1,3-dioxolane

Repeated dose toxicity: inhalation

Administrative data

Endpoint:

sub-chronic toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From March 1989 to August 1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

The study report does not indicate that a guideline was followed. However, the study was performed using a procedure comparable to OECD 413 (version before 9 October 2017) with acceptable restrictions indicated in the test guideline description.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1990

Report date:

1990

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 413 (Subchronic Inhalation Toxicity: 90-Day Study)

Deviations:

yes

Remarks:

Parameters not recorded: food consumption, mean corpuscular haemoglobin, mean corpuscular volume, mean corpuscular haemoglobin concentration, reticulocytes, clotting potential parameter. Tissues not collected: gallbladder, olfactory bulb, ureter, urethra

GLP compliance:

yes

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: Grant Chemical CO., Baton rouge LA - lot #8313

Test animals

Species:

rat

Strain:

Fischer 344

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories, Inc, Kingston, NY
- Females nulliparous and non-pregnant: no data
- Age at study initiation: 7 weeks (purchased at 4 weeks and acclimated for 3 weeks prior to exposure)
- Weight at study initiation: males: 146.5 - 149.8 g; females: 106.0 - 108.7 g
- Fasting period before study: no
- Housing: housed into groups of 10
- Diet: ad libitum, Purina Certified Rodent Chow #5002 (Ralston Purina Co, St Louis, MO) except during test item exposure
- Water: ad libitum, municipal water supply, except during test item exposure
- Acclimation period: 3 weeks

DETAILS OF FOOD AND WATER QUALITY: Analysis of Purina Certified Rodent Chow was supplied by the Ralston Purina Company to confirm that the diet provided adequate nutrition and to quantify the levels of selected contaminants associated with the formulation processes. Analysis of tap water was performed in accordance with Standard Operating Procedures of the laboratory.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): no data
- Humidity (%): no data
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): no data

IN-LIFE DATES: From: To: no data

Administration / exposure

Route of administration:

inhalation: vapour

Type of inhalation exposure:

whole body

Vehicle:

air

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: rats were exposed to test material vapour in 4 cubic meter stainless steel and glass chambers (152 cm wide x 143 cm high x 152 cm deep with a pyramidal top and bottom) under dynamic air flow conditions
- Generating system: vapours of the test material were generated using a glass J-tube method. Liquid test material was metered into the J-tube. Nitrogen heated with a flameless torch to the minimum extent necessary to volatilize the test material passed through the J-tube. Nitrogen flow through the J-tubes was maintained at approximately 30-35 L/minute.
- Source and rate of air: dynamic airflow conditions
- Method of conditioning air: no data
- Temperature, humidity: 20-23°C, 52-56%
- Air flow rate: 30-35 liters/minute
- Air change rate: Chamber air flow will be maintained at approximately 800 liters/minute.
- Treatment of exhaust air: no data

TEST ATMOSPHERE
- Brief description of analytical method used: The nominal concentration of the test material in each chamber was calculated for each exposure period based on the amount of test material used and the approximate air flow through the chamber.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The analytical concentration of 1,3-dioxolane in each chamber was determined at least 10 times/exposure period with a MIRAN 1A infrared spectrophotometer (IR).

Duration of treatment / exposure:

6 hours/day, 5 days/week, for 13 weeks.

Frequency of treatment:

daily

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 ppm (nominal)

Remarks:

Control

Doses / concentrations 2

Dose / conc.:

300 ppm (nominal)

Remarks:

298 ppm (measured)

Doses / concentrations 3

Dose / conc.:

1 000 ppm (nominal)

Remarks:

1000 ppm (measured)

Doses / concentrations 4

Dose / conc.:

3 000 ppm (nominal)

Remarks:

3010 ppm (measured)

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Dose selection was based on previous studies. CNS depression was the primary effect when 5 rats/sex were exposed for 4 or 6 hours to 13,050 or 7,700 ppm 1,3-dioxolane, respectively. All rats survived these acute exposures. In a subsequent two-week inhalation toxicity study, 5 rats/sex received 9 exposures to targeted concentrations of 500, 2000 or 5000 ppm 1,3-dioxolane for 6 hours/day. Mean body weight was decreased in male (12%) and female rats (4%) exposed to 5000 ppm 1,3-dioxolane, and all rats of this group had decreased white blood cell counts.
- Rationale for animal assignment: rats were weighed, randomized by body weight into groups of 10.
- Fasting period before blood sampling for clinical biochemistry: all rats are fasted on the day after the final exposure
- Rationale for selecting satellite groups: satellite group are held to evaluate recovery at 28 days post-exposure
- Post-exposure recovery period in satellite groups: 8 weeks

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations checked in table were included.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: weekly

BODY WEIGHT: Yes
- Time schedule for examinations: day 1 (prior to exposure) and weekly thereafter

OPHTHALMOSCOPIC EXAMINATION: Yes - pen-light ophthalmologic examination and in situ examination of the eyes with a glass side under fluorescent illumination
- Time schedule for examinations: prior to exposure and at necropsy, respectively
- Dose groups that were examined: all rats in the control and highest 1,3-dioxolane exposure group.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: on the day after the final exposure for the main group and at 4 and 13 weeks of exposure and again 4 and 8 weeks into the post-recovery period for the satellite group rats.
- Anaesthetic used for blood collection: Yes, animals slightly anesthetized with methoxyflurane
- Animals fasted: Yes
- How many animals: all animals
- Parameters examined: packed cell volume (PCV), hemoglobin (HGB), erythrocyte count (RBC), total leukocyte (WBC), platelet count (PLAT), differential WBC, RBC morphology and absolute numbers of lymphocytes

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: collected simultaneously with hematology samples, main group: week 13, satellite group: weeks 4, 13 plus post-exposure week 4
- Animals fasted: Yes
- How many animals: all animals
- Parameters examined: urea nitrogen (BUN), creatinine (CREAT), alanine aminotransferase activity (ALT), aspartate aminotransferase actvity (AST), alkaline phosphatase activity (AP), glucose (GLUC), total protein (TP), albumin (ALB), globulin (GLOB, calculated), total bilirubin (TBILI), cholesterol (CHOL), triglycerides (TRIG), phosphatus (PHOS) and calcium (CALC), sodium, potassium and chloride.
URINALYSIS: Yes
- Time schedule for collection of urine: conducted on all rats during the final week of exposure, repeated on the main study group of animals prior to necropsy and conducted again on the satellite group at 4 and 8 weeks post exposure.
- Metabolism cages used for collection of urine: Not specified
- Parameters examined: color, appearance, bilirubin, glucose, ketones, blood, pH, protein and urobilinogen, specific gravity and examination of urine sediment.

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
Necropsy performed on the day after the final exposure for the main study rats and at the end of the 8-week recovery period for the satellite group rats.
HISTOPATHOLOGY: Yes

Other examinations:

Main study group rats were examined in a functional observation battery (FOB) during the sixth and last week of the study. This FOB consisted of observations for any unusual conditions with respect to: pupil size, respiration, movement, skin and haircoat, salivation, lacrimation, urine staining, fecal staining, locomotor behavior and responsiveness to touch, noise and tail pinch

Statistics:

Descriptive statistics (mean and standard deviation) were used to report chamber concentrations, temperature, and relative humidity.
Body weights, absolute and relative organ weights, clinical chemistry and appropriate hematology data for animals to be terminated after 13 weeks will be evaluated by Bartlett's test for equality of variances. Clinical chemistry and appropriate hematology data for animals to be terminated after 4 weeks will be evaluated in the same manner. Based on the outcome of Bartlett's test, exploratory data analysis will be performed by a parametric or non-parametric analysis of variance (ANOVA), followed respectively by Dunnett's test or the Wilcoxon Rank-Sum test with a Bonferroni correction for multiple comparisons. Statistical outliers will be identified by a sequential test and excluded from analysis only for documented, scientifically sound reasons, which are unrelated to exposure.

Results and discussion

Results of examinations

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

Some male and female rats had notable clinical observations including red swollen eyes, perineal soiling, diarrhea and darkened crusty material around the eyes and/or nares. One male rat exposed to 1000 ppm has persistent redness, swelling and/or crusty material around one eye. However, this condition was resolved by study termination.The orbital sinus blood sampling procedure probably resulted in some of the observations of crusty material around the eyes. Because of a lack of a dose-response relationship and persistence in any of the observations, none were considered to be exposure-related or toxicologically significant.

