Registration Dossier

Administrative data

Endpoint:
sub-chronic toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1989-08 to 1989-12
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

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:
OECD Guideline 411 (Subchronic Dermal Toxicity: 90-Day Study)
Deviations:
yes
Remarks:
purity, storage conditions not reported
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
Name of test material: Trimethylolpropanepoly(oxypropylene)triamine
EC no.: 500-105-6
CAS no.: 39423-51-3
Physical state: clear colourless liquid
Specific details on test material used for the study:
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Stability under test conditions: responsibility of the Sponsor

OTHER SPECIFICS:
- Name of test material (as cited in study report): 6398-10-1

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Wilmington, Massachusetts
- Age at study initiation: 47 days old
- Weight at study initiation: approximately 135 to 215 g
- Fasting period before study: no
- Housing: vertical cage positioning in stainless steel 1/2" wire mesh cages. Cage sizes were in accordance w i t h the "Guide for the Care and Use of Laboratory Animals" of the Institute of Laboratory Animal Resources, National Research Council (NIH 86-23, 1985)
- Diet (e.g. ad libitum): Purina Certified Rodent Lab Meal , ad libitum
- Water (e.g. ad libitum): fresh tap water, ad libitum
- Acclimation period: 11 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C±3°C
- Humidity (%): 30-70%
- Photoperiod (hrs dark / hrs light): 12h dark/12h light

Administration / exposure

Type of coverage:
occlusive
Vehicle:
water
Details on exposure:
TEST SITE
- Area of exposure: clipped dorsal area
- Type of wrap if used: a gauze patch (2x2 cm) wrapped with an elastic bandage which was taped to the animal
- Time intervals for shavings or clipplings: weekly or when needed

REMOVAL OF TEST SUBSTANCE: no washing

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 5 ml/kg
- Constant volume or concentration used: yes

VEHICLE: deionized water

USE OF RESTRAINERS FOR PREVENTING INGESTION: no data
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
approximately 6 hours
Frequency of treatment:
once daily, five days per week for a period of 90 days
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day (nominal)
Dose / conc.:
16 mg/kg bw/day (nominal)
Remarks:
low dose group
Dose / conc.:
50 mg/kg bw/day (nominal)
Remarks:
mid dose group
Dose / conc.:
160 mg/kg bw/day (nominal)
Remarks:
high dose group
No. of animals per sex per dose:
- Group I - 5 males + 5 females: control recovery group
- Groups II, III, and IV - 15 males + 15 females: low , mid and high dose groups
- Groups V and VI - 10 males + 10 females: control-recovery and high recovery dose groups
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Based upon a 28 Day Dermal Toxicity Study in Rats:
The test substance was administered in a solution by dermal application to five groups of 10 rats (five males and five females per group) for 6 hours/day, 5 days/week for 28 days. The dose groups were 10, 50, 100, 250, and 500 mg/kg. An additional group of 10 rats (five male, five female) received the vehicle (deionized water) and served as a vehicle control group. No clinical signs were observed in the vehicle control group or the 10 and 50 mg/kg dose groups.
Clinical signs observed in the 100, 250 and 500 mg/kg treatment groups included very slight to severe erythema and extreme sloughing of skin, fissuring of skin, 15% to 75% necrosis of dose area, and scattered necrosis of dose area. The severity of these observations was dose dependent. Three rats died on study (one female in the high dose group on Day15 and two females in the high dose group on Day 17). Necropsy of the animals dying on study revealed mottled lungs and/or pale and prominent lobular patter of the liver. There were no statistically significant differences found in the group mean body weights, weight gains or daily food consumption. Terminal necropsy revealed mottled or pale lungs, red foci throughout lungs, tan foci and yellow-brown discoloration of the medial lobe of the liver, adhered lobes of the liver and mottled kidneys in the treatment groups. Terminal necropsy of the vehicle control group revealed mottled lungs, red foci throughout lungs and/or mottled kidneys. Based up on the results of this dose-range finding study, the suggested doses for the 90 -day study in rats were 16, 50 and 160 mg/kg.

- A negative control recovery group and a high dose recovery group remained on test untreated for an additional 28 days to determine reversibility, persistence, or delayed occurrence of toxic effects.


