Registration Dossier

Toxicological information

Repeated dose toxicity: oral

Currently viewing:

Administrative data

sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Start: December 2008, End: October 2009
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Standard OECD Guideline study, GLP

Data source

Reference Type:
study report

Materials and methods

Test guideline
according to
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
GLP compliance:
yes (incl. certificate)
Limit test:

Test material

Details on test material:
Test Material Name
Chemical Name
Supplier, City, State (Lot, Reference Number)
ANGUS Chemical Company, a wholly owned subsidiary of The Dow Chemical Company, Buffalo Grove, Illinois (lot # CEC-200803388-4)
Purity/Characterization (Method of Analysis and Reference)
The purity of the test material was determined to be 98.4% on an anhydrous basis by gas chromatography with identification by gas chromatography mass spectroscopy and nuclear magnetic resonance (Roy et al., 2009).

Test animals

Details on test animals and environmental conditions:
Species and Sex
Rats, male and female (nulliparous and nonpregnant)
Strain and Justification
Crl:CD(SD) and Crl:WI(Han) rats were selected because of their general acceptance and suitability for toxicity testing, the conflicting results from previous toxicity studies on 3-amino-4-octanol using these strains, availability of historical control background data, and the reliability of the commercial supplier.
Supplier and Location
Crl:WI(Han) - Charles River Laboratories Inc. (Raleigh, North Carolina)
Crl:CD(SD) - Charles River Laboratories Inc. (Portage, Michigan)
Age at Study Start
Crl:WI(Han) - 7 weeks
Crl:CD(SD) – 7 weeks
Physical and Acclimation
Each animal was evaluated by a laboratory veterinarian, or a trained animal/toxicology technician under the direct supervision of a laboratory veterinarian, to determine the general health status and acceptability for study purposes upon arrival at the laboratory (fully accredited by the Association for Assessment and Accreditation of Laboratory Animal Care International - AAALAC International). The animals were housed two-three per cage in stainless steel cages, in rooms designed to maintain adequate conditions (temperature, humidity, and photocycle), and acclimated to the laboratory for at least one week prior to the start of the study.
After assignment, animals were housed two per cage in stainless steel cages. All rats were single housed on test day nine due to the large amount of feed consumed by the rats while double housed and the necessity to measure feed consumption weekly. Cages had wire mesh floors and were suspended above absorbent paper. Non-woven gauze was placed in the cages to provide a cushion from the flooring for rodent feet. The non-woven gauze and a stainless steel object placed in the cage also provided environmental enrichment. Cages contained a feed crock and a pressure activated lixit valve-type watering system. The following environmental conditions were maintained:
Temperature: 22°C with a tolerance of ± 1°C (and a maximum permissible excursion of ± 3°C)
Humidity: 40-70%
Air Changes: 12-15 times/hour
Photoperiod: 12-hour light/dark (on at 6:00 a.m. and off at 6:00 p.m.)

Randomization and Identification
Animals were stratified by body weight and then randomly assigned to treatment groups using a computer program designed to increase the probability of uniform group mean weights and standard deviations prior the start of the study. Animals that were placed on study were uniquely identified via subcutaneously implanted transponders (BioMedic Data Systems, Seaford, Delaware) that were correlated to unique alphanumeric identification numbers.

Feed and Water
Animals were provided LabDiet Certified Rodent Diet #5002 (PMI Nutrition International, St. Louis, Missouri) in meal form. Feed and municipal water were provided ad libitum. Analyses of the feed were performed by PMI Nutrition International to confirm the diet provided adequate nutrition and to quantify the levels of selected contaminants. Drinking water obtained from the municipal water source was periodically analyzed for chemical parameters and biological contaminants by the municipal water department. In addition, specific analyses for chemical contaminants were conducted at periodic intervals by an independent testing facility. Copies of these analyses are maintained in the study record. There were no appreciable amounts of contaminants in feed and water that could have adversely impacted the study.
Animal Welfare
In accordance with the U.S. Department of Agriculture's rules on animal welfare, 9 CFR Parts 1-4, the animal care and use activities required for conduct of this study were reviewed and approved by the Institutional Animal Care and Use Committee (IACUC). The IACUC determined that the proposed Activities were in full accordance with these Final Rules. The IACUC-approved Animal Care and Use Activities used for this study were Subchronic/Chronic Tox 01, DCO 01, Humane Endpoints 01, Neurotox 01, DART 01, and Animal ID 01.