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

Male rat body weights were not affected by exposure. A statistical difference in 1000 ppm exposed males on day 31 was not part of an exposure response pattern. Female rat body weights from the 3000 ppm 1,3-dioxolane group were slightly lower than controls during the 13-week exposure period. This effect was statistically identified on days 5, 11, 18, 25, 31 and 67. There were no statistical differences in mean body weights of 1,3-dioxolane exposed rats relative to controls during the 8-week post exposure period.

Ophthalmological findings:

no effects observed

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

Mean WBC counts of male and female rats exposed to 3000 ppm were significantly lower than for controls during the 13-week exposure period; slight decreases in WBC counts also occurred in rats exposed to 1000 ppm but were never below the lower limits of the historical control values.
Evaluation of absolute numbers of lymphocytes and segmented neutrophils derived from the differential leukocyte counts revealed decreased mean values at several intervals, primarily from rats exposed to 1000 or 3000 ppm 1,3-dioxolane. By 8 weeks post exposure, the mean lymphocyte numbers of rats exposed to 3000 ppm were comparable to control values. The reduced lymphocyte numbers in peripheral blood were accompanied by a slight reduction in the number of myeloid cells in the bone marrow of male and female rats exposed to 3000 ppm . However, no myeloid cytotoxicity or any evidence of altered marrow environment were observed. There was also no microscopic indication of structural alteration in lymphoid organs or increased body demand for WBC (decreased M/E ratio but similar to that of controls after 8 weeks post exposure).
The % eosinophils in 3000 ppm 1,3-dioxolane exposed rats were increased relative to controls after 13 weeks of exposure.
Small increases in mean platelet counts occurred during the exposure period in some of the 1000 and 3000 ppm groups. There was a single observation of increased platelets in females exposed to 300 ppm at 4 weeks, however, there was no apparent exposure response relationship and no difference in platelet counts between control and 300 ppm females at 13 weeks. The minimal statistical platelet elevations in 1000 and 3000 ppm exposed rats were within the range of available historical control data; hence these differences were considered to be of minor biological significance.
Red blood cell counts were statistically increased in females exposed to 1000 (main group only) and 3000 ppm at 13 weeks. These increases were minimal and were well within the available historical control range; therefore, these were considered to be of no toxicological significance.
During the post-exposure period, slight (<3%) decreases in RBC (at 4 weeks only) and HCT were statistically identified in 3000 ppm exposed males. These mean values were within the available historical control range.

Clinical biochemistry findings:

effects observed, non-treatment-related

Description (incidence and severity):

There were a number of statistically identified differences in various clinical chemistry parameters in 1,3-dioxolane exposed male or female rats when compared to their respective controls, but there was no clear relationship to exposure. Since these parameters were generally within the historical control ranges or were not associated with corresponding gross or microscopic effects, these clinical chemistry changes were not considered to be diagnostic of a target organ effect. However, increased ALT in 3000 ppm exposed rats at 4 weeks, and, in females only at 13 weeks, could be related to the small increases in liver weight. Histologic changes were also noted in the livers of 3000 ppm exposed males.
Serum electrolytes revealed no clear exposure related effects. Although there were a number of statistically identified differences between control and exposed groups, the lack of a dose response and variability of control values raises doubt regarding the biological significance of the statistical differences. All statistically identified electrolyte values in this study were within the range of control group values seen at the 4, 13 or 4-week post-exposure samples.

Urinalysis findings:

effects observed, treatment-related

Description (incidence and severity):

Male and female rats exposed to 3000 ppm 1,3-dioxolane for 13-week (main study and satellite groups) had statistically decreased urine specific gravity. Repeat urinalyses revealed similar results. The specific gravity alteration was not associated with a morphologic indication of nephrotoxicity. It may represent a physiologic response to repeated exposure to 1,3-dioxolane.

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

The mean absolute spleen weight of female rats exposed to 3000 ppm of 1,3-dioxolane was statistically decreased, as were the mean relative spleen weights of males exposed to 3000 ppm and females exposed to 1000 and 3000 ppm. The toxicological significance of this observation is uncertain because there were no morphologic alterations observed in the spleen from any animal in the study. The lymphocyte populations in B- and T-cell dependant areas appeared similar in all treated and control animals. In addition, the morphology of cells comprising the red pulp region appeared similar to controls. It is possible that the decreased relative spleen weight was associated with reduced WBC counts, though there was no morphologic indication of toxicity to the spleen.
The mean absolute and relative liver weight of male and female rats exposed to 3000 ppm 1,3-dioxolane were statistically increased when compared to control values, as was the mean relative liver weight of females exposed to 1000 ppm. Male rats had minimal microscopic liver changes while females had no morphologic liver alterations. The liver weight increases may have been an adaptive physiologic response following repeated exposures to 1,3-dioxolane.
The mean absolute brain weight of female rats exposed to 3000 ppm 1,3-dioxolane was statistically decreased from the female mean control value. The mean relative brain weight was not identified as different although the mean terminal body weights were virtually identical, and there was no indication of a difference in brain weight at 8 weeks post-exposure. There were no morphologic changes in the brain. Therefore, the statistical difference in brain weight was not considered to be toxicologically significant. The statistical increase in absolute (but not relative) testes weight in 1000 ppm exposed males at 13 weeks was not considered to be exposure-related because of the absence of an effect on testes weights in 3000 ppm group males. The mean absolute (but not relative) lung weight of males exposed to 3000 ppm was increased by approximately 6% after 13 weeks of exposure and 8 weeks post exposure. Since no microscopic changes were observed in their lungs, this finding did not appear to represent an injurious effect.
Examinations of other organ and body weight data from rats 8 weeks post exposure to1,3-dioxolane indicated no toxicologically significant findings. In particular, spleen and liver weights of exposed and control rats were comparable. Mean absolute heart weights were statistically increased in females exposed to 1000 ppm, though the responsa did not consistently increase with exposure concentration.

Gross pathological findings:

no effects observed

Description (incidence and severity):

There were no exposure-related gross or microscopic changes in male or female rats exposed to 300, 1000 or 3000 ppm 1,3-dioxolane, except for minimal liver changes. In male rats exposed to 3000 ppm 1,3-dioxolane, hepatocytes in centrilobular regions of lobules were slightly larger and had slightly more cytoplasmic eosinophilia than comparable centrilobular hepatocytes of control males. Microscopic examination of bone marrow from exposed and control rats following 13 weeks of exposure to 1,3-dioxolane indicated that myeloid and erythroid cells were approximately similar in number, had normal morphology and apparently normal maturation sequences. In addition, there was no cytotoxicity of the microenvironment at the light microscopic level.
Hence, the decreased in WBC counts were not associated with observable histopathologic changes in the bone marrow.
After 8 weeks post exposure, there were no exposure-related gross or microscopic changes. Microscopic examination of bone marrow from control and high exposure group rats following 8 weeks post exposure was similar to the bone marrow obtained after 13 weeks of exposure to 1,3-dioxolane.

