1,3-dioxolane

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Information not provided in the study report

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1991

Report date:

1991

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Deviations:

yes

Remarks:

Deviations included in "Details on study design"

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

1,3-dioxolane

EC Number:

211-463-5

EC Name:

1,3-dioxolane

Cas Number:

646-06-0

Molecular formula:

C3H6O2

IUPAC Name:

1,3-dioxolane

Test material form:

liquid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch number of test material: Celanese Hoechst corp

Test animals

Species:

rat

Strain:

Sprague-Dawley

Remarks:

Crl:CD (SD) BR

Details on species / strain selection:

The Charles River Crl:CD (SD)BR rat was selected for evaluation of 1,3-Dioxolane because: 1) it is one mammalian species standardly accepted for use in developmental toxicity (embryo-fetal toxicity/teratogenicity) and reproductive toxicity studies; 2) this strain of rat has been demonstrated to be sensitive to developmental toxins; 3) the strain has been widely used throughout industry in nonclinical studies for potential reproductive toxicity; and 4) historical data and experience exist

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories, Inc. Raleigh, North California
- Age at study initiation: minimum of 42 days of age at receipt
- Weight at study initiation: expected to weight from 151 to 175 at receipt
- Fasting period before study: no fasting period described in the study report
- Housing: individual housing
- Diet: Certified Rodent Chow 5002 (Ralston Purina); ad libitum
- Water: local source; ad libitum
- Acclimation period: not specified

DETAILS OF FOOD AND WATER QUALITY: no analyses other than those routinely performed on food by the feed supplier; water is analysed for possible bacterial contamination monthly and for possible chemical,contamination annually.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.1 - 24.4°C
- Humidity (%): 35 - 65%
- Air changes: minimum of 10 changes per hour of 100% fresh air, that has been passed through 99.97% HEPA filters.
- Photoperiod (hrs dark / hrs light): 12 hours dark/12 hours light

IN-LIFE DATES: From: October 31, 1989 To: November 15, 1989

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: The test substance will be added directly to the vehicle in a graduated cylinder. Additional vehicle will be added to the solution to achieve a total volume of 250 mL.

VEHICLE
- Justification for use and choice of vehicle (if other than water): not provided
- Concentration in vehicle: not reported
- Amount of vehicle (if gavage): 5 mL/kg

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

14 days

Frequency of treatment:

Once daily as closely as possible to the same time each day

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 mg/kg bw/day (nominal)

Remarks:

deionized water

Doses / concentrations 2

Dose / conc.:

0 mg/kg bw/day (nominal)

Remarks:

vehicle corn oil

Doses / concentrations 3

Dose / conc.:

75 mg/kg bw/day (nominal)

Doses / concentrations 4

Dose / conc.:

250 mg/kg bw/day (nominal)

Doses / concentrations 5

Dose / conc.:

750 mg/kg bw/day (nominal)

Doses / concentrations 6

Dose / conc.:

2 000 mg/kg bw/day (nominal)

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

other: Reverse osmosis membrane processed deionized water

Details on study design:

- Dose selection rationale: Dosages were selected on the basis of previous evaluations conducted by the Sponsor.
- Rationale for animal assignment: random
- Fasting period before blood sampling for clinical biochemistry: no fasting period described in the study report

Deviations from the OECD TG 407 (1995):
- Justification for use of a 14-day exposure period not provided
- The females should be nulliparous and non-pregnant but this information was not reported
- No measurement of blood clotting time/potential reported
- No clinical biochemistry determinations performed
- All organs listed in the OECD TG 407 (1995) were weighted with the exception of the brain and the heart
- The following organs were not subjected to full histopathological examination on all animals in the control and high dose groups or were not examined at all:
o Not examined on each animal: urinary bladder, trachea, stomach, heart
o Not examined at all: small and large intestines, thyroid, lymph nodes, peripheral nerve, bone marrow, brain, spinal cord
- No sensory reactivity to stimuli of different types was examined. However, the OECD TG 407 indicates that it should be performed in the fourth exposure week.

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: Not specified

BODY WEIGHT: Yes
- Time schedule for examinations: daily during the dosage period and on the day of scheduled sacrifice

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Time schedule for examinations: weekly

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: approximately 24 hours after the last dosage
- Anaesthetic used for blood collection: Yes (identity of the anaesthetic not provided)
- Animals fasted: No
- How many animals: 10 male and ten female rats from each group (surviving animals)
- Parameters checked: red blood cell count, hematocrit, hemoglobin, mean corpuscular volume, white blood cell count, white blood cell differential count, red blood indices for mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration, reticulocyte counts (absolute), red blood cell morphological assessment and platelet counts (absolute)

CLINICAL CHEMISTRY: No

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes; examined for gross lesions of the thoracic, abdominal and pelvic viscera. Organ weights were recorded for the liver, kidneys, adrenals, spleen, pancreas, lungs, pituitary, thymus, as well as the reproductive organs (male rats - testes, epididymides and accessory sex glands; female rats - ovaries and uterus)

HISTOPATHOLOGY: Yes

Statistics:

All hematology data were statistically compared to both control groups using a one-way analysis of variance test and then Dunnett's test, to determine the statistical significance of the individual groups from each of the control groups.

All body weight, feed consumption and organ weights data were statistically compared against both control groups using the following statistical tests: analysis of variance, Dunnett's test, Bartlett's test of homogeneity, Kruskal-Wallis test and Dunn's test.

Results and discussion

Results of examinations

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

In males, clinical observations attributable to the 2000 mg/kg/day dosage of the test substance included hypotonia, excess salivation, ataxia, decreased motor activity, rales, impaired and lost righting reflex, labored breathing, red penile exudate and urine-stained abdominal fur.
In females, clinical observations attributable to the 750 and 2000 mg/kg/day dosages of the test substance included excess salivation (750 and 2000 mg/kg/day), hypotonia, ataxia, decreased motor activity, impaired and lost righting reflex, gasping, bradypnea, oral exudate, rales, lacrimation and urine-stained abdominal fur (2000 mg/kg/day).

Mortality:

mortality observed, treatment-related

Description (incidence):

Three high dosage group male rats were found dead on days 4, 9 and 14 of the study. Deaths were preceded by clinical observations, and two of these rats had severe weight losses. Necropsy of one rat revealed gastric erosions. No other deaths occurred during the conduct of the study.
Four 2000 mg/kg/day dosage group female rats died. These deaths occurred on days 3, 8, 9 and 10 of the study and were preceded by clinical observations. These rats had moderate, although sometimes transient, weight losses occur, and necropsy of one rat revealed reddened areas in the stomach. Two additional high dosage group deaths occurred on days 10 and 13 of the study; these deaths were related to intubation accidents. No other deaths occurred during the conduct of the study.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Males treated with 250, 750 and 2000 mg/kg/day showed reduced body weight gains, as compared with both control group values. The 250 and 750 mg/kg/day dosage groups gained only 88.5% and 76.97%, respectively, of the corn oil control group value, and the 2000 mg/kg/day dosage group had weight loss (18.3 g) on days 1 to 15 of the study. Average body weights were significantly (P≤0.05 to P≤0.01) reduced only for the 2000 mg/kg/day dosage group for days 5 to 15 of the dosage period, when compared to the corn oil control group. Mean final body weight for the other dosage groups were similar to controls.
Females treated with 250 mg/kg/day and higher showed reduced body weight gains during the first week of intubation. During this period, body weight gains of the 75, 250 and 750 mg/kg/day dosage groups were 133.7%, 60.5% and 48.8% of the corn oil control group value, and the 2000 mg/kg/day dosage group had a 10.1 g weight loss. Only the 2000 mg/kg/day dosage group had reduced body weight gains for the entire dosage period; body weight gain for this group was 12.5% of the corn oil control group value on days 1 to 15 of the study. The average body weight was significantly (P≤0.05 to P≤0.01) reduced for the 2000 mg/kg/day dosage group on days 3, 4, 5, 6 and 9 of the dosage period, when compared to the corn oil control group.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

Groups of males given 250 mg/kg/day and higher dosages of the test substance had marginally to reduced feed consumption values, as compared with the corn oil control group values. In the 75, 250, 750 and 2000 mg/kg/day dosage groups respectively, absolute feed consumption values averaged 100%, 94.7%, 88.5% and 60.2%.
Groups of females given 250 mg/kg/day and higher dosages of the test substance had marginally reduced feed consumption values, as compared with the corn oil control group values. In the 75, 250, 750 and 2000 mg/kg/day dosage groups, respectively, absolute feed consumption values averaged 102.0%, 96.6%, 96.6% and 84.2% of the corn oil control group value on days 1 to 15 of the study.

