.7



9.7



8.4



9.7



10.3



10.7



10.3



12.6



11.6



11.8



14.4



12.4



11.9



13.9



Group 4



N



15



15



15



15



15



15



15



15



15



15



15



15



15



15



 



%Diff



3.1



5.7



3.7



5.0



5.4



3.7



3.2



5.2



4.0



3.4



2.7



3.3



1.9



1.5



[G] - Anova & Dunnett


[G1] - Kruskal-Wallis & Dunn


 


Table 7: Body weigths (gram) summary (Males) -Recovery
















































































Sex: Male



Day(s) Relative to Start Date



92



99



106



113



120



0



Mean



419.0



425.6



432.4



437.0



440.8



mg/kg/day



SD



31.8



30.6



30.8



33.4



30.9



Group 1



N



5



5



5



5



5



1000



Mean



414.4



424.2



433.6



441.2



443.2



mg/kg/day



SD



33.6



33.7



32.6



34.3



35.5



Group 4



N



5



5



5



5



5



 



%Diff



-1.1



-0.3



0.3



1.0



0.5



 


Table 8: Body weigths (gram) summary (Females) -Recovery
















































































Sex: Female



Day(s) Relative to Start Date



92



99



106



113



120



0



Mean



254.0



254.2



260.0



259.8



259.0



mg/kg/day



SD



11.0



9.0



10.8



10.6



13.3



Group 1



N



5



5



5



5



5



1000



Mean



242.0



244.2



249.0



244.6



244.4



mg/kg/day



SD



12.4



16.7



17.0



15.4



15.9



Group 4



N



5



5



5



5



5



 



%Diff



-4.7



-3.9



-4.2



-5.9



-5.6



 


Table 9: Body weight gain (%) summary (Males) -Main










































































































































































































































Sex: Male



Day(s) Relative to Start Date



1 → 8 [G]



8 → 15 [G]



15 → 22 [G]



22 → 29 [G]



29 → 36 [G]



36 → 43 [G1]



43 → 50 [G]



50 → 57



57 → 64



64 → 71



71 → 78



78 → 85



85 → 91



1 → 91



0


mg/kg/day


Group 1



Mean



42.4



34.3



26.1



20.9



21.4



11.8



16.6



10.5



10.0



8.3



6.1



9.9



4.1



222.2



SD



4.0



5.9



4.8



4.2



4.4



4.9



3.9



5.8



3.8



4.4



3.7



3.3



2.4



23.9



N



15



15



15



15



15



15



15



15



15



15



15



15



15



15



100


mg/kg/day


Group 2



Mean



42.1



33.4



24.5



21.7



20.8



14.8



17.8



9.2



10.1



10.6



7.6



8.5



3.0



224.1



SD



7.5



4.9



3.6



3.0



3.0



3.1



5.4



3.6



3.7



5.5



4.1



4.1



3.1



29.0



N



10



10



10



10



10



10



10



10



10



10



10



10



10



10



300


mg/kg/day


Group 3



Mean



39.2



31.0



24.0



20.3



22.2



16.1



14.6



12.0



9.2



11.3



8.9



7.4



4.6



220.8



SD



4.6



6.0



9.7



4.6



3.4



2.9



6.2



3.2



3.2



2.2



2.8



3.8



4.4



32.0



N



10



10



10



10



10



10



10



10



10



10



10



10



10



10



1000


mg/kg/day


Group 4



Mean



40.3



33.0



28.2



20.3



19.6



12.3



15.0



7.9



8.1



9.1



8.1



2.6**



7.0



211.6



SD



5.0



5.8



5.6



7.7



4.5



5.5



4.6



4.5



11.3



5.8



5.4



3.2



4.8



30.7



N



15



15



15



15



15



15



15



15



15



15



15



15



15



15



[G] - Anova & Dunnett


[G1] - Kruskal-Wallis & Dunn


 


Table 10: Body weight gain (%) summary (females) -Main










































































































































































































































Sex: Female



Day(s) Relative to Start Date



1 → 8 [G]



8 → 15 [G]



15 → 22 [G1]



22 → 29 [G]



29 → 36 [G]



36 → 43 [G]



43 → 50 [G]



50 → 57



57 → 64



64 → 71



71 → 78



78 → 85



85 → 91



1 → 91



0


mg/kg/day


Group 1



Mean



16.1



21.5



14.1



11.9



11.3



6.1



7.3



5.9



3.1



4.3



2.8



4.5



0.7



109.7



SD



4.3



4.9



4.6



5.3



4.5



4.4



4.6



5.4



4.3



5.8



4.2



4.3



4.0



8.5



N



15



15



15



15



15



15



15



15



15



15



15



15



15



15



100


mg/kg/day


Group 2



Mean



19.6



16.0



17.6



11.9



12.2



5.5



9.4



3.7



4.9



2.3



4.0



2.2



0.9



110.2



SD



3.9



3.1



4.4



4.1



5.0



3.7



5.2



4.6



3.1



3.7



4.1



4.3



5.1



11.8



N



10



10



10



10



10



10



10



10



10



10



10



10



10



10



300


mg/kg/day


Group 3



Mean



15.7



18.8



16.3



11.2



10.1



6.7



7.9



2.5



4.4



3.4



4.1



4.1



0.6



105.8



SD



4.6



5.4



1.5



3.5



5.0



3.3



3.0



3.9



4.3



5.1



3.7



3.7



7.0



12.9



N



10



10



10



10



10



10



10



10



10



10



10



10



10



10



1000


mg/kg/day


Group 4



Mean



20.5*



19.4



16.9



13.3



8.3



5.2



12.1



3.5



1.7



2.9



4.3



1.3



-0.2



109.1



SD



5.1



5.7



4.5



4.0



6.5



5.3



6.9



6.4



4.8



9.4



5.4



4.1



5.8



11.6



N



15



15



15



15



15



15



15



15



15



15



15



15



15



15



[G] - Anova & Dunnett: * = p ≤ 0.05
[G1] - Kruskal-Wallis & Dunn


 


Table 11: Body weight gain (%) summary (males) -Recovery



































































Sex: Male



Day(s) Relative to Start Date



92 → 99 [G]



99 → 106 [G]



106 → 113 [G1]



113 → 120 [G]



92 → 120 [G]



0


mg/kg/day


Group 1



Mean



6.6



6.8



4.6



3.8



21.8



SD



1.9



1.3



4.8



2.9



5.0



N



5



5



5



5



5



1000


mg/kg/day


Group 4



Mean



9.8



9.4



7.6



2.0



28.8*



SD



2.6



2.3



2.5



1.6



4.0



N



5



5



5



5



5



[G] - Anova & Dunnett: * = p ≤ 0.05


 


Table 12: Body weight gain (%) summary (females) -Recovery



































































Sex: Female



Day(s) Relative to Start Date



92 → 99



99 → 106



106 → 113



113 → 120



92 → 120



0


mg/kg/day


Group 1



Mean



0.2



5.8



-0.2



-0.8



5.0



SD



3.1



3.5



4.3



4.5



3.7



N



5



5



5



5



5



1000


mg/kg/day


Group 4



Mean



2.2



4.8



-4.4



-0.2



2.4



SD



6.2



3.0



2.9



4.8



3.6



N



5



5



5



5



5



 


Table 13: Haematology summary (males) - Day 92 (end of treatment)






























































































































































































































































































































Sex: Male



Reporting Hematology



WBC (10^9/L)


[G]



NEUT (10^9/L)


[G]



LYMPH (10^9/L)


[G]



MONO (10^9/L)


[G]



EOS (10^9/L)


[G]



BASO (10^9/L)


[G]



LUC (10^9/L)


[G]



RBC (10^12/L)


[G]



RETIC (10^9/L)


[G]



RDWG (%)


[G]



HGB


 


(g/L)


 


[G1]



HCT (L/L)


[G]



MCV (fL)


[G]



MCH


 


(pg)


 


[G]



MCHC


 


(g/L)


 


[G]



PLT


(10^9/L)


[G]



0


mg/kg/day


Group 1



Mean



6.008



1.074



4.658



0.096



0.116



0.008



0.053



9.302



206.11



12.67



160.9



0.4814



51.75



17.30



334.2



723.0



SD



1.263



0.331



1.013



0.031



0.023



0.004



0.013



0.224



27.72



0.54



4.7



0.0186



1.07



0.42



6.7



35.2



N



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



100


mg/kg/day


Group 2



Mean



5.138



1.106



3.785



0.079



0.128



0.005



0.033



9.142



188.16



12.23



159.9



0.4724



51.70



17.53



339.0



643.8



SD



1.661



0.366



1.683



0.021



0.031



0.007



0.015



0.549



25.76



0.35



7.2



0.0267



0.94



0.44



7.3



80.9



N



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



 



tCtrl



0.86



1.03



0.81



0.82



1.10



0.63



0.62



0.98



0.91



0.97



0.99



0.98



1.00



1.01



1.01



0.89



300


mg/kg/day


Group 3


 



Mean



6.502



1.235



4.931



0.107



0.160



0.007



0.062



9.086



202.13



12.44



161.7



0.4768



52.52



17.83



339.6



674.3



SD



1.688



0.259



1.532



0.042



0.040



0.005



0.022



0.645



38.11



0.44



7.5



0.0263



1.43



0.61



6.3



97.1



N



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



tCtrl



1.08



1.15



1.06



1.11



1.38



0.88



1.17



0.98



0.98



0.98



1.00



0.99



1.01



1.03



1.02



0.93



1000


mg/kg/day


Group 4


 