Effect levels

Key result

Dose descriptor:

NOAEC

Effect level:

298 ppm (analytical)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

haematology

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

Table 1: Analytical and nominal concentrations

Groups		Analytical	Nominal
Control	Concentration	0	0
	SD	0	0
	N	67	67
300 ppm	Concentration	298	315
	SD	18	17
	N	67	67
1000 ppm	Concentration	1000	1031
	SD	44	57
	N	67	67
3000 ppm	Concentration	3010	3119
	SD	104	219
	N	67	67

 

Table 2: Body weight – males (g)

		Days on test
Conc. (ppm)		1	5	11	18	25	31	39	46	53	60	67	74	81
0	Mean	149.8	157.7	185.3	204.0	222.7	231.6	242.7	253.2	263.4	272.8	280.5	288.1	292.6
	SD	11.0	11.9	11.3	9.3	11.6	11.9	11.4	11.3	13.3	13.4	13.7	14.2	14.0
	N	20	20	20	20	20	20	20	20	20	20	20	20	20
300	Mean	149.5	159.2	185.1	204.1	222.0	229.9	239.9	252.6	264.1	272.5	281.1	287.5	292.3
	SD	9.9	10.8	11.9	13.2	13.0	13.8	14.5	16.2	17.0	16.6	17.7	18.4	19.2
	N	20	20	20	20	20	20	20	20	20	20	20	20	20
1000	Mean	147.2	154.3	178.5	196.2	213.6	222.3*	232.8	247.1	256.9	268.1	275.8	283.3	290.3
	SD	12.7	12.4	13.3	13.2	12.7	11.2	15.1	14.5	14.4	14.4	13.9	13.7	14.4
	N	20	20	20	20	20	20	20	20	20	20	20	20	20
3000	Mean	146.5	155.1	182.9	203.1	221.3	230.5	241.1	256.8	267.0	278.2	285.1	286.9	299.2
	SD	8.7	8.5	8.9	9.7	10.3	10.8	12.8	11.7	11.8	12.4	12.6	15.5	13.5
	N	20	20	20	20	20	20	20	20	20	20	20	20	20

 

Table 2 (cont): Body weight – males (g)

 

		Days on test
Conc. (ppm)		86	96	102	109	116	123	131	139	144	152
0	Mean	299.1	293.6	309.3	318.2	325.3	327.3	320.5	325.4	332.7	322.6
	SD	13.8	13.6	15.6	14.9	15.1	16.5	16.8	14.5	15.6	15.3
	N	20	10	10	10	10	10	10	10	10	10
300	Mean	299.9	294.3	304.8	313.1	318.9	319.9	316.0	320.5	325.7	314.2
	SD	19.9	14.5	13.8	12.5	13.1	13.8	15.0	16.6	17.1	16.6
	N	20	10	10	10	10	10	10	10	10	10
1000	Mean	298.0	287.4	299.8	307.8	314.5	316.5	313.3	318.4	321.0	310.6
	SD	13.9	16.2	17.4	17.1	17.6	18.2	18.9	20.2	21.2	20.9
	N	20	10	10	10	10	10	10	10	10	10
3000	Mean	306.2	302.8	316.3	326.2	331.6	329.9	329.3	334.5	340.3	328.5
	SD	14.6	13.4	14.4	14.1	14.7	17.3	19.0	19.1	17.5	17.6
	N	20	10	10	10	10	10	10	10	10	10

 

Table 3: Body weight – females (g)

		Days on test
Conc. (ppm)		1	5	11	18	25	31	39	46	53	60	67	74	81
0	Mean	108.7	114.7	130.3	138.9	148.8	153.1	157.9	163.8	168.2	172.1	175.7	180.3	181.4
	SD	5.3	5.4	6.3	6.9	7.4	7.7	8.8	8.6	8.9	8.7	8.6	9.7	10.0
	N	20	20	20	20	20	20	20	20	20	20	20	20	20
300	Mean	107.7	114.5	130.2	140.4	150.0	154.3	159.0	164.5	168.9	175.6	177.9	182.2	182.6
	SD	5.3	4.7	5.4	6.0	5.8	6.6	7.7	8.4	7.8	6.4	9.4	9.9	9.8
	N	20	20	20	20	20	20	20	20	20	20	20	20	20
1000	Mean	108.0	111.4	126.2	135.4	144.6	148.1	155.0	160.5	166.9	170.6	174.6	178.8	179.5
	SD	7.1	7.8	7.3	6.9	7.0	8.2	7.7	7.1	8.4	9.2	7.9	8.4	9.2
	N	20	20	20	20	20	20	20	20	20	20	20	20	20
3000	Mean	106.0	109.9*	124.1*	132.6*	142.7*	147.1*	153.8	158.4	162.8	167.4	168.9*	175.6	176.1
	SD	6.0	5.2	4.9	5.2	5.1	4.8	5.6	5.8	6.7	6.6	5.6	6.6	7.7
	N	20	20	20	20	20	20	20	20	20	20	20	20	20

 

Table 3 (cont): Body weight – females (g)

		Days on test
Conc. (ppm)		86	96	102	109	116	123	131	139	144	152
0	Mean	182.1	179.8	185.6	191.7	192.4	192.3	189.5	193.1	196.4	187.1
	SD	9.6	10.4	9.2	9.7	10.4	10.5	11.1	11.3	10.8	11.1
	N	20	10	10	10	10	10	10	10	10	10
300	Mean	183.8	180.0	184.5	189.1	191.2	190.6	188.9	191.0	193.3	185.9
	SD	10.0	13.4	12.0	14.1	12.9	13.2	7.0	12.4	13.0	11.7
	N	20	10	10	10	10	10	10	10	10	10
1000	Mean	181.8	182.7	189.1	194.2	196.1	193.3	191.3	196.0	199.5	189.9
	SD	8.4	6.5	5.6	6.5	6.8	7.1	6.5	7.0	7.0	6.7
	N	20	10	10	10	10	10	10	10	10	10
3000	Mean	179.2	173.3	181.6	186.6	188.8	188.2	185.6	189.3	194.0	197.5
	SD	6.5	6.2	6.6	8.2	9.2	8.0	8.4	7.8	8.6	6.5
	N	20	10	10	10	10	10	10	10	10	10

Table 4: Weekly clinical examination – males

		Day
Observation	Concentration	1-7	8-14	15-21	22-28	29-35	36-42	43-49	50-56	57-63	64-70	71-77	78-84	85-91
Crust or staining around eyes or nares	0 ppm	0	0	0	0	0	0	1	0	0	0	0	0	1
	300 ppm	0	0	0	0	1	0	0	0	0	0	0	0	1
	1000 ppm	0	0	0	0	0	1	1	1	1	1	1	0	0
	3000 ppm	0	0	0	0	0	1a	0	0	0	1	0	1	1
Diarrhea	0 ppm	0	0	0	0	0	0	0	0	0	0	0	0	0
	300 ppm	0	0	0	0	0	0	0	0	0	0	0	0	0
	1000 ppm	0	0	0	0	0	0	0	0	0	0	0	0	1
	3000 ppm	0	0	0	0	0	0	0	0	0	0	0	0	0

a: swollen eye

Table 5: Weekly clinical examination – females

		Day
Observation	Concentration	1-7	8-14	15-21	22-28	29-35	36-42	43-49	50-56	57-63	64-70	71-77	78-84	85-91
Crust or staining around eyes or nares	0 ppm	0	0	0	0	0	2	0	1	0	0	0	0	0
	300 ppm	0	0	0	0	1	1	0	0	0	0	0	0	0
	1000 ppm	0	0	0	0	0	0	0	0	0	0	0	0	1
	3000 ppm	1	0	0	0	0	0	0	0	0	0	0	1	0
Perineal soiling or staining	0 ppm	0	2	0	0	0	1	0	1	0	1	1	0	0
	300 ppm	0	0	0	0	0	1	0	0	1	0	0	0	1
	1000 ppm	0	0	0	0	0	0	0	0	0	0	0	0	1
	3000 ppm	0	0	0	0	2	1	0	0	0	0	0	0	1

 

Table 6: Organ and organ/body weights – males – 13 weeks

		Final body weight (g)	Adrenals		Brain		Heart		Kidneys		Liver		Lungs
Concentration (ppm)			(g)	(g/100)	(g)	(g/100)	(g)	(g/100)	(g)	(g/100)	(g)	(g/100)	(g)	(g/100)
0	Mean	261.1	0.047	0.0182	1.873	0.718	0.835	0.320	1.874	0.718	6.819	2.611	0.967	0.371
	SD	12.9	0.005	0.0024	0.054	0.025	0.045	0.015	0.098	0.016	0.479	0.105	0.040	0.018
	N	10	10	10	10	10	10	10	10	10	10	10	10	10
300	Mean	260.6	0.047	0.0182	1.878	0.725	0.849	0.326	1.848	0.711	6.716	2.574	0.954	0.367
	SD	24.5	0.005	0.0024	0.046	0.059	0.090	0.025	0.160	0.037	0.789	0.109	0.067	0.016
	N	10	10	10	10	10	10	10	10	10	10	10	10	10
1000	Mean	270.8	0.048	0.0178	1.883	0.696	0.841	0.311	1.901	0.702	7.171	2.649	0.998	0.369
	SD	6.7	0.004	0.0012	0.026	0.018	0.064	0.028	0.072	0.020	0.251	0.073	0.044	0.017
	N	10	10	10	10	10	10	10	10	10	10	10	10	10
3000	Mean	271.5	0.046	0.0170	1.857	0.685	0.870	0.321	1.926	0.710	7.808*	2.874*	1.028*	0.379
	SD	7.6	0.003	0.0013	0.032	0.018	0.084	0.033	0.079	0.026	0.325	0.082	0.041	0.018
	N	10	10	10	10	10	10	10	10	10	10	10	10	10

