Registration Dossier

Administrative data

Description of key information

Repeated dose toxicity - oral: A repeated dose (31-day) oral toxicity study via feeding was conducted in rats at dose levels of 93 and 239 mg/kg/day (supporting study, Klimisch 4; American Cyanamid Company, 1968). The NOAEL was deemed to be 239 mg/kg/day based on the lack of effects reported at this dose level.  

Repeated dose toxicity - dermal: A key, high quality 90-day dermal toxicity study (equivalent to OECD 411; Pharmakon research International Inc., 1990) and a high quality, supporting 28-day dermal toxicity study (Pharmakon Research International Inc., 1989) are available.  In the 90-day dermal study, no systemic toxicity was observed at the highest dose level tested (250 mg/kg/day). Therefore, the NOAEL for systemic toxicity was determined to be at least 250 mg/kg/day. However, significant irritation was observed in the high dose group. Therefore, the NOAEL for local, dermal effects was deemed to be 80 mg/kg.

Repeated dose toxicity - inhalation: No reliable study is available for this route of exposure. However, no further testing is needed since reliable studies are available for repeated dose toxicity via the oral or dermal route (REACH regulation, Column 2 adaptation, Annex VIII).

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Referenceopen allclose all

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
no data
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Remarks:
Limited information on study designs and methodology. Deviations from OECD Guideline 407: no data regarding observation period, test substance or animal husbandry; only 2 test groups were dosed compared to 3 test groups as recommended in the guideline; hematology, clinical biochemistry were not performed; no detailed data provided on gross autopsy results.
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)
Deviations:
yes
Remarks:
No data provided regarding observation period, test substance or animal husbandry, only 2 test groups were dosed compared to 3 test groups as recommended in the guideline, hematology, clinical biochemistry were not performed.
GLP compliance:
not specified
Limit test:
no
Specific details on test material used for the study:
No test item information
Species:
rat
Strain:
other: Albino (MR Wistar)
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Mean weight at study initiation:
0% concentration in diet
113.3 g

0.083% concentration in diet
113.3 g

0.208% concentration in diet
113.4 g
Route of administration:
oral: feed
Vehicle:
not specified
Details on oral exposure:
No data on feeding conditions.
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
No data
Duration of treatment / exposure:
31 days
Frequency of treatment:
daily
Dose / conc.:
0 other: % nominal in diet
Dose / conc.:
0.083 other: % nominal in diet
Dose / conc.:
0.208 other: % nominal in diet
No. of animals per sex per dose:
5 animals/sex/dose
Control animals:
yes
Details on study design:
No data
Positive control:
No data
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: No data

DETAILED CLINICAL OBSERVATIONS: No data

BODY WEIGHT: Yes
- Time schedule for examinations: No data

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No data

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data

OPHTHALMOSCOPIC EXAMINATION: No data

HAEMATOLOGY: No data

CLINICAL CHEMISTRY: No data

URINALYSIS: No data

NEUROBEHAVIOURAL EXAMINATION: No data
Sacrifice and pathology:
GROSS PATHOLOGY: The animals were sacrificed at the end of the feeding period and were subjected to a thorough gross autopsy. Autopsy results were not provided.
HISTOPATHOLOGY: No data
Other examinations:
Food intake; weight gain measured.
Statistics:
No data
Clinical signs:
no effects observed
Description (incidence and severity):
The overall appearance of the treated and control animals was good.
Mortality:
no mortality observed
Description (incidence):
The overall appearance of the treated and control animals was good.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Since weight gain depends, in part, upon the amount of food consumed, the variances and covariance of body weight and food intake were analyzed. Male ate and gained significantly more than the females. There were no significant interactions between sex and treatment, indicating that the responses of males and females to the treatments were similar. Therefore, observations on males and females were combined for comparison of the treatment groups with the control group. There were no significant differences between the 0.208% group or the 0.083% group and the control group in mean food intake or mean weight gain, even after allowances were made for variations in food consumption.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Since weight gain depends, in part, upon the amount of food consumed, the variances and covariance of body weight and food intake were analyzed. Male ate and gained significantly more than the females. There were no significant interactions between sex and treatment, indicating that the responses of males and females to the treatments were similar. Therefore, observations on males and females were combined for comparison of the treatment groups with the control group. There were no significant differences between the 0.208% group or the 0.083% group and the control group in mean food intake or mean weight gain, even after allowances were made for variations in food consumption.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
no effects observed
Description (incidence and severity):
No relevant gross pathology was noted at autopsy in animals of either sex.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Dose descriptor:
NOEL
Effect level:
>= 0.208 other: %
Based on:
other: dietary concentrations
Sex:
male/female
Basis for effect level:
other: overall effects mortality; body weight; food consumption; gross pathology
Critical effects observed:
not specified

Summary of results of thirty-one daily doses in the diets of male and female albino rats

Concentration in diet (%)

0

0.083

0.208

Number of rats (M +F)

5+5

5+5

5+5

Mean dosage (g/kg/day)

-

0.093

0.239

Mean initial weight (g)

113.3

113.3

113.4

Mean food intake (g)

618.4

606.9

615.1

Mean weight gain (g)

119.9

124.6

118.0

Mean weight gain, adjusted for food intake (g)

119.0

124.2

118.7

 

Conclusions:
Male and remale rats were exposed to 0.083 and 0.208% of the test substance incorporated into the diet in a 30 day repeated dose study. This 30 day exposure did not produce any mortality or evidence of systemic toxicity. No changes were observed for food intake, or body weight gain for the study animals. There were no histopathological findings noted on any of the study animals at necropsy. The no observable effect level (NOEL) for systemic toxicity was the highest dose level tested, 0.208%.
Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
2015-08-31 to 2015-10-02
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
Preliminary toxicity study, with supporting results, not according to OECD and EC guidelines but based on OECD 407 and GLP principles.
Principles of method if other than guideline:
The preliminary toxicity study was performed based on the principles described in OECD guideline 407.
GLP compliance:
no
Remarks:
The work was performed following GLP principles, no claim is made.
Limit test:
no
Specific details on test material used for the study:
- Name as cited in study report: POPDA, reaction products of propane-1,2-diol, propoxylated by animation of the terminal hydroxyl groups
- Color: yellow
- Analytical purity: 100%
- Impurities: water 0.12%
- Lot/Batch number: DR66700414
- Expiry date: 2016-12-05
- Stability: satisfactory stability was documented at 10 and 200 mg/ml following refrigerated (nominally 4-8°C) or ambient (nominally 21°C) storage for 15 days
- Storage: dark
- Total correction factor: none
-Ppurity/weighing factor: none
Species:
rabbit
Strain:
New Zealand White
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: 6 female New Zealand White rabbits, obtained from stock
- Age at start of study (day 1): 12 to 40 weeks old
- Weight at start of study (day 1): 3.00 to 3.44 kg
- Fasting period before study: no data
- Housing: individually housed in suspended plastic cages fitted with perforated floor panels and mounted in batteries. Undertrays lined with absorbent paper were changed at least 3 times a week.
- Diet (e.g. ad libitum): restricted availability (200 g/animal/day), Harlan High Fiber 2931C diet
- Water (e.g. ad libitum): ad libitum, potable water from the public supply via polycarbonate bottles with sipper tubes
- Acclimation period: 3 days before commencement of treatment

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16-20 °C
- Humidity (%): 40-70%
- Air changes (per hr): filtered fresh air which was passed to atmosphere and not recirculated, no further data
- Photoperiod (hrs dark / hrs light): 12/12, artificial lighting

IN-LIFE DATES:
- group 1: from 2015-09-03 to 2015-09-12
- group 2: from 2015-09-21 to 2015-10-02
Route of administration:
oral: gavage
Vehicle:
water
Remarks:
purified
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
- An appropriate volume of vehicle was added to the test substance and mixed by magnetic stirring until dissolution was achieved. A series of solutions at the required concentrations were prepared by serial dilution of the primary solution with the vehicle. Formulations were stirred using a magnetic stirrer before and throughout the dosing procedure.
- Frequency of preparation: weekly
- Storage of formulation: ambient temperature (nominally 21°C)
- Detailed records of compound usage were maintained. The amount of test substance necessary to prepare the formulations and the amount actually used were determined on each occasion. The difference between these amounts was checked before the formulations were dispensed.


