Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

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

Diss Factsheets

Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.

REACH

Acetic acid, chloro-, sodium salt, reaction products with 4,5-dihydro-2-undecyl-1H-imidazole-1-ethanol and sodium hydroxide

EC number: 271-794-6 | CAS number: 68608-66-2

Life Cycle description
Uses advised against

Toxicological information

Repeated dose toxicity: oral

Currently viewing:

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 all close all

Test guideline 1

Qualifier:: according to guideline
Guideline:: OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Version / remarks:: June 2018
Deviations:: no

Test guideline 2

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

Test guideline 3

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

Test material information

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 all close all

Doses / concentrations 1

Dose / conc.:: 100 mg/kg bw/day (actual dose received)
Remarks:: group 2 - low dose group

Doses / concentrations 2

Dose / conc.:: 300 mg/kg bw/day (actual dose received)
Remarks:: group 3 - mid dose group

Doses / concentrations 3

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
Sex: Male		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
Sex: Female		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
Sex: Male		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
Sex: Female		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
Sex: Male		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
Sex: Female		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
Sex: Male		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
Sex: Female		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
Sex: Male		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
Sex: Female		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
Sex: Male		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
Sex: Female		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
Sex: Male		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	0.324	37.52	0.1327
	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	0.059	5.28	0.0737
	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	0.316	36.09	0.1350
	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	0.071	7.27	0.1308
	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	0.98	0.96	1.02
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	0.331	41.68	0.1354
	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	0.070	5.78	0.0650
	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	1.02	1.11	1.02
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	36.79	0.0863
	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	0.082	5.52	0.0568
	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	1.08	0.98	0.65

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

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

Sex: Female		Reporting Biochemistry
Sex: Female		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	0.328	150.8	3.76	111.6	2.629	1.533	0.479	24.72	0.1075
	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	0.050	1.0	0.20	1.4	0.082	0.200	0.107	5.03	0.0759
	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	0.306	150.3	3.90	112.0	2.604	1.104**	0.495	30.04	0.0829
	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	0.060	1.2	0.36	1.5	0.078	0.192	0.106	8.18	0.0525
	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	0.93	1.00	1.04	1.00	0.99	0.72	1.03	1.22	0.77
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	0.287	147.2**	3.85	109.5*	2.671	1.299	0.414	24.36	0.0687
	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	0.075	2.6	0.21	2.3	0.192	0.296	0.051	6.66	0.0504
	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	0.88	0.98	1.02	0.98	1.02	0.85	0.86	0.99	0.64
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	0.387	146.8**	3.81	108.5**	2.705	1.406	0.544	33.98*	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	0.084	0.8	0.27	1.4	0.122	0.203	0.115	7.40	0.0400
	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	1.18	0.97	1.01	0.97	1.03	0.92	1.14	1.37	0.88

[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
Sex: Male		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
Sex: Female		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	4615	4448	3870	2739*
	ST.DEV	1238	1689	954	754
	N	5	5	5	5
Ambulations	MEAN	1095	968	1018	66
	ST.DEV	250	382	298	298
	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.

Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.

Registration Dossier

Registration Dossier

Data platform availability banner - registered substances factsheets

Diss Factsheets

Acetic acid, chloro-, sodium salt, reaction products with 4,5-dihydro-2-undecyl-1H-imidazole-1-ethanol and sodium hydroxide

Repeated dose toxicity: oral

Administrative data

Data source

Reference

Materials and methods

Test guidelineopen allclose all

Test guideline 1

Test guideline 2

Test guideline 3

Test material

Test material information

Constituent 1

Test animals

Administration / exposure

Doses / concentrationsopen allclose all

Doses / concentrations 1

Doses / concentrations 2

Doses / concentrations 3

Examinations

Results and discussion

Results of examinations

Effect levels

Target system / organ toxicity

Any other information on results incl. tables

Applicant's summary and conclusion

Test guidelineopen all close all

Doses / concentrationsopen all close all