 

Table 6 (cont): Organ and organ/body weights – males – 13 weeks

		Final body weight (g)	Spleen		Testes		Thymus
Concentration (ppm)			(g)	(g/100)	(g)	(g/100)	(g)	(g/100)
0	Mean	261.1	0.552	0.211	3.077	1.180	0.169	0.065
	SD	12.9	0.048	0.011	0.093	0.043	0.027	0.011
	N	10	10	10	10	10	10	10
300	Mean	260.6	0.566	0.218	2.978	1.146	0.164	0.063
	SD	24.5	0.045	0.013	0.272	0.088	0.026	0.008
	N	10	10	10	10	10	10	10
1000	Mean	270.8	0.571	0.211	3.184*	1.177	0.169	0.063
	SD	6.7	0.027	0.012	0.079	0.043	0.013	0.004
	N	10	10	10	10	10	10	10
3000	Mean	271.5	0.523	0.193*	3.156	1.163	0.162	0.060
	SD	7.6	0.036	0.007	0.224	0.063	0.018	0.007
	N	10	10	10	10	10	10	10

 

Table 7: Organ and organ/body weights – females – 13 weeks

		Final body weight (g)	Adrenals		Brain		Heart		Kidneys		Liver		Lungs
Concentration (ppm)			(g)	(g/100)	(g)	(g/100)	(g)	(g/100)	(g)	(g/100)	(g)	(g/100)	(g)	(g/100)
0	Mean	159.1	0.048	0.0304	1.761	1.110	0.593	0.373	1.187	0.747	4.182	2.630	0.785	0.495
	SD	9.2	0.003	0.0028	0.037	0.062	0.025	0.022	0.062	0.024	0.244	0.068	0.039	0.037
	N	10	10	10	10	10	10	10	10	10	10	10	10	10
300	Mean	163.4	0.050	0.0309	1.791	1.098	0.616	0.377	1.229	0.753	4.286	2.624	0.796	0.487
	SD	6.5	0.003	0.0015	0.025	0.048	0.030	0.023	0.046	0.039	0.155	0.087	0.037	0.025
	N	10	10	10	10	10	10	10	10	10	10	10	10	10
1000	Mean	155.5	0.050	0.0321	1.743	1.122	0.598	0.384	1.164	0.747	4.285	2.754*	0.783	0.503
	SD	7.8	0.006	0.0041	0.051	0.033	0.063	0.035	0.092	0.027	0.306	0.121	0.067	0.027
	N	10	10	10	10	10	10	10	10	10	10	10	10	10
3000	Mean	159.5	0.049	0.0309	1.715*	1.076	0.620	0.389	1.194	0.749	4.496*	2.819*	0.784	0.491
	SD	6.3	0.004	0.002	0.030	0.036	0.039	0.025	0.064	0.024	0.184	0.069	0.033	0.014
	N	10	10	10	10	10	10	10	10	10	10	10	10	10

 

Table 7 (cont): Organ and organ/body weights – females – 13 weeks

		Final body weight (g)	Spleen		Thymus
Concentration (ppm)			(g)	(g/100)	(g)	(g/100)
0	Mean	159.1	0.488	0.306	0.173	0.109
	SD	9.2	0.056	0.023	0.018	0.009
	N	10	10	10	10	10
300	Mean	163.4	0.474	0.290	0.189	0.116
	SD	6.5	0.041	0.023	0.014	0.008
	N	10	10	10	10	10
1000	Mean	155.5	0.421	0.271*	0.171	0.110
	SD	7.8	0.051	0.028	0.030	0.017
	N	10	10	10	10	10
3000	Mean	159.5	0.358*	0.225*	0.171	0.107
	SD	6.3	0.016	0.007	0.018	0.009
	N	10	10	10	10	10

 

Table 8: Organ and organ/body weights – males – 8-week recovery

		Final body weight (g)	Adrenals		Brain		Heart		Kidneys		Liver		Lungs
Concentration (ppm)			(g)	(g/100)	(g)	(g/100)	(g)	(g/100)	(g)	(g/100)	(g)	(g/100)	(g)	(g/100)
0	Mean	300.7	0.045	0.0148	1.970	0.656	0.852	0.284	2.014	0.670	7.348	2.444	1.066	0.355
	SD	14.9	0.007	0.0025	0.041	0.030	0.049	0.019	0.108	0.027	0.459	0.094	0.062	0.018
	N	10	10	10	10	10	10	10	10	10	10	10	10	10
300	Mean	292.5	0.045	0.0153	1.991	0.682	0.809	0.277	2.016	0.690	7.243	2.478	1.041	0.357
	SD	17.0	0.006	0.0019	0.051	0.039	0.057	0.011	0.123	0.029	0.404	0.090	0.022	0.018
	N	10	10	10	10	10	10	10	10	10	10	10	10	10
1000	Mean	289.9	0.046	0.0158	1.948	0.674	0.825	0.286	1.962	0.677	6.909	2.381	1.012	0.349
	SD	19.9	0.004	0.0022	0.049	0.040	0.034	0.021	0.139	0.029	0.632	0.090	0.064	0.014
	N	10	10	10	10	10	10	10	10	10	10	10	10	10
3000	Mean	305.9	0.047	0.0154	1.957	0.641	0.855	0.280	2.089	0.684	7.586	2.478	1.136*	0.372
	SD	17.4	0.007	0.0023	0.41	0.029	0.060	0.015	0.175	0.059	0.637	0.132	0.066	0.015
	N	10	10	10	10	10	10	10	10	10	10	10	10	10

 

Table 8 (cont): Organ and organ/body weights – males – 8-week recovery

		Final body weight (g)	Spleen		Testes		Thymus
Concentration (ppm)			(g)	(g/100)	(g)	(g/100)	(g)	(g/100)
0	Mean	300.7	0.552	0.183	3.215	1.070	0.121	0.040
	SD	14.9	0.043	0.008	0.271	0.087	0.013	0.004
	N	10	10	10	10	10	10	10
300	Mean	292.5	0.536	0.183	3.121	1.068	0.111	0.038
	SD	17.0	0.034	0.007	0.159	0.044	0.022	0.007
	N	10	10	10	10	10	10	10
1000	Mean	289.9	0.527	0.182	2.994	1.036	0.116	0.040
	SD	19.9	0.048	0.014	0.301	0.118	0.016	0.005
	N	10	10	10	10	10	10	10
3000	Mean	305.9	0.552	0.180	3.201	1.049	0.128	0.042
	SD	17.4	0.045	0.007	0.109	0.049	0.020	0.005
	N	10	10	10	10	10	10	10

 

Table 9: Organ and organ/body weights – females – 8-week recovery

		Final body weight (g)	Adrenals		Brain		Heart		Kidneys		Liver		Lungs
Concentration (ppm)			(g)	(g/100)	(g)	(g/100)	(g)	(g/100)	(g)	(g/100)	(g)	(g/100)	(g)	(g/100)
0	Mean	172.3	0.053	0.0304	1.825	1.062	0.591	0.343	1.241	0.721	4.339	2.517	0.854	0.496
	SD	10.1	0.007	0.0031	0.041	0.058	0.037	0.016	0.074	0.033	0.347	0.113	0.067	0.016
	N	10	10	10	10	10	10	10	10	10	10	10	10	10
300	Mean	171.0	0.053	0.0310	1.839	1.079	0.609	0.357	1.273	0.745	4.391	2.569	0.851	0.498
	SD	12.0	0.005	0.0031	0.067	0.065	0.043	0.020	0.084	0.022	0.321	0.076	0.072	0.029
	N	10	10	10	10	10	10	10	10	10	10	10	10	10
1000	Mean	175.2	0.053	0.0319	1.839	1.051	0.636*	0.363	1.292	0.738	4.535	2.588	0.867	0.495
	SD	5.5	0.005	0.0026	0.024	0.031	0.043	0.026	0.071	0.046	0.303	0.124	0.055	0.026
	N	10	10	10	10	10	10	10	10	10	10	10	10	10
3000	Mean	171.3	0.053	0.0306	1.799	1.052	0.621	0.363	1.245	0.727	4.343	2.533	0.855	0.499
	SD	6.6	0.005	0.0022	0.040	0.056	0.029	0.021	0.050	0.026	0.279	0.091	0.046	0.024
	N	10	10	10	10	10	10	10	10	10	10	10	10	10