VEHICLE
- Concentration in vehicle: 60 mg/ml (300 mg/kg) and 120 mg/ml (600 mg/kg)
- Amount of vehicle (if gavage): 5 ml/kg
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
group 1: 10 days (group prematurely terminated on day 10 of study)
group 2: 11 days (group prematurely terminated on day 12 of study, with animals having received 11 consecutive daily doses)
Frequency of treatment:
group 1: single doses on days 1 and 8 - third planned dose on day 15 was not administered
group 2: repeated daily dosing (11 occasions)
Dose / conc.:
300 mg/kg bw/day (actual dose received)
Remarks:
group 1 day 1, group 2
Dose / conc.:
600 mg/kg bw/day (actual dose received)
Remarks:
group 1, day 8
No. of animals per sex per dose:
3 females/group; 2 groups
Control animals:
no
Details on study design:
- Dose selection rationale: the starting dose of 300 mg/kg/day was selected by the sponsor. The subsequent dose levels for group 1 was based upon the apparent lack of a reaction observed, following a single dose at 300 mg/kg/day. Due to marked effects in the stomach of group 1 animals following a single dose at 600 mg/kg/day, the dose level for group 2 animals for repeated daily dosing was set at 300 mg/kg/day.
Positive control:
no
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least twice daily (at least once daily during acclimatization period)
- Animals were inspected visually for evidence of ill-health or reaction to treatment. Cages were inspected for evidence of animal ill-health amongst the occupant.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: at least twice daily:
- on dosing days: predose observation, upon completion of dosing, 30 minutes to 1 hour after completion of dosing, 2 to 3 hours after completion of dosing, as late as possible in the working day;
-on non-dosing days: in the morning, as late as possible in the working day.

BODY WEIGHT: Yes
- Time schedule for examinations: weekly during acclimatization, daily from 3 days prior to the start of treatment (Day -3), on the day that treatment commenced (Day 1), daily throughout the study and before necropsy.

FOOD CONSUMPTION:
- The weight of food supplied to each cage, that remaining and an estimate of any spilled was recorded daily from 3 days prior to the start of treatment (day -3) and throughout the study.

FOOD EFFICIENCY: No

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: No

CLINICAL CHEMISTRY: No

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

OTHER:
MORTALITY:
- A viability check was performed near the start and end of each working day.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
- All animals were subject to a detailed necropsy. After a review of the history of each animal, a full macroscopic examination of the tissues was performed. All external features and orifices were examined visually. Any abnormality in the appearance or size of any organ and tissue (external and cut surface) was recorded and the required tissue samples preserved in appropriate fixative. In light of the macropathology findings for the Group 1 decedent animal, the stomach was also retained.
- The retained tissues were checked before disposal of the carcass.
- Schedule: Group 1 animals: day 10 (2 days after last dose); Group 2 animals: day 12 (1 day after last dose)
- Method of kill: Intravenous injection of sodium pentobarbitone.

HISTOPATHOLOGY: No
Statistics:
No statitical analysis performed.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
- Following a single dose at 300 mg/kg/day in group 1, there were no marked clinical signs to Day 8 of study. A single dose of PODPA at 300 mg/kg/day was well tolerated.
- Following a single dose at 600 mg/kg in group 1, one female (number 2) showed grinding of teeth the day after dosing (day 9). On Day 10 of study (2 days after dosing at 600 mg/kg/day) female 2 was found dead.
- Repeat dosing of 300 mg/kg/day (group 2) elicited clinical signs of little hay eaten and few faeces for the two most affected females (numbers 4 and 6). Clinical signs of female 5 were restricted to little hay eaten on one occasion.
Mortality:
mortality observed, treatment-related
Description (incidence):
- Female 4 (group 2) was found dead on day 12, having received 11 consecutive daily doses at 300 mg/kg/day.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
- Following a single dose at 300 mg/kg/day in group 1, there were no adverse effects upon body weight performance to Day 8 of study. A single dose of PODPA at 300 mg/kg/day was well tolerated.
- Repeat dosing of 300 mg/kg/day (group 2) elicited a marked reduction in food intake in 2 out of 3 females (number 4 and 6), with subsequent body weight loss of between 210 and 220 g. Female 5 showed no clear or marked effect of treatment upon food consumption. Female 5 and overall body weight loss of just 20 g was recorded. The reduction in food intake was considered treatment related, with secondary effects manifest as body weight loss and clinical signs consistent with inappetance.
Food consumption and compound intake (if feeding study):
not examined
Description (incidence and severity):
- Following a single dose at 300 mg/kg/day in group 1, there were no adverse effects upon food consumption to Day 8 of study. A single dose of PODPA at 300 mg/kg/day was well tolerated.
- Following a single dose at a dose level of 600 mg/kg in group 1, all animals showed reduced food intake from the day of dosing.
- Repeat dosing of 300 mg/kg/day (group 2) elicited a marked reduction in food intake in 2 out of 3 females (number 4 and 6), with subsequent body weight loss of between 210 and 220 g. Female 5 showed no clear or marked effect of treatment upon food consumption. Female 5 and overall body weight loss of just 20 g was recorded. The reduction in food intake was considered treatment related, with secondary effects manifest as body weight loss and clinical signs consistent with inappetance.

Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
- Macroscopic examination of female 2 (group 1) revealed red fluid in the abdomen, thickened omental adipose tissue with poorly defined dark areas, dark areas on the jejunum which contained yellow viscous fluid, a pale liver, and a pyloric perforation and several depression(s) in the pylorus of the stomach.
- Macroscopic examination of females 1 and 3 revealed findings including pale liver and/or duodenum, depressions on the pylorus and corpus, thickened pylorus, a distended stomach with liquefied or clear gelatinous contents and yellow viscous fluid in the jejunum. These findings were considered treatment-related.
- Macroscopic examination of female 4 (group 2) revealed dark areas on the mucosal aspect of the caecum, which contained liquified contents, multiple pylorus depressions on the stomach which was also distended and contained clear, viscous fluid and a thickened pylorus, and a depression of the stomach fundus. Due to the similarity of the findings of the GI tract in this decedent to those observed at 600 mg/kg/day, treatment at 300 mg/kg/day was subsequently suspended with surviving females killed for reasons of animal welfare on day 12 of their treatment period, having received 11 consecutive daily doses.
- Macroscopic examination of female 6 revealed a distended caecum, with liquified contents, a pale liver, multiple pylorus depressions and dark stomach contents.
- There were no macroscopic findings observed in female 5.
- The consistent macroscopic findings relating to the GI tract observed in two out of three females following repeat daily dosing at 300 mg/kg/day were considered treatment related.
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Description (incidence and severity):
- Due to the marked macroscopic findings in female 2 of group 1, which were considered treatment related, and the reduced food intake and body weight loss observed in surviving females, which was also considered treatment related, with the associated clinical sign of little hay eaten, this dose group (group 1) was terminated and a dose of 600 mg/kg/day was considered unsuitable for any further investigation. Surviving females 1 and 3 were subsequently killed for reasons of animal welfare.
Critical effects observed:
not specified
Conclusions:
Dose levels of 300 mg/kg/day or above are clearly unsuitable for any further repeat dosing investigations in the female New Zealand White Rabbit.
Due to the marked nature of the macroscopic necropsy findings observed in the stomach and gastrointestinal tract, a recommended high dose level for the first phase of a two-phased preliminary embryo-fetal study in the New Zealand White Rabbit would be 150 mg/kg/day.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
239 mg/kg bw/day
Study duration:
subchronic
Species:
rat