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

In males, hematologic evaluations indicated dosage-dependent decreases in platelet counts. These decreases were statistically significant (P≤0.01) for the 250, 750 and 2000 mg/kg/day dosage groups as compared with the water control group value. Significant decreases (P≤0.01) for platelet counts also occurred for the 750 and 2000 mg/kg/day dosage groups, as compared with the corn oil control group value.
In females, hematologic evaluation indicated that the high dosage (2000 mg/kg/day) of the test substance significantly reduced (P≤0.01) platelets, as compared to values for either control group.
Significant decreases (P≤0.05 to P≤0.01) in lymphocyte counts occurred for the 250, 750 and 2000 mg/kg/day dosage groups, as compared with the corn oil control group value. Reticulocyte count was significantly decreased (P≤0.01) for the 250 mg/kg/day dosage group as compared with the corn oil control group value. These observations may be effects of the test substance, although the values were not dosage-dependent.

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

Reflecting the decrease in body weight, the relative weights of the epididymides, testes and seminal vesicles (with and without fluid) were increased for the 2000 mg/kg/day dosage group, as compared with both control group values.
The 750 and 2000 mg/kg/day dosage groups had increased relative liver and lung weights and a reduced relative thymus weight, as compared with the values for both control groups. The 2000 mg/kg/day dosage group also had an increased relative kidney weight and a reduced relative spleen weight, as compared with both control group values, as well as significant changes (P≤0.05 to P≤0.01) in the relative weights of the liver, spleen, lungs and thymus, as compared with the water control group values. These relative organ weight changes were interrelated with the reduced weights of the 750 and 2000 mg/kg/day dosage groups and the relative conservation and/or possible increase in a specific organ weight when exposed to a toxicant and in conditions of weight loss. There were no dosage-dependent differences in the absolute or relative weights for the pituitary, adrenals or pancreas, as compared with values for either control group.
There were no differences among the groups in the average weights of the ovaries or the nongravid uterus.
Groups of females given 750 to 2000 mg/kg/day dosages of the test substance had dosage-dependent increased relative liver weights and reduced relative spleen and thymus weights. The 2000 mg/kg/day dosage group also had increased relative kidney weights and a reduced relative pancreas weight, as compared to both control group values. Statistically significant changes (P≤0.05 to P≤0.01) occurred in the relative liver weights for the 75, 750 and 2000 mg/kg/day dosage groups, for the relative spleen weights for the 750 and 2000 mg/kg/day dosage groups, and for the relative thymus weights for the 2000 mg/kg/day dosage group, as compared with the water control group values. Differences in the 75 mg/kg/day dosage group were not considered biologically important, because the averages were comparable to the corn oil control group. The observations for the spleen and thymus were possibly the result of specific sensitivity of these organs to the test substance, in combination with reduced body weight gains that were caused by the test substance; the observations for the pancreas and kidney were interrelated with the reduced body weight of the high dosage group.

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

No treatment-related microscopic changes were observed in any of the male rats given 75, 250 or 750 mg/kg/day of 1,3-Dioxolane. Treatment-related microscopic changes were observed in the liver, kidneys, thymus, and testes of rats given 2000 mg/kg/day of 1,3-Dioxolane. The treatment-related changes consisted of centrilobular hepatocellular hypertrophy and midzonal hepatocellular vacuolation, thymic atrophy, renal cortical tubular basophilia and dilatation and accumulations of birefringent intratubular crystals, subacute renal pyelitis and multifocal testicular degeneration.
No treatment-related microscopic changes were observed in any of the female rats given 75, 250, or 750 mg/kg/day of 1,3-Dioxolane. Treatment related microscopic changes were observed in the liver, kidneys and thymus of female rats given the 2000 mg/kg/day dosage of 1,3-Dioxolane. The treatment related changes consisted of centrilobular hepatocellular hypertrophy and midzonal hepatocellular vacuolation, thymic atrophy and subacute renal pyelitis.

Effect levels

Dose descriptor:

NOAEL

Effect level:

75 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

clinical signs

food consumption and compound intake

gross pathology

haematology

histopathology: non-neoplastic

mortality

organ weights and organ / body weight ratios

Remarks on result:

other: The study report states that the NOAEL for females is 250 mg/kg/day. However, due to significant observation in this treated group, a NOAEL of 75 for both males and females was defined.

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

Table 1 : Clinical observations - males

Dosage Group Dosage (mg/kg bw/day)	I (Water) 0	II (Corn oil) 0	III 75	IV 250	V 750	VI 2000
Maximum possible incidence (a)	150/10	150/10	150/10	150/10	150/10	132/10
Found dead	0	0	0	0	0	3
Hypotonia	0/0	0/0	0/0	0/0	0/0	11/8 (b,c)
Excess salivation	0/0	0/0	1/1	0/0	1/1	16/7 (b,d)
Ataxia	0/0	0/0	0/0	0/0	0/0	12/6 (b,c)
Decreased motor activity	0/0	0/0	0/0	0/0	0/0	10/6 (b,c)
Rales	0/0	0/0	0/0	0/0	0/0	7/5 (b,e)
Impaired righting reflex	0/0	0/0	0/0	0/0	0/0	4/3 (b)
Loss of righting reflex	0/0	0/0	0/0	0/0	0/0	3/3 (c)
Labored breathing	0/0	0/0	0/0	0/0	0/0	4/2
Red exudate: penis	0/0	0/0	0/0	0/0	0/0	2/1
Urine-stained abdominal fur	0/0	0/0	0/0	0/0	0/0	1/1
Chromorhinorrhea	1/1	1/1	3/2	0/0	0/0	1/1 (b)
Chromodacryorrhea	7/1	4/1	5/1	0/0	0/0	0/0
Incisor(s): misaligned, broken and/or missing	7/1	10/1	3/1	0/0	0/0	0/0
Lesion (on the back)	1/1	0/0	0/0	0/0	0/0	0/0

(a) Maximum possible incidence = (days x rats)/number of rats examined per group

(b) Occurred in rat that was found dead on day 9 of study

(c) Occurred in rat that was found dead on day 4 of study

(d) Occurred in rat that was found dead on day 14 of study

Table 2: Clinical observations – females

Dosage Group Dosage (mg/kg bw/day)	I (Water) 0	II (Corn oil) 0	III 75	IV 250	V 750	VI 2000
Maximum possible incidence (a)	150/10	150/10	150/10	150/10	150/10	113/10
Found dead	0	0	0	0	0	6
Excess salivation	0/0	3/1	1/1	1/1	6/3	22/10 (b, c, d, e, f, g)
Hypotonia	0/0	0/0	0/0	0/0	0/0	14/8 (b, d, f, g)
Ataxia	0/0	0/0	0/0	0/0	0/0	13/8 (b, e, f, g)
Decreased motor activity	0/0	0/0	0/0	0/0	0/0	6/5 (d,f)
Impaired righting reflex	0/0	0/0	0/0	0/0	0/0	4/4 (d)
Loss of righting reflex	0/0	0/0	0/0	0/0	0/0	4/4 (f)
Gasping	0/0	0/0	0/0	0/0	0/0	4/3 (f,g)
Bradypnea	0/0	0/0	0/0	0/0	0/0	2/2 (d, g)
Oral exudate	0/0	0/0	0/0	0/0	0/0	3/2 (f)
Rales	0/0	0/0	0/0	0/0	0/0	2/2 (g)
Lacrimation	0/0	0/0	0/0	0/0	0/0	1/1 (d)
Urine-stained abdominal fur	0/0	0/0	0/0	0/0	0/0	1/1 (d)
Urine: dark yellow	0/0	0/0	0/0	0/0	0/0	1/1 (d)
Alopecia	0/0	0/0	0/0	0/0	0/0	5/1 (c)

(a) Maximum possible incidence = (days x rats)/number of rats examined per group

(b) Occurred in rat that was found dead on day 13 of study; death was attributed to an intubation injury

(c) Occurred in rat that was found dead on day 8 of study

(d) Occurred in rat that was found dead on day 10 of study; death was attributed to an intubation injury

(e) Occurred in rat that was found dead on day 10 of study

(f) Occurred in rat that was found dead on day 3 of study

(g) Occurred in rat that was found dead on day 9 of study

Table 3: Necropsy observations – male rats

Dosage Group Dosage (mg/kg bw/day)		I (Water) 0	II (Corn oil) 0	III 75	IV 250	V 750	VI 2000
Rats examined	N	10	10	10	10	10	10
Found dead	N	0	0	0	0	0	3
Kidneys: dilated pelvis	N	0	0	0	1	0	0
Urinary bladder; contained calculi	N	0	0	0	0	0	1
Stomach: erosions	N	0	0	0	0	0	1