 

Table 9 (cont): Organ and organ/body weights – females – 8-week recovery

		Final body weight (g)	Spleen		Thymus
Concentration (ppm)			(g)	(g/100)	(g)	(g/100)
0	Mean	172.3	0.440	0.255	0.133	0.077
	SD	10.1	0.046	0.019	0.015	0.007
	N	10	10	10	10	10
300	Mean	171.0	0.451	0.264	0.136	0.080
	SD	12.0	0.038	0.016	0.013	0.005
	N	10	10	10	10	10
1000	Mean	175.2	0.439	0.251	0.133	0.076
	SD	5.5	0.033	0.013	0.012	0.006
	N	10	10	10	10	10
3000	Mean	171.3	0.456	0.266	0.142	0.083
	SD	6.6	0.028	0.015	0.013	0.006
	N	10	10	10	10	10

 

Table 10: Urine specific gravity - males

Exposure concentration ppm		13-week main group	13-week main group repeat	13-week satellite	4-week recovery satellite	8-week recovery satellite
0	Mean	1.054	1.058	1.057	1.063	1.058
	SD	0.012	0.006	0.005	0.002	0.004
	N	10	10	10	10	10
300	Mean	Data missing	1.056	1.058	1.056*	1.058
	SD	Data missing	0.005	0.005	0.008	0.004
	N	10	10	10	10	10
1000	Mean	1.047	1.056	1.047	1.055*	1.055
	SD	0.012	0.004	0.017	0.004	0.009
	N	10	10	10	10	10
3000	Mean	1.038*	1.049*	1.048*	1.055*	1.052
	SD	0.010	0.008	0.009	0.008	0.007
	N	10	10	10	10	10

 

Historical control summary

	Urinary specific gravity
Number of control groups	21
Total number of animals	192
Minimum control group mean	1.044
Maximum control group mean	1.067
Mean of control group means	1.060
SD of control group mean	0.006
Average control group SD	0.010

 

Table 11: Urine specific gravity - females

Exposure concentration ppm		13-week main group	13-week main group repeat	13-week satellite	4-week recovery satellite	8-week recovery satellite
0	Mean	1.035	1.048	1.042	1.051	1.042
	SD	0.013	0.014	0.010	0.007	0.009
	N	10	10	10	10	10
300	Mean	1.043	1.053	1.049	1.051	1.050*
	SD	0.015	0.010	0.009	0.009	0.006
	N	10	10	10	10	10
1000	Mean	1.033	1.038	1.040	1.051	1.051*
	SD	0.012	0.013	0.014	0.007	0.008
	N	10	10	10	10	10
3000	Mean	1.016*	1.018*	1.025*	1.048	1.046
	SD	0.010	0.010	0.007	0.006	0.007
	N	10	10	10	10	10

 

Historical control summary

	Urinary specific gravity
Number of control groups	20
Total number of animals	191
Minimum control group mean	1.035
Maximum control group mean	1.069
Mean of control group means	1.055
SD of control group mean	0.009
Average control group SD	0.011

 

Table 12: Clinical chemistry – males – 4-week satellite

Dose Ppm		UN (mg/dL)	ALT (mU/mL)	AP (mU/mL)	AST (mU/mL)	GLUC (mg/dL)	TP (g/dL)	ALB (g/dL)	GLB (g/dL)	CHOL (mg/dL)	TRIG (mg/dL)	TBILI (mg/dL)	CREAT (mg/dL)
0	Mean	19	43	150	94	116	7.2	4.2	3.1	67	73	0.2	0.7
	SD	2	5	11	14	9	0.4	0.2	0.2	9	13	0.0	0.1
	N	10	10	10	10	10	10	10	10	10	10	10	10
300	Mean	19	40	145	82*	114	7.2	4.1	3.1	68	66	0.2	0.7
	SD	2	6	10	10	9	0.3	0.1	0.2	8	20	0.1	0.1
	N	10	10	10	10	10	10	10	10	10	10	10	10
1000	Mean	19	42	146	86	110	7.2	4.2	3.1	66	67	0.2	0.7
	SD	2	3	13	8	6	0.3	0.1	0.2	5	27	0.1	0.1
	N	10	10	10	10	10	10	10	10	10	10	10	10
3000	Mean	18	48*	135*	96	113	7.4	4.2	3.2	69	46*	0.2	0.8
	SD	1	3	11	11	7	0.3	0.2	0.2	5	17	0.0	0.1
	N	10	10	10	10	10	10	10	10	10	10	10	10

 

 

Table 13: Clinical chemistry – females – 4-week satellite

Dose Ppm		UN (mg/dL)	ALT (mU/mL)	AP (mU/mL)	AST (mU/mL)	GLUC (mg/dL)	TP (g/dL)	ALB (g/dL)	GLB (g/dL)	CHOL (mg/dL)	TRIG (mg/dL)	TBILI (mg/dL)	CREAT (mg/dL)
0	Mean	24	38	127	96	88	7.5	4.4	3.1	114	58	0.2	0.8
	SD	2	3	12	7	10	0.4	0.2	0.3	12	29	0.1	0.1
	N	10	10	10	10	10	10	10	10	10	10	10	10
300	Mean	23	37	125	90	97*	7.9	4.5	3.4	114	55	0.2	0.7
	SD	2	4	12	7	5	0.4	0.3	0.2	12	20	0.1	0.2
	N	10	10	10	10	10	10	10	10	10	10	10	10
1000	Mean	23	41	129	95	89	7.5	4.4	3.2	104	56	0.2	0.8
	SD	3	6	13	16	8	0.3	0.2	0.3	9	21	0.1	0.1
	N	10	10	10	10	10	10	10	10	10	10	10	10
3000	Mean	23	48*	110*	107	88	7.6	4.4	3.2	109	45	0.2	0.8
	SD	3	7	11	18	6	0.3	0.2	0.2	6	12	0.0	0.1
	N	10	10	10	10	10	10	10	10	10	10	10	10

 

Table 14: Clinical chemistry – males – 13-week main group

Dose Ppm		UN (mg/dL)	ALT (mU/mL)	AP (mU/mL)	AST (mU/mL)	GLUC (mg/dL)	TP (g/dL)	ALB (g/dL)	GLB (g/dL)	CHOL (mg/dL)	TRIG (mg/dL)	TBILI (mg/dL)	CREAT (mg/dL)
0	Mean	19	62	87	101	108	7.1	4.2	2.9	80	79	0.2	0.7
	SD	2	18	8	23	9	0.2	0.1	0.2	6	18	0.1	0.1
	N	10	10	10	10	10	10	10	10	10	10	10	10
300	Mean	20	58	84	85	106	7.1	4.2	2.9	81	70	0.2	0.6
	SD	2	11	6	12	11	0.2	0.2	0.2	7	16	0.1	0.1
	N	10	10	10	10	10	10	10	10	10	10	10	10
1000	Mean	21	64	85	107	115	7.3	4.2	3.0	80	60*	0.1	0.7
	SD	2	12	5	38	13	0.3	0.1	0.2	4	16	0.0	0.1
	N	10	10	10	10	10	10	10	10	10	10	10	10
3000	Mean	23*	69	79*	93	114	7.4*	4.3	3.1	92*	61*	0.2	0.7
	SD	3	13	3	11	16	0.2	0.1	0.2	9	11	0.1	0.0
	N	10	10	10	10	10	10	10	10	10	10	10	10

 