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: inhalation
Data waiving:
exposure considerations
Justification for data waiving:
a short-term toxicity study does not need to be conducted because a reliable sub-chronic (90 days) or chronic toxicity study is available, conducted with an appropriate species, dosage, solvent and route of administration
other:
Critical effects observed:
not specified
Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: inhalation
Data waiving:
exposure considerations
Justification for data waiving:
a short-term toxicity study does not need to be conducted because a reliable sub-chronic (90 days) or chronic toxicity study is available, conducted with an appropriate species, dosage, solvent and route of administration
other:
Critical effects observed:
not specified
Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records

Referenceopen allclose all

Endpoint:
sub-chronic toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1990-08-07 to 1990-04-27
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
Well documented study with significant information on study design and methodology to evaluate results according to OECD Guideline 411 Subchronic Dermal Toxicity: 90 Day Study. Deviations from OECD Guideline: females rats are below the suggested weight range, number of animals used was below the suggested 20 for each dose group; details on test substance analysis were lacking.
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 411 (Subchronic Dermal Toxicity: 90-Day Study)
Deviations:
yes
Remarks:
females rats are below the suggested weight range, number of animals are below the suggested 20 for each dose group.
GLP compliance:
yes
Limit test:
no
Specific details on test material used for the study:
- Name of test material (as cited in study report): 6398-9-20
- Molecular weight: 230
- Substance type: Clear liquid
- Physical state: There was no apparent change in the physical state of the test substance during administration.
- Analytical purity: The purity, identify, strength and stability of the test substance were the responsibility of the sponsor.
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Wilmington, Massachusetts
- Age at study initiation: 47 days old
- Weight at study initiation:
Males (Mean weight in grams)
Control: 204.5
50 mg/kg: 203.9
80 mg/kg: 206.7
250 mg/kg: 204.2
Control recovery: 202.2
250 mg/kg recovery: 203.5

Females (Mean weight in grams)
Control: 147.9
50 mg/kg: 142.3
80 mg/kg: 144.6
250 mg/kg: 146.3
Control recovery: 146.4
250 mg/kg recovery: 141.9

- Housing:Stainless steel 1/2 inch wire mesh cages. Cage sizes were in accordance with the Guide for the /care and Use of Laboratory Animals. of the Institute of Laboratory Resources, National Research Council (NIH 86-23, 1985).
- Diet (e.g. ad libitum): Purina Certified Rodent Lab Meal, ad libitum. Feeders are designed to reduce soiling, bridging and scattering.
- Water (e.g. ad libitum): Fresh tap water, ad libitum
- Acclimation period: 11 days prior to study initiation. During acclimation, animals were weighed and observed for clinical signs of disease and weight gain and were found to be normal at study initiation.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 deg C +/- 3 deg C (63 deg F to 74 deg F)
- Humidity (%): 30 to 70%
- Photoperiod (hrs dark / hrs light): 12 hours dark/12 hours light

IN-LIFE DATES: From: To: August 7, 1089-December 4, 1989
Type of coverage:
semiocclusive
Vehicle:
water
Details on exposure:
TEST SITE
- Area of exposure: Dorsal
- Type of wrap if used: Gauze patch (2 x 2) and wrapped with an elastic bandage. The wrapping was then taped to the animal.
- Time intervals for shavings or clipplings: Clipping was employed weekly or as needed.


TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 5 ml/kg
- Concentration (if solution): 0, 50, 80, 250 mg/kg
- Constant volume or concentration used: constant concentration


VEHICLE
- Justification for use and choice of vehicle (if other than water): deionized water
- Amount(s) applied (volume or weight with unit): 5 ml/kg
- Concentration (if solution): 5 ml/kg


USE OF RESTRAINERS FOR PREVENTING INGESTION: no
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
No data
Duration of treatment / exposure:
The duration of exposure was approximately 6 hours.
Frequency of treatment:
Once daily, five days per week for a period of 90 days.
Dose / conc.:
0 mg/kg bw/day (nominal)
Dose / conc.:
50 mg/kg bw/day (nominal)
Dose / conc.:
80 mg/kg bw/day (nominal)
Dose / conc.:
250 mg/kg bw/day (nominal)
No. of animals per sex per dose:
Pretest (Baseline data for hematology and clinical chemistries): 5 males, 5 females
0, 50, 80, 250 mg/kg : 15 males, 15 females
0, 250 mg/kg recovery group: 10 males, 10 females.
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Doses were selected by the sponsor and based upon a 28 Day Dermal Toxicity Study in Rats
- Rationale for animal assignment (if not random): All rats were weighed, ranked according to body weight and assigned to treatment groups using a table for random numbers so that each treatment group had a similar distribution according to body weight.
- Rationale for selecting satellite groups: A negative control recovery group and a high dose recovery groups remained on test, untreated , for an additional 28 days to determine reversibility, persistence, or delayed occurrence of toxic effects.
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations: Daily observations to document onset, degree and duration of: changes in skin and fur, eyes and mucous membranes, respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behavior pattern. Mortality checks were made daily and recorded.
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Scheduled sacrifice periods of the study ( 30, 90 and 118 days)

DERMAL IRRITATION (if dermal study): Yes
- Time schedule for examinations: Study did not specify

BODY WEIGHT: Yes
- Time schedule for examinations: Weeky

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

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No

WATER CONSUMPTION: No
- Time schedule for examinations: N/A

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: Pretest, and on Days 25, 88 and 116
- Dose groups that were examined: all animals

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Scheduled sacrifice periods of the study ( 30, 90 and 118 days)
- Anaesthetic used for blood collection: No data
- Animals fasted: Yes, all animals were fasted overnight prior to euthanasia
- How many animals: 10 rats (5/sex)
- Parameters examined: hemoglobin, hematocrit, erythrocyte count, total and differential leukocyte count, platelet count, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Scheduled sacrifice periods of the study ( 30, 90 and 118 days)
- Animals fasted: Yes, all animals were fasted overnight prior to euthanasia
- How many animals: 10 rats (5/sex)
- Parameters examined: calcium, phosphorus, chloride, sodium, potassium, fasting glucose, serum alamine aminotransferase, serum aspartate aminotransferase, gamma glutamyl transpeptidase, urea nitrogen, albumin, globulin, blood creatinine, total bilirubin, total serum protein measurements

URINALYSIS: No
- Time schedule for collection of urine: N/A
- Metabolism cages used for collection of urine: N/A
- Animals fasted: N/A
- Parameters checked in table [No.?] were examined. N/A

NEUROBEHAVIOURAL EXAMINATION: No
- Time schedule for examinations: N/A
- Dose groups that were examined: N/A
- Battery of functions tested: sensory activity / grip strength / motor activity / other: N/A

OTHER:
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
Necropsy was performed on all animals and included examination of the external surface of the body, all orifices, and the cranial, thoracic, abdominal and pelvis cavities and their contents.
HISTOPATHOLOGY: Yes
The following organs and tissues were fixed and preserved in 10% neutral buffered formal for possible histopathological examination: gross lesions, brain-including section 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 organ (epididymides, prostate, and if present, seminal vesicles), aorta, skin (treated and untreated areas), esophagus, nasal turbinates, stomach, duodenum, jejunum, ileum, cecum, colon, rectum, urinary bladder, representative lymph nodes (submandibular), mammary gland, thigh musculature, peripheral nerve, eyes, femur-including articular surface, spinal cord at three levels- cervical, midthoracic and lumbar, exorbital lachrymal glands.
Other examinations:
The following organs were weighed: liver, adrenal glands, kidneys, gonads, brain.
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 ere indicated, Dunnett¿s or Student¿s t test was used to determine significant differences from control means.
Clinical signs:
no effects observed
Dermal irritation:
effects observed, treatment-related
Mortality:
no mortality observed
Description (incidence):
Two rats died on test. One male from the high dose group died on Day 1 and one female from the recovery control group died on Day 11. The deaths were judged coincidental and not related to the administration of the test substance.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
No statistically significant differences in body weight were detected between males of the control and low and mid dose groups or females of the control and low and high dose groups. Statistically significant reductions in body weights were observed in mid dose females on Day 14 when compared to the control group. The male high dose group mean body weights were significantly reduced on Days 7, 14 ,and 28.