Mean



5.481



0.992



4.216



0.088



0.130



0.006



0.049



8.558 **



216.19



12.04 **



155.7



0.4581



53.47 *



18.19 **



340.2



726.4



SD



2.253



0.199



2.146



0.028



0.043



0.005



0.038



0.563



26.23



0.38



12.1



0.0398



2.05



0.76



7.5



69.1



N



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



tCtrl



0.91



0.92



0.91



0.92



1.12



0.75



0.92



0.92



1.05



0.95



0.97



0.95



1.03



1.05



1.02



1.00



[G] - Anova & Dunnett: ** = p ≤ 0.01
[G1] - Kruskal-Wallis & Dunn


 


Table 14: Haematology summary (Females) - Day 93 (end of treatment)






























































































































































































































































































































Sex: Female



Reporting Hematology



WBC (10^9/L)


[G]



NEUT (10^9/L)


[G]



LYMPH (10^9/L)


[G]



MONO (10^9/L)


[G]



EOS (10^9/L)


[G]



BASO (10^9/L)


[G1]



LUC (10^9/L)


[G]



RBC (10^12/L)


[G]



RETIC (10^9/L)


[G]



RDWG (%)


[G]



HGB


 


(g/L)


 


[G]



HCT (L/L)


[G]



MCV (fL)


[G]



MCH


 


(pg)


 


[G]



MCHC


 


(g/L)


 


[G]



PLT


(10^9/L)


[G]



0


mg/kg/day


Group 1



Mean



3.863



0.430



3.239



0.061



0.088



0.005



0.039



7.942



222.87



10.97



149.6



0.4435



55.86



18.86



338.0



723.9



SD



0.933



0.165



0.733



0.031



0.034



0.005



0.021



0.363



24.72



0.29



5.9



0.0183



1.25



0.65



7.0



110.8



N



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



100


mg/kg/day


Group 2



Mean



3.189



0.591



2.431



0.052



0.089



0.000 *



0.027



8.269



218.39



11.01



149.4



0.4477



54.17 **



18.08 **



333.7



755.7



SD



0.724



0.239



0.546



0.023



0.034



0.000



0.008



0.290



25.11



0.35



5.1



0.0123



1.09



0.56



4.6



84.2



N



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



 



tCtrl



0.83



1.37



0.75



0.85



1.01



0.00



0.69



1.04



0.98



1.00



1.00



1.01



0.97



0.96



0.99



1.04



300


mg/kg/day


Group 3


 



Mean



3.715



0.815 *



2.664



0.077



0.117



0.000 *



0.039



8.222



212.56



11.18



152.7



0.4513



54.93



18.59



338.5



735.2



SD



1.399



0.505



0.822



0.046



0.076



0.000



0.022



0.269



25.28



0.35



4.9



0.0150



1.27



0.59



6.2



57.6



N



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



9



tCtrl



0.96



1.90



0.82



1.26



1.33



0.00



1.00



1.04



0.95



1.02



1.02



1.02



0.98



0.99



1.00



1.02



1000


mg/kg/day


Group 4


 



Mean



4.430



0.745 *



3.462



0.077



0.097



0.005



0.043



8.106



215.62



10.83



149.5



0.4425



54.60 *



18.45



338.1



801.0



SD



1.372



0.369



1.159



0.029



0.025



0.007



0.026



0.347



45.04



0.29



5.3



0.0190



0.74



0.37



4.0



92.3



N



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



tCtrl



1.15



1.73



1.07



1.26



1.10



1.00



1.10



1.02



0.97



0.99



1.00



1.00



0.98



0.98



1.00



1.11



[G] - Anova & Dunnett


 


Table 15: Haematology summary (males) - Day 121 (end of recovery)



































































































































































Sex: Male



Reporting Hematology



WBC (10^9/L)


[G]



NEUT (10^9/L)


[G]



LYMPH (10^9/L)


[G]



MONO (10^9/L)


[G]



EOS (10^9/L)


[G]



BASO (10^9/L)


[G]



LUC (10^9/L)


[G]



RBC (10^12/L)


[G]



RETIC (10^9/L)


[G]



RDWG (%)


[G]



HGB


 


(g/L)


 


[G]



HCT (L/L)


[G]



MCV (fL)


[G]



MCH


 


(pg)


 


[G]



MCHC


 


(g/L)


 


[G]



PLT


(10^9/L)


[G]



0


mg/kg/day


Group 1



Mean



4.578



0.928



3.414



0.090



0.114



0.008



0.024



8.780



186.72



12.38



158.4



0.4568



52.14



18.14



347.6



751.0



SD



1.457



0.335



1.192



0.033



0.040



0.004



0.009



0.892



14.08



0.59



11.6



0.0420



1.20



1.01



16.2



83.3



N



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



4



1000


mg/kg/day


Group 4


 



Mean



5.108



0.894



3.978



0.096



0.112



0.008



0.022



8.662



172.64



11.68 *



160.8



0.4676



54.02



18.60



344.4



730.0



SD



2.131



0.347



1.732



0.063



0.023



0.004



0.013



0.557



31.43



0.26



4.9



0.0229



1.73



0.97



9.9



132.8



N



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



tCtrl



1.12



0.96



1.17



1.07



0.98



1.00



0.92



0.99



0.92



0.94



1.02



1.02



1.04



1.03



0.99



0.97



[G] - Anova & Dunnett: * = p ≤ 0.05


 


Table 16: Haematology summary (Females) - Day 121 (end of recovery)



































































































































































Sex: Female



Reporting Hematology



WBC (10^9/L)


[G]



NEUT (10^9/L)


[G1]



LYMPH (10^9/L)


[G]



MONO (10^9/L)


[G1]



EOS (10^9/L)


[G]



BASO (10^9/L)


[G1]



LUC (10^9/L)


[G1]



RBC (10^12/L)


[G]



RETIC (10^9/L)


[G]



RDWG (%)


[G1]



HGB


 


(g/L)


 


[G]



HCT (L/L)


[G]



MCV (fL)


[G]



MCH


 


(pg)


 


[G]



MCHC


 


(g/L)


 


[G]



PLT


(10^9/L)


[G]



0


mg/kg/day


Group 1



Mean



2.426



0.602



1.670



0.050



0.086



0.004



0.008



7.516



181.82



10.54



140.0



0.4116



54.78



18.64



340.2



710.2



SD



0.783



0.466



0.754



0.016



0.015



0.005



0.004



0.726



12.28



0.27



12.5



0.0396



1.42



0.43



5.2



80.4



N



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



1000


mg/kg/day


Group 4


 



Mean



1.698



0.310



1.270



0.030



0.076



0.004



0.008



8.260



153.40



10.40



149.6



0.4410



53.42



18.12



339.2



723.6



SD



0.294



0.067



0.274



0.012



0.038



0.005



0.004



0.250



44.49



0.32



3.0



0.0124



0.90



0.29



4.1



40.8



N



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



tCtrl



0.70



0.51



0.76



0.60



0.88



1.00



1.00



1.10



0.84



0.99



1.07



1.07



0.98



0.97



1.00



1.02



[G] - Anova & Dunnett
[G1] - Anova & Dunnett


 


Table 17: Clinical biochemistry summary (males) - Day 92 (end of treatment)













































































































































































































































































































































































































 Sex: Male



Reporting Biochemistry



ALT (U/L)


[G]



AST (U/L)


[G]



ALP (U/L)


[G1]



TPROT


 


(g/L)


 


[G]



ALB


 


(g/L)


 


[G]



TBIL


(umol/L) [G]



UREA


(mmol/L) [G1]



CREAT


(umol/L) [G]



GLUC


(mmol/L) [G]



CHOL


(mmol/L) [G]



TRIG


(mmol/L) [G]



HDL


(mmol/L) [G]



LDL


(mmol/L) [G]



NA


(mmol/L) [G]



K


(mmol/L) [G]



CL


(mmol/L) [G]



CA


(mmol/L) [G]



PHOS


(mmol/L) [G]



T3


(ng/mL) [G]



T4


(ng/mL) [G]



TSH


 


(mU/L)


 


[G1]



0


mg/kg/day


Group 1



Mean



51.7



71.7



102.8



65.23



40.96



1.59



5.38



28.2



9.830



1.875



0.956



1.266



0.340



146.3



4.15



107.4



2.562



1.617


0.32437.520.1327

SD



8.2



7.9



18.5



2.46



0.97



0.22



0.74



3.3



1.323



0.436



0.371



0.243



0.073



0.9



0.50



1.0



0.140



0.109


0.0595.280.0737

N



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



100


mg/kg/day


Group 2


 



Mean



49.2



75.0



85.4



63.59



40.15



1.94



5.37



29.6



10.179



1.892



0.959



1.274



0.333



146.5



4.07



107.3



2.597



1.494


0.31636.090.1350

SD



12.3



25.7



13.3



1.73



1.26



0.26



0.44



2.0



1.700



0.192



0.309



0.116



0.042



1.4



0.30



1.6



0.168



0.183


0.0717.270.1308

N



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



tCtrl



0.95



1.05



0.83



0.97



0.98



1.22



1.00



1.05



1.04



1.01



1.00



1.01



0.98



1.0



0.98



1.0



1.01



0.92


0.980.961.02

300


mg/kg/day


Group 3


 



Mean



51.3



69.7



95.0



65.35



41.17



1.59



5.46



29.0



9.169



1.853



1.025



1.258



0.321



146.6



3.88



107.3



2.642



1.600


0.33141.680.1354

SD



9.8



5.8



31.1



2.41



1.41



0.32



0.50



3.0



1.359



0.202



0.336



0.165



0.040



1.2



0.29



1.6



0.105



0.182


0.0705.780.0650

N



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



tCtrl



0.99



0.97



0.92



1.00



1.01



1.00



1.01



1.03



0.93



0.99



1.07



0.99



0.94



1.0



0.93



1.0



1.03



0.99


1.021.111.02

1000


mg/kg/day


Group 4


 