Table 15: Clinical chemistry – females – 13-week main group

Dose Ppm		UN (mg/dL)	ALT (mU/mL)	AP (mU/mL)	AST (mU/mL)	GLUC (mg/dL)	TP (g/dL)	ALB (g/dL)	GLB (g/dL)	CHOL (mg/dL)	TRIG (mg/dL)	TBILI (mg/dL)	CREAT (mg/dL)
0	Mean	22	44	69	122	94	6.9	4.0	2.9	107	35	0.2	0.7
	SD	3	14	13	55	7	0.3	0.2	0.2	17	6	0.0	0.1
	N	10	10	10	10	10	10	10	10	10	10	10	10
300	Mean	25	57	72	127	107*	7.2	4.1	3.1	115	32	0.2	0.8
	SD	5	16	13	35	13	0.3	0.1	0.2	11	9	0.0	0.1
	N	10	10	10	10	10	10	10	10	10	10	10	10
1000	Mean	25	56	68	119	109*	7.3*	4.2*	3.1	115	28	0.2	0.7
	SD	2	20	8	30	10	0.3	0.1	0.3	17	10	0.0	0.0
	N	10	10	10	10	10	10	10	10	10	10	10	10
3000	Mean	23	72*	64	140	111*	7.3*	4.2	3.1	121	29	0.2	0.7
	SD	3	27	4	50	7	0.3	0.1	0.3	14	8	0.0	0.1
	N	10	10	10	10	10	10	10	10	10	10	10	10

 

Table 16: Clinical chemistry – males – 4-week recovery satellite

Dose Ppm		UN (mg/dL)	ALT (mU/mL)	AP (mU/mL)	AST (mU/mL)	GLUC (mg/dL)	TP (g/dL)	ALB (g/dL)	GLB (g/dL)	CHOL (mg/dL)	TRIG (mg/dL)	TBILI (mg/dL)	CREAT (mg/dL)
0	Mean	17	67	82	148	75	6.3	3.6	2.8	68	82	0.3	0.6
	SD	1	14	12	34	10	0.2	0.2	0.2	8	19	0.1	0.1
	N	10	10	10	10	10	10	10	10	10	10	10	10
300	Mean	17	63	78	116*	81	6.3	3.5	2.8	65	76	0.2	0.6
	SD	3	10	8	22	5	0.1	0.1	0.1	6	17	0.0	0.1
	N	10	10	10	10	10	10	10	10	10	10	10	10
1000	Mean	17	69	80	138	80	6.4	3.6	2.8	70	86	0.2	0.6
	SD	2	12	7	20	8	0.2	0.1	0.2	6	24	0.0	0.1
	N	10	10	10	10	10	10	10	10	10	10	10	10
3000	Mean	19	70	82	140	79	6.5	3.6	2.9	75*	76	0.2	0.6
	SD	4	15	9	31	7	0.2	0.1	0.1	5	28	0.0	0.0
	N	10	10	10	10	10	10	10	10	10	10	10	10

 

Table 17: Clinical chemistry – males – 4-week recovery satellite

Dose Ppm		UN (mg/dL)	ALT (mU/mL)	AP (mU/mL)	AST (mU/mL)	GLUC (mg/dL)	TP (g/dL)	ALB (g/dL)	GLB (g/dL)	CHOL (mg/dL)	TRIG (mg/dL)	TBILI (mg/dL)	CREAT (mg/dL)
0	Mean	22	43	73	100	67	6.7	3.9	2.8	106	52	0.2	0.6
	SD	2	6	9	11	9	0.3	0.1	0.4	10	27	0.1	0.1
	N	10	10	10	10	10	10	10	10	10	10	10	10
300	Mean	22	47	68	98	70	6.6	3.8	2.8	104	43	0.3	0.6
	SD	3	9	7	11	8	0.3	0.2	0.2	11	7	0.2	0.1
	N	10	10	10	10	10	10	10	10	10	10	10	10
1000	Mean	22	44	73	99	75	6.7	3.8	2.8	105	37	0.2	0.6
	SD	3	8	12	10	8	0.2	0.2	0.2	11	7	0.0	0.0
	N	10	10	10	10	10	10	10	10	10	10	10	10
3000	Mean	24	48	76	101	67	6.6	3.8	2.8	107	56	0.2	0.6
	SD	6	12	14	17	7	0.2	0.2	0.2	9	32	0.1	0.0
	N	10	10	10	10	10	10	10	10	10	10	10	10

 

Table 18: Clinical chemistry – males – 90 days – Historical control summary

	UN (mg/dL)	ALT (mU/mL)	AP (mU/mL)	AST (mU/mL)	GLUC (mg/dL)	Protein (g/dL)	ALB (g/dL)	GLB (g/dL)	CHOL (mg/dL)	TRIG (mg/dL)
Number of control groups	22	22	22	11	21	13	13	13	3	3
Total number of animals	212	212	212	106	202	125	125	125	26	26
Minimum control group mean	14	18	62	77	120	5.7	3.2	2.2	40	53
Maximum control group mean	22	92	94	175	182	6.6	4.0	2.8	43	67
Mean of control group means	17	39	71	116	148	6.1	3.6	2.5	41	61
SD of control group mean	2	19	7	29	17	0.3	0.2	0.2	1	6
Average control group SD	2	19	9	39	17	0.3	0.2	0.2	5	11

 

Table 18 (cont): Clinical chemistry – males – 90 days – Historical control summary

	TBILI (mg/dL)	CK (mU/mL)	CREAT (mg/dL)	CA (mmol/L)	P (mmol/L)	Na (mmol/L)	K (mmol/L)	Cl (mmol/L)
Number of control groups	9	0	4	5	5	1	1	1
Total number of animals	87	0	40	50	50	10	10	10
Minimum control group mean	0.1	No data	0.6	9.5	6.6	140	5.1	104
Maximum control group mean	0.4	No data	0.6	11.2	11.2	140	5.1	104
Mean of control group means	0.2	No data	0.6	10.0	7.9	140	5.1	104
SD of control group mean	0.1	No data	0.0	0.6	1.7	0	0.0	0
Average control group SD	0.1	No data	0.1	0.2	0.9	2	0.3	2

 

Table 19: Clinical chemistry – females – 90 days – Historical control summary

	UN (mg/dL)	ALT (mU/mL)	AP (mU/mL)	AST (mU/mL)	GLUC (mg/dL)	Protein (g/dL)	ALB (g/dL)	GLB (g/dL)	CHOL (mg/dL)	TRIG (mg/dL)
Number of control groups	21	21	21	10	20	12	12	12	3	3
Total number of animals	204	204	204	98	194	114	114	114	26	26
Minimum control group mean	14	14	32	58	101	5.4	3.2	2.0	57	37
Maximum control group mean	21	49	74	148	152	6.7	4.0	2.9	59	39
Mean of control group means	17	27	54	104	130	5.9	3.6	2.3	58	38
SD of control group mean	2	13	10	30	13	0.4	0.2	0.3	1	1
Average control group SD	2	6	8	22	11	0.3	0.2	0.2	8	5

 

Table 19 (cont): Clinical chemistry – females – 90 days – Historical control summary

	TBILI (mg/dL)	CK (mU/mL)	CREAT (mg/dL)	CA (mmol/L)	P (mmol/L)	Na (mmol/L)	K (mmol/L)	Cl (mmol/L)
Number of control groups	9	0	4	5	5	1	1	1
Total number of animals	88	0	40	48	48	8	8	8
Minimum control group mean	0.2	No data	0.5	9.2	6.3	141	5.7	107
Maximum control group mean	0.4	No data	0.7	11.0	11.1	141	5.7	107
Mean of control group means	0.3	No data	0.6	9.9	7.9	141	5.7	107
SD of control group mean	0.1	No data	0.1	0.6	1.8	0	0.0	0
Average control group SD	0.1	No data	0.1	0.3	0.9	1	0.4	1

 

Table 20: Hematology – males – 4-week interim satellite

Dose ppm		RBC X10E6 /CU/MM	HGB G/dL	HCT %	PLAT X10E3 /CU/MM	WBC X10E3 /CU/MM
0	Mean	8.04	17.5	48.4	983	10.7
	SD	0.12	0.3	0.7	47	2.1
	N	10	10	10	10	10
300	Mean	8.14	17.8	48.9	1032	9.4
	SD	0.22	0.6	1.5	57	1.2
	N	10	10	10	10	10
1000	Mean	8.15	17.7	48.6	1046*	8.2*
	SD	0.15	0.4	0.8	41	0.8
	N	10	10	10	10	10
3000	Mean	8.06	17.2	47.8	1117*	5.5*
	SD	0.27	0.5	1.4	54	1.0
	N	10	10	10	10	10