Daily body weight gain was comparable between control groups and test substance treated groups except on four occasions. The male group mean daily body weight gains were significantly increased in the low dose group on Day 30 (fasted weight) and significantly reduced in the high dose group on Days 7 and 14. The female high dose daily body weight gains were significantly increased on Day 21.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
No statistically significant differences in food consumption were detected between the non-recovery control group and non-recovery test substance treated group. The male high dose recovery group mean daily food consumption was significantly reduced on Day 112 when compared to the recovery control group. Female high dose recovery group mean daily food consumption was significantly reduced on Days 42 and 112.
Food efficiency:
not examined
Description (incidence and severity):
not applicable
Water consumption and compound intake (if drinking water study):
not examined
Description (incidence and severity):
not applicable
Ophthalmological findings:
no effects observed
Description (incidence and severity):
There were no ocular findings identified in this study that were attributed to test substance administration.
Haematological findings:
no effects observed
Description (incidence and severity):
A significant decrease was observed in segmented neutrophils on Day 90 for the mid dose females. This decrease was considered not to be biologically significant. There were no additional statistical significant differences in hematological values on Day 30, 90 or 118.
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
Evaluation of the clinical chemistry data just prior to the Day 30 interim sacrifice revealed significantly higher male values in calcium for the mid dose group, phosphorus for the mid and high dose groups and blood urea nitrogen for the high dose group. Significant lower male total protein values were observed for the low dose group and higher chloride values were noted for each treated male dose group. Significantly higher female phosphorus values were noted for the mid dose group. Significantly lower female values were noted in potassium for the mid and high dose groups and blood urea nitrogen for each treated female dose group. These differences were not dose dependent, are within normal limits and are considered not biologically significant. Evaluation of the clinical chemistry data at the Day 90 sacrifice revealed significantly lower male potassium for the low dose group and creatinine for mid and high dose groups. Significantly higher male chloride values were noted for the mid dose group. Female chloride and phosphorus values were significantly higher for the mid dose group and lower blood urea nitrogen values for the mid and high dose groups. These differences are within normal limits and are considered not biologically significant. Evaluation of the clinical chemistry data for the male high dose recovery group revealed significantly higher creatinine and blood urea nitrogen values when compared to the recovery control group. These values are within normal limits and are considered not biologically significant. There were no significant differences in the clinical chemistry values for the high dose recovery females.
Urinalysis findings:
not examined
Description (incidence and severity):
not applicable
Behaviour (functional findings):
not examined
Description (incidence and severity):
not applicable
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
- There were no statistically significant differences in absolute organ weights between the control and treated groups. A statistically significant decrease was observed in relative liver to body weight ratios for the male high dose group on Day 30 wen compared to the control group. There were no statistically significant differences in relative organ to brain weight ratios between the control and treated groups. The differences were not dose dependent and not considered biologically significant.
- Relative Organ to Body Weights Ratios:A statistically significant decrease was observed in relative liver to body weight ratios for the male high dose group on Day 30 when compared to the control group. There were no significant differences noted for the female groups or the high dose recovery group.
- Relative Organ to Brain Weights Ratios: There were no statistically significant differences in relative organ to brain weight ratios between the control groups and treated groups.
Gross pathological findings:
no effects observed
Description (incidence and severity):
No statistically significant, biologically relevant effects were noted. The majority of gross findings at the Day 30 interim necropsy, Day 90 terminal necropsy and after 28 day recovery (day 118) were non-specific and low in incidence. Based on the histopathology, these lesions were not considered to be related to administration of the test substance.
Description (incidence and severity):
not applicable
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
When compared to the male and female rats in the control group, except for mild irritation of the skin at the treatment site of some of the rats, there were no histomorphologic changes present in the tissues evaluated which were related to the dermal exposure to the test substance. The skin irritation was characterized by a minimal to slight hyperkeratosis and/or slight epidermal hyperplasia in five males and seven females. Five of these seven females had multifocal areas of minimal to moderate acute inflammation of the epidermis of the skin (multifocal epidermatitis).
Histopathological findings: neoplastic:
no effects observed
Description (incidence and severity):
not applicable
Key result
Dose descriptor:
NOAEL
Remarks:
systemic toxicity
Effect level:
>= 250 mg/kg bw/day (nominal)
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects
Key result
Dose descriptor:
NOAEL
Remarks:
local (dermal) effects
Effect level:
80 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: erythema, edema, necrosis, fissuring/sloughing of skin and alopecia (hair loss).
Key result
Critical effects observed:
no

No definitive evidence of test substance related dermal irritation for the low or mid dose males groups. Infrequent dermal irritation was noted for the low dose females during a few day of the study. Several mid dose females displayed test substance related dermal irritation toward the latter part of the treatment interval. In the high dose group, clinical findings indicative of dermal irritation at the application site were noted and included some or all of the following observations: erythema, edema, scattered areas of necrosis, 25-50% necrosis at the application site, scab formation, fissuring and /or sloughing of skin, and alopecia. The incidence and severity of dermal effects varied throughout the study period, but generally increased with duration of treatment. Clinical observations indicative of dermal irritation in the high dose groups on Day 90 were no longer evident for the recovery group following the end of the 28 day reversibility period (Day 118).

Conclusions:
Based upon the results of the Subchronic 90 Day Dermal Toxicity Study in rats with test substance, there were no clinical signs of systemic toxicity which were attributable to the administration of the vehicle control or test substance. The test substance related mild skin irritation was primarily observed in the high dose group. The skin irritation was reversible after discontinuation of treatment during the recovery period. Two non-treatment related deaths occurred during this study. The deaths were judged coincidental. Significant differences in body weight, daily body weight gain and daily food consumption were detected during the few days of the study. There were no statistically significant differences in the absolute organ weights, or relative organ to brain weight ratios. One significant difference was noted for the male relative to liver to body weight data. Several significant differences were noted in the clinical chemistry data. There was one significant difference noted in the hematological data. These differences were not dose dependent, are within limits and are considered not biologically significant or attributable to the administration of the test substance. Except for the mild skin irritation at the treatment site of the high dose animals, there were not histomorphological alterations attributable to the administration of the test substance. Based upon these findings, dermal application of the test substance to rats did not produce a systemic toxicity when administered five days per week for 30 and 90 days at doses 50, 80 or 250 mg/kg. The no observable effect level (NOEL) for systemic toxicity was the highest dose level tested, 250 mg/kg/day. The NOEL for dermal effects was 80 mg/kd/day.
Endpoint:
short-term repeated dose toxicity: dermal
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1989-06-26 to 1989-09-28
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study without detailed documentation
Remarks:
initial body weight slightly less than prescribed by guideline; haematology, clinical chemistry, urinalysis, and histopathology not performed. No information was provided on the test substance (e.g. purity).
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 410 (Repeated Dose Dermal Toxicity: 21/28-Day Study)
Deviations:
yes
Remarks:
initial body weight slightly less than prescribed by guideline; Haematology, clinical chemistry, urinalysis, and histopathology not performed. No information was provided on the test substance.
GLP compliance:
not specified
Limit test:
no
Specific details on test material used for the study:
- Name as cited in study report: Poly[oxy(methyl-1,2-ethanediyl)], alpha-(2-aminomethylethyl)-omega-(2-aminomethylethoxy)-
- Molecular weight: 230
- Substance type: Active
- Color: clear
- Lot/Batch number: 89-006
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Wilmington, Massachusetts
- Age at study initiation: 6-8 weeks old
- Weight at study initiation: Mean weight of all 5 treatment groups combined was 191.4 grams
- Fasting period before study: no data
- Housing: Individually housed in stainless steel ½¿ wire mesh cages
- Diet (e.g. ad libitum): Purina Certified Rodent Meal, ad libitum
- Water (e.g. ad libitum): Fresh tap water, ad libitum
- Acclimation period: 10 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3
- Humidity (%): 30-70%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES (Repeat Dose-28 day study): From: 1989-06-26 To: 1989-07-24
IN-LIFE DATES (Pilot Dose-Range-Study): From: 1989-06-08 To: 1989-06-12
Type of coverage:
not specified
Vehicle:
other: deionized water
Details on exposure:
TEST SITE
- Area of exposure: dorsal area, shaved
- % coverage: No data
- Type of wrap if used: 2 X 2 inch gauze patch and wrapped with an elastic bandage and then taped to the animal
- Time intervals for shavings or clipplings: Weekly or as needed basis