 

Table 4: Necropsy observations – female rats

Dosage Group Dosage (mg/kg bw/day)		I (Water) 0	II (Corn oil) 0	III 75	IV 250	V 750	VI 2000
Rats examined	N	10	10	10	10	10	10
Found dead	N	0	0	0	0	0	6
Uterus: dilated	N	0	0	0	0	1	0
Stomach: reddened arena	N	0	0	0	0	0	1
Lungs: congestion	N	0	0	0	0	0	1
Pericardium: contained material	N	0	0	0	0	0	1
Esophagus: hole	N	0	0	0	0	0	2

 

Table 5: Body weight – male rats

Dosage Group Dosage (mg/kg bw/day)		I (Water) 0	II (Corn oil) 0	III 75	IV 250	V 750	VI 2000
Rats tested	N	10	10	10	10	10	10
Body weight (g)
Day 1	Mean ± SD	341.6 ± 16.6	338.2 ± 27.7	336.6 ± 21.8	341.6 ± 23.1	342.1 ± 21.4	346.4 ± 18.4
Day 2	Mean ± SD	350.7 ± 15.6	344.4 ± 29.8	339.0 ± 28.5	341.1 ± 27.3	343.3 ± 19.5	346.1 ± 16.8
Day 3	Mean ± SD	352.0 ± 15.9	348.3 ± 33.4	341.8 ± 33.6	342.4 ± 32.9	345.2 ± 20.4	335.4 ± 21.6
Day 4	Mean ± SD	358.2 ± 17.0	351.9 ± 39.3	345.4 ± 39.5	352.8 ± 26.7	351.1 ± 22.7	330.8 ± 26.2
Day 5	Mean ± SD	363.7 ± 18.6	355.6 ± 42.9	357.8 ± 28.3	358.0 ± 26.2	354.3 ± 21.8	324.4 ± 32.8*
Day 6	Mean ± SD	367.6 ± 18.3	357.9 ± 48.4	364.7 ± 25.8	362.6 ± 26.3	358.2 ± 23.8	322.7 ± 30.6**
Day 7	Mean ± SD	365.3 ± 29.3	361.2 ± 39.7	355.2 ± 32.4	363.6 ± 28.9	359.7 ± 24.0	316.9 ± 21.7**
Day 8	Mean ± SD	371.1 ± 22.5	366.7 ± 38.2	364.1 ± 29.8	368.3 ± 28.6	362.6 ± 22.4	316.6 ± 22.3**
Day 9	Mean ± SD	378.8 ± 23.8	373.4 ± 38.9	372.3 ± 30.4	375.2 ± 28.9	369.7 ± 24.3	324.4 ± 22.8**
Day 10	Mean ± SD	380.8 ± 21.8	376.9 ± 40.0	377.0 ± 31.2	377.4 ± 28.2	372.7 ± 24.1	326.2 ± 27.0**
Day 11	Mean ± SD	382.9 ± 20.7	382.3 ± 38.5	382.0 ± 32.5	381.1 ± 30.2	376.9 ± 24.5	320.4 ± 34.0**
Day 12	Mean ± SD	388.5 ± 21.9	388.2 ± 38.1	387.7 ± 31.4	386.6 ± 30.5	380.4 ± 25.2	317.9 ± 32.9**
Day 13	Mean ± SD	390.7 ± 21.8	394.5 ± 42.7	392.7 ± 31.6	388.9 ± 30.6	384.2 ± 26.9	314.5 ± 33.8**
Day 14	Mean ± SD	396.4 ± 20.4	396.8 ± 41.8	396.2 ± 31.3	393.5 ± 31.2	387.6 ± 28.2	319.0 ± 33.3**
Day 15	Mean ± SD	404.9 ± 20.5	405.2 ± 42.8	403.2 ± 30.7	400.9 ± 32.8	393.6 ± 28.2	323.1 ± 36.8**

 

Table 6: Body weight – female rats

Dosage Group Dosage (mg/kg bw/day)		I (Water) 0	II (Corn oil) 0	III 75	IV 250	V 750	VI 2000
Rats tested	N	10	10	10	10	10	10
Body weight (g)
Day 1	Mean ± SD	223.0 ± 11.4	225.8 ± 14.8	223.3 ± 13.6	226.5 ± 9.7	223.9 ± 15.2	224.8 ± 10.2
Day 2	Mean ± SD	221.2 ± 16.0	224.6 ± 14.0	223.7 ± 15.2	222.8 ± 9.4	228.4 ± 7.7	212.7 ± 15.6
Day 3	Mean ± SD	224.9 ± 11.3	227.9 ± 14.5	228.2 ± 14.2	224.8 ± 9.9	226.0 ± 9.0	211.1 ± 15.4*
Day 4	Mean ± SD	226.9 ± 10.5	232.1 ± 15.2	230.0 ± 15.6	227.6 ± 10.7	225.6 ± 9.3	211.7 ± 15.0**
Day 5	Mean ± SD	229.6 ± 11.5	231.7 ± 17.9	229.4 ± 14.6	230.0 ± 11.1	227.9 ± 10.0	212.7 ± 16.5*
Day 6	Mean ± SD	229.3 ± 13.1	229.7 ± 18.8	228.1 ± 15.0	226.3 ± 11.0	229.3 ± 11.7	214.4 ± 16.2
Day 7	Mean ± SD	232.6 ± 12.5	233.0 ± 18.0	230.4 ± 15.8	229.8 ± 12.2	227.8 ± 9.8	215.2 ± 13.6
Day 8	Mean ± SD	233.3 ± 11.6	234.4 ± 18.5	234.8 ± 18.5	231.7 ± 12.7	228.1 ± 9.4	212.0 ± 12.1**
Day 9	Mean ± SD	231.2 ± 13.3	234.9 ± 16.8	234.6 ± 17.2	230.7 ± 12.4	231.0 ± 12.6	212.2 ± 11.3**
Day 10	Mean ± SD	233.3 ± 15. 1	233.3 ± 17.7	231.8 ± 19.1	228.8 ± 12.8	232.3 ± 14.8	212.3 ± 14.0
Day 11	Mean ± SD	235.4 ± 13.3	233.4 ± 18.5	234.2 ± 19.4	232.9 ± 13.4	232.7 ± 14.8	211.6 ± 19.2
Day 12	Mean ± SD	237.6 ± 12.8	236.8 ± 17.5	236.7 ± 18.6	234.3 ± 14.6	234.2 ± 13.7	214.4 ± 20.0
Day 13	Mean ± SD	237.3 ± 11.6	240.3 ± 16.4	238.5 ± 18.9	235.8 ± 14.6	238.6 ± 11.0	218.4 ± 18.3
Day 14	Mean ± SD	238.8 ± 12.9	242.3 ± 18.2	239.7 ± 18.1	234.9 ± 13.7	239.4 ± 10.2	226.8 ± 11.1
Day 15	Mean ± SD	242.8 ± 16.8	245.8 ± 20.2	244.3 ± 19.1	242.5 ± 14.7	244.1 ± 11.8	229.5 ± 7.6

 

Table 7: Body weight data – male rats

Dosage Group Dosage (mg/kg bw/day)		I (Water) 0	II (Corn oil) 0	III 75	IV 250	V 750	VI 2000
Rats tested	N	10	10	10	10	10	10
Body weight change (g)
Days 1-8	Mean ± SD	+29.5 ± 20.3	+28.5 ± 13.6	+27.5 ± 14.2	+26.7 ± 13.5	+20.5 ± 10.1	-29.0 ± 29.6
Days 8-15	Mean ± SD	+33.8 ± 14.6	+38.5 ± 12.9	+37.4 ± 9.2	+32.6 ± 8.6	+31.0 ± 10.3	+9.3 ± 22.4
Days 1-15	Mean ± SD	+63.3 ± 23.9	+67.0 ± 18.1	+66.2 ± 12.1	+59.3 ± 16.7	+51.5 ± 14.2	-18.3 ± 37.6

 