 

Table 21: Hematology – females – 4-week interim satellite

Dose ppm		RBC X10E6 /CU/MM	HGB G/dL	HCT %	PLAT X10E3 /CU/MM	WBC X10E3 /CU/MM
0	Mean	7.43	17.0	46.9	1064	7.3
	SD	0.31	0.6	1.7	43	1.4
	N	10	10	10	10	10
300	Mean	7.63	17.6	48.1	1129*	8.7*
	SD	0.25	0.6	1.5	42	1.6
	N	10	10	10	10	10
1000	Mean	7.59	17.2	47.4	1149*	6.2
	SD	0.34	0.8	2.2	59	0.8
	N	10	10	10	10	10
3000	Mean	7.68	17.2	47.5	1159*	49*
	SD	0.21	0.5	1.3	67	0.9
	N	10	10	10	10	10

 

Table 22: Hematology – males – 13-week main group

Dose ppm		RBC X10E6 /CU/MM	HGB G/dL	HCT %	PLAT X10E3 /CU/MM	WBC X10E3 /CU/MM
0	Mean	8.36	17.1	48.2	833	9.1
	SD	0.15	0.3	1.2	41	1.3
	N	10	10	10	10	10
300	Mean	8.41	17.3	48.6	862	9.6
	SD	0.20	0.4	1.3	47	1.4
	N	10	10	10	10	10
1000	Mean	8.48	17.2	48.3	917*	8.6
	SD	0.16	0.4	0.9	42	1.0
	N	10	10	10	10	10
3000	Mean	8.47	16.8	47.5	1114*	6.0*
	SD	0.12	0.3	1.0	38	1.0
	N	10	10	10	10	10

 

Table 23: Hematology – males – 13-week satellite

Dose ppm		RBC X10E6 /CU/MM	HGB G/dL	HCT %	PLAT X10E3 /CU/MM	WBC X10E3 /CU/MM
0	Mean	8.79	17.8	49.8	898	8.8
	SD	0.15	0.4	0.8	57	1.1
	N	10	10	10	10	10
300	Mean	8.69	17.6	49.5	896	9.3
	SD	0.23	0.5	1.8	36	1.2
	N	10	10	10	10	10
1000	Mean	8.94	18.0	50.0	946*	8.5
	SD	0.12	0.3	1.2	22	1.2
	N	10	10	10	10	10
3000	Mean	8.80	17.3	48.5	1159*	6.2*
	SD	0.30	0.7	1.7	46	0.8
	N	10	10	10	10	10

 

Table 24: Hematology – females – 13-week main group

Dose ppm		RBC X10E6 /CU/MM	HGB G/dL	HCT %	PLAT X10E3 /CU/MM	WBC X10E3 /CU/MM
0	Mean	7.78	16.9	47.8	916	9.2
	SD	0.24	0.5	1.5	47	0.9
	N	10	10	10	10	10
300	Mean	7.95	17.3	48.5	927	9.1
	SD	0.18	0.4	1.1	39	1.1
	N	10	10	10	10	10
1000	Mean	8.03*	17.0	48.2	994*	7.8*
	SD	0.17	0.4	0.9	46	0.5
	N	10	10	10	10	10
3000	Mean	8.07*	16.7	47.5	1139*	5.1*
	SD	0.18	0.4	0.7	91	0.6
	N	10	10	10	10	10

 

Table 25: Hematology – females – 13-week satellite

Dose ppm		RBC X10E6 /CU/MM	HGB G/dL	HCT %	PLAT X10E3 /CU/MM	WBC X10E3 /CU/MM
0	Mean	7.92	16.9	49.5	965	7.3
	SD	0.35	0.9	2.3	62	1.0
	N	10	10	10	10	10
300	Mean	7.99	17.2	49.5	963	7.1
	SD	0.22	0.5	1.5	33	0.9
	N	10	10	10	10	10
1000	Mean	8.13	17.2	49.5	1045*	5.6*
	SD	0.18	0.5	1.2	48	0.9
	N	10	10	10	10	10
3000	Mean	8.29*	17.2	49.6	1150*	4.3*
	SD	0.18	0.3	1.2	85	0.6
	N	10	10	10	10	10

 

Table 26: Hematology – males – 4-week recovery satellite

Dose ppm		RBC X10E6 /CU/MM	HGB G/dL	HCT %	PLAT X10E3 /CU/MM	WBC X10E3 /CU/MM
0	Mean	8.51	17.4	50.7	888	10.2
	SD	0.21	0.6	1.1	52	1.0
	N	10	10	10	10	10
300	Mean	8.51	17.3	50.1	914	8.2*
	SD	0.16	0.4	1.3	70	0.8
	N	10	10	10	10	10
1000	Mean	8.49	17.2	50.2	886	8.7*
	SD	0.20	0.3	1.1	42	1.5
	N	10	10	10	10	10
3000	Mean	8.38	16.9*	49.3*	969*	8.9*
	SD	0.25	0.4	1.2	31	0.9
	N	10	10	10	10	10

 

Table 27: Hematology – females – 4-week recovery satellite

Dose ppm		RBC X10E6 /CU/MM	HGB G/dL	HCT %	PLAT X10E3 /CU/MM	WBC X10E3 /CU/MM
0	Mean	7.76	17.4	49.7	918	7.2
	SD	0.19	0.6	1.3	53	1.3
	N	10	10	10	10	10
300	Mean	7.76	17.0	49.5	889	6.4
	SD	0.20	0.6	1.3	65	1.1
	N	10	10	10	10	10
1000	Mean	7.85	17.3	49.9	914	6.1
	SD	0.32	0.6	1.9	50	0.9
	N	10	10	10	10	10
3000	Mean	7.77	17.0	49.2	913	6.0*
	SD	0.15	0.5	0.9	55	1.0
	N	10	10	10	10	10

 

Table 28: Hematology – males – 8-week recovery satellite

Dose ppm		RBC X10E6 /CU/MM	HGB G/dL	HCT %	PLAT X10E3 /CU/MM	WBC X10E3 /CU/MM
0	Mean	8.88	18.2	51.6	910	10.0
	SD	0.23	0.5	1.1	36	1.2
	N	10	10	10	10	10
300	Mean	8.78	17.9	50.8	880	9.1
	SD	0.22	0.4	1.4	45	1.1
	N	10	10	10	10	10
1000	Mean	8.88	18.3	51.8	838*	8.4*
	SD	0.22	0.4	1.0	91	0.5
	N	10	10	10	10	10
3000	Mean	8.64*	17.5	50.1*	913	8.6*
	SD	0.15	0.4	0.9	72	1.2
	N	10	10	10	10	10

 

Table 29: Hematology – females – 8-week recovery satellite

Dose ppm		RBC X10E6 /CU/MM	HGB G/dL	HCT %	PLAT X10E3 /CU/MM	WBC X10E3 /CU/MM
0	Mean	7.85	17.6	49.5	926	6.4
	SD	0.20	0.4	1.1	43	1.1
	N	10	10	10	10	10
300	Mean	7.90	17.6	49.0	940	6.7
	SD	0.19	0.3	3.2	134	0.8
	N	10	10	10	10	10
1000	Mean	7.86	17.5	49.6	918	6.0
	SD	0.16	0.4	1.1	55	0.6
	N	10	10	10	10	10
3000	Mean	7.85	17.5	49.6	912	5.8
	SD	0.19	0.4	1.2	43	0.6
	N	10	10	10	10	10

 

Table 30: Mean WBC – males

Exposure conc. ppm		4-week satellite WBC X10E3 / CU MM	13-week WBC X10E3 /CU MM	13-week satellite WBC X10E3 /CU MM	4-week recovery satellite WBC X10E3 /CU MM	8-week recovery satellite WBC X 10E3 /CU MM
0	Mean	10.7	9.1	8.8	10.2	10.0
	SD	2.1	1.3	1.1	1.0	1.2
	N	10	10	10	10	10
300	Mean	9.4	9.6	9.3	8.2*	9.1
	SD	1.2	1.4	1.2	0.8	1.1
	N	10	10	10	10	10
1000	Mean	8.2*	8.6	8.5	8.7*	8.4*
	SD	0.8	1.0	1.2	1.5	0.5
	N	10	10	10	10	10
3000	Mean	5.5*	6.0*	6.2*	8.9*	8.6*
	SD	1.0	1.0	0.8	0.9	1.2
	N	10	10	10	10	10