REMOVAL OF TEST SUBSTANCE
- Washing (if done): No data although the study states that exposure was exactly 6 hours implying that washing occurred after 6 hours after application.
- Time after start of exposure: No data

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 5 ml/kg
- Concentration (if solution): 50, 100, 250, 500, and 1000 mg/kg
- Constant volume or concentration used: No data
- For solids, paste formed: Not applicable

VEHICLE
- Justification for use and choice of vehicle (if other than water): Not applicable
- Amount(s) applied (volume or weight with unit): No data
- Concentration (if solution): No data
- Lot/batch no. (if required): No data
- Purity: No data

USE OF RESTRAINERS FOR PREVENTING INGESTION: No data
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
No data
Duration of treatment / exposure:
6 hours/day, 5 days/week, for 4 weeks (28 days)
Frequency of treatment:
once daily
Dose / conc.:
50 mg/kg bw/day (nominal)
Dose / conc.:
100 mg/kg bw/day (nominal)
Dose / conc.:
250 mg/kg bw/day (nominal)
Dose / conc.:
500 mg/kg bw/day (nominal)
Dose / conc.:
750 mg/kg bw/day (nominal)
No. of animals per sex per dose:
5/sex/dose
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: In the Pilot Dose-Range-Study, the dose levels administered were 100, 1000, and 2000 mg/kg. Three groups of six rats (3 male and 3 female per group) were dermally administered the test substance for approximately 6 hours at the respective dose levels. The rats were treated daily for four days and observed at the time of dosing and after unwrapping for clinical signs and mortality. Body weights were recorded at initiation and termination or when found dead. At the 100 mg/kg dose level, slight erythema was observed. At both the 1000 and 2000 mg/kg dose levels, light to severe erythema was observed, as well as, areas of scattered necrosis at the application site and necrosis of the skin at the application site. None of the animals died at the 100 or 1000 mg/kg dose levels. Two of the six animals died at the 2000 mg/kg dose level. Due to the severe erythema and necrosis observed at the 2000 mg/kg dose level, the animals were terminated from study prior to Day 3 of treatment. Based on these results, dose levels were selected for the repeat dose dermal toxicity study in rats ¿ 28 days based on the most recent body weights.
- Rationale for animal assignment (if not random): Before technical initiation, all rats were weighed, ranked according to body weight and assigned to treatment groups using a table of random numbers so that each treatment group had a similar distribution according to body weight.
- Rationale for selecting satellite groups: Not applicable
- Post-exposure recovery period in satellite groups: Not applicable
- Section schedule rationale (if not random): Not applicable
Positive control:
Not applicable
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: No data
- Cage side observations: Cage side observations included but were not limited to: changes in skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous systems, somatomotor activity and behavior patterns.

DETAILED CLINICAL OBSERVATIONS: No
- Time schedule: Not applicable

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

BODY WEIGHT: Yes
- Time schedule for examinations: Initial, weekly, and final

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

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No

WATER CONSUMPTION: No
- Time schedule for examinations: Not applicable

OPHTHALMOSCOPIC EXAMINATION: No
- Time schedule for examinations: Not applicable
- Dose groups that were examined: Not applicable

HAEMATOLOGY: No
- Time schedule for collection of blood: Not applicable
- Anaesthetic used for blood collection:Not applicable
- Animals fasted: Not applicable
- How many animals: Not applicable
- Parameters checked: Not applicable

CLINICAL CHEMISTRY: No
- Time schedule for collection of blood: Not applicable
- Animals fasted: Not applicable
- How many animals: Not applicable
- Parameters checked: Not applicable

URINALYSIS: No
- Time schedule for collection of urine: Not applicable
- Metabolism cages used for collection of urine: Not applicable
- Animals fasted: Not applicable
- Parameters checked: Not applicable

NEUROBEHAVIOURAL EXAMINATION: No
- Time schedule for examinations: Not applicable
- Dose groups that were examined: Not applicable
- Battery of functions tested: sensory activity / grip strength / motor activity / other: Not applicable

OTHER: Not applicable
Sacrifice and pathology:
GROSS PATHOLOGY: Yes, examination of the external surface of the body, all orifices, and the cranial, thoracic, abdominal and pelvic cavities and their contents.
HISTOPATHOLOGY: No
Other examinations:
Not applicable
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 test was used to determine significant differences from control means. Analysis of discrete data, where appropriate, was conducted using non-parametric procedures.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
There were no clinical signs observed in the vehicle control group or the 50 and 100 mg/kg dose groups.
Clinical signs observed in the 250 mg/kg dose groups were very slight to well-defined erythema, very slight edema, fissuring of skin, sloughing of skin and scattered necrosis of the dose area.
Clinical signs observed in the 500 mg/kg dose group included very slight to severe erythema and edema, fissuring of skin, sloughing of skin, 20% of dose area necrotic and scattered necrosis of dose area.
In the 750 mg/kg dose group, clinical signs observed were very slight to severe erythema and edema, fissuring of skin, sloughing of skin, 20%, 50%, and 75% of dose area necrotic and scattered necrosis of the dose area. The severity of these signs were dose-dependent.
Dermal irritation:
effects observed, treatment-related
Description (incidence and severity):
see below
Mortality:
mortality observed, treatment-related
Description (incidence):
None of the animals died during the study.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
There were no statistically significant differences observed in the group mean body weights in both males and females.
A statistically significant decrease was observed in the group mean daily body weight gains in the high-dose males on Day 21. This was considered incidental and not biologically significant. There were no statistically significant differences observed in the group mean daily body weight gains in the females.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
There were no statistically significant differences observed in the group mean daily food consumption in both males and females.
Food efficiency:
not examined
Description (incidence and severity):
not applicable
Water consumption and compound intake (if drinking water study):
not examined
Description (incidence and severity):
not applicable
Ophthalmological findings:
not examined
Description (incidence and severity):
not applicable
Haematological findings:
not examined
Description (incidence and severity):
not applicable
Clinical biochemistry findings:
not examined
Description (incidence and severity):
not applicable
Urinalysis findings:
not examined
Description (incidence and severity):
not applicable
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Description (incidence and severity):
not applicable
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
Terminal necropsy revealed mottled lungs, red foci throughout the lungs, yellow discoloration of the left lateral lobe of the liver, tan foci on the medial lobe of the liver and mottled kidneys in the test substance treated groups. Terminal necropsy of the vehicle control group revealed small granular, yellow-brown discoloration of the left lateral liver lobe in one animal.
Neuropathological findings:
not examined
Description (incidence and severity):
not applicable
Histopathological findings: non-neoplastic:
not examined
Description (incidence and severity):
Not applicable
Histopathological findings: neoplastic:
not examined
Description (incidence and severity):
Not applicable
Other effects:
not specified
Dose descriptor:
LOAEL
Effect level:
250 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: (local effects) clinical signs including: well-defined erythema, very slight edema, fissuring of skin, sloughing of skin and scattered necrosis of the dose area. (LOAEL for local effects.)
Dose descriptor:
NOAEL
Effect level:
100 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No local effects were seen at this dose. (NOAEL for local effects).
Dose descriptor:
LOAEL
Effect level:
50 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: (Systemic effects) Animals exhibited mottled organs (lungs/liver/kidneys) at necropsy. Doses were recommended for 90 day study.
Critical effects observed:
not specified
Conclusions:
During the OECD 410 guideline (28 day) dermal study, the LOAEL for local effects for males and females was 250 mg/kg bw based on well-defined erythema, very slight edema, fissuring of skin, sloughing of skin and scattered necrosis of the dose area. The LOAEL for systemic effects was 50 mg/kg bw based on effects on necropsy.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
250 mg/kg bw/day
Study duration:
subchronic
Species:
rat