Table 8: Body weight data – female rats

Dosage Group Dosage (mg/kg bw/day)		I (Water) 0	II (Corn oil) 0	III 75	IV 250	V 750	VI 2000
Rats tested	N	10	10	10	10	10	10
Body weight change (g)
Days 1-8	Mean ± SD	+11.0 ± 7.5	+8.6 ± 8.3	+11.5 ± 10.5	+5.2 ± 3.8	+4.2 ± 8.1	-10.1 ± 6.5
Days 8-15	Mean ± SD	+12.6 ± 6.5	+11.4 ± 9.5	+9.5 ± 3.5	+10.8 ± 5.1	+16.0 ± 5.0	+11.0 ± 11.0
Days 1-15	Mean ± SD	+19.8 ± 16.0	+20.0 ± 9.5	+21.0 ± 10.5	+16.0 ± 6.1	+20.2 ± 9.4	+2.5 ± 7.0

 

Table 9: Feed consumption – male rats

Dosage Group Dosage (mg/kg bw/day)		I (Water) 0	II (Corn oil) 0	III 75	IV 250	V 750	VI 2000
Rats tested	N	10	10	10	10	10	10
Feed consumption (g/day)
Days 1-8	Mean ± SD	26.7 ± 2.8	23.8 ± 2.6	22.9 ± 36	22.2 ± 3.6	20.8 ± 1.7	13.1 ± 4.6
Days 8-15	Mean ± SD	25.2 ± 32	22.3 ± 3.3	23.0 ± 2.8	21.1 ± 2.4	19.4 ± 2.3	13.5 ± 3.9
Days 1-15	Mean ± SD	25.7 ± 2.4	22.6 ± 2.7	22.6 ± 2.6	21.4 ± 2.7	20.0 ± 1.8	13.6 ± 3.4
Feed consumption (g/kg/day)
Days 1-8	Mean ± SD	74.2 ± 7.5	65.9 ± 2.0	63.8 ± 6.9	62.2 ± 6.8	59.0 ± 2.4	39.5 ± 13.0
Days 8-15	Mean ± SD	65.1 ± 9.5	57.9 ± 9.1	60.0 ± 7.4	54.8 ± 5.8	51.4 ± 4.9	41.8 ± 9.4
Days 1-15	Mean ± SD	68.4 ± 5.7	59.6 ± 1.9	60.2 ± 2.7	57.5 ± 4.0	54.4 ± 2.5	41.2 ± 8.5

 

Table 10: Feed consumption – female rats

Dosage Group Dosage (mg/kg bw/day)		I (Water) 0	II (Corn oil) 0	III 75	IV 250	V 750	VI 2000
Rats tested	N	10	10	10	10	10	10
Feed consumption (g/day)
Days 1-8	Mean ± SD	16.7 ± 3.6	14.9 ± 1.1	15.2 ± 2.0	14.3 ± 1.3	13.7 ± 1.1	11.3 ± 1.7
Days 8-15	Mean ± SD	16.3 ± 1.8	14.3 ± 1.6	14.7 ± 1.8	13.8 ± 1.2	14.5 ± 1.1	13.4 ± 2.6
Days 1-15	Mean ± SD	6.7 ± 1.8	14.6 ± 1.5	14.9 ± 1.9	14.1 ± 1.2	14.1 ± 1.0	12.3 ± 1.6
Feed consumption (g/kg/day)
Days 1-8	Mean ± SD	73.6 ± 16.7	64.4 ± 6.4	66.1 ± 7.2	62.5 ± 3.9	60.4 ± 4.1	51.6 ± 7.3
Days 8-15	Mean ± SD	68.6 ± 4.2	59.6 ± 5.4	61.4 ± 7.3	58.3 ± 2.5	61.6 ± 3.5	59.8 ± 12.2
Days 1-15	Mean ± SD	71.4 ± 8.2	62.0 ± 4.4	63.7 ± 6.9	60.3 ± 2.6	60.8 ± 3.5	54.6 ± 7.6

 

Table 11: Final body weights and organ weights at necropsy – male rats

Dosage Group Dosage (mg/kg bw/day)		I (Water) 0	II (Corn oil) 0	III 75	IV 250	V 750	VI 2000
Rats tested	N	10	10	10	10	10	10
Final body weight	Mean ± SD	404.9 ± 20.5	405.2 ± 42.8	403.2 ± 30.7	400.9 ± 32.8	393.6 ± 28.2	323.1 ± 36.8**
Epididymis left	Mean ± SD	0.5314 ± 0.0663	0.5323 ± 0.0663	0.5567 ± 0.0614	0.5826 ± 0.0774	0.5803 ± 0.0550	0.5109 ± 0.0932
Epididymis right	Mean ± SD	0.5243 ± 0.1124	0.5566 ± 0.0651	0.5602 ± 0.0742	0.5884 ± 0.0490	0.5963 ± 0.0613	0.5455 ± 0.0676
Testis left	Mean ± SD	1.5996 ± 0.1570	1.5265 ± 0.1880	1.6815 ± 0.1698	1.6972 ± 0.1094	1.6714 ± 0.1398	1.4368 ± 0.2129
Testis right	Mean ± SD	1.6344 ± 0.1600	1.5508 ± 0.1700	1.6753 ± 0.1873	1.6973 ± 0.1295	1.6774 ± 0.1362	1.4664 ± 0.1507
Prostate	Mean ± SD	1.0849 ± 0.2557	1.1511 ± 0.1390	1.0752 ± 0.1870	1.0535 ± 0.1528	1.0208 ± 0.1289	0.8415 ± 0.1335**
Seminal vesicles with fluid	Mean ± SD	1.558 ± 0.3908	1.2765 ± 0.3371	1.2674 ± 0.2580	1.4560 ± 0.2274	1.5105 ± 0.2953	1.5221 ± 0.2922
Seminal vesicles without fluid	Mean ± SD	0.8030 ± 0.1700	0.7099 ± 0.1055	0.7470 ± 0.1056	0.7015 ± 0.1088	0.8035 ± 0.1598	0.6953 ± 0.1656

 

Table 11 (cont): Final body weights and organ weights at necropsy – male rats

Dosage Group Dosage (mg/kg bw/day)		I (Water) 0	II (Corn oil) 0	III 75	IV 250	V 750	VI 2000
Rats tested	N	10	10	10	10	10	10
Final body weight	Mean ± SD	404.9 ± 20.5	405.2 ± 42.8	403.2 ± 30.7	400.9 ± 32.8	393.6 ± 28.2	323.1 ± 36.8**
Pituitary	Mean ± SD	0.0145 ± 0.0043	0.0115 ± 0.0048	0.0145 ± 0.0064	0.0098 ± 0.0042	0.0100 ± 0.0053	0.0090 ± 0.0045
Liver	Mean ± SD	16.33 ± 1.03	15.90 ± 2.60	15.63 ± 2.74	16.34 ± 2.56	17.15 ± 2.16	16.51 ± 2.65
Kidneys	Mean ± SD	3.35 ± 0.23	3.41 ± 0.50	3.24 ± 0.41	3.34 ± 0.34	3.32 ± 0.24	2.96 ± 0.45
Adrenals	Mean ± SD	0.0684 ± 0.0099	0.0637 ± 0.0101	0.0756 ± 0.0179	0.0601 ± 0.0153	0.0669 ± 0.0099	0.0652 ± 0.0161
Spleen	Mean ± SD	0.8362 ± 0.1533	0.7679 ± 0.0947	0.7733 ± 0.1460	0.7320 ± 0.0994	0.7035 ± 0.0977*	0.5265 ± 0.1200**
Lungs	Mean ± SD	1.91 ± 0.18	2.09 ± 0.41	1.85 ± 0.34	1.97 ± 0.23	2.25 ± 0.60	1.90 ± 0.31
Pancreas	Mean ± SD	0.8303 ± 0.2044	0.6409 ± 0.1783	0.7591 ± 0.1654	07057 ± 0.2089	0.5373 ± 0.1779*	0.6033 ± 0.2169*
Thymus	Mean ± SD	0.6110 ± 0.1009	0.5798 ± 0.1350	0.6898 ± 0.2072	0.5628 ± 0.1467	0.5163 ± 0.1535	0.2951 ± 0.1300**

 

Table 12: Ratio (%) of organ weights to final body weight at necropsy – male rats