Administration / exposure

Route of administration:
oral: gavage
propylene glycol
Details on oral exposure:
All dosing solutions were prepared by mixing the test material in propylene glycol at concentrations of 0, 2.5, 10, or 25 mg/ml and administered at a dose volume of 6 ml/kg body weight to achieve the targeted dose levels. Dose solutions were not corrected for purity. Dose volumes were adjusted at least weekly based on individual body weights. The control rats were dosed with propylene glycol at 6 ml/kg body weight.
Analytical verification of doses or concentrations:
Details on analytical verification of doses or concentrations:
Dose Confirmation and Homogeneity
Dose confirmation and homogeneity analyses of all dose solutions were determined pre-exposure, near the middle and end of the study. The method used for analyzing the test material in propylene glycol was high performance liquid chromatography with positive ion electrospray ionization and mass spectrometry detection (HPLC/+ESI/MS). Quantitation was performed incorporating an isotopically labeled internal standard.
3-amino-4-octanol was stable for at least 29 days in propylene glycol at concentrations ranging from 0.25 to 250 mg/mL (Ito, 2008). The established concentration range and duration spanned those used in this study.
Retainer Samples
A sample of each lot number of the bulk test material was retained. No samples of dose solutions were retained.
Duration of treatment / exposure:
At least 90 days, 7 days per week
Frequency of treatment:
Doses / concentrationsopen allclose all
Doses / Concentrations:
Crl:CD(SD) at dose levels of 0, 15, 60, or 150 mg/kg body weight/day
actual ingested
Doses / Concentrations:
Crl:WI(Han) rats at dose levels of 0 or 150 mg/kg body weight/day
actual ingested
No. of animals per sex per dose:
Crl:CD(SD) = 10 males and 10 females per dose group
Crl:WI(Han) - 10 male rats per dose group
Control animals:
yes, concurrent vehicle