Repeated dose toxicity: dermal - local effects

Link to relevant study records

Referenceopen allclose all

Endpoint:
sub-chronic toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1990-08-07 to 1990-04-27
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
Well documented study with significant information on study design and methodology to evaluate results according to OECD Guideline 411 Subchronic Dermal Toxicity: 90 Day Study. Deviations from OECD Guideline: females rats are below the suggested weight range, number of animals used was below the suggested 20 for each dose group; details on test substance analysis were lacking.
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 411 (Subchronic Dermal Toxicity: 90-Day Study)
Deviations:
yes
Remarks:
females rats are below the suggested weight range, number of animals are below the suggested 20 for each dose group.
GLP compliance:
yes
Limit test:
no
Specific details on test material used for the study:
- Name of test material (as cited in study report): 6398-9-20
- Molecular weight: 230
- Substance type: Clear liquid
- Physical state: There was no apparent change in the physical state of the test substance during administration.
- Analytical purity: The purity, identify, strength and stability of the test substance were the responsibility of the sponsor.
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Wilmington, Massachusetts
- Age at study initiation: 47 days old
- Weight at study initiation:
Males (Mean weight in grams)
Control: 204.5
50 mg/kg: 203.9
80 mg/kg: 206.7
250 mg/kg: 204.2
Control recovery: 202.2
250 mg/kg recovery: 203.5

Females (Mean weight in grams)
Control: 147.9
50 mg/kg: 142.3
80 mg/kg: 144.6
250 mg/kg: 146.3
Control recovery: 146.4
250 mg/kg recovery: 141.9

- Housing:Stainless steel 1/2 inch wire mesh cages. Cage sizes were in accordance with the Guide for the /care and Use of Laboratory Animals. of the Institute of Laboratory Resources, National Research Council (NIH 86-23, 1985).
- Diet (e.g. ad libitum): Purina Certified Rodent Lab Meal, ad libitum. Feeders are designed to reduce soiling, bridging and scattering.
- Water (e.g. ad libitum): Fresh tap water, ad libitum
- Acclimation period: 11 days prior to study initiation. During acclimation, animals were weighed and observed for clinical signs of disease and weight gain and were found to be normal at study initiation.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 deg C +/- 3 deg C (63 deg F to 74 deg F)
- Humidity (%): 30 to 70%
- Photoperiod (hrs dark / hrs light): 12 hours dark/12 hours light

IN-LIFE DATES: From: To: August 7, 1089-December 4, 1989
Type of coverage:
semiocclusive
Vehicle:
water
Details on exposure:
TEST SITE
- Area of exposure: Dorsal
- Type of wrap if used: Gauze patch (2 x 2) and wrapped with an elastic bandage. The wrapping was then taped to the animal.
- Time intervals for shavings or clipplings: Clipping was employed weekly or as needed.


TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 5 ml/kg
- Concentration (if solution): 0, 50, 80, 250 mg/kg
- Constant volume or concentration used: constant concentration


VEHICLE
- Justification for use and choice of vehicle (if other than water): deionized water
- Amount(s) applied (volume or weight with unit): 5 ml/kg
- Concentration (if solution): 5 ml/kg


USE OF RESTRAINERS FOR PREVENTING INGESTION: no
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
No data
Duration of treatment / exposure:
The duration of exposure was approximately 6 hours.
Frequency of treatment:
Once daily, five days per week for a period of 90 days.
Dose / conc.:
0 mg/kg bw/day (nominal)
Dose / conc.:
50 mg/kg bw/day (nominal)
Dose / conc.:
80 mg/kg bw/day (nominal)
Dose / conc.:
250 mg/kg bw/day (nominal)
No. of animals per sex per dose:
Pretest (Baseline data for hematology and clinical chemistries): 5 males, 5 females
0, 50, 80, 250 mg/kg : 15 males, 15 females
0, 250 mg/kg recovery group: 10 males, 10 females.
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Doses were selected by the sponsor and based upon a 28 Day Dermal Toxicity Study in Rats
- Rationale for animal assignment (if not random): All rats were weighed, ranked according to body weight and assigned to treatment groups using a table for random numbers so that each treatment group had a similar distribution according to body weight.
- Rationale for selecting satellite groups: A negative control recovery group and a high dose recovery groups remained on test, untreated , for an additional 28 days to determine reversibility, persistence, or delayed occurrence of toxic effects.
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations: Daily observations to document onset, degree and duration of: changes in skin and fur, eyes and mucous membranes, respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behavior pattern. Mortality checks were made daily and recorded.
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Scheduled sacrifice periods of the study ( 30, 90 and 118 days)

DERMAL IRRITATION (if dermal study): Yes
- Time schedule for examinations: Study did not specify

BODY WEIGHT: Yes
- Time schedule for examinations: Weeky

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

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No

WATER CONSUMPTION: No
- Time schedule for examinations: N/A

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: Pretest, and on Days 25, 88 and 116
- Dose groups that were examined: all animals

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Scheduled sacrifice periods of the study ( 30, 90 and 118 days)
- Anaesthetic used for blood collection: No data
- Animals fasted: Yes, all animals were fasted overnight prior to euthanasia
- How many animals: 10 rats (5/sex)
- Parameters examined: hemoglobin, hematocrit, erythrocyte count, total and differential leukocyte count, platelet count, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Scheduled sacrifice periods of the study ( 30, 90 and 118 days)
- Animals fasted: Yes, all animals were fasted overnight prior to euthanasia
- How many animals: 10 rats (5/sex)
- Parameters examined: calcium, phosphorus, chloride, sodium, potassium, fasting glucose, serum alamine aminotransferase, serum aspartate aminotransferase, gamma glutamyl transpeptidase, urea nitrogen, albumin, globulin, blood creatinine, total bilirubin, total serum protein measurements

URINALYSIS: No
- Time schedule for collection of urine: N/A
- Metabolism cages used for collection of urine: N/A
- Animals fasted: N/A
- Parameters checked in table [No.?] were examined. N/A

NEUROBEHAVIOURAL EXAMINATION: No
- Time schedule for examinations: N/A
- Dose groups that were examined: N/A
- Battery of functions tested: sensory activity / grip strength / motor activity / other: N/A

OTHER:
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
Necropsy was performed on all animals and included examination of the external surface of the body, all orifices, and the cranial, thoracic, abdominal and pelvis cavities and their contents.
HISTOPATHOLOGY: Yes
The following organs and tissues were fixed and preserved in 10% neutral buffered formal for possible histopathological examination: gross lesions, brain-including section 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 organ (epididymides, prostate, and if present, seminal vesicles), aorta, skin (treated and untreated areas), esophagus, nasal turbinates, stomach, duodenum, jejunum, ileum, cecum, colon, rectum, urinary bladder, representative lymph nodes (submandibular), mammary gland, thigh musculature, peripheral nerve, eyes, femur-including articular surface, spinal cord at three levels- cervical, midthoracic and lumbar, exorbital lachrymal glands.
Other examinations:
The following organs were weighed: liver, adrenal glands, kidneys, gonads, brain.
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 ere indicated, Dunnett¿s or Student¿s t test was used to determine significant differences from control means.
Clinical signs:
no effects observed
Dermal irritation:
effects observed, treatment-related
Mortality:
no mortality observed
Description (incidence):
Two rats died on test. One male from the high dose group died on Day 1 and one female from the recovery control group died on Day 11. The deaths were judged coincidental and not related to the administration of the test substance.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
No statistically significant differences in body weight were detected between males of the control and low and mid dose groups or females of the control and low and high dose groups. Statistically significant reductions in body weights were observed in mid dose females on Day 14 when compared to the control group. The male high dose group mean body weights were significantly reduced on Days 7, 14 ,and 28.