Dosage Group Dosage (mg/kg bw/day)		I (Water) 0	II (Corn oil) 0	III 75	IV 250	V 750	VI 2000
Rats tested	N	10	10	10	10	10	10
Final body weight	Mean ± SD	404.9 ± 20.5	405.2 ± 42.8	403.2 ± 30.7	400.9 ± 32.8	393.6 ± 28.2	323.1 ± 36.8**
Epididymis left	Mean ± SD	0.13 ± 0.02	0.13 ± 0.02	0.11 ± 0.02	0.15 ± 0.02	0.15 ± 0.02	0.16 ± 0.04
Epididymis right	Mean ± SD	0.13 ± 0.03	0.14 ± 0.02	0.15 ± 0.02	0.15 ± 0.02	0.15 ± 0.02	0.17 ± 0.02**
Testis left	Mean ± SD	0.40 ± 0.04	0.38 ± 0.05	0.42 ± 0.03	0.43 ± 0.04	0.43 ± 0.04	0.45 ± 0.10
Testis right	Mean ± SD	0.40 ± 0.05	0.38 ± 0.05	0.42 ± 0.03	0.42 ± 0.04	0.43 ± 0.04	0.46 ± 0.09
Prostate	Mean ± SD	0.27 ± 0.06	0.28 ± 0.04	0.27 ± 0.03	0.26 ± 0.04	0.26 ± 0.04	0.26 ± 0.04
Seminal vesicles with fluid	Mean ± SD	0.38 ± 0.08	0.32 ± 0.08	0.31 ± 0.08	0.36 ± 0.05	0.38 ± 0.08	0.47 ± 0.10**
Seminal vesicles without fluid	Mean ± SD	0.20 ± 0.04	0.17 ± 0.02	0.19 ± 0.03	0.18 ± 0.02	0.20 ± 0.04	0.22 ± 0.05*

 

Table 12 (cont): Ratio (%) of organ weights to final body weight at necropsy – male rats

Dosage Group Dosage (mg/kg bw/day)		I (Water) 0	II (Corn oil) 0	III 75	IV 250	V 750	VI 2000
Rats tested	N	10	10	10	10	10	10
Final body weight	Mean ± SD	404.9 ± 20.5	405.2 ± 42.8	403.2 ± 30.7	400.9 ± 32.8	393.6 ± 28.2	323.1 ± 36.8**
Pituitary	Mean ± SD	3.58 ± 1.14	2.82 ± 106	3.52 ± 1.69	2.46 ± 1.09	2.58 ± 1.42	2.74 ± 1.26
Liver	Mean ± SD	4.04 ± 0.24	3.90 ± 0.34	4.03 ± 0.35	4.05 ± 0.36	4.34 ± 0.29*	5.09 ± 0.38**
Kidneys	Mean ± SD	0.83 ± 0.07	0.84 ± 0.10	0.81 ± 0.08	0.83 ± 0.05	0.84 ± 0.05	0.92 ± 0.17
Adrenals	Mean ± SD	0.02 ± 0.00	0.02 ± 0.00	0.02 ± 0.00	0.02 ± 0.00	0.02 ± 0.00	0.02 ± 0.01
Spleen	Mean ± SD	0.02 ± 0.03	0.19 ± 0.03	0.20 ± 0.02	0.18 ± 0.02	0.18 ± 0.02	0.16 ± 0.03**
Lungs	Mean ± SD	0.47 ± 0.04	0.51 ± 0.07	0.48 ± 0.08	0.49 ± 0.05	0.58 ± 0.16	0.59 ± 0.07*
Pancreas	Mean ± SD	0.20 ± 0.05	0.16 ± 0.04	0.19 ± 0.03	0.18 ± 0.06	0.13 ± 0.04**	0.18 ± 0.05
Thymus	Mean ± SD	0.15 ± 0.02	0.14 ± 0.06	0.18 ± 0.04	0.14 ± 0.03	0.13 ± 0.04	0.09 ± 0.03**

 

Table 13: Final body weights and organ weights at necropsy – female rats

Dosage Group Dosage (mg/kg bw/day)		I (Water) 0	II (Corn oil) 0	III 75	IV 250	V 750	VI 2000
Rats tested	N	10	10	10	10	10	10
Final body weight	Mean ± SD	242.8 ± 16.8	245.8 ± 20.2	244.3 ± 19.1	242.5 ± 14.7	244.1 ± 11.8	229.5 ± 7.6
Pituitary	Mean ± SD	0.0149 ± 0.0038	0.0126 ± 0.0034	0.0163 ± 0.0029	0.0142 ± 0.0038	0.0182 ± 0.0030**	0.0130 ± 0.0038
Liver	Mean ± SD	9.53 ± 1.39	10.01 ± 1.43	10.54 ± 1.56	10.40 ± 1.41	10.51 ± 1.07	10.53 ± 0.62
Kidneys	Mean ± SD	2.00 ± 0.15	2.01 ± 0.33	1.97 ± 0.28	1.92 ± 0.22	1.90 ± 0.15	2.08 ± 0.21
Adrenals	Mean ± SD	0.0852 ± 0.0197	0.0829 ± 0.0136	0.0789 ± 0.0099	0.0753 ± 0.0195	0.0756 ± 0.0126	0.0840 ± 0.0107
Spleen	Mean ± SD	0.5578 ± 0.0780	0.5598 ± 0.1093	0.5195 ± 0.0784	0.5081 ± 0.0587	0.4702 ± 0.0469*	0.3613 ± 0.0502**
Lungs	Mean ± SD	1.54 ± 0.13	1.53 ± 0.31	1.58 ± 0.44	1.57 ± 0.30	0.150 ± 0.19	1.41 ± 0.20
Pancreas	Mean ± SD	0.4932 ± 0.2	0.5888 ± 0.1885	0.7065 ± 0.1650	0.5454 ± 0.2382	0.5373 ± 0.1243	0.4186 ± 0.1986
Thymus	Mean ± SD	0.5083 ± 0.0858	0.4824 ± 0.1021	0.4709 ± 0.1493	0.4665 ±0.0895	0.4178 ± 0.0700	0.3114 ±0.0762**
Ovary left	Mean ± SD	0.0623 ± 0.0216	0.0625 ± 0.0095	0.0560 ± 0.0102	0.0636 ± 0.0152	0.0578 ± 0.0125	0.0643 ± 0.0092
Ovary right	Mean ± SD	0.0724 ± 0.0182	0.0698 ± 0.0156	0.0668 ± 0.0136	0.0606 ± 0.0142	0.0606 ± 0.0150	0.0620 ± 0.0257
Uterus non-gravid	Mean ± SD	0.3878 ± 0.0540	0.6623 ± 0.3116	0.6197 ± 0.2099**	0.4912 ± 0.1488	0.7166 ± 0.1773**	0.5485 ± 0.2408

 

Table 14: Ratio (%) of organ weights to final body weight at necropsy – female rats

Dosage Group Dosage (mg/kg bw/day)		I (Water) 0	II (Corn oil) 0	III 75	IV 250	V 750	VI 2000
Rats tested	N	10	10	10	10	10	10
Final body weight	Mean ± SD	242.8 ± 16.8	245.8 ± 20.2	244.3 ± 19.1	242.5 ± 14.7	244.1 ± 11.8	229.5 ± 7.6
Pituitary	Mean ± SD	6.11 ± 1.43	5.17 ± 1.52	6.69 ± 1.06	5.84 ± 1.54**	7.48 ± 1.33	5.65 ± 1.66
Liver	Mean ± SD	3.91 ± 0.36	4.06 ± 0.31	4.30 ± 0.40*	4.27 ± 0.33	4.31 ± 0.41*	4.58 ± 0.15**
Kidneys	Mean ± SD	0.82 ± 0.05	0.81 ± 0.09	0.81 ± 0.10	0.79 ± 0.07	0.78 ± 0.06	0.90 ± 0.10
Adrenals	Mean ± SD	0.04 ± 0.01	0.03 ± 0.00	0.03 ± 0.00	0.03 ± 0.01	0.03 ± 0.00	0.04 ± 0.00
Spleen	Mean ± SD	0.23 ± 0.02	0.23 ± 0.03	0.21 ± 0.03	0.21 ± 0.02	0.19 ± 0.02**	0.16 ± 0.03**
Lungs	Mean ± SD	0.63 ± 0.05	0.62 ± 0.10	0.64 ± 0.16	0.65 ± 0.13	0.61 ± 0.08	0.62 ± 0.09
Pancreas	Mean ± SD	0.20 ± 0.07	0.24 ± 0.07	0.29 ± 0.07	0.23 ± 0.10	0.22 ± 0.04	0.18 ± 0.08
Thymus	Mean ± SD	0.21 ± 0.03	0.20 ± 0.04	0.19 ± 0.06	0.19 ± 0.03	0.17 ± 0.03	0.13 ± 0.03**
Ovary left	Mean ± SD	0.02 ± 0.01	0.02 ± 0.00	0.02 ± 0.00	0.03 ± 0.01	0.02 ± 0.01	0.03 ± 0.00
Ovary right	Mean ± SD	0.03 ± 0.01	0.03 ± 0.00	0.03 ± 0.00	0.02 ± 0.00	0.02 ± 0.01	0.03 ± 0.01
Uterus non-gravid	Mean ± SD	0.16 ± 0.02	0.27 ± 0.14	0.25 ± 0.08*	0.20 ± 0.07	0.30 ± 0.08**	0.24 ± 0.10