Mean



53.4



87.5



92.5



64.27



41.05



1.82



5.86



29.6



10.106



2.010



1.338



1.362


145.8

145.8



4.17



106.9



2.613



1.791



0.349


36.790.0863

SD



10.0



55.3



22.8



3.24



2.15



0.49



1.34



4.6



1.409



0.190



0.438



0.131



0.046



1.1



0.37



1.5



0.135



0.238


0.0825.520.0568

N



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



tCtrl



1.03



1.22



0.90



0.99



1.00



1.14



1.09



1.05



1.03



1.07



1.40



1.08



1.06



1.0



1.0



1.0



1.02



1.11


1.080.980.65

[G] - Anova & Dunnett
[G1] - Kruskal-Wallis & Dunn


 


Table 18: Clinical biochemistry summary (females) - Day 93 (end of treatment)













































































































































































































































































































































































































 Sex: Female



Reporting Biochemistry



ALT (U/L)


[G]



AST (U/L)


[G1]



ALP (U/L)


[G]



TPROT


 


(g/L)


 


[G]



ALB


 


(g/L)


 


[G]



TBIL


(umol/L) [G]



UREA


(mmol/L) [G1]



CREAT


(umol/L) [G]



GLUC


(mmol/L) [G]



CHOL


(mmol/L) [G]



TRIG


(mmol/L) [G]



HDL


(mmol/L) [G]



LDL


(mmol/L) [G]



NA


(mmol/L) [G]



K


(mmol/L) [G]



CL


(mmol/L) [G]



CA


(mmol/L) [G1]



PHOS


(mmol/L) [G]



T3


(ng/mL) [G]



T4


(ng/mL) [G]



TSH


 


(mU/L)


 


[G1]



0


mg/kg/day


Group 1



Mean



56.4



102.6



38.1



71.08



45.71



2.36



6.85



35.3



8.201



1.628



0.840



1.111


0.328150.83.76111.62.6291.5330.47924.720.1075

SD



17.9



58.1



11.8



2.19



1.64



0.34



0.71



3.2



0.851



0.300



0.317



0.192


0.0501.00.201.40.0820.2000.1075.030.0759

N



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



100


mg/kg/day


Group 2


 



Mean



50.7



74.8



36.1



71.40



45.63



2.01



6.56



37.5



8.172



1.477



0.903



0.995


0.306150.33.90112.02.6041.104**0.49530.040.0829

SD



8.6



6.8



6.9



2.28



1.77



0.36



0.67



3.4



0.652



0.279



0.360



0.189


0.0601.20.361.50.0780.1920.1068.180.0525

N



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



tCtrl



0.90



0.73



0.95



1.00



1.00



0.85



0.96



1.06



1.00



0.91



1.08



0.90


0.931.001.041.000.990.721.031.220.77

300


mg/kg/day


Group 3


 



Mean



46.1



73.4



38.6



67.05 *



42.96 *



1.93



6.60



34.2



7.832



1.408



1.013



0.950


0.287147.2**3.85109.5*2.6711.2990.41424.360.0687

SD



9.1



11.3



18.4



4.85



2.56



0.36



1.02



3.3



1.440



0.358



0.501



0.250


0.0752.60.212.30.1920.2960.0516.660.0504

N



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



tCtrl



0.82



0.72



1.01



0.94



0.94



0.82



0.96



0.97



0.96



0.86



1.21



0.86


0.880.981.020.981.020.850.860.990.64

1000


mg/kg/day


Group 4


 



Mean



54.3



67.3 *



41.3



69.92



44.71



2.25



6.65



32.8



8.691



1.796



0.924



1.254


0.387146.8**3.81108.5**2.7051.4060.54433.98*0.0941

SD



13.0



5.4



11.7



2.83



2.07



0.53



0.82



2.9



1.021



0.335



0.388



0.230


0.0840.80.271.40.1220.2030.1157.400.0400

N



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



10



tCtrl



0.96



0.66



1.08



0.98



0.98



0.95



0.97



0.93



1.06



1.10



1.10



1.13


1.180.971.010.971.030.921.141.370.88

[G] - Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01
[G1] - Kruskal-Wallis & Dunn


 


Table 19: Clinical biochemistry summary (males) - Day 121 (end of recovery)











































































































































































































 Sex: Male



Reporting Biochemistry



ALT (U/L)


[G]



AST (U/L)


[G]



ALP (U/L)


[G]



TPROT


 


(g/L)


 


[G]



ALB


 


(g/L)


 


[G]



TBIL


(umol/L) [G]



UREA


(mmol/L) [G1]



CREAT


(umol/L) [G]



GLUC


(mmol/L) [G]



CHOL


(mmol/L) [G]



TRIG


(mmol/L) [G]



HDL


(mmol/L) [G]



LDL


(mmol/L) [G]



NA


(mmol/L) [G1]



K


(mmol/L) [G]



CL


(mmol/L) [G]



CA


(mmol/L) [G]



PHOS


(mmol/L) [G]



T3


(ng/mL) [G]



T4


(ng/mL) [G1]



TSH


 


(mU/L)


 


[G1]



0


mg/kg/day


Group 1



Mean



46.5



69.3



87.8



62.74



39.14



1.94



4.93



28.7



8.630



1.844



1.044



1.278



0.348



144.6



3.76



106.8



2.590



1.468



0.328



37.74



0.2830



SD



9.6



9.2



20.5



2.36



1.34



0.33



0.50



1.4



0.482



0.338



0.541



0.208



0.047



0.5



0.25



1.1



0.082



0.104



0.070



3.13



0.2264



N



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



1000


mg/kg/day


Group 4


 



Mean



42.0



62.4



86.9



63.42



39.10



1.92



5.41



29.3



9.218



1.790



1.176



1.246



0.338



145.0



3.86



107.6



2.542



1.532



0.286



40.78



0.2070



SD



4.5



8.7



16.4



3.40



1.15



0.40



0.35



0.8



0.686



0.170



0.786



0.130



0.059



1.4



0.22



1.3



0.072



0.102



0.067



10.51



0.0911



N



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



tCtrl



0.90



0.90



0.99



1.01



1.00



0.99



1.10



1.02



1.07



0.97



1.13



0.97



0.97



1.00



1.03



1.01



0.98



1.04



0.87



1.08



0.73



[G] - Anova & Dunnett
[G1] - Kruskal-Wallis & Dunn


 


Table 20: Clinical biochemistry summary (females) - Day 121 (end of recovery)











































































































































































































 Sex: Female



Reporting Biochemistry



ALT (U/L)


[G]



AST (U/L)


[G]



ALP (U/L)


[G1]



TPROT


 


(g/L)


 


[G]



ALB


 


(g/L)


 


[G]



TBIL


(umol/L) [G]



UREA


(mmol/L) [G1]



CREAT


(umol/L) [G]



GLUC


(mmol/L) [G]



CHOL


(mmol/L) [G]



TRIG


(mmol/L) [G]



HDL


(mmol/L) [G]



LDL


(mmol/L) [G]



NA


(mmol/L) [G]



K


(mmol/L) [G]



CL


(mmol/L) [G1]



CA


(mmol/L) [G]



PHOS


(mmol/L) [G]



T3


(ng/mL) [G]



T4


(ng/mL) [G]



TSH


 


(mU/L)


 


[G]



0


mg/kg/day


Group 1



Mean



56.4



142.6



46.0



66.08



42.30



3.18



6.46



33.4



7.566



1.630



0.710



1.178



0.314



143.8



3.63



106.8



2.622



1.478



0.402



26.26



0.0754



SD



30.7



155.3



12.3



2.79



2.06



0.66



0.69



3.5



0.732



0.374



0.247



0.261



0.103



1.5



0.41



1.3



0.098



0.172



0.038



3.65



0.0422



N



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



1000


mg/kg/day


Group 4


 



Mean



40.2



77.5



30.0 *



61.66 *



39.62



1.94 **



6.62



35.8



8.112



1.310



0.576



0.906



0.248



143.8



3.64



108.2



2.592



1.494



0.448



25.18



0.0398



SD



8.6



15.4



5.8



2.05



1.90



0.23



1.02



5.3



0.340



0.311



0.227



0.242



0.062



0.4



0.23



0.4



0.146



0.311



0.060



5.00



0.0295



N



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



5



tCtrl



0.71



0.54



0.65



0.93



0.94



0.61



1.03



1.07



1.07



0.80



0.81



0.77



0.79



1.00



1.00



1.01



0.99



1.01



1.11



0.96



0.53



[G] - Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01
[G1] - Kruskal-Wallis & Dunn


 


Table 21: Functional observations (males)





























































































































































































































































































 



GROUP 1 CONTROL



GROUP 2


100 MG/KG



GROUP 3


300 MG/KG



GROUP 4


1000 MG/KG



 


AT WEEK 13


HEARING



 


MEDIAN



 


0



 


0



 


0



 


0



SCORE 0/1



N



5



5



5



5



PUPIL L



MEDIAN



0



0



0



0



SCORE 0/1



N



5



5



5



5



PUPIL R



MEDIAN



0



0



0



0



SCORE 0/1



N



5



5



5



5



STATIC R



MEDIAN



0



0



0



0



SCORE 0/1



N



5



5



5



5



GRIP FORE



MEAN



1836



1753



1587



1620



GRAM



ST.DEV



144



126



272



180



 



N



5



5



5



5



GRIP HIND



MEAN



1236



1025



1083



1061



GRAM



ST.DEV



215



104



193



131



 



