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Toxicological information

Repeated dose toxicity: inhalation

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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

Constituent 1
Chemical structure
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 / concentrationsopen allclose all
Dose / conc.:
0 ppm (nominal)
Remarks:
Control
Dose / conc.:
300 ppm (nominal)
Remarks:
298 ppm (measured)
Dose / conc.:
1 000 ppm (nominal)
Remarks:
1000 ppm (measured)
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.