 

Table 15: Evaluation of hematologic data – male rats

Dosage Group Dosage (mg/kg bw/day)		I (Water) 0	II (Corn oil) 0	III 75	IV 250	V 750	VI 2000	Reference range
Rats tested	N	10	10	10	10	10	10
WBC THSN/UL	Mean ± SD	24.1 ± 14.58	15.85 ± 4.33	15.78 ± 4.49	12.63 ± 2.42	13.47 ± 12.17	34.53 ± 38.94	9.9-14.9
RBC MILL/UL	Mean ± SD	6.81 ± 0.674	6.72 ± 0.502	6.58 ± 0.425	6.85 ± 0.348	6.85 ± 0.477	6.06 ± 1.03	6.2-9
HGB GM/DL	Mean ± SD	13.9 ± 0.74	14.61 ± 0.75	14.33 ± 0.89	14.55 ± 0.56	14.63 ± 0.76	13.17 ± 1.3	13.4-16.4
HCT (%)	Mean ± SD	40.53 ± 6.31	40.18 ± 2.2	39.21 ± 2.63	39.81 ± 1.33	40.24 ± 1.98	37.2 ± 4.25	40-49
MCV FL	Mean ± SD	59.26 ± 3.39	59.84 ± 2	59.61 ± 2.28	58.21 ± 2.14	58.96 ± 4.56	61.8 ± 4.34	52-66
MCH PICO GRA	Mean ± SD	20.57 ± 2.11	21.75 ± 0.75	21.79 ± 0.9	21.28 ± 0.76	21.41 ± 1.69	21.9 ± 1.91	17.7-19.1
MCHC (%)	Mean ± SD	34.94 ± 4.96	36.39 ± 0.57	36.59 ± 0.76	36.55 ± 0.44	36.34 ± 0.58	35.43 ± 0.67	32-33.5
PLATELET THSN/UL	Mean ± SD	1130.1 ± 265.6	956.8 ± 92.8	978.2 ± 99.1	844.1 ± 143	698.9 ± 114.3	482.3 ± 109.1	780-1400
BANDS THSN/UL	Mean ± SD	0.08 ± 0.212	0.05 ± 0.095	0.01 ± 0.032	0 ± 0	0.02 ± 0.045	0 ± 0	0-0.06
SEGS THSN/UL	Mean ± SD	3.92 ± 3.519	5.1 ± 3.22	6.82 ± 2.178	4.66 ± 1.051	5.07 ± 4.711	18.08 ± 19.58	0.58-6.3

 

WBC THSN/UL: White blood cells (leukocytes) in thousands per microliter

RBC MILL/UL: Red blood cells (erythrocytes) in millions per microliter

HGB GM/DL: Hemoglobin in grams per deciliter

HCT (%): Hematocrit (Packed Cell Volume) in percent

MCV FL: Mean corpuscular volume, the volume of the average erythrocyte, in femtoliters

MCH PICO GRA: Mean corpuscular hemoglobin, the amount of hemoglobin in the average erythrocyte in picograms

MCHC (%): Mean corpuscular hemoglobin concentration, the proportion of the erythrocyte mass composed of hemoglobin in percent

PLATELET THSN/UL: Platelets in thousands per microliter

BANDS THSN/UL: Band neutrophils (non-segmented neutrophils) in thousands per microliter

SEGS THSN/UL: Segmented neutrophils in thousands per microliter

 

Table 15 (cont): Evaluation of hematologic data – male rats

Dosage Group Dosage (mg/kg bw/day)		I (Water) 0	II (Corn oil) 0	III 75	IV 250	V 750	VI 2000	Reference range
Rats tested	N	10	10	10	10	10	10
LYMPHS THSN/UL	Mean ± SD	19.371 ± 10.385	10.621 ± 1.628	8.88 ± 3.921	7.84 ± 2.586	8.305 ± 7.623	16.457 ± 19.416	3.78-15.32
MONOS THSN/UL	Mean ± SD	0.02 ± 0.063	0 ± 0	0.03 ± 0.055	0.02 ± 0.045	0 ± 0	0 ± 0	0.02-1.2
EOSN THSN/UL	Mean ± SD	0.47 ± 0.578	0.09 ± 0.126	0.04 ± 0.071	0.08 ± 0.105	0.07 ± 0.063	0 ± 0	0-0.31
BASOS THSN/UL	Mean ± SD	/	/	/	/	/	/	0-0.1
ATLYMPH THSN/UL	Mean ± SD	/	/	/	/	/	/	0-0
METAS THSN/UL	Mean ± SD	0.08 ± 0.212	/	/	0.031 ± 0.063	0.08	/	0-0
MYELOS	Mean ± SD	0.16 ± 0.42	/	/	/	/	/	0-0
PROS THSN/UL	Mean ± SD	/	/	/	/	/	/	0-0
BLASTS THSN/UL	Mean ± SD	/	/	/	/	/	/	0-0
NRBC/100 WBC	Mean ± SD	0 ± 0	0.6 ± 0.8	0.9 ± 1.1	1.4 ± 0.7	0.7 ± 0.9	0.1 ± 0.4	0-0
RETIC (%)	Mean ± SD	2.57 ± 0.86	1.59 ± 0.47	1.64 ± 0.52	2.19 ± 1.19	1.24 ± 0.68	0.41 ± 0.55	0.1-4

 

LYMPHS THSN/UL: Lymphocytes in thousands per microliter

MONOS THSN/UL: Monocytes in thousands per microliter

EOSN THSN/UL: Eosinophils in thousands per microliter

BASOS THSN/UL: Basophils in thousands per microliter. No entry indicates none seen

ATLYMPH THSN/UL: Atypical lymphocytes in thousands per microliter. No entry indicates none seen

METAS THSN/UL: Metamyelocytes in thousands per microliter. No entry indicates none seen

MYELOS: Myelocytes in thousands per microliter. No entry indicates none seen

PROS THSN/UL: Promyelocytes in thousands per microliter. No entry indicates none seen

BLASTS THSN/UL: Blast cells in thousands per microliter. No entry indicates none seen

NRBC/100 WBC: Nucleated red blood cells (nucleated erythrocytic precursors) seen per 100 white blood cells

RETIC (%): Reticulocytes expressed a percent of total red blood cells

Table 16: Evaluation of hematologic data – female rats

Dosage Group Dosage (mg/kg bw/day)		I (Water) 0	II (Corn oil) 0	III 75	IV 250	V 750	VI 2000	Reference range
Rats tested	N	10	10	10	10	10	10
WBC THSN/UL	Mean ± SD	17.2 ± 4.12	14.53 ± 3.99	11.75 ± 0.83	11.42 ± 1.9	10.67 ± 3.72	8.9 ± 3.25	9.4-14.9
RBC MILL/UL	Mean ± SD	6.69 ± 0.17	6.79 ± 0.277	6.91 ± 0.226	6.92 ± 0.27	6.73 ± 0.473	7.19 ± 0.318	6.2-9
HGB GM/DL	Mean ± SD	14.14 ± 037	14.34 ± 0.47	14 ± 0.39	14.37 ± 0.48	13.98 ± 0.68	14.35 ± 0.35	13.4-16.4
HCT (%)	Mean ± SD	39.6 ± 1.62	40.26 ± 1.45	39.98 ± 1.16	40.43 ± 1.52	39.65 ± 2.36	40.5 ± 1.13	40-49
MCV FL	Mean ± SD	59.21 ± 1.44	59.31 ± 0.99	57.88 ± 0.87	58.48 ± 2.32	58.92 ± 1.33	56.45 ± 0.92	52-66
MCH PICO GRA	Mean ± SD	21.14 ± 0.73	21.13 ± 0.56	20.25 ± 0.34	20.78 ± 0.84	20.8 ± 0.77	20 ± 0.42	17.7-19.1
MCHC (%)	Mean ± SD	35.74 ± 1.31	35.63 ± 1.01	35.02 ± 0.32	35.54 ± 0.51	35.28 ± 1.03	35.45 ± 0.07	32-33.5
PLATELET THSN/UL	Mean ± SD	1060.8 ± 182.4	1024.3 ± 190.8	1030.3 ± 160.1	960.8 ± 77.7	909.3 ± 136.5	657.5 ± 17.7	780-1400
BANDS THSN/UL	Mean ± SD	0.15 ± 0.217	0.07 ± 0.089	0.14 ± 0.126	0 ± 0	0.03 ± 0.045	0 ± 0	0-0.06
SEGS THSN/UL	Mean ± SD	3.93 ± 2.71	3.08 ± 2.931	2.51 ± 2.341	5.92 ± 2.109	4.11 ± 1.952	5.7 ± 3.026	0.58-6.3