Observations and examinations performed and frequency:
Daily Observations
A cage-side examination was conducted twice daily, generally at the same time each day. This examination was typically performed with the animals in their cages and was designed to detect significant clinical abnormalities that were clearly visible upon a limited examination, and to monitor the general health of the animals. The animals were not hand-held for these observations unless deemed necessary. Significant abnormalities that could have been observed included, but were not limited to: decreased/increased activity, repetitive behavior, vocalization, incoordination/limping, injury, neuromuscular function (convulsion, fasciculation, tremor, and twitches), altered respiration, blue/pale skin and mucous membranes, severe eye injury (rupture), alterations in fecal consistency, and fecal/urinary quantity. In addition, all animals were observed for morbidity, mortality, and the availability of feed and water at least twice daily.
Clinical Observations
Animals were observed approximately one hour after dosing. Clinical observations included a careful evaluation of abnormalities in the eyes, urine, feces, gastrointestinal tract, extremities, movement, posture, reproductive system, respiration, skin/hair-coat, and mucous membranes, as well as an assessment of general behavior, injuries, or palpable mass/swellings.
Detailed Clinical Observations
Detailed clinical observations (DCO) were conducted on all Crl:CD(SD) rats pre-exposure and once per week throughout the study. The DCO was conducted at approximately the same time each examination day, according to an established format. The examination included cage-side, hand-held and open-field observations, which were recorded categorically or using explicitly defined scales (ranks). Details of these observations can be found in Table 2 and Appendix A. DCO were not conducted on Crl:WI(Han) rats.
Functional Tests
The functional tests (sensory evaluation, rectal temperature, grip performance and motor activity) were conducted pre-exposure and during the last week of the treatment period for all Crl:CD(SD) rats. Details of these observations can be found in Appendix B. Functional tests were not conducted on Crl:WI(Han) rats.
Sensory Evaluation
The sensory evaluation included a test for nociception (responsiveness to tail pinch) and for startle response (responsiveness to sharp noise). The evaluation was conducted in a clear plastic box. Details of the methods for each test and specific definitions of the ranks can be found in Appendix B.
Rectal Temperature
Rectal temperature was measured by carefully placing a rectal thermistor (Physitemp, Clifton, New Jersey) approximately 4 cm into the rectum for approximately 10 seconds. Temperature was recorded. The thermistor was validated at 37C before, during, and after the study. The instrument was re-calibrated if the temperature recordings differed from the reference thermometer by more than  0.5C.
Grip Performance
Hindlimb grip performance was tested according to the procedure described by Mattsson et al. (1986). Briefly, the observer placed the rat’s forelegs on a plastic bench and the hindfeet were set on a horizontal screen attached to an electronic strain gauge (Chatillon, Greensboro, North Carolina). The observer then smoothly but firmly pulled backward on the tail until the rat’s grip on the screen was broken (Appendix C). An electronic strain gauge was used to record the rat’s resistance to the pull in grams. The average of three trials was used for statistical analysis. Forelimb grip performance was similarly tested. In this application, a bench was not used, and the rats were placed so that the forefeet were on the screen and the hindfeet were suspended approximately 10 cm above the plastic platform.
Instrument Validation: A standard 500-gram weight attached to a fine-gauge wire was suspended from the load cell and was checked just before and just after testing (a 1% tolerance, i.e., 500  5 grams, is acceptable).
Motor Activity
An automated system was used for motor activity (MA) data collection (Appendix D). No entry into the MA test room was allowed during the testing period. Each test session consisted of eight 8-minute epochs, totaling 64 minutes of testing per animal per test session. This duration was chosen based on the results of a validation study indicating that performance of control animals approached asymptote in 50-60 minutes in CD rats (Marty and Andrus, 2007). Activity counts for each epoch were recorded.
Motor Activity Cage Calibration: Cages used for testing were calibrated prior to testing each day. Calibration was performed with a rod attached to a rotary motor that broke the infrared beam at a constant speed. The duration of each beam break was calculated to ensure equivalence across chambers.
Motor Activity Cage Allocation: Rats were allocated to the motor activity cages in such a way that the counterbalancing of treatment groups and sexes across cages and test times was maximized.
The eyes of all animals were examined by a veterinarian pre-exposure and prior to the scheduled necropsy using indirect ophthalmoscopy. One drop of 0.5% tropicamide ophthalmic solution was instilled topically in each eye to produce mydriasis prior to the indirect ophthalmic examinations. Eyes were also examined by a prosector during the necropsy using a moistened glass slide pressed to the cornea.