Positive control:
no

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Cage side observations checked in table [No.?] were included. cage side observation included, but were not limited to, changes in skin and fur, eyes and mucous membranes, respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

DERMAL IRRITATION (if dermal study): Yes
- Time schedule for examinations: daily

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION: Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes weekly

FOOD EFFICIENCY: No

WATER CONSUMPTION: No data

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: prior to treatment and during treatment at day 25, day 88 and day 116
- Dose groups that were examined: all animals

HAEMATOLOGY: Yes
- Time schedule for collection of blood: prior to initiation of the study and at sacrifice period
- Anaesthetic used for blood collection: No data
- Animals fasted: Yes, overnight
- How many animals: 10 (5/sex)
- Parameters examined: hemoglobin, hematocrit, erythrocyte count, total and differential leucocyte counts, platelet count, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: prior to initiation of the study and at sacrifice period
- Animals fasted: Yes, overnight
- How many animals: 10 (5/sex)
- Parameters examined: calcium, phosphorus, chloride, sodium, potassium, fasting glucose, serum alanine aminotransferase, serum aspartate aminotransferase, gamma glutamyl transpeptidase, urea nitrogen, albumin, globulin, blood creatinine, total bilirubin, total serum protein measurements

URINALYSIS: No data

NEUROBEHAVIOURAL EXAMINATION: No data


Sacrifice and pathology:
GROSS PATHOLOGY: Yes
- on all animals.
- examination of the external surface of the body, all orifices, and the cranial, thoracic, abdaminal and pelvic cavities and their contents.
- the following organs were weighed: liver, gonads, kidneys, adrenal glands, brain

HISTOPATHOLOGY: Yes
- the following organs and tissues were fixed and preserved for histopathological examination: gross lesions, brain-including sections of medulla/pons, cerebellar cortex and cerebral cortex, pituitary, thyroid/parathyroid, thymus, lungs (intact), trachea, heart, sternum with bone marrow, salivary glands, liver, spleen, kidneys/adrenals, pancreas, gonads, uterus, accessory genital organs (epididymides, prostate, and if present, seminal vesicles), aorta, skin (treated and uncreaced areas), esophagus, nasal turbinates, stomach, duodenum, jejunum, ileum, cecum, colon, rectum, urinary bladder, representative lymph node (submandibular), mammary gland, thigh musculature, peripheral nerve, eyes, femur-including arricular surface, spinal cord at three levels - cervical, midthoracic and lumbar, exorbital lachrymal glands
Statistics:
Evaluation of equality of means was made by the one way analysis of variance using the F distribution to assess significance. If significant differences among the means were indicated, Dunnett's or Student's t-test was used to determine significant differences from control means.