 

WBC THSN/UL: White blood cells (leukocytes) in thousands per microliter

RBC MILL/UL: Red blood cells (erythrocytes) in millions per microliter

HGB GM/DL: Hemoglobin in grams per deciliter

HCT (%): Hematocrit (Packed Cell Volume) in percent

MCV FL: Mean corpuscular volume, the volume of the average erythrocyte, in femtoliters

MCH PICO GRA: Mean corpuscular hemoglobin, the amount of hemoglobin in the average erythrocyte in picograms

MCHC (%): Mean corpuscular hemoglobin concentration, the proportion of the erythrocyte mass composed of hemoglobin in percent

PLATELET THSN/UL: Platelets in thousands per microliter

BANDS THSN/UL: Band neutrophils (non-segmented neutrophils) in thousands per microliter

SEGS THSN/UL: Segmented neutrophils in thousands per microliter

 

Table 16 (cont): Evaluation of hematologic data – female rats

Dosage Group Dosage (mg/kg bw/day)		I (Water) 0	II (Corn oil) 0	III 75	IV 250	V 750	VI 2000	Reference range
Rats tested	N	10	10	10	10	10	10
LYMPHS THSN/UL	Mean ± SD	12.858 ± 3.864	11.241 ± 3.36	8.902 ± 1.678	5.374 ± 1.439	6.402 ± 3.375	3.11 ± 0.2	3.78-15.32
MONOS THSN/UL	Mean ± SD	0.02 ± 0.045	0.03 ± 0.077	0.13 ± 0.221	0.01 ± 0.032	0.05 ± 0.071	0 ± 0	0.02-1.2
EOSN THSN/UL	Mean ± SD	0.13 ± 0.114	0.11 ± 0.145	0.08 ± 0.095	0.11 ± 0.084	0.03 ± 0.045	0.09 ± 0.032	0-031
BASOS THSN/UL	Mean ± SD	/	/	/	/	/	/	0-0.01
ATLYMPH THSN/UL	Mean ± SD	/	/	/	/	/	/	0-0
METAS THSN/UL	Mean ± SD	0.118 ± 0.134	/	/	/	0.057 ± 0.089	/	0-0
MYELOS	Mean ± SD	/	/	/	/	/	/	0-0
PROS THSN/UL	Mean ± SD	/	/	/	/	/	/	0-0
BLASTS THSN/UL	Mean ± SD	/	/	/	/	/	/	0-0
NRBC/100 WBC	Mean ± SD	7.6 ± 5.6	5.2 ± 3.2	3.5 ± 2.9	5.2 ± 1.9	3.5 ± 2	2 ± 1.4	0-0
RETIC (%)	Mean ± SD	1.63 ± 0.45	2.17 ± 0.8	1.7 ± 0.59	0.81 ± 0.57	1.27 ± 0.88	2.8 ± 2.03	0.1-4

 

LYMPHS THSN/UL: Lymphocytes in thousands per microliter

MONOS THSN/UL: Monocytes in thousands per microliter

EOSN THSN/UL: Eosinophils in thousands per microliter

BASOS THSN/UL: Basophils in thousands per microliter. No entry indicates none seen

ATLYMPH THSN/UL: Atypical lymphocytes in thousands per microliter. No entry indicates none seen

METAS THSN/UL: Metamyelocytes in thousands per microliter. No entry indicates none seen

MYELOS: Myelocytes in thousands per microliter. No entry indicates none seen

PROS THSN/UL: Promyelocytes in thousands per microliter. No entry indicates none seen

BLASTS THSN/UL: Blast cells in thousands per microliter. No entry indicates none seen

NRBC/100 WBC: Nucleated red blood cells (nucleated erythrocytic precursors) seen per 100 white blood cells

RETIC (%): Reticulocytes expressed a percent of total red blood cells

Table 17: Incidence of histomorphologic observations

Dose level (mg/kg/day):	0	0	75	250	750	2000	0	0	75	250	750	2000
Sex:	M	M	M	M	M	M	F	F	F	F	F	F
Number of animals/group:	10	10	10	10	10	10	10	10	10	10	10	10
Adrenals glands
No. examined	10	10	0	0	0	10	10	10	0	0	0	10
No. normal	10	9	0	0	0	10	10	10	0	0	0	10
Hypertrophy, zona glomerulosa	0	1	0	0	0	0	0	0	0	0	0	0
Epididymides
No. examined	10	10	0	0	0	10
No. normal	9	10	0	0	0	8
Hypospermia	0	0	0	0	0	1
Inflammation, interstitial	1	0	0	0	0	1
Esophagus
No. examined	0	0	0	0	0	3	0	0	0	0	0	6
No. normal	0	0	0	0	0	0	0	0	0	0	0	0
Hemorrhage, periesophageal	0	0	0	0	0	0	0	0	0	0	0	5
Inflammation, periesophageal	0	0	0	0	0	2	0	0	0	0	0	3
Necrosis, muscular wall, focal	0	0	0	0	0	3	0	0	0	0	0	3
Heart
No. examined	0	0	0	0	0	2	0	0	0	0	0	5
No. normal	0	0	0	0	0	1	0	0	0	0	0	1
Hemorrhage, pericardial	0	0	0	0	0	1	0	0	0	0	0	1
Pericarditis	0	0	0	0	0	1	0	0	0	0	0	3
Kidneys
No. examined	10	10	10	10	10	10	10	10	10	10	10	10
No. normal	9	9	9	6	9	6	8	7	10	8	9	7
Basophilia, cortical tubules, multifocal	0	0	0	0	0	3	0	0	0	0	0	0
Crystals, intratubular	0	0	0	0	0	2	0	0	0	0	0	0
Cystic tubules, cortex	0	0	0	0	0	0	0	0	0	1	0	0
Dilatation, pelvis	1	1	1	4	1	0	1	2	0	0	0	0
Dilatation, tubular, multifocal	0	0	0	0	0	3	0	0	0	0	0	0
Hyaline droplets, cortical tubules	0	0	0	0	0	1	0	0	0	0	0	0
Infiltration, mononuclear-cell, focal	0	0	0	0	0	0	1	0	0	0	0	0
Mineralization, multifocal	0	0	0	0	0	0	0	1	0	0	1	1
Nephritis, chronic, focal	0	0	0	0	0	0	0	0	0	0	0	1
Pyelitis, chronic	0	0	0	0	0	2	0	0	0	0	0	2
Pyelonephritis, chronic	0	0	0	0	0	0	0	0	0	1	0	0
Liver
No. examined	10	10	10	10	10	10	10	10	10	10	10	10
No. normal	5	6	8	9	9	3	6	8	7	8	10	5
Cellular alteration, focus/area, clear-cell	0	1	0	0	0	0	0	0	0	0	0	0
Congestion	0	0	0	0	0	0	0	0	0	0	0	1
Hematopoiesis, extramedullary	0	0	0	1	0	0	0	0	0	1	0	0
Hypertrophy, hepatocellular, centrilobular	0	0	0	0	0	6	0	0	0	0	0	2
Infiltration, mononuclear-cell, multifocal	5	3	2	1	1	1	4	1	3	2	0	2
Necrosis, focal	0	0	0	0	0	0	1	0	0	0	0	0
Vacuolation, hepatocellular, midzonal	0	0	0	0	0	5	0	0	0	0	0	3
Vacuolation, hepatocellular, multifocal	0	0	0	0	0	0	0	1	0	0	0	0

 