N



5



5



5



5



Total movements



MEAN



3194



2383



2374



2863



 



ST.DEV



919



521



634



400



 



N



5



5



5



5



Ambulations



MEAN



517



389



371



306



 



ST.DEV



155



123



123



108



 



N



5



5



5



5



 


END OF RECOVERY


HEARING



 


MEDIAN



 


0



 



 



 


0



SCORE 0/1



N



5



 



 



5



PUPIL L



MEDIAN



0



 



 



0



SCORE 0/1



N



5



 



 



5



PUPIL R



MEDIAN



0



 



 



0



SCORE 0/1



N



5



 



 



5



STATIC R



MEDIAN



0



 



 



0



SCORE 0/1



N



5



 



 



5



GRIP FORE



MEAN



1385



 



 



1318



GRAM



ST.DEV



101



 



 



108



 



N



5



 



 



5



GRIP HIND



MEAN



621



 



 



610



GRAM



ST.DEV



90



 



 



46



 



N



5



 



 



5



 


Table 22: Functional observations (females)





























































































































































































































































































 



GROUP 1 CONTROL



GROUP 2


100 MG/KG



GROUP 3


300 MG/KG



GROUP 4


1000 MG/KG



 


AT WEEK 13


HEARING



 


MEDIAN



 


0



 


0



 


0



 


0



SCORE 0/1



N



5



5



5



5



PUPIL L



MEDIAN



0



0



0



0



SCORE 0/1



N



5



5



5



5



PUPIL R



MEDIAN



0



0



0



0



SCORE 0/1



N



5



5



5



5



STATIC R



MEDIAN



0



0



0



0



SCORE 0/1



N



5



5



5



5



GRIP FORE



MEAN



1318



1379



1385



1309



GRAM



ST.DEV



100



122



260



298



 



N



5



5



5



5



GRIP HIND



MEAN



753



757



758



731



GRAM



ST.DEV



112



124



108



55



 



N



5



5



5



5



Total movements



MEAN


4615444838702739*

 



ST.DEV


12381689954754

 



N



5



5



5



5



Ambulations



MEAN


1095968101866

 



ST.DEV


250382298298

 



N



5



5



5



5



 


END OF RECOVERY


HEARING



 


MEDIAN



 


0



 



 



 


0



SCORE 0/1



N



5



 



 



5



PUPIL L



MEDIAN



0



 



 



0



SCORE 0/1



N



5



 



 



5



PUPIL R



MEDIAN



0



 



 



0



SCORE 0/1



N



5



 



 



5



STATIC R



MEDIAN



0



 



 



0



SCORE 0/1



N



5



 



 



5



GRIP FORE



MEAN



1141



 



 



1118



GRAM



ST.DEV



51



 



 



143



 



N



5



 



 



5



GRIP HIND



MEAN



498



 



 



420



GRAM



ST.DEV



68



 



 



48



 



N



5



 



 



5



* indicates a p-value <0.05


 


Table 23: organ weights (males) - end of treatment













































































































































































































































































































































































































































































































































































































































































































































































































































































































Sex: Male


 


Day(s) Relative to Start Date



0


mg/kg/day Group 1



100


mg/kg/day Group 2



300


mg/kg/day Group 3



1000


mg/kg/day Group 4



Terminal Body Weight (g) - [G]



Mean



390.2



397.5



392.7



379.9



 



SD



34.3



31.1



35.0



38.2



 



N



10



10



10



10



 



%Diff



-



1.9



0.6



-2.6



Brain Weight (g) - [G]



Mean



2.0935



2.1146



2.0818



2.0894



 



SD



0.0440



0.0863



0.0783



0.0921



 



N



10



10



10



10



 



%Diff



-



1.0079



-0.5589



-0.1958



Brain (%bw) - [G]



Mean



0.54047



0.53454



0.53342



0.55451



 



SD



0.05097



0.04181



0.04581



0.05655



 



N



10



10



10



10



 



%Diff



-



-1.09685



-1.30435



2.59825



Epididymis Weight (g) - [G]



Mean



1.1058



1.2115



1.1425



1.1034



 



SD



0.2424



0.1529



0.0569



0.0814



 



N



10



10



10



10



 



%Diff



-



9.5587



3.3189



-0.2170



Epididymis (%bw) - [G]



Mean



0.28625



0.30616



0.29311



0.29259



 



SD



0.06911



0.04314



0.03155



0.03184



 



N



10



10



10



10



 



%Diff



-



6.95598



2.39837



2.21606



Gland, Adrenal Weight (g) - [G]



Mean



0.05769



0.05761



0.05602



0.04911



 



SD



0.00851



0.00549



0.00635



0.01015



 



N



10



10



10



10



 



%Diff



-



-0.13867



-2.89478



-14.87259



Gland, Adrenal (%bw) - [G]



Mean



0.01484



0.01454



0.01438



0.01305



 



SD



0.00216



0.00149



0.00222



0.00291



 



N



10



10



10



10



 



%Diff



-



-2.04209



-3.15592



-12.06055



Gland, Pituitary Weight (g) [G]



Mean



0.00824



0.00858



0.00785



0.00821



 



SD



0.00105



0.00140



0.00140



0.00071



 



N



10



10



10



10



 



%Diff



-



4.12621



-4.73301



-0.36408



Gland, Pituitary (%bw) [G1]



Mean



0.00211



0.00215



0.00200



0.00217



 



SD



0.00021



0.00027



0.00035



0.00014



 



N



10



10



10



10



 



%Diff



-



1.94015



-5.12563



2.58658



Gland, Prostate Weight (g) [G]



Mean



0.9458



0.9291



0.8763



0.8380



 



SD



0.1549



0.1875



0.1589



0.1562



 



N



10



10



10



10



 



%Diff



-



-1.7657



-7.3483



-11.3978



Gland, Prostate (%bw) [G]



Mean



0.24348



0.23503



0.22478



0.22299



 



SD



0.04065



0.05206



0.04463



0.04970



 



N



10



10



10



10



 



%Diff



-



-3.47163



-7.68092



-8.41815



Seminal Vesicle Weight (g) - [G]



Mean



1.2683



1.3261



1.2883



1.2644



 



SD



0.1735



0.2229



0.1849



0.2585



 



N



10



10



10



10



 



%Diff



-



4.5573



1.5769



-0.3075



Seminal Vesicle (%bw) - [G]



Mean



0.32660



0.33502



0.32943



0.33592



 



SD



0.04811



0.05917



0.04807



0.07580



 



N



10



10



10



10



 



%Diff



-



2.57825



0.86792



2.85589



Thyroid/Parathyroid Weight (g) - [G1]



Mean



0.01561



0.01824



0.01915



0.01891



 



SD



0.00394



0.00529



0.00305



0.00751



 



N



10



10



10



10



 



%Diff



-



16.84817



22.67777



21.14029



Thyroid/Parathyroid (%bw) - [G]



Mean



0.00399



0.00460



0.00492



0.00491



 



SD



0.00094



0.00133



0.00090



0.00171



 



N



10



10



10



10



 



%Diff



-



15.30492



23.42286



23.16273



Heart Weight (g) - [G]



Mean



1.0040



1.0086



1.0097



0.9653



 



SD



0.0791



0.1207



0.0761



0.0657



 



N



10



10



10



10



 



%Diff



-



0.4582



0.5677



-3.8546



Heart (%bw) - [G]



Mean



0.25866



0.25349



0.25822



0.25538



 



SD



0.02636



0.01924



0.02185



0.01998



 



N



10



10



10



10



 



%Diff



-



-1.99783



-0.17065



-1.26674



Kidney Weight (g) - [G]



Mean



2.3623



2.4359



2.4542



2.2696



 



SD



0.2188



0.3319



0.1431



0.2500



 



N



10



10



10



10



 



%Diff



-



3.1156



3.8903



-3.9241



Kidney (%bw) - [G]



Mean



0.60731



0.61216



0.62858



0.59839



 



SD



0.05371



0.05901



0.05800



0.04842



 



N



10



10



10



10



 



%Diff



-



0.79814



3.50222



-1.46983



Liver Weight (g) - [G]



Mean



8.7411



8.7143



8.7788



8.9585



 



SD



0.9853



0.9765



1.0136



0.8868



 



N



10



10



10



10



 



%Diff



-



-0.3066



0.4313



2.4871



Liver (%bw) - [G]



Mean



2.23748



2.18929



2.23487



2.36098



 



SD



0.10434



0.11829



0.15663



0.11509



 



N



10



10



10



10



 



%Diff



-



-2.15404



-0.11678



5.51955



Spleen Weight (g) - [G1]



Mean



0.5104



0.5840



0.5480



0.5271



 



SD



0.0500



0.1114



0.0702



0.0474



 



N



10



10



10



10



 



%Diff



-



14.4201



7.3668



3.2719



Spleen (%bw) - [G]



Mean



0.13119



0.14625



0.14010



0.13916



 



SD



0.01222



0.02020



0.01873



0.00983



 



N



10



10



10



10



 



%Diff



-



11.47773



6.78977



6.07030



Testis Weight (g) - [G]



Mean



3.2215



3.7755



3.6595



3.5242



 



SD



0.9363



0.3880



0.3237



0.1842



 



N



10



10



10



10



 



%Diff



-



17.1970



13.5962



9.3962



Testis (%bw) - [G]



Mean



0.83486



0.95425



0.93987



0.93357



 



SD



0.25480



0.11391



0.12907



0.07621



 



N



10



10



10



10



 



%Diff



-



14.30168



12.57932



11.82419



Thymus Weight (g) - [G]



Mean



0.2759



0.2790



0.3239



0.3061



 