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Description (incidence and severity):
- no clinical signs of systematic toxicity observed during the study
- additional clinical observation: noted during the study and considered not to be related to test article
Dermal irritation:
effects observed, treatment-related
Description (incidence and severity):
- test article related moderate to severe skin irritation was observed in the mid and high dose groups. The skin irritation was reversible after discontinuation of treatment during the recovery period
- control or low dose group for either sex: no evidence of dermal irritation; in mid dose males and females at day 33 and day 34 respectively: dermal irritation observed; in high dose males and females: day 2 and day 5 dermal irritation observed;
- incidence and severity of dermal irritation increased with both the dose and duration of treatment and included the following observation for both mid and high dose animals: erythema, edema, sloughing of skin, scattered areas of necrosis, 25% of the area necrotic, and alopecia
- high dose recovery males and females: dermal irritation of similar incidence and severity occured during the 90 day treatment period, however, there was no evidence of delayed toxicity for either sex during the reversibility period, except for very slight erythema in one of ten high dose recovery females dermal irritation was no longer evident after the 28 day reversibility period (Day 118)
Mortality:
no mortality observed
Description (incidence):
- 2 non-treatment related deaths occured during study (judged as coincidental)
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
- statistically significant decreases in bw detected: in the male high dose group on days 7, 14, 21 and 28 when compared to control values. After day 35 there was a dose-related decrease in bw in males through day 91; however, these differences were not statistically significant.
- no statistically significant differences observed in female body weights. No statistically significant differences were detected between recovery control and recovery high dose groups during the 90 day treatment or 28 day recovery period.
- a statistically significant decrease in the group mean body weight gains for high dose males on days 7 and 14 and for mid dose males on day 91 when compared to che control group.
- significant decreases in bw gains were observed for the male high recovery groups on days 7 and 56 and a significant increase on day 21 when compared to the recovery control group. A significant decrease was observed for the female high dose recovery group on day 7 and an increase on day 77 when compared to the recovery control group.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
- a statistically significant decrease observed: in the group mean daily food consumption for the male high dose on Day 30. No other statistically significant differences were detected between non-recovery test article treated and control groups. Male high dose recovery group mean daily food
consumption was significantly increased on day 21 and day 84 when compared to the recovery control group. Female high dose recovery group mean daily food consumprion was significantly increased on days 91 and 118.
Food efficiency:
not specified
Description (incidence and severity):
No data
Water consumption and compound intake (if drinking water study):
not specified
Description (incidence and severity):
No data
Ophthalmological findings:
no effects observed
Description (incidence and severity):
- no ocular findings identified that were attributed to test article administration
- the lesions identified were consistent with sequela from a sialodacryoadenitis virus infection (striate retinopathy, keratitis and anterior synechia) or a congenial defect (disc coloboma)
- the lesions noted occurred in the following animals: On day 25, striate retinopathy was noted in Group II males (#9233-left eye; 9237 and 9239-right eye) and Group V males (#9321 and 9329-left eye); a disc coloboma was noted in a Group II male (#9239-left eye) and nasal keratitis occurred in Group II female (#9377-both eyes). On day 88, striate retinopathy was noted in a Group I female (#9224-left eye), a Group II male (#9239-right eye), a
Group IV male (#9300-right eye), two Group V males (#9329-left eye,; #9321-right eye) and a Group VI male (#9350-left eye). In addition, a group II male had anterior synechia (#9239-right eye) and a Group III female had keratitis (#9388-both eyes). On Day 118 a Group V male had striate retinopathy (#9329-left eye)
Haematological findings:
no effects observed
Description (incidence and severity):
- a few significant differences were noted in the hematological data (not dose dependent, within historical limits, not biologically significant or attributable to the administration of the test article)
- on day 30 there was a statistically significant decrease in male values for eosinophils (low and mid dose groups), lymphocytes, white blood cells (low, mid and high dose groups) and platelets (high dose group). There were no significant differences between control and test article treated females on day 30. On day 90, there w ere no significant differences between control and test article treated males. Statistically significant increases were noted for female hemoglobin and mean corpuscular hemoglobin concentration (high dose group) and red blood cell numbers (low, mid and high dose groups). On day 118, except for statistically significant decrease in platelets for high dose recovery females and a statistically significant decrease in eosinophils for high dose recovery males, there were no additional significant difference in the hematology values for recovery animals of either sex.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
- several significant differences were noted in the clinical chemistry data
- evaluation of the clinical chemistry data just prior to the day 30 interim sacrifice revealed significantly lower male values for total protein (mid dose) and globulin (mid and high dose); lower female values for GGPT (low dose), total bilirubin (low and mid dose), blood urea nitrogen, AST and sodium (mid dose); and higher female values for calcium (mid dose).
- on Day 90, significantly lower male values for creatinine, total protein, albumin (low, mid and high dose), sodium and calcium (high dose) ; higher male chloride (low, mid and high dose), male phosphorus (mid and high dose) and GGPT (high dose). Significantly lower female values for blood urea nitrogen (mid and high dose) and creatinine (mid dose); higher female values for ALT, AST and sodium (low dose), phosphorus (low, mid and high dose) and potassium (high dose).
- clinical chemistry data evaluated just prior to the day 118 recovery necropsy revealed a significantly higher male potassium value for the high dose recovery males. There was significantly higher AST for the high dose recovery females
Urinalysis findings:
not specified
Description (incidence and severity):
No data
Behaviour (functional findings):
not specified
Description (incidence and severity):
No data
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
- no statistically significant differences in absolute organ weights between the control and treated groups after 30 and 90 days of treatment or 28 days of recovery for either sex.
- a statistically significant decrease in fasted body weight was noted for the high dose males at the day 90 necropsy.
Gross pathological findings:
no effects observed
Description (incidence and severity):
- The majority of the gross findings at the day 30 (interim), day 90 (terminal) and day 118 (recovery) necropsies were non-specific and low incidence. Based upon histomorphologic evaluation of the selected tissues, there was no apparent relationship of these gross lesions to test article administration.
- At the day 30 necropsy, one high dose male, one mid dose female and two high dose females exhibited scattered or small scabs on the treatment site. At the day 90 necropsy, two high dose males exhibited multiple scabs and ulcers on treated skin and one high dose female exhibited multiple scabs on treated skin. At the day 118 (recovery) necropsy there were no gross lesions detected in the treated or untreated skin sites for either sex.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
- except for the skin irritation at the treatment site of the mid and high dose , there were no histomorphological alterations attributable to the administration of the test article
Histopathological findings: neoplastic:
not specified
Description (incidence and severity):
No data
Other effects:
not specified

Effect levels

Key result
Dose descriptor:
NOAEL
Effect level:
> 160 mg/kg bw/day
Sex:
male/female
Basis for effect level:
other: no clinical signs of systemic toxicity were attributable to the administration of the test article (160 mg/kg/d or less)
Remarks on result:
not determinable
Remarks:
no NOAEL identified

Target system / organ toxicity

Critical effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
Based upon the results of this subchronic 90 day dermal toxicity study in rats, the test substance did not produce systemic toxicity when administered dermally five days per week for 30 and 90 days at doses of 16, 50 and 160 mg/kg. Therefore, the NOAEL was established as greater than 160 mg/kg body weight/day.