Table 17 (cont): Incidence of histomorphologic observations

Dose level (mg/kg/day):	0	0	75	250	750	2000	0	0	75	250	750	2000
Sex:	M	M	M	M	M	M	F	F	F	F	F	F
Number of animals/group:	10	10	10	10	10	10	10	10	10	10	10	10
Lungs
No. examined	10	10	0	0	0	10	10	10	0	0	0	10
No. normal	6	7	0	0	0	5	5	4	0	0	0	3
Bronchopneumonia, necrotizing	0	0	0	0	0	0	0	0	0	0	0	1
Congestion	0	0	0	0	0	2	0	0	0	0	0	2
Congestion/edema	0	0	0	0	0	0	0	0	0	0	0	1
Inflammation, interstitial, focal	1	2	0	0	0	2	0	1	0	0	0	1
Macrophages, alveoli	0	0	0	0	0	1	0	0	0	0	0	0
Necrosis, bronchus	0	0	0	0	0	0	0	0	0	0	0	1
Pleuritis	0	0	0	0	0	0	0	0	0	0	0	2
Proliferation, lymphoid, peribronchial/perivascular	3	3	0	0	0	3	5	5	0	0	0	2
Ovaries
No. examined							10	10	0	0	0	10
No. normal							10	10	0	0	0	10
Pancreas
No. examined	10	10	0	0	0	10	10	10	0	0	0	10
No. normal	10	9	0	0	0	10	10	10	0	0	0	10
Atrophy, acinar, focal	0	1	0	0	0	0	0	0	0	0	0	0
Pituitary
No. examined	10	9	0	0	0	9	10	10	0	0	0	9
No. normal	8	9	0	0	0	8	8	10	0	0	0	9
Cyst(s)	2	0	0	0	0	1	2	0	0	0	0	0
Prostate
No. examined	10	10	0	0	0	10
No. normal	10	10	0	0	0	8
Necrosis, focal	0	0	0	0	0	1
Prostatitis, interstitial, chronic	0	0	0	0	0	1
Seminal vesicle(s)
No. examined	10	10	0	0	0	10
No. normal	10	10	0	0	0	10
Spleen
No. examined	10	10	0	0	0	10	10	10	0	0	0	10
No. normal	10	10	0	0	0	9	10	10	0	0	0	8
Atrophy	0	0	0	0	0	0	0	0	0	0	0	2
Hyperplasia, red pulp	0	0	0	0	0	1	0	0	0	0	0	0
Stomach
No. examined	0	0	0	0	0	1	0	0	0	0	0	1
No. normal	0	0	0	0	0	0	0	0	0	0	0	0
Edema, limiting ridge	0	0	0	0	0	0	0	0	0	0	0	1
Edema, submucosa	0	0	0	0	0	0	0	0	0	0	0	1
Edema/inflammation, submucosa	0	0	0	0	0	1	0	0	0	0	0	0
Necrosis, superficial epithelium, focal	0	0	0	0	0	1	0	0	0	0	0	0
Ulcer, glandular mucosa	0	0	0	0	0	1	0	0	0	0	0	1

 

Table 17 (cont): Incidence of histomorphologic observations

Dose level (mg/kg/day):	0	0	75	250	750	2000	0	0	75	250	750	2000
Sex:	M	M	M	M	M	M	F	F	F	F	F	F
Number of animals/group:	10	10	10	10	10	10	10	10	10	10	10	10
Testes
No. examined	10	10	10	10	10	10
No. normal	8	9	9	8	10	8
Atrophy, bilateral, multifocal	1	1	0	1	0	0
Atrophy, unilateral, multifocal	1	1	0	1	0	0
Degeneration, testicular	0	0	0	0	0	2
Multinucleated cells	0	0	0	0	0	2
Thoracic cavity
No. examined	0	0	0	0	0	0	0	0	0	0	0	1
No. normal	0	0	0	0	0	0	0	0	0	0	0	0
Abscess	0	0	0	0	0	0	0	0	0	0	0	1
Thymus
No. examined	10	10	10	10	10	10	10	10	10	10	10	9
No. normal	10	10	10	10	7	4	8	9	7	7	7	4
Abscess	0	0	0	0	0	0	0	0	0	0	0	1
Atrophy	0	0	0	0	0	5	0	0	0	0	0	3
Congestion	0	0	0	0	3	2	2	1	3	2	3	3
Hemorrhage, focal	0	0	0	0	0	0	0	0	0	0	0	2
Necrosis, focal	0	0	0	0	0	0	0	0	0	1	0	1
Trachea
No. examined	0	0	0	0	0	3	0	0	0	0	0	6
No. normal	0	0	0	0	0	2	0	0	0	0	0	3
Congestion	0	0	0	0	0	0	0	0	0	0	0	1
Inflammation, peritracheal	0	0	0	0	0	0	0	0	0	0	0	1
Necrosis	0	0	0	0	0	1	0	0	0	0	0	2
Urinary bladder
No. examined	0	0	0	0	0	1	0	0	0	0	0	0
No. normal	0	0	0	0	0	0	0	0	0	0	0	0
Cystisis, hyperplastic	0	0	0	0	0	1	0	0	0	0	0	0
Microcalculi	0	0	0	0	0	1	0	0	0	0	0	0
Uterus
No. examined							10	10	0	0	1	10
No. normal							9	5	0	0	0	8
Distention, lumen							1	5	0	0	1	2

 

Applicant's summary and conclusion

Conclusions:

Administration of 1,3-Dioxolane for 14 days to male rats at dosages of 250, 750 and 2000 mg/kg/day caused dosage-dependent decreases in platelet counts and reduced body weight gains and feed consumption values. The 750 and 2000 mg/kg/day dosage groups had increased average relative weights of the liver and lungs and reduced averages for the relative weight of the thymus.
The 2000 mg/kg/day dosage group had increased relative kidney weights and reduced relative spleen weight. The 2000 mg/kg/day dosage of the test substance also produced mortality, clinical and necropsy observations and histopathologic changes in the liver, kidneys, thymus and testes.
In female rats, administration of 1,3-Dioxolane for 14 days at dosages of 750 and 2000 mg/kg/day caused increases in averages for the relative liver weights and reduced relative weights of the spleen and thymus. The 2000 mg/kg/day dosage group also had increased relative kidney weights and a reduced relative pancreas weight. Dosages of 250 mg/kg/day and higher reduced
body weight gains, feed consumption values, lymphocyte counts and reticulocyte count (250 mg/kg/day dosage group only). Dosages of 750 and 2000 mg/kg/day caused clinical observations, and the 2000 mg/kg/day dosage also caused mortality, a gross lesion, reduced platelet count and histopathologic changes in the liver, kidneys and thymus.
Based on this 14-day evaluation, the no-observable-adverse-effect level (NOAEL) for 1,3-Dioxolane in male and female rats was 75 mg/kg/day.

Executive summary:

Introduction

The purpose of this repeated dose oral toxicity study was to provide information on the toxicity of 1,3-Dioxolane in male and female rats and to provide information for use in the selection of dosages of 1,3-Dioxolane to be used in the developmental toxicity (embryo-fetal toxicity and teratogenic potential) study.

Methods

Each dosage group consisted of ten male and ten female Crl:CD(SD)BR rats. The test substance, 1,3-Dioxolane, was administered orally via gavage to the male and female rats as a solution in corn oil for 14 consecutive days at dosages of 75, 250, 750 and 2000 mg/kg/day. There were two control groups, one given reverse osmosis membrane processed deionized water (R.O. deionized water) and the other given the vehicle (corn oil).

Results

Three males and four females of the high dosage groups were found dead during the study. No other death was reported.

Clinical observations were reported at the high dosage groups only and included hypotonia, excess salivation, ataxia, decreased motor activity, rales, impaired and lost righting reflex, labored breathing, red penile exudate, urine-stained abdominal fur, gasping, bradypnea, oral exudate and lacrimation. Only excess salivation was observed at 750 mg/kg/day in female rats.

Reduced body weight gains were observed in the 250, 750 and 2000 mg/kg/day groups, both in male and female rats.

Groups given 250 mg/kg/day and higher dosages of the test substance had marginally reduced feed consumption values, as compared with the corn oil control group values.

Male rats exposed to 750 and 2000 mg 1,3-Dioxolane/kg/day showed increased relative liver and lung weights and a reducted relative thymus weight. An increased relative kidney weight and a reduced relative spleen weight were also observed in male rats of the highest dosage level.

Female rats exposed to 750 and 2000 mg 1,3-Dioxolane/kg/day showed increased relative liver weight and reduced relative spleen and thymus weights. The 2000 mg/kg/day dosage group also had increased relative kidney weights and a reduced relative pancreas weight.

Significant decreases in platelet counts were observed in males of the 250, 750 and 2000 mg/kg/day dosage groups and in females of the 2000 mg/kg/day dosage group. Significant decreases in lymphocyte counts also occured in females of the 250, 750 and 2000 mg/kg/day dosage groups as well as a decreased in reticulocyte count in females of the 250 mg/kg/day dosage group.

No treatment-related microscopic changes were observed in any of the male and female rats given 75, 250 or 750 mg/kg/day of 1,3-Dioxolane. Treatment-related microscopic changes were observed in the liver, kidneys, thymus in males and females and in testes of male rats given 2000 mg/kg/day of 1,3-Dioxolane.