SD



0.0834



0.0786



0.0605



0.0959



 



N



10



10



10



10



 



%Diff



-



1.1236



17.3976



10.9460



Thymus (%bw) - [G]



Mean



0.06975



0.06991



0.08280



0.07997



 



SD



0.01722



0.01779



0.01446



0.02148



 



N



10



10



10



10



 



%Diff



-



0.23340



18.70625



14.65698



[G] - Anova & Dunnett
[G1] - Kruskal-Wallis & Dunn: * = p ≤ 0.05


 


Table 24: organ weights (females) - end of treatment













































































































































































































































































































































































































































































































































































































































































































































































Sex: Female


 


Day(s) Relative to Start Date



0


mg/kg/day Group 1



100


mg/kg/day Group 2



300


mg/kg/day Group 3



1000


mg/kg/day Group 4



Terminal Body Weight (g) - [G]



Mean



220.6



226.2



213.3



231.0



 



SD



13.7



15.3



15.3



13.0



 



N



10



10



10



10



 



%Diff



-



2.5



-3.3



4.7



Brain Weight (g) - [G1]



Mean



1.8958



1.9140



1.8589



1.8968



 



SD



0.0612



0.0492



0.0271



0.0563



 



N



10



10



10



10



 



%Diff



-



0.9600



-1.9464



0.0527



Brain (%bw) - [G]



Mean



0.86149



0.84923



0.87619



0.82264



 



SD



0.04436



0.05478



0.07360



0.03431



 



N



10



10



10



10



 



%Diff



-



-1.42301



1.70715



-4.50976



Gland, Adrenal Weight (g) - [G]



Mean



0.05907



0.06330



0.06361



0.06358



 



SD



0.00971



0.01033



0.00735



0.00694



 



N



10



10



10



10



 



%Diff



-



7.16100



7.68580



7.63501



Gland, Adrenal (%bw) - [G]



Mean



0.02672



0.02794



0.02982



0.02754



 



SD



0.00369



0.00378



0.00273



0.00268



 



N



10



10



10



10



 



%Diff



-



4.59109



11.62243



3.06385



Gland, Pituitary Weight (g) [G1]



Mean



0.01478



0.01014



0.01116



0.01126



 



SD



0.00532



0.00255



0.00296



0.00167



 



N



10



10



10



10



 



%Diff



-



-31.39378



-24.49256



-23.81597



Gland, Pituitary (%bw) [G1]



Mean



0.00666



0.00448 *



0.00523



0.00488



 



SD



0.00223



0.00103



0.00130



0.00073



 



N



10



10



10



10



 



%Diff



-



-32.71386



-21.48404



-26.68346



Thyroid/Parathyroid Weight (g) - [G]



Mean



0.01239



0.01440



0.01277



0.01443



 



SD



0.00220



0.00284



0.00367



0.00354



 



N



10



10



10



10



 



%Diff



-



16.22276



3.06699



16.46489



Thyroid/Parathyroid (%bw) - [G]



Mean



0.00564



0.00638



0.00600



0.00630



 



SD



0.00111



0.00132



0.00174



0.00182



 



N



10



10



10



10



 



%Diff



-



13.19007



6.42890



11.71956



Heart Weight (g) - [G]



Mean



0.6792



0.6821



0.6207



0.7181



 



SD



0.0507



0.0757



0.0536



0.0448



 



N



10



10



10



10



 



%Diff



-



0.4270



-8.6131



5.7273



Heart (%bw) - [G]



Mean



0.30853



0.30129



0.29126



0.31131



 



SD



0.02475



0.02313



0.01832



0.01892



 



N



10



10



10



10



 



%Diff



-



-2.34775



-5.59609



0.90247



Kidney Weight (g) - [G]



Mean



1.4672



1.5831



1.4736



1.7272 **



 



SD



0.0838



0.1193



0.1311



0.1965



 



N



10



10



10



10



 



%Diff



-



7.8994



0.4362



17.7208



Kidney (%bw) - [G1]



Mean



0.66594



0.70065



0.69076



0.74781 **



 



SD



0.03215



0.04251



0.03323



0.07497



 



N



10



10



10



10



 



%Diff



-



5.21116



3.72597



12.29292



Liver Weight (g) - [G]



Mean



5.3753



5.4101



5.0887



6.4360 **



 



SD



0.3872



0.5482



0.4642



0.5899



 



N



10



10



10



10



 



%Diff



-



0.6474



-5.3318



19.7329



Liver (%bw) - [G]



Mean



2.43901



2.39304



2.38706



2.79153 **



 



SD



0.14303



0.20287



0.15493



0.27667



 



N



10



10



10



10



 



%Diff



-



-1.88477



-2.12999



14.45347



Ovary Paired (g) - [G]



Mean



0.1361



0.1468



0.1421



0.1377



 



SD



0.0210



0.0210



0.0243



0.0172



 



N



10



10



10



10



 



%Diff



-



7.8619



4.4085



1.1756



Ovary Paired (%bw) - [G]



Mean



0.06164



0.06522



0.06703



0.05966



 



SD



0.00831



0.01101



0.01308



0.00699



 



N



10



10



10



10



 



%Diff



-



5.80168



8.74297



-3.22236



Spleen Weight (g) - [G1]



Mean



0.4167



0.4165



0.3773



0.4617



 



SD



0.0763



0.0388



0.0371



0.0357



 



N



10



10



10



10



 



%Diff



-



-0.0480



-9.4552



10.7991



Spleen (%bw) - [G]



Mean



0.18857



0.18411



0.17756



0.20006



 



SD



0.03104



0.01172



0.02108



0.01443



 



N



10



10



10



10



 



%Diff



-



-2.36212



-5.83761



6.09656



Thymus Weight (g) - [G1]



Mean



0.2929



0.2894



0.2506



0.2886



 



SD



0.0820



0.0706



0.0479



0.0574



 



N



10



10



10



10



 



%Diff



-



-1.1949



-14.4418



-1.4681



Thymus (%bw) - [G1]



Mean



0.13204



0.12701



0.11827



0.12521



 



SD



0.03288



0.02467



0.02659



0.02594



 



N



10



10



10



10



 



%Diff



-



-3.81140



-10.42849



-5.17288



Uterus/Cervix (g) - [G]



Mean



0.6704



0.6153



0.4746



0.5312



 



SD



0.2816



0.2422



0.0977



0.1136



 



N



10



10



10



10



 



%Diff



-



-8.2190



-29.2064



-20.7637



Uterus/Cervix (%bw) - [G]



Mean



0.30787



0.27105



0.22254



0.23051



 



SD



0.13656



0.09880



0.04277



0.05042



 



N



10



10



10



10



 



%Diff



-



-11.96024



-27.71662



-25.12699



[G] - Anova & Dunnett: ** = p ≤ 0.01
[G] - Anova & Dunnett


 


Table 25: organ weights (males) - end of recovery



















































































































































































































































































































































































































































































































































































































































































Sex: Male


 


Day(s) Relative to Start Date



0


mg/kg/day Group 1



1000


mg/kg/day Group 4



Terminal Body Weight (g) - [G]



Mean



416.2



417.8



 



SD



30.4



33.6



 



N



5



5



 



%Diff



-



0.4



Brain Weight (g) - [G]



Mean



2.1002



2.0828



 



SD



0.1227



0.1062



 



N



5



5



 



%Diff



-



-0.8285



Brain (%bw) - [G]



Mean



0.50524



0.50009



 



SD



0.01655



0.03204



 



N



5



5



 



%Diff



-



-1.01955



Epididymis Weight (g) - [G]



Mean



1.2254



1.2330



 



SD



0.1105



0.1300



 



N



5



5



 



%Diff



-



0.6202



Epididymis (%bw) - [G]



Mean



0.29438



0.29543



 



SD



0.01556



0.02716



 



N



5



5



 



%Diff



-



0.35575



Gland, Adrenal Weight (g) - [G]



Mean



0.05264



0.05538



 



SD



0.00576



0.00582



 



N



5



5



 



%Diff



-



5.20517



Gland, Adrenal (%bw) - [G]



Mean



0.01266



0.01333



 



SD



0.00117



0.00178



 



N



5



5



 



%Diff



-



5.26824



Gland, Pituitary Weight (g) [G1]



Mean



0.00922



0.01498



 



SD



0.00154



0.00829



 



N



5



5



 



%Diff



-



62.47289



Gland, Pituitary (%bw) [G1]



Mean



0.00221



0.00353



 



SD



0.00027



0.00185



 



N



5



5



 



%Diff



-



59.90894



Gland, Prostate Weight (g) [G]



Mean



1.0258



0.8896



 



SD



0.1418



0.1802



 



N



5



5



 



%Diff



-



-13.2774



Gland, Prostate (%bw) [G]



Mean



0.24663



0.21396



 



SD



0.03056



0.04640



 



N



5



5



 



%Diff



-



-13.24820



Seminal Vesicle Weight (g) - [G]



Mean



1.3982



1.5228



 



SD



0.2755



0.3414



 



N



5



5



 



%Diff



-



8.9115



Seminal Vesicle (%bw) - [G]



Mean



0.33727



0.36373



 



SD



0.06901



0.06780



 



N



5



5



 



%Diff



-



7.84566



Thyroid/Parathyroid Weight (g) - [G1]



Mean



0.01610



0.01556



 



SD



0.00533



0.00319



 



N



5



5



 



%Diff



-



-3.35404



Thyroid/Parathyroid (%bw) - [G1]



Mean



0.00388



0.00370



 



SD



0.00129



0.00054



 



N



5



5



 



%Diff



-



-4.55710



Heart Weight (g) - [G]