Body Weights/Body Weight Gains
All rats were weighed pre-exposure and at least weekly during the dosing period. Body weight gains were calculated relative to day 1.
Feed Consumption
Feed consumed was determined at least weekly for all animals by weighing feed containers at the start and end of a measurement cycle. Feed consumption was calculated using the following equation:
Feed consumption (g/day) = (initial weight of feed container - final weight of feed container)
(# of days in measurement cycle) (# of animals per cage)
Sacrifice and pathology:
Clinical Pathology
Animals were fasted overnight prior to blood collection. Blood samples were obtained from the orbital sinus following anesthesia with CO2/O2 at the scheduled necropsy. Blood was not obtained from animals that died prior to their scheduled necropsy. Blood was not collected from Crl:WI(Han) rats.
Sample Preparation
Blood samples for a complete blood count were mixed with ethylenediamine-tetraacetic acid (EDTA). Blood smears were prepared, stained with Wright-Giemsa stain, cover-slipped and archived for potential future evaluation, if warranted.
Hematologic parameters were assayed using the Bayer Advia 120 Hematology Analyzer (Siemens Medical Solutions Diagnostics, Tarrytown, New York).
Hematocrit (HCT)
Hemoglobin (HGB) concentration
Red blood cell (RBC) count
Total white blood cell (WBC) count
Differential WBC count
Platelet (PLT) count
Reticulocyte (RET) count
RBC indices:
Mean Corpuscular Hemoglobin (MCH)
Mean Corpuscular Volume (MCV)
Mean Corpuscular Hemoglobin Concentration (MCHC)
Sample Preparation
Blood samples were collected in sodium citrate tubes, centrifuged, plasma collected, and assayed using the Instrumentation Laboratory ACL9000 Analyzer (Beckman Coulter, Inc., Miami, Florida).
Prothrombin time (PT)
Clinical Chemistry
Sample Preparation
Blood samples were collected and serum was separated from cells as soon as was possible. The following serum parameters were measured using the Hitachi 912 Clinical Chemistry Analyzer (Roche Diagnostics, Indianapolis, Indiana).
Enzyme Activities of:
Alkaline phosphatase (ALP)
Alanine aminotransferase (ALT)
Aspartate aminotransferase (AST)
Gamma glutamyl transpeptidase (GGT)
Concentrations of:
Albumin (ALB)
Albumin/Globulin Ratio (A/G) (calculated)
Cholesterol (CHOL)
Creatinine (CREA)
Electrolytes (NA, K, PHOS, CL and CA)
Globulin (calculated)
Glucose (GLUC)
Total bilirubin (TBIL)
Total protein (TP)
Triglycerides (TRIG)
Urea nitrogen (UN)
Urine samples were obtained from Crl:CD(SD) rats the week prior to the scheduled necropsy. Animals were housed in metabolism cages and the urine was collected overnight (approximately 16 hours). Feed and water were available during this procedure. Urine was not obtained from Crl:WI(Han) rats.
Color, appearance, specific gravity (refractometer) and urine volume
Semiquantitative analysis of the following was conducted using Multistix Reagent Strips (Bayer Corporation, Elkhardt, Indiana) on the HealthCare Clinitek Advantus Analyzer (Siemens Medical Solutions Diagnostics, Tarrytown, New York):
Microscopic Examination
Urine samples were also collected from each animal (Crl:CD(SD) rats only) by manual compression of the urinary bladder. The urine samples were pooled from each group, and the microsediment was characterized microscopically.
Anatomic Pathology
Fasted rats submitted alive for necropsy were anesthetized by the inhalation of CO2/O2, weighed, and blood samples (Crl:CD(SD) rats only) obtained from the orbital sinus. Their tracheas were exposed and clamped, and the animals were euthanized by decapitation. A complete necropsy was conducted on all animals by a veterinary pathologist assisted by a team of trained individuals. The necropsy included an examination of the external tissues and all orifices. The head was removed, the cranial cavity opened and the brain, pituitary and adjacent cervical tissues were examined. The eyes were examined in situ by application of a moistened microscope slide to each cornea. The skin was reflected from the carcass, the thoracic and abdominal cavities were opened and the viscera examined. All visceral tissues were dissected from the carcass, re-examined and selected tissues were incised. The nasal cavity was flushed via the nasopharyngeal duct (Crl:CD(SD) rats only) and the lungs were distended to an approximately normal inspiratory volume with neutral, phosphate-buffered 10% formalin using a hand-held syringe and blunt needle.
For the Crl:WI(Han) rats the liver, kidneys, testes, and epididymides were trimmed and weighed immediately. The ratios of organ weight to terminal body weight were calculated. For the Crl:CD(SD) rats the brain, liver, kidneys, heart, adrenals, testes, epididymides, uterus, ovaries, thymus and spleen were trimmed and weighed immediately. The ratios of organ weight to terminal body weight were calculated.
Representative samples of tissues listed in Table 3 were collected and preserved in neutral, phosphate-buffered 10% formalin, except that both testes and the right epididymis were preserved in Bouin’s. Transponders were removed and placed in jars with the tissues. Similar necropsy procedures were followed for animals found dead, except that body weights, organ weights, and blood samples were not obtained.