 

Table 31: Mean WBC – females

Exposure conc. ppm		4-week satellite WBC X10E3 / CU MM	13-week WBC X10E3 /CU MM	13-week satellite WBC X10E3 /CU MM	4-week recovery satellite WBC X10E3 /CU MM	8-week recovery satellite WBC X 10E3 /CU MM
0	Mean	7.3	9.2	7.3	7.2	6.4
	SD	1.4	0.9	1.0	1.3	1.1
	N	10	10	10	10	10
300	Mean	8.7*	9.1	7.1	6.4	6.7
	SD	1.6	1.1	0.9	1.1	0.8
	N	10	10	10	10	10
1000	Mean	6.2	7.8*	5.6*	6.1	6.0
	SD	0.8	0.5	0.9	0.9	0.5
	N	10	10	10	10	10
3000	Mean	4.9*	5.1*	4.3*	6.0*	5.8
	SD	0.9	0.6	0.6	1.0	0.5
	N	10	10	10	10	10

 

Table 32: Hematology – males – historical control summary

	RBC X10E6 /CU MM	HGB (G/dL)	HCT %	PLAT. X10E3 /CU MM	WBC X10E3 /CU MM
Number of control groups	22	22	22	12	20
Total number of animals	203	203	203	120	187
Minimum control group mean	7.83	15.7	42.2	502	4.9
Maximum control group mean	9.41	18.0	52.9	1281	19.7
Mean of control group means	8.72	16.8	48.2	833	10.2
SD of control group mean	0.41	0.6	2.6	191	4.2
Average control group SD	0.35	0.6	1.8	77	1.9

 

Table 33: Hematology – females – historical control summary

	RBC X10E6 /CU MM	HGB (G/dL)	HCT %	PLAT. X10E3 /CU MM	WBC X10E3 /CU MM
Number of control groups	21	21	21	11	21
Total number of animals	191	191	191	109	191
Minimum control group mean	7.57	15.7	42.4	582	5.7
Maximum control group mean	8.63	17.5	50.6	1317	15.1
Mean of control group means	8.06	16.5	47.1	846	8.7
SD of control group mean	0.31	0.5	1.9	182	3.3
Average control group SD	0.30	0.5	2.0	87	1.8

 

Table 34: Mean absolute lymphocytes – males

Exposure conc. ppm		4-week satellite	13-week	13-week satellite	4-week recovery satellite	8-week recovery satellite
0	Mean	7982.60	6450.00	5960.00	7316.00	6555.40
	SD	1465.89	989.53	888.65	1054.86	738.47
	N	10	10	10	10	10
300	Mean	7211.00	6949.80	6581.20	5846.80*	6247.20
	SD	909.82	1254.39	786.52	486.27	837.66
	N	10	10	10	10	10
1000	Mean	5972.20*	5820.20	5072.60*	6029.60*	5524.40*
	SD	715.95	788.45	592.60	1014.74	390.67
	N	10	10	10	10	10
3000	Mean	3564.80*	3645.00*	3816.60*	6179.80*	5908.40
	SD	629.22	899.30	571.21	864.17	966.02
	N	10	10	10	10	10

 

Table 35: Mean absolute segmented neutrophils – females

Exposure conc. ppm		4-week satellite	13-week	13-week satellite	4-week recovery satellite	8-week recovery satellite
0	Mean	1339.60	1785.20	2108.80	1674.20	1452.00
	SD	452.08	515.10	651.71	557.30	540.03
	N	10	10	10	10	10
300	Mean	1564.80	1597.80	1859.80	1516.60	1402.00
	SD	434.61	415.99	254.44	375.42	156.43
	N	10	10	10	10	10
1000	Mean	1305.80	1608.80	1607.20*	1411.60	1390.80
	SD	294.71	277.63	334.10	204.15	299.15
	N	10	10	10	10	10
3000	Mean	1295.80	1043.60*	1469.20*	1593.20	1209.00
	SD	575.83	307.49	416.88	695.71	222.22
	N	10	10	10	10	10

 

Table 36: Mean absolute lymphocytes – females

Exposure conc. ppm		4-week satellite	13-week	13-week satellite	4-week recovery satellite	8-week recovery satellite
0	Mean	5662.80	7105.40	4956.40	5290.00	4747.00
	SD	1164.92	657.44	563.10	941.32	785.90
	N	10	10	10	10	10
300	Mean	6800.00*	7144.80	4977.20	4680.40	5023.00
	SD	1354.17	967.32	592.94	849.77	617.74
	N	10	10	10	10	10
1000	Mean	4690.80	5908.60	3816.00*	4487.80	4390.00
	SD	752.39	458.09	749.67	692.09	504.94
	N	10	10	10	10	10
3000	Mean	3456.80*	3868.40*	2618.20*	4120.60*	4390.40
	SD	431.66	503.36	292.44	791.87	552.94
	N	10	10	10	10	10

 

Applicant's summary and conclusion

Conclusions:

The primary effect of exposure to 1,3-dioxolane at 1000 and 3000 ppm was a reduction in white blood cell (WBC) counts, primarily due to decreased numbers of lymphocytes. Despite the decreased WBC counts, rats appeared healthy and there were only minimal morphological effects in those exposed to 3000 ppm 1,3-dioxolane. There was recovery of WBC values during the 8-week post-exposure period to normal values. The reduced WBC counts were also accompanied by a slight reduction in myeloid cells of the bone marrow in 3000 ppm exposed rats. There was no histologic indication of myeloid cytotoxicity.
Urinary specific gravity was reduced in 3000 ppm 1,3-dioxolane exposed rats, although there were no morphologic indications of nephrotoxicity. Exposure-related pathologic effects were limited to slight enlargement of centrilobular hepatocytes in 3000 ppm 1,3-dioxolane exposed males.
There was no indication of testicular toxicity based on careful light microscopic examination. The results of the study reported herein indicate that there is not a significant potential for testicular toxicity despite repeated exposures to concentrations as high as 3000 ppm.

Executive summary:

Groups of 10 rats/sex were exposed to 0, 300, 1000 or 3000 ppm of 1,3-dioxolane for 6 hours/day, 5 days/week for 13 weeks. Satellite groups of 10 rats/sex were also exposed to the same concentrations of 1,3-dioxolane as listed above for 13 weeks to allow multiple clinical pathology assessments and an 8-week post-exposure evaluation period. 20 rats/sex/group were consequently exposed to 1,3-dioxolane.

The primary effect of exposure to 1,3-dioxolane was a reduction in white blood cell (WBC) counts, primarily due to decreased numbers of lymphocytes; the reductions in WBC counts were apparent in animals exposed to 1000 ppm or 3000 ppm 1,3-dioxolane. There was recovery of WBC values during the 8-week post-exposure period to normal values. The reduced WBC counts were also accompanied by a slight reduction in myeloid cells of the bone marrow in 3000 ppm exposed rats, but there was no histologic indication of myeloid cytotoxicity. Body weights were slightly reduced relative to controls in females exposed to 3000 ppm 1,3-dioxolane. Mean relative spleen weights were reduced in 1000 ppm (females only) and 3000 ppm 1,3-dioxolane exposed rats. Mean liver weights were slightly increased in 3000 ppm 1,3-dioxolane exposed rats. Clinical observations revealed only a transient reduction in responsiveness of animals at the end of exposure to 3000 ppm 1,3-dioxolane. Statistical differences in clinical chemistry values were identified, but with the exception of alanine aminotransferase, no clear relationship to exposure could be established. Exposure-related pathologic effects were limited to slight enlargement of centrilobular hepatocytes in 3000 ppm 1,3-dioxolane exposed males at 13 weeks only. Urinary specific gravity was reduced in 3000 ppm 1,3-dioxolane exposed rats, although there were no morphologic indications of nephrotoxicity.

Despite the decreased WBC counts, rats appeared healthy and there were only minimal morphological effects in those exposed to 3000 ppm 1,3-dioxolane. There were no observed adverse effects in rats exposed to 300 ppm 1,3-dioxolane.