Mean



1.0272



1.0704



 



SD



0.0534



0.0746



 



N



5



5



 



%Diff



-



4.2056



Heart (%bw) - [G]



Mean



0.24756



0.25665



 



SD



0.01706



0.01272



 



N



5



5



 



%Diff



-



3.66822



Kidney Weight (g) - [G]



Mean



2.3472



2.5980



 



SD



0.2748



0.3611



 



N



5



5



 



%Diff



-



10.6851



Kidney (%bw) - [G]



Mean



0.56367



0.62134



 



SD



0.04690



0.06600



 



N



5



5



 



%Diff



-



10.23148



Liver Weight (g) - [G]



Mean



8.8570



9.6144



 



SD



1.0204



1.5403



 



N



5



5



 



%Diff



-



8.5514



Liver (%bw) - [G]



Mean



2.12282



2.29077



 



SD



0.09992



0.21010



 



N



5



5



 



%Diff



-



7.91134



Spleen Weight (g) - [G]



Mean



0.6252



0.6066



 



SD



0.0440



0.1345



 



N



5



5



 



%Diff



-



-2.9750



Spleen (%bw) - [G1]



Mean



0.15038



0.14470



 



SD



0.00762



0.02485



 



N



5



5



 



%Diff



-



-3.77737



Testis Weight (g) - [G]



Mean



3.6980



3.7970



 



SD



0.1267



0.1574



 



N



5



5



 



%Diff



-



2.6771



Testis (%bw) - [G]



Mean



0.89239



0.91195



 



SD



0.07222



0.05657



 



N



5



5



 



%Diff



-



2.19240



Thymus Weight (g) - [G]



Mean



0.2932



0.2612



 



SD



0.0699



0.0988



 



N



5



5



 



%Diff



-



-10.9141



Thymus (%bw) - [G]



Mean



0.07044



0.06192



 



SD



0.01514



0.02120



 



N



5



5



 



%Diff



-



-12.09428



[G] - Anova & Dunnett
[G1] - Kruskal-Wallis & Dunn


 


Table 26: organ weights (females) - end of recovery



















































































































































































































































































































































































































































































































































































Sex: Female


 


Day(s) Relative to Start Date



0


mg/kg/day Group 1



1000


mg/kg/day Group 4



Terminal Body Weight (g) - [G]



Mean



239.8



227.6



 



SD



9.6



15.5



 



N



5



5



 



%Diff



-



-5.1



Brain Weight (g) - [G1]



Mean



1.9122



1.9748



 



SD



0.0511



0.1385



 



N



5



5



 



%Diff



-



3.2737



Brain (%bw) - [G]



Mean



0.79809



0.86811 **



 



SD



0.02740



0.03459



 



N



5



5



 



%Diff



-



8.77419



Gland, Adrenal Weight (g) - [G]



Mean



0.06100



0.07034



 



SD



0.00904



0.01447



 



N



5



5



 



%Diff



-



15.31148



Gland, Adrenal (%bw) - [G]



Mean



0.02552



0.03068



 



SD



0.00446



0.00444



 



N



5



5



 



%Diff



-



20.21525



Gland, Pituitary Weight (g) [G1]



Mean



0.01114



0.01256



 



SD



0.00414



0.00060



 



N



5



5



 



%Diff



-



12.74686



Gland, Pituitary (%bw) [G]



Mean



0.00460



0.00554



 



SD



0.00161



0.00042



 



N



5



5



 



%Diff



-



20.31781



Thyroid/Parathyroid Weight (g) - [G]



Mean



0.01410



0.01654



 



SD



0.00103



0.00277



 



N



5



5



 



%Diff



-



17.30496



Thyroid/Parathyroid (%bw) - [G]



Mean



0.00589



0.00732



 



SD



0.00057



0.00157



 



N



5



5



 



%Diff



-



24.25956



Heart Weight (g) - [G]



Mean



0.7156



0.6652



 



SD



0.0564



0.0945



 



N



5



5



 



%Diff



-



-7.0430



Heart (%bw) - [G]



Mean



0.29813



0.29128



 



SD



0.01336



0.02578



 



N



5



5



 



%Diff



-



-2.29654



Kidney Weight (g) - [G]



Mean



1.5562



1.5080



 



SD



0.1430



0.1578



 



N



5



5



 



%Diff



-



-3.0973



Kidney (%bw) - [G]



Mean



0.64965



0.66137



 



SD



0.06221



0.02983



 



N



5



5



 



%Diff



-



1.80335



Liver Weight (g) - [G]



Mean



5.8176



5.3962



 



SD



0.5446



0.6612



 



N



5



5



 



%Diff



-



-7.2435



Liver (%bw) - [G]



Mean



2.42291



2.36682



 



SD



0.14767



0.20047



 



N



5



5



 



%Diff



-



-2.31522



Ovary Paired (g) - [G]



Mean



0.1388



0.1400



 



SD



0.0169



0.0182



 



N



5



5



 



%Diff



-



0.8646



Ovary Paired (%bw) - [G]



Mean



0.05802



0.06134



 



SD



0.00809



0.00477



 



N



5



5



 



%Diff



-



5.72474



Spleen Weight (g) - [G1]



Mean



0.4208



0.3914



 



SD



0.0223



0.0478



 



N



5



5



 



%Diff



-



-6.9867



Spleen (%bw) - [G]



Mean



0.17580



0.17162



 



SD



0.01300



0.01342



 



N



5



5



 



%Diff



-



-2.37705



Thymus Weight (g) - [G]



Mean



0.2656



0.2180



 



SD



0.0429



0.0323



 



N



5



5



 



%Diff



-



-17.9217



Thymus (%bw) - [G]



Mean



0.11060



0.09586



 



SD



0.01639



0.01350



 



N



5



5



 



%Diff



-



-13.32804



Uterus/Cervix (g) - [G]



Mean



0.6696



0.5598



 



SD



0.2254



0.1107



 



N



5



5



 



%Diff



-



-16.3978



Uterus/Cervix (%bw) - [G1]



Mean



0.27920



0.24616



 



SD



0.09299



0.04749



 



N



5



5



 



%Diff



-



-11.83234



[G] - Anova & Dunnett: ** = p ≤ 0.01
[G1] - Kruskal-Wallis & Dunn


 


Table 27: microscopic pathology (males & females) - end of treatment































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































































Removal Reason(s): TERMINAL EUTHANASIA


Summary: Incidence


 


 


 


 


Number of Animals:



 


 


0


 


mg/kg/ day Group 1



Male


 


100         300


 


mg/kg/                mg/kg/ day     day Group                Group


2              3



 


 


1000


 


mg/kg/ day Group 4



 


 


0


 


mg/kg/ day Group 1



Female


 


100         300


 


mg/kg/ mg/kg/ day                day Group                Group


2              3



 


 


1000


 


mg/kg/ day Group 4



10



10



10



10



10



10



10



10



ARTERY, AORTA



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



10



10



.



.



10



BONE MARROW, STERNUM



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



10



10



.



.



10



BONE, STERNUM



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



10



10



.



.



10



BRAIN



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



10



10



.



.



10



CERVIX



 



 



 



 



 



 



 



 



Examined



.



.



.



.



10



0



0



10



No Visible Lesions



.



.



.



.



10



.



.



10



EPIDIDYMIS



 



 



 



 



 



 



 



 



Examined



10



0



0



10



.



.



.



.



No Visible Lesions



9



.



.



9



.



.



.



.



Sperm, decreased; lumen



1



.



.



0



.



.



.



.



.... marked



1



.



.



0



.



.



.



.



Infiltration, mononuclear cell;



0



.



.



1



.



.



.



.



interstitial



 



 



 



 



 



 



 



 



.... minimal



0



.



.



1



.



.



.



.



ESOPHAGUS



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



10



10



.



.



10



EYE



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



9



10



.



.



9



Hemorrhage; acute



0



.



.



1



0



.



.



1



.... minimal



0



.



.



1



0



.



.



1



GALT



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



10



10



.



.



10



GLAND, ADRENAL



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



9



10



.



.



10



Vacuolation; zona fasciculata



0



.



.



1



0



.



.



0



.... minimal



0



.



.



1



0



.



.



0



GLAND, CLITORAL



 



 



 



 



 



 



 



 



Examined



.



.



.



.



0



1



1



0



Dilatation; ductular



.



.



.



.



.



1



1



.



.... minimal



.



.



.



.



.



1



1



.



GLAND, COAGULATING



 



 



 



 



 



 



 



 



Examined



10



0



0



10



.



.



.



.



No Visible Lesions



10



.



.



10



.



.



.



.



GLAND, MAMMARY



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



10



10



.



.



10



GLAND, PARATHYROID



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



10



10



.



.



10



GLAND, PITUITARY



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



10



10



.



.



10



GLAND, PREPUTIAL



 



 



 



 



 



 



 



 



Examined



0



0



1



0



.



.



.



.



Dilatation; acinar



.



.



1



.



.



.



.



.



.... minimal



.



.



1



.



.



.



.



.



GLAND, PROSTATE



 



 



 



 



 



 



 



 



Examined



10



0



0



10



.



.



.



.



No Visible Lesions



8



.



.



10



.



.



.



.



Infiltration, mononuclear cell;



2



.



.



0



.



.



.



.



interstitial



 



 



 



 



 



 



 



 



.... minimal



2



.



.



0



.



.



.



.



GLAND, SALIVARY,



 



 



 



 



 



 



 



 



MANDIBULAR



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



10



10



.



.



10



GLAND, SALIVARY,



 



 



 



 



 



 



 



 



SUBLINGUAL



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



10



10



.



.