Moribund animals that were not expected to survive until the next observation period were humanely euthanized. These animals, as well as any animals found dead, were necropsied on that day during the normal workweek. However, animals euthanized or found dead after routine working hours or on weekends, and holidays were refrigerated until the next scheduled workday, at which time they were necropsied.
Sperm Analysis
Sperm parameters were evaluated in all males at termination. The left and right epididymides and testes were allocated as follows: right epididymis-motility and histopathology: left epididymis-counts; right and left testis-histopathology.
Immediately after euthanasia of males and isolation of their epididymides, a small sample of sperm from the right cauda epididymis was expressed into a dish containing SpermPrep Medium (ZDL, Lexington, Kentucky) and was incubated at room temperature for approximately 2-3 minutes. An aliquot of the incubated sperm suspension was placed in a chamber of the HTM Integrated Visual Optical System (IVOS; Hamilton-Thorne Research, Beverly, Massachusetts) for the determination of total percent motile (showing any motion) and percent progressively motile (showing net forward motion) sperm. Images from the motility analyses were recorded on CD-R and were archived with the study file. After sperm were released, the epididymis was placed in Bouin’s fixative and examined following histologic preparation.
The left cauda epididymis was weighed and then frozen at approximately -20˚C for subsequent determination of the number of homogenization-resistant spermatids and sperm per cauda epididymis and per gram of epididymal tissue. The epididymis was minced, diluted and stained with a fluorescent DNA-binding dye (HTM-IDENT, Hamilton-Thorne Research, Beverly, Massachusetts) and the spermatid or sperm count was determined from an aliquot loaded into the IVOS analyzer as described by Stradler et al. (1996). Samples from the high-dose and control animals were evaluated.
An aliquot of sperm suspension was also placed on a slide, and a smear prepared and air-dried for subsequent evaluation of sperm morphology. At least 200 sperm per male were evaluated and were classified as normal or abnormal as described by Filler (1993). Morphological evaluation of sperm from control and high-dose males was conducted.
Histologic examination of the tissues indicated in Table 3 was conducted on all Crl:CD(SD) control and high-dose rats. Histologic slide preparation was performed at MPI Research, Mattawan, Michigan for all tissues with the exception of the testes. Histologic examination of the testes was conducted for all Crl:WI(Han) rats. Examination of tissues from the remaining groups was limited to relevant gross lesions. Paraffin embedded tissues were sectioned approximately 6 µm thick, stained with hematoxylin and eosin and examined by a veterinary pathologist using a light microscope.
Histopathological examination of the testes included a qualitative assessment of stages of spermatogenesis. A cross section through the approximate center of both testes of control and high-dose males was embedded in paraffin, sectioned at 5 μm and stained with modified periodic acid Schiffs-hematoxylin. The presence and integrity of the stages of spermatogenesis was qualitatively evaluated following the criteria and guidance of Russell et al. (1990). Microscopic evaluation included a qualitative assessment of the relationships between spermatogonia, spermatocytes, spermatids, and spermatozoa seen in cross sections of the seminiferous tubules. The progression of these cellular associations defined the cycle of spermatogenesis. In addition, sections of both testes were examined for the presence of degenerative changes (e.g., vacuolation of the germinal epithelium, a preponderance of Sertoli cells, sperm stasis, inflammatory changes, mineralization, and fibrosis).
Selected histopathologic findings were graded to reflect the severity of specific lesions to evaluate: 1) the contribution of a specific lesion to the health status of an animal, 2) exacerbation of common naturally occurring lesions as a result of the test material, and 3) dose-response relationships for treatment-related effects. Very slight and slight grades were used for conditions that were altered from the normal textbook appearance of an organ/tissue, but were of minimal severity and usually with less than 25% involvement of the parenchyma. This type of change would neither be expected to significantly affect the function of the specific organ/tissue nor have a significant effect on the overall health of the animal. A moderate grade was used for conditions that were of sufficient severity and/or extent (up to 50% of the parenchyma) that the function of the organ/tissue may have been adversely affected, but not to the point of organ failure. The health status of the animal may or may not be affected, depending on the organ/tissue involved, but generally lesions graded as moderate would not be life threatening. A severe grade was used for conditions that were extensive enough to cause significant organ/tissue dysfunction or failure. This degree of change in a critical organ/tissue may be life threatening.
A complete set of tissues (listed in Table 3) was examined from rats found dead or moribund. Histological examination was conducted in a similar manner as described above.
Statistical significance of all findings assessed using appropriate methods