Daily body weight gain was comparable between control groups and test substance treated groups except on four occasions. The male group mean daily body weight gains were significantly increased in the low dose group on Day 30 (fasted weight) and significantly reduced in the high dose group on Days 7 and 14. The female high dose daily body weight gains were significantly increased on Day 21.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
No statistically significant differences in food consumption were detected between the non-recovery control group and non-recovery test substance treated group. The male high dose recovery group mean daily food consumption was significantly reduced on Day 112 when compared to the recovery control group. Female high dose recovery group mean daily food consumption was significantly reduced on Days 42 and 112.
Food efficiency:
not examined
Description (incidence and severity):
not applicable
Water consumption and compound intake (if drinking water study):
not examined
Description (incidence and severity):
not applicable
Ophthalmological findings:
no effects observed
Description (incidence and severity):
There were no ocular findings identified in this study that were attributed to test substance administration.
Haematological findings:
no effects observed
Description (incidence and severity):
A significant decrease was observed in segmented neutrophils on Day 90 for the mid dose females. This decrease was considered not to be biologically significant. There were no additional statistical significant differences in hematological values on Day 30, 90 or 118.
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
Evaluation of the clinical chemistry data just prior to the Day 30 interim sacrifice revealed significantly higher male values in calcium for the mid dose group, phosphorus for the mid and high dose groups and blood urea nitrogen for the high dose group. Significant lower male total protein values were observed for the low dose group and higher chloride values were noted for each treated male dose group. Significantly higher female phosphorus values were noted for the mid dose group. Significantly lower female values were noted in potassium for the mid and high dose groups and blood urea nitrogen for each treated female dose group. These differences were not dose dependent, are within normal limits and are considered not biologically significant. Evaluation of the clinical chemistry data at the Day 90 sacrifice revealed significantly lower male potassium for the low dose group and creatinine for mid and high dose groups. Significantly higher male chloride values were noted for the mid dose group. Female chloride and phosphorus values were significantly higher for the mid dose group and lower blood urea nitrogen values for the mid and high dose groups. These differences are within normal limits and are considered not biologically significant. Evaluation of the clinical chemistry data for the male high dose recovery group revealed significantly higher creatinine and blood urea nitrogen values when compared to the recovery control group. These values are within normal limits and are considered not biologically significant. There were no significant differences in the clinical chemistry values for the high dose recovery females.
Urinalysis findings:
not examined
Description (incidence and severity):
not applicable
Behaviour (functional findings):
not examined
Description (incidence and severity):
not applicable
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
- There were no statistically significant differences in absolute organ weights between the control and treated groups. A statistically significant decrease was observed in relative liver to body weight ratios for the male high dose group on Day 30 wen compared to the control group. There were no statistically significant differences in relative organ to brain weight ratios between the control and treated groups. The differences were not dose dependent and not considered biologically significant.
- Relative Organ to Body Weights Ratios:A statistically significant decrease was observed in relative liver to body weight ratios for the male high dose group on Day 30 when compared to the control group. There were no significant differences noted for the female groups or the high dose recovery group.
- Relative Organ to Brain Weights Ratios: There were no statistically significant differences in relative organ to brain weight ratios between the control groups and treated groups.
Gross pathological findings:
no effects observed
Description (incidence and severity):
No statistically significant, biologically relevant effects were noted. The majority of gross findings at the Day 30 interim necropsy, Day 90 terminal necropsy and after 28 day recovery (day 118) were non-specific and low in incidence. Based on the histopathology, these lesions were not considered to be related to administration of the test substance.
Description (incidence and severity):
not applicable
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
When compared to the male and female rats in the control group, except for mild irritation of the skin at the treatment site of some of the rats, there were no histomorphologic changes present in the tissues evaluated which were related to the dermal exposure to the test substance. The skin irritation was characterized by a minimal to slight hyperkeratosis and/or slight epidermal hyperplasia in five males and seven females. Five of these seven females had multifocal areas of minimal to moderate acute inflammation of the epidermis of the skin (multifocal epidermatitis).
Histopathological findings: neoplastic:
no effects observed
Description (incidence and severity):
not applicable
Key result
Dose descriptor:
NOAEL
Remarks:
systemic toxicity
Effect level:
>= 250 mg/kg bw/day (nominal)
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects
Key result
Dose descriptor:
NOAEL
Remarks:
local (dermal) effects
Effect level:
80 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: erythema, edema, necrosis, fissuring/sloughing of skin and alopecia (hair loss).
Key result
Critical effects observed:
no

No definitive evidence of test substance related dermal irritation for the low or mid dose males groups. Infrequent dermal irritation was noted for the low dose females during a few day of the study. Several mid dose females displayed test substance related dermal irritation toward the latter part of the treatment interval. In the high dose group, clinical findings indicative of dermal irritation at the application site were noted and included some or all of the following observations: erythema, edema, scattered areas of necrosis, 25-50% necrosis at the application site, scab formation, fissuring and /or sloughing of skin, and alopecia. The incidence and severity of dermal effects varied throughout the study period, but generally increased with duration of treatment. Clinical observations indicative of dermal irritation in the high dose groups on Day 90 were no longer evident for the recovery group following the end of the 28 day reversibility period (Day 118).

Conclusions:
Based upon the results of the Subchronic 90 Day Dermal Toxicity Study in rats with test substance, there were no clinical signs of systemic toxicity which were attributable to the administration of the vehicle control or test substance. The test substance related mild skin irritation was primarily observed in the high dose group. The skin irritation was reversible after discontinuation of treatment during the recovery period. Two non-treatment related deaths occurred during this study. The deaths were judged coincidental. Significant differences in body weight, daily body weight gain and daily food consumption were detected during the few days of the study. There were no statistically significant differences in the absolute organ weights, or relative organ to brain weight ratios. One significant difference was noted for the male relative to liver to body weight data. Several significant differences were noted in the clinical chemistry data. There was one significant difference noted in the hematological data. These differences were not dose dependent, are within limits and are considered not biologically significant or attributable to the administration of the test substance. Except for the mild skin irritation at the treatment site of the high dose animals, there were not histomorphological alterations attributable to the administration of the test substance. Based upon these findings, dermal application of the test substance to rats did not produce a systemic toxicity when administered five days per week for 30 and 90 days at doses 50, 80 or 250 mg/kg. The no observable effect level (NOEL) for systemic toxicity was the highest dose level tested, 250 mg/kg/day. The NOEL for dermal effects was 80 mg/kd/day.
Endpoint:
short-term repeated dose toxicity: dermal
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1989-06-26 to 1989-09-28
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study without detailed documentation
Remarks:
initial body weight slightly less than prescribed by guideline; haematology, clinical chemistry, urinalysis, and histopathology not performed. No information was provided on the test substance (e.g. purity).
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 410 (Repeated Dose Dermal Toxicity: 21/28-Day Study)
Deviations:
yes
Remarks:
initial body weight slightly less than prescribed by guideline; Haematology, clinical chemistry, urinalysis, and histopathology not performed. No information was provided on the test substance.
GLP compliance:
not specified
Limit test:
no
Specific details on test material used for the study:
- Name as cited in study report: Poly[oxy(methyl-1,2-ethanediyl)], alpha-(2-aminomethylethyl)-omega-(2-aminomethylethoxy)-
- Molecular weight: 230
- Substance type: Active
- Color: clear
- Lot/Batch number: 89-006
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Wilmington, Massachusetts
- Age at study initiation: 6-8 weeks old
- Weight at study initiation: Mean weight of all 5 treatment groups combined was 191.4 grams
- Fasting period before study: no data
- Housing: Individually housed in stainless steel ½¿ wire mesh cages
- Diet (e.g. ad libitum): Purina Certified Rodent Meal, ad libitum
- Water (e.g. ad libitum): Fresh tap water, ad libitum
- Acclimation period: 10 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3
- Humidity (%): 30-70%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES (Repeat Dose-28 day study): From: 1989-06-26 To: 1989-07-24
IN-LIFE DATES (Pilot Dose-Range-Study): From: 1989-06-08 To: 1989-06-12
Type of coverage:
not specified
Vehicle:
other: deionized water
Details on exposure:
TEST SITE
- Area of exposure: dorsal area, shaved
- % coverage: No data
- Type of wrap if used: 2 X 2 inch gauze patch and wrapped with an elastic bandage and then taped to the animal
- Time intervals for shavings or clipplings: Weekly or as needed basis