10



GLAND, SEMINAL VESICLE



 



 



 



 



 



 



 



 



Examined



10



0



0



10



.



.



.



.



No Visible Lesions



10



.



.



10



.



.



.



.



GLAND, THYROID



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



8



.



.



7



7



.



.



7



Cyst; ultimobranchial



0



.



.



0



2



.



.



2



.... minimal



0



.



.



0



2



.



.



2



Hypertrophy; follicular cell



1



.



.



2



0



.



.



0



.... minimal



1



.



.



2



0



.



.



0



Ectopia; thymic



1



.



.



1



1



.



.



1



.... minimal



1



.



.



1



1



.



.



1



HEART



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



9



10



.



.



10



Infiltration, mononuclear cell;



0



.



.



1



0



.



.



0



myocardial



 



 



 



 



 



 



 



 



.... minimal



0



.



.



1



0



.



.



0



KIDNEY



 



 



 



 



 



 



 



 



Examined



10



0



3



10



10



0



0



10



No Visible Lesions



1



.



2



5



8



.



.



7



Basophilia; tubular



6



.



0



2



1



.



.



1



.... minimal



6



.



0



2



1



.



.



1



Accumulation; hyaline droplets



7



.



0



2



0



.



.



0



.... minimal



5



.



0



2



0



.



.



0



.... mild



2



.



0



0



0



.



.



0



Infiltration, mononuclear cell;



3



.



0



1



1



.



.



3



interstitial



 



 



 



 



 



 



 



 



.... minimal



3



.



0



1



1



.



.



3



Dilatation; cortical, tubular



0



.



0



1



0



.



.



0



.... minimal



0



.



0



1



0



.



.



0



Dilatation; pelvis



0



.



1



0



0



.



.



0



.... minimal



0



.



1



0



0



.



.



0



LARGE INTESTINE, CECUM



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



10



10



.



.



10



LARGE INTESTINE, COLON



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



10



10



.



.



10



LARGE INTESTINE, RECTUM



 



 



 



 



 



 



 



 



Examined



10



10



10



10



10



0



0



10



No Visible Lesions



10



10



10



7



10



.



.



10



Hyperplasia; goblet cell



0



0



0



3



0



.



.



0



.... minimal



0



0



0



3



0



.



.



0



LIVER



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



5



.



.



5



8



.



.



10



Infiltration, mononuclear cell



5



.



.



5



2



.



.



0



.... minimal



5



.



.



5



2



.



.



0



Atrophy



2



.



.



0



0



.



.



0



.... minimal



1



.



.



0



0



.



.



0



.... mild



1



.



.



0



0



.



.



0



LUNG



 



 



 



 



 



 



 



 



Examined



10



0



1



10



10



0



0



10



No Visible Lesions



9



.



0



7



9



.



.



8



Aggregate; alveolar, macrophage



0



.



1



1



0



.



.



0



.... minimal



0



.



0



1



0



.



.



0



.... moderate



0



.



1



0



0



.



.



0



Infiltration, mixed cell;



1



.



0



1



1



.



.



2



perivascular



 



 



 



 



 



 



 



 



.... minimal



1



.



0



1



1



.



.



2



Metaplasia; osseous



0



.



0



1



0



.



.



0



.... minimal



0



.



0



1



0



.



.



0



LYMPH NODE,



 



 



 



 



 



 



 



 



MANDIBULAR



 



 



 



 



 



 



 



 



Examined



10



2



0



10



10



1



0



10



No Visible Lesions



8



0



.



9



7



0



.



10



Hemorrhage; acute



2



2



.



1



3



1



.



0



.... minimal



2



2



.



1



2



1



.



0



.... mild



0



0



.



0



1



0



.



0



LYMPH NODE, MESENTERIC



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



10



10



.



.



10



MUSCLE, SKELETAL



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



5



.



.



9



9



.



.



9



Degeneration; myofiber



5



.



.



0



1



.



.



1



.... minimal



5



.



.



0



1



.



.



1



Infiltration, mononuclear cell



1



.



.



1



0



.



.



0



.... minimal



1



.



.



1



0



.



.



0



NERVE, OPTIC



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



10



10



.



.



10



NERVE, SCIATIC



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



10



10



.



.



10



OVARY



 



 



 



 



 



 



 



 



Examined



.



.



.



.



10



0



1



10



No Visible Lesions



.



.



.



.



10



.



1



10



PANCREAS



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



10



10



.



.



9



Atrophy; acinar



0



.



.



0



0



.



.



1



.... minimal



0



.



.



0



0



.



.



1



SKIN



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



10



10



.



.



10



SMALL INTESTINE,



 



 



 



 



 



 



 



 



DUODENUM



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



10



10



.



.



10



SMALL INTESTINE, ILEUM



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



10



10



.



.



10



SMALL INTESTINE,



 



 



 



 



 



 



 



 



JEJUNUM



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



10



10



.



.



10



SPINAL CORD



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



10



10



.



.



10



SPLEEN



 



 



 



 



 



 



 



 



Examined



10



1



0



10



10



0



0



10



No Visible Lesions



1



1



.



3



0



.



.



0



Pigment



9



0



.



6



10



.



.



10



.... minimal



8



0



.



6



9



.



.



9



.... mild



1



0



.



0



1



.



.



1



Extramedullary hematopoiesis



3



0



.



2



6



.



.



5



.... minimal



2



0



.



0



4



.



.



4



.... mild



1



0



.



2



2



.



.



1



STOMACH



 



 



 



 



 



 



 



 



Examined



10



0



1



10



10



10



10



10



No Visible Lesions



8



.



1



5



7



10



8



4



Hyperplasia; basal cell; glandular



0



.



0



1



0



0



0



1



.... minimal



0



.



0



1



0



0



0



1



Hyperplasia; squamous cell;



0



.



0



0



0



0



1



3



non-glandular



 



 



 



 



 



 



 



 



.... minimal



0



.



0



0



0



0



1



2



.... mild



0



.



0



0



0



0



0



1



Vacuolation; epithelial, limiting



0



.



0



0



2



0



0



2



ridge



 



 



 



 



 



 



 



 



.... minimal



0



.



0



0



2



0



0



2



Erosion; non-glandular



0



.



0



0



0



0



0



1



.... minimal



0



.



0



0



0



0



0



1



Infiltration, lymphocytic;



2



.



0



3



1



0



1



1



submucosal, glandular



 



 



 



 



 



 



 



 



.... minimal



2



.



0



3



1



0



1



1



Infiltration, lymphocytic; limiting



0



.



0



0



0



0



0



1



ridge



 



 



 



 



 



 



 



 



.... minimal



0



.



0



0



0



0



0



1



Infiltration, mixed cell;



1



.



0



1



0



0



0



1



submucosal, glandular



 



 



 



 



 



 



 



 



.... minimal



1



.



0



1



0



0



0



1



TESTIS



 



 



 



 



 



 



 



 



Examined



10



0



0



10



.



.



.



.



No Visible Lesions



6



.



.



8



.



.



.



.



Degeneration/atrophy; tubular



4



.



.



2



.



.



.



.



.... minimal



2



.



.



2



.



.



.



.



.... mild



1



.



.



0



.



.



.



.



.... marked



1



.



.



0



.



.



.



.



THYMUS



 



 



 



 



 



 



 



 



Examined



10



1



0



10



10



0



0



10



No Visible Lesions



10



0



.



10



10



.



.



10



Hemorrhage; acute



0



1



.



0



0



.



.



0



.... minimal



0



1



.



0



0



.



.



0



TRACHEA



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



10



10



.



.



10



URINARY BLADDER



 



 



 



 



 



 



 



 



Examined



10



0



0



10



10



0



0



10



No Visible Lesions



10



.



.



10



10



.



.



10



UTERUS



 



 



 



 



 



 



 



 



Examined



.



.



.



.



10



2



0



10



No Visible Lesions



.



.



.



.



10



2



.



10



VAGINA



 



 



 



 



 



 



 



 



Examined



.



.



.



.



10



0



0



10



Diestrus



.



.



.



.



5



.



.



7



Estrus



.



.



.



.



2



.



.



3



Proestrus



.



.



.



.



3



.



.



0



 


Table 28: microscopic pathology (males & females) - end of recovery



































































































































































































































Removal Reason(s): RECOVERY EUTHANASIA Summary: Incidence


 


 


 


 


Number of Animals:



Male


0          1000


 


mg/kg/ mg/kg/ day         day Group    Group


1              4



Female


0          1000


 


mg/kg/ mg/kg/ day         day Group    Group


1              4



5



5



5



5



KIDNEY



 



 



 



 



Examined



0



1



0



1



No Visible Lesions



.



0



.



1



Cyst; cortical



.



1



.



0



.... mild



.



1



.



0



LARGE INTESTINE, RECTUM



 



 



 



 



Examined



5



5



.



.



No Visible Lesions



5



5



.



.



LIVER



 



 



 



 



Examined



1



0



.



.



Atrophy



1



.



.



.



.... minimal



1



.



.



.



LUNG



 



 



 



 



Examined



.



.



1



0



Hemorrhage; acute, alveolar



.



.



1



.



.... minimal



.



.



1



.



STOMACH



 



 



 



 



Examined



0



1



5



5



No Visible Lesions



.



1



4



3



Vacuolation; epithelial, limiting ridge



.



0



1



1



.... minimal



.



0



1



1



Erosion; glandular



.



0



0



1



.... minimal



.



0



0



1



THYMUS



 



 



 



 



Examined



.



.



0



1



Hemorrhage; acute



.



.



.



1



.... minimal



.



.



.



1



UTERUS



 



 



 



 



Examined



.



.