Results and discussion

Results of examinations

Clinical signs:
no effects observed
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
no effects observed
Water consumption and compound intake (if drinking water study):
no effects observed
Ophthalmological findings:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Details on results:
Wistar rats:

Groups of 10 male Crl:WI(Han) rats were administered 0 or 150 mg/kg/day 3-amino-4-octanol by gavage. There were treatment-related decreases in body weight (4.3 – 7.0%) and body weight gain (7.0 – 16.8%), relative to controls, throughout the study. The body weight decrements reached statistical significance on test day (TD) 8, 15, 22, 29, 57, 64, 71, 78, and 85. There was a slight, but statistically significant, 5.2-6.3% decrease in feed consumption, relative to controls, between TD 1-8 and 9-15. Feed consumption in the treated group was comparable to controls after TD 22 for the remainder of the study.
Treatment-related clinical observations, noted in six of ten animals, included reflux of test material, noisy respiration, slow respiration, labored respiration (without mouth breathing), and/or blood coming from the nasal cavity. Observations unrelated to treatment included one animal (#288) with decreased feces from TD 55-56 and pre-exposure ophthalmic findings of cloudy cornea of the left eye in one animal (#281) and one animal (#286) with pale fundus of the left eye, both of which resolved before the terminal ophthalmic examination.
All of the above effects were attributed to the irritation caused by the test material and were not considered to be signs of systemic toxicity.

There were no treatment-related effects on any epididymal sperm counts, motility, and morphology.

Crl:CD(SD) Results
All male and female rats survived the 90-day test period, except for two females (#251 and #252) in the 15 mg/kg/day group and three (#265, #270, and #272) in the 150 mg/kg/day group that died prior to scheduled necropsy due to gavage errors.

Detailed Clinical and Cage-Side Observations

There were no treatment-related clinical observations in male and female rats at any dose level tested. There was one female (#265 on TD 48) and one male (#225 on TD 38) in the 150 mg/kg/day group, and two males (#216 on TD 82 and #221 on TD 82) in the 60 mg/kg/day group with a single occurrence of test material reflux after gavage dosing. Animal #221 also had labored respiration with mouth breathing associated with the reflux on TD 82 and animal #225 had noisy respiration not associated with reflux on TD 59. These observations were considered not toxicologically significant due to low incidence, short duration (lasting only one day), and lack of a dose response. Other clinical observations noted which were unrelated to treatment included maloccluded incisors, dermatitis, apparent mechanical injury, and excessive hairloss.
For the scored portion of the DCO, all rats were within normal limits at each of the weekly intervals.

Functional Tests
Sensory Evaluation
Examinations performed on males and females revealed no statistically significant or treatment-related findings.

Rectal Temperature
There were no treatment-related effects in rectal temperature. The treatment by sex interaction was not significant (p = 0.5435), which indicated that there were no treatment-related sex differences in rectal temperature. The main effect of treatment was also not significant (p = 0.1613), which indicated that treatment did not affect rectal temperature in rats.

Grip Performance
There were no treatment-related effects in grip performance. The treatment by sex interaction was not significant for hindlimb (p = 0.8781) or forelimb (p = 0.6150) grip performance, which indicated that there were no treatment-related sex differences in grip performance. The main effect of treatment was also not significant for hindlimb (p = 0.7586) or forelimb (p = 0.8086) grip performance, which indicated that treatment did not affect grip performance in rats.

Motor Activity
There were no treatment-related effects in motor activity. The treatment by time by sex interaction was not significant (p = 0.8629), which indicated that there was not a treatment-related sex difference in motor activity at either time point. The treatment by time interaction was also not significant (p = 0.5443), which indicated that exposure did not affect motor activity in rats. The treatment by time by epoch interaction was not significant (p = 0.5898), which indicated that treatment did not affect the within session distribution of motor activity counts at either time interval.

Pre-exposure examination on all rats placed on study indicated all rats were suitable for study assignment; however, one animal in the control group (#195) had anterior synechia of the right eye with pale fundus (resolved before terminal ophthalmic examination) and one animal in the 150 mg/kg/day group (#230) had pale fundus. Pale fundus was noted prior to study termination in two animals (#198-control group and #230-150 mg/kg/day group). These observations were interpreted to be unrelated to treatment due to low incidence and/or lack of a dose response.
Body Weights/Body Weight Gains

There was no treatment-related effect on body weights of male or female rats in any dose group. Male body weights in the 15, 60, and 150 mg/kg/day groups were slightly lower than controls; however, this was deemed spurious and unrelated to treatment as it did not reach statistical significance and there was no clear dose-response relationship. Body weight and body weight gain of females at all dose levels were similar to controls.
Feed Consumption

There were no statistically identified differences in the feed consumption of male or female rats when compared to their respective controls and no differences related to treatment.

Clinical Pathology
There were no statistically significant differences in any of the hematologic parameters for male and female rats at any dose level. There was one male rat (#205) in the 15 mg/kg/day group with a higher reticulocyte count, higher MCV, and lower erythrocyte counts, suggesting regenerative anemia, and lower leukocyte counts than the remainder of the dose group. Terminal body weight of this animal was ~100 g less than others in this dose group. The cause of these findings are unknown; however, due to the isolated incidence of these observations to a single animal, a lack of dose response, and no treatment-related hematologic or histopathologic alterations in animals at higher dose levels, this finding was considered spurious and unrelated to treatment.

Prothrombin Time
There were no treatment-related alterations in the prothrombin times of male and female rats at any dose level.

Clinical Chemistry
There were no treatment-related alterations in any of the clinical chemistry parameters for male and female rats at any dose level. The urea nitrogen in the 60 mg/kg/day group and the alkaline phosphatase in the 150 mg/kg/day group of males and females were statistically identified as higher compared to controls (Text Tables 5 and 6; Table 50); however, these effects were considered unrelated to treatment due to lack of dose response relationship. Historical control data for urea nitrogen and alkaline phosphatase are presented, but concurrent control values for these parameters were outside historical control or at equal to the high end value.
There were no toxicologically significant alterations in the urinalysis parameters for male and female rats. Urine volume was minimally increased in males and more significantly in females with a corresponding decrease in urine specific gravity (Text Tables 7 and 8); however, as there were no other indications of renal toxicity (e.g., kidney histopathologic findings) this finding is likely a consequence of the alcohol in the test material having a diuretic effect and considered not toxicologically significant. Historical control data for urine volume and specific gravity are presented, but concurrent control values for these parameters are outside historical control (above or below). In addition, there was a slight dose-related increase in the incidence of higher urine pH that was attributed to excretion of the alkaline test material.
Anatomic Pathology
Organ Weights
Female rats in the 150 mg/kg/day group did have a treatment-related statistically identified 14% increase in mean absolute liver weight compared to control; however, there were no correlating treatment-related histological liver effects and was therefore considered not toxicologically significant. Absolute brain weights were statistically identified as higher in the 15 mg/kg/day group for males and females however, were deemed unrelated to treatment as there was no dose response and no change in relative brain weights.

Sperm Parameters
There were no treatment-related observations in any epididymal sperm counts, motility, or morphology at any dose level.