2



1



No Visible Lesions



.



.



2



1



 


Table 29: clinical observations (males & females) - Main














































































































































































































































































































































































































































































































































































































































































































Observation Type: All Types


From Day 1 (Start Date) to 93 (Start Date)



 


0


mg/kg/day Group 1



Male


100                     300


mg/kg/day mg/kg/day Group 2                     Group 3



 


1000


mg/kg/day Group 4



 


0


mg/kg/day Group 1



Female


100                     300


mg/kg/day mg/kg/day Group 2                     Group 3



 


1000


mg/kg/day Group 4



Retching



 



 



 



 



 



 



 



 



Number of Animals Affected



0



1



0



6



0



1



3



9



Number of Times Recorded



0



1



0



8



0



1



10



13



% of Affected Animals



0



10



0



40



0



10



30



60



First to Last seen



-



44 - 44



-



43 - 48



-



44 - 44



44 - 49



43 - 48



Salivation



 



 



 



 



 



 



 



 



Number of Animals Affected



0



0



10



15



0



0



4



15



Number of Times Recorded



0



0



96



1223



0



0



25



1205



% of Affected Animals



0



0



100



100



0



0



40



100



First to Last seen



-



-



12 - 91



7 - 91



-



-



12 - 89



7 - 92



Breathing, Deep



 



 



 



 



 



 



 



 



Number of Animals Affected



0



0



0



0



0



0



1



0



Number of Times Recorded



0



0



0



0



0



0



2



0



% of Affected Animals



0



0



0



0



0



0



10



0



First to Last seen



-



-



-



-



-



-



91 - 92



-



Breathing, Labored



 



 



 



 



 



 



 



 



Number of Animals Affected



0



0



0



0



0



0



0



1



Number of Times Recorded



0



0



0



0



0



0



0



1



% of Affected Animals



0



0



0



0



0



0



0



7



First to Last seen



-



-



-



-



-



-



-



81 - 81



Breathing, Abnormal Sounds



 



 



 



 



 



 



 



 



Number of Animals Affected



0



3



3



10



1



1



5



6



Number of Times Recorded



0



7



10



36



11



3



15



20



% of Affected Animals



0



30



30



67



7



10



50



40



First to Last seen



-



9 - 91



8 - 91



8 - 91



61 - 90



74 - 76



42 - 92



3 - 91



Breathing, Shallow



 



 



 



 



 



 



 



 



Number of Animals Affected



0



2



0



1



0



0



0



1



Number of Times Recorded



0



6



0



1



0



0



0



1



% of Affected Animals



0



20



0



7



0



0



0



7



First to Last seen



-



89 - 91



-



89 - 89



-



-



-



89 - 89



Hunched Posture



 



 



 



 



 



 



 



 



Number of Animals Affected



0



0



1



1



0



0



0



1



Number of Times Recorded



0



0



13



2



0



0



0



1



% of Affected Animals



0



0



10



7



0



0



0



7



First to Last seen



-



-



23 - 35



28 - 29



-



-



-



81 - 81



Fur, Erected



 



 



 



 



 



 



 



 



Number of Animals Affected



0



0



2



4



0



0



1



1



Number of Times Recorded



0



0



7



9



0



0



2



1



% of Affected Animals



0



0



20



27



0



0



10



7



First to Last seen



-



-



23 - 35



42 - 67



-



-



91 - 92



81 - 81



Fur, Loss



 



 



 



 



 



 



 



 



Number of Animals Affected



0



0



1



0



0



0



0



0



Number of Times Recorded



0



0



75



0



0



0



0



0



% of Affected Animals



0



0



10



0



0



0



0



0



First to Last seen



-



-



26 - 92



-



-



-



-



-



Skin, Lesion, Dorsal Cervical



 



 



 



 



 



 



 



 



Number of Animals Affected



0



0



1



0



0



0



0



0



Number of Times Recorded



0



0



9



0



0



0



0



0



% of Affected Animals



0



0



10



0



0



0



0



0



First to Last seen



-



-



12 - 29



-



-



-



-



-



Skin, Scab, Dorsal Cervical



 



 



 



 



 



 



 



 



Number of Animals Affected



0



0



1



0



0



0



0



0



Number of Times Recorded



0



0



8



0



0



0



0



0



Skin, Scab, Dorsal Cervical (Continued...)



 



 



 



 



 



 



 



 



% of Affected Animals



0



0



10



0



0



0



0



0



First to Last seen



-



-



18 - 25



-



-



-



-



-



Ploughing



 



 



 



 



 



 



 



 



Number of Animals Affected



0



0



1



10



0



0



0



12



Number of Times Recorded



0



0



2



33



0



0



0



36



% of Affected Animals



0



0



10



67



0



0



0



80



First to Last seen



-



-



43 - 44



43 - 47



-



-



-



43 - 47



 


Table 30: clinical observations (males & females) - Recovery




















































































































Observation Type: All Types


From Day 92 (Start Date) to 121 (Start Date)



Male


0                  1000


mg/kg/day mg/kg/day Group 1                     Group 4



Female


0                  1000


mg/kg/day mg/kg/day Group 1                     Group 4



Breathing, Labored



 



 



 



 



Number of Animals Affected



0



0



0



1



Number of Times Recorded



0



0



0



1



% of Affected Animals



0



0



0



20



First to Last seen



-



-



-



106 - 106



Breathing, Abnormal Sounds



 



 



 



 



Number of Animals Affected



0



0



0



1



Number of Times Recorded



0



0



0



4



% of Affected Animals



0



0



0



20



First to Last seen



-



-



-



106 - 121



Fur, Erected



 



 



 



 



Number of Animals Affected



0



0



1



0



Number of Times Recorded



0



0



1



0



% of Affected Animals



0



0



20



0



First to Last seen



-



-



121 - 121



-



 

Applicant's summary and conclusion

Conclusions:
A 90-day repeated dose toxicity study was performed according to OECD TG 408 and in accordance with GLP principles. In conclusion, administration of the substance by once daily oral gavage was tolerated in Wistar Han rats at levels up to 1000 mg/kg bw/day. There were no adverse test material related alterations observed.
Based on these results, the No Observed Adverse Effect Level (NOAEL) was considered to be at least 1000 mg/kg bw/day in males and females.
Executive summary:

A 90-day repeated dose toxicity study was performed according to OECD TG 408 and in accordance with GLP principles.


The objectives of this study were to determine the potential toxicity of the substance, when given orally by gavage for 90 days to Wistar Han rats and to evaluate the potential reversibility of any findings. In addition, a No Observed Adverse Effect Level (NOAEL) was evaluated. Ten animals per sex per dose received 0 (vehicle), 100, 300 or 1000 mg/kg bw/day. An additional 5 animals per sex per dose in the recovery group received 0 or 1000 mg/kg bw/day.


Chemical analyses of formulations were conducted on Weeks 1, 6 and 12 on Groups 1-3 to assess accuracy and homogeneity. Formulation analyses confirmed that formulations were prepared accurately and homogenously for all occasions. The following parameters and end points were evaluated in this study: mortality, clinical signs, functional observation tests, body weights, food consumption, ophthalmology, stage of estrous, clinical pathology parameters (hematology, coagulation, and clinical chemistry), macroscopic examination, organ weights and microscopic examination.


No mortality occurred during the study.
At 100 mg/kg bw/day, abnormal breathing sounds, shallow breathing and retching were noted in males and/or females, which were considered to be non-adverse.
At 300 mg/kg bw/day, non-adverse abnormal breathing sounds, deep breathing, retching, hunched posture, erected fur were noted in males and/or females. Histopathological changes consisted of non-adverse squamous cell hyperplasia in the stomach in females, which was often accompanied by hyperkeratosis.
At 1000 mg/kg bw/day, abnormal breathing sounds, labored breathing, shallow breathing, retching, hunched posture and erected fur were noted in males and/or females and were considered to be non-adverse. Non-adverse decreased total movements and ambulations were noted for females. Hematological findings comprised non-adverse decreased red blood cell count and red blood cell distribution width and increased mean corpuscular volume and mean corpuscular hemoglobin in males. Red blood cell count was recovered, while red blood cell distribution width, mean corpuscular volume and mean corpuscular hemoglobin were not recovered at the end of the Recovery Period. It is noted that all reported changes on Hematology parameters were within historical control for this rat strain at the site. Nonadverse clinical chemistry findings consisted of an increase in triglyceride concentration in males, which partly recovered at the end of the Recovery Period. Hormone analysis showed an increased T4 levels in females compared to the concurrent control group however, the mean values still remained within the historical control range and levels recovered at the end of the Recovery Period. Furthermore, no histopathological changes were observed in the thyroid. Overall, the observed increase in the study was considered non-adverse and not toxicologically relevant. Histopathological changes consisted of non-adverse squamous cell hyperplasia often accompanied by hyperkeratosis in the stomach in females and goblet cell hyperplasia of the rectum in a few males. In addition, a higher kidney and liver weight was noted in females. As kidney and liver weights remained within the historical control data and changes were no longer observed after recovery and in the absence of concomitant histopathological changes, the effects on organ weights were considered non-adverse. All histopathological and organ weight changes were fully recovered at the end of the Recovery Period.


No test material-related changes were noted in any of the remaining parameters investigated in this study (i.e., mortality, body weight, food consumption, ophthalmoscopy, coagulation and macroscopic pathology).
In conclusion, administration of the substance by once daily oral gavage was tolerated in Wistar Han rats at levels up to 1000 mg/kg bw/day. There were no adverse test material related alterations observed.
Based on these results, the No Observed Adverse Effect Level (NOAEL) was considered to be at least 1000 mg/kg bw/day in males and females.