Gross Pathology
Two females given 15 mg/kg/day (#251 – Day 5; #252 – Day 10) and three females given 150 mg/kg/day (#265 – Day 8; #270 – Day 9; #272 – Day 6) died spontaneously. Observations in females #252, 265 and 270 consisted of hydrothorax (clear or serosanguineous), congestion (liver, lungs and/or kidneys) and a dark lung lobe. One of these females (#272) also had fractured/missing upper incisors, hemorrhage involving the muzzle and lip trauma. An additional female given 150 mg/kg/day (#271) that survived until the end of the study had roughened kidneys, pelvic dilatation, calculi in the bladder and a thickened bladder wall. None of these gross pathologic observations were interpreted to be treatment related. The hydrothorax and congestion were attributed as secondary to probable gavage error. Kidney and bladder lesions were interpreted to be spontaneous lesions that are not uncommon in rats of this strain and age. All gross pathologic observations (skin inflammation and alopecia, kidney mass, pale focus liver, congestion in liver, dark cervical lymph nodes) were considered to be spontaneous alterations or iatrogenic (thymic hemorrhage) and were unassociated with exposure to the test material.
All observations were considered to be spontaneous alterations unassociated with exposure or were induced by mechanical administration of the test material by the gavage procedure (esophageal trauma – inflammation; subacute to chronic, and inadvertent administration of the test material into the trachea - tracheal/laryngeal necrosis with inflammation). Based on gross and histopathologic observations the cause of death for all five females was attributed to gavage error.

Testis Histopathology
A focus of this study was to resolve discrepencies between a marginal testis histopathologic finding in an OECD 421 study in Sprague Dawley rats and a 28-day study with no testis histopathologic findings in Wistar Han rats (see Previous Toxicity section). As the current 90-day study would allow for a full wave of spermatogenesis, approximately 56 days in Sprague Dawley rats (Russell et al., 1990), the testis histopathologic results from this study was considered definitive. There were no treatment-related testis histopathologic findings in Crl:CD(SD) and Crl:WI(Han) rats given 150 mg/kg/day 3-amino-4-octanol.

Effect levels

Dose descriptor:
Effect level:
150 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: see 'Remark'

Target system / organ toxicity

Critical effects observed:
not specified

Applicant's summary and conclusion

The no effect level for the repeated dose toxicity of 3-amino-4-octanol is 150 mg/kg bw for systemic toxicity. The testicular effects observed in the OECD 422 study were not corroborated by this longer and more detailed study and so are not considered to be an adverse effect associated with exposure to 3-amino-4-octanol.
Executive summary:

3-amino-4-octanol was administered by gavage, at 6 ml/kg body weight dose volume in propylene glycol vehicle, to groups of ten male and ten female Crl:CD(SD) at dose levels of 0, 15, 60, or 150 mg/kg body weight/day and to groups of ten male Crl:WI(Han) rats at dose levels of 0 or 150 mg/kg body weight/day for at least 90 days to evaluate the potential for systemic toxicity. Parameters evaluated were daily cage-side observations, daily clinical observations, weekly detailed clinical observations, ophthalmic examinations, functional tests, body weights, feed consumption, hematology, prothrombin time, urinalysis, clinical chemistry, selected organ weights, epididymal sperm analyses (motility, counts, and morphology), and gross and histopathologic examinations.

There were no treatment-related effects in clinical signs, functional tests, body weights, feed consumption, ophthalmic, hematology, prothrombin time, or clinical chemistry parameters in Crl:CD(SD) rats of either sex. Male Crl:WI(Han) rats had treatment-related decreases in body weight, body weight gain, and feed consumption with treatment-related clinical observations including reflux of test material, noisy respiration, slow respiration, labored respiration (without mouth breathing), and/or blood coming from the nasal cavity in the 150 mg/kg/day. There were no treatment-related epididymal sperm parameters (motility, counts, or morphology), gross or histopathologic observations and no toxicologically significant effects in urinalysis parameters or organ weight effects in the Crl:CD(SD) or Crl:WI(Han) rats.

The no-observed-effect level (NOEL) for male reproductive effects in either strain was 150 mg/kg/day, the highest dose level tested. The no-observed-adverse-effect level (NOAEL) for Crl:CD(SD) rats of either sex was 150 mg/kg/day 3-amino-4-octanol.