REMOVAL OF TEST SUBSTANCE
- Washing (if done): No data although the study states that exposure was exactly 6 hours implying that washing occurred after 6 hours after application.
- Time after start of exposure: No data

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 5 ml/kg
- Concentration (if solution): 50, 100, 250, 500, and 1000 mg/kg
- Constant volume or concentration used: No data
- For solids, paste formed: Not applicable

VEHICLE
- Justification for use and choice of vehicle (if other than water): Not applicable
- Amount(s) applied (volume or weight with unit): No data
- Concentration (if solution): No data
- Lot/batch no. (if required): No data
- Purity: No data

USE OF RESTRAINERS FOR PREVENTING INGESTION: No data
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
No data
Duration of treatment / exposure:
6 hours/day, 5 days/week, for 4 weeks (28 days)
Frequency of treatment:
once daily
Dose / conc.:
50 mg/kg bw/day (nominal)
Dose / conc.:
100 mg/kg bw/day (nominal)
Dose / conc.:
250 mg/kg bw/day (nominal)
Dose / conc.:
500 mg/kg bw/day (nominal)
Dose / conc.:
750 mg/kg bw/day (nominal)
No. of animals per sex per dose:
5/sex/dose
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: In the Pilot Dose-Range-Study, the dose levels administered were 100, 1000, and 2000 mg/kg. Three groups of six rats (3 male and 3 female per group) were dermally administered the test substance for approximately 6 hours at the respective dose levels. The rats were treated daily for four days and observed at the time of dosing and after unwrapping for clinical signs and mortality. Body weights were recorded at initiation and termination or when found dead. At the 100 mg/kg dose level, slight erythema was observed. At both the 1000 and 2000 mg/kg dose levels, light to severe erythema was observed, as well as, areas of scattered necrosis at the application site and necrosis of the skin at the application site. None of the animals died at the 100 or 1000 mg/kg dose levels. Two of the six animals died at the 2000 mg/kg dose level. Due to the severe erythema and necrosis observed at the 2000 mg/kg dose level, the animals were terminated from study prior to Day 3 of treatment. Based on these results, dose levels were selected for the repeat dose dermal toxicity study in rats ¿ 28 days based on the most recent body weights.
- Rationale for animal assignment (if not random): Before technical initiation, all rats were weighed, ranked according to body weight and assigned to treatment groups using a table of random numbers so that each treatment group had a similar distribution according to body weight.
- Rationale for selecting satellite groups: Not applicable
- Post-exposure recovery period in satellite groups: Not applicable
- Section schedule rationale (if not random): Not applicable
Positive control:
Not applicable
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: No data
- Cage side observations: Cage side observations included but were not limited to: changes in skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous systems, somatomotor activity and behavior patterns.

DETAILED CLINICAL OBSERVATIONS: No
- Time schedule: Not applicable

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

BODY WEIGHT: Yes
- Time schedule for examinations: Initial, weekly, and final

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

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No

WATER CONSUMPTION: No
- Time schedule for examinations: Not applicable

OPHTHALMOSCOPIC EXAMINATION: No
- Time schedule for examinations: Not applicable
- Dose groups that were examined: Not applicable

HAEMATOLOGY: No
- Time schedule for collection of blood: Not applicable
- Anaesthetic used for blood collection:Not applicable
- Animals fasted: Not applicable
- How many animals: Not applicable
- Parameters checked: Not applicable

CLINICAL CHEMISTRY: No
- Time schedule for collection of blood: Not applicable
- Animals fasted: Not applicable
- How many animals: Not applicable
- Parameters checked: Not applicable

URINALYSIS: No
- Time schedule for collection of urine: Not applicable
- Metabolism cages used for collection of urine: Not applicable
- Animals fasted: Not applicable
- Parameters checked: Not applicable

NEUROBEHAVIOURAL EXAMINATION: No
- Time schedule for examinations: Not applicable
- Dose groups that were examined: Not applicable
- Battery of functions tested: sensory activity / grip strength / motor activity / other: Not applicable

OTHER: Not applicable
Sacrifice and pathology:
GROSS PATHOLOGY: Yes, examination of the external surface of the body, all orifices, and the cranial, thoracic, abdominal and pelvic cavities and their contents.
HISTOPATHOLOGY: No
Other examinations:
Not applicable
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 test was used to determine significant differences from control means. Analysis of discrete data, where appropriate, was conducted using non-parametric procedures.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
There were no clinical signs observed in the vehicle control group or the 50 and 100 mg/kg dose groups.
Clinical signs observed in the 250 mg/kg dose groups were very slight to well-defined erythema, very slight edema, fissuring of skin, sloughing of skin and scattered necrosis of the dose area.
Clinical signs observed in the 500 mg/kg dose group included very slight to severe erythema and edema, fissuring of skin, sloughing of skin, 20% of dose area necrotic and scattered necrosis of dose area.
In the 750 mg/kg dose group, clinical signs observed were very slight to severe erythema and edema, fissuring of skin, sloughing of skin, 20%, 50%, and 75% of dose area necrotic and scattered necrosis of the dose area. The severity of these signs were dose-dependent.
Dermal irritation:
effects observed, treatment-related
Description (incidence and severity):
see below
Mortality:
mortality observed, treatment-related
Description (incidence):
None of the animals died during the study.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
There were no statistically significant differences observed in the group mean body weights in both males and females.
A statistically significant decrease was observed in the group mean daily body weight gains in the high-dose males on Day 21. This was considered incidental and not biologically significant. There were no statistically significant differences observed in the group mean daily body weight gains in the females.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
There were no statistically significant differences observed in the group mean daily food consumption in both males and females.
Food efficiency:
not examined
Description (incidence and severity):
not applicable
Water consumption and compound intake (if drinking water study):
not examined
Description (incidence and severity):
not applicable
Ophthalmological findings:
not examined
Description (incidence and severity):
not applicable
Haematological findings:
not examined
Description (incidence and severity):
not applicable
Clinical biochemistry findings:
not examined
Description (incidence and severity):
not applicable
Urinalysis findings:
not examined
Description (incidence and severity):
not applicable
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Description (incidence and severity):
not applicable
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
Terminal necropsy revealed mottled lungs, red foci throughout the lungs, yellow discoloration of the left lateral lobe of the liver, tan foci on the medial lobe of the liver and mottled kidneys in the test substance treated groups. Terminal necropsy of the vehicle control group revealed small granular, yellow-brown discoloration of the left lateral liver lobe in one animal.
Neuropathological findings:
not examined
Description (incidence and severity):
not applicable
Histopathological findings: non-neoplastic:
not examined
Description (incidence and severity):
Not applicable
Histopathological findings: neoplastic:
not examined
Description (incidence and severity):
Not applicable
Other effects:
not specified
Dose descriptor:
LOAEL
Effect level:
250 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: (local effects) clinical signs including: well-defined erythema, very slight edema, fissuring of skin, sloughing of skin and scattered necrosis of the dose area. (LOAEL for local effects.)
Dose descriptor:
NOAEL
Effect level:
100 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No local effects were seen at this dose. (NOAEL for local effects).
Dose descriptor:
LOAEL
Effect level:
50 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: (Systemic effects) Animals exhibited mottled organs (lungs/liver/kidneys) at necropsy. Doses were recommended for 90 day study.
Critical effects observed:
not specified
Conclusions:
During the OECD 410 guideline (28 day) dermal study, the LOAEL for local effects for males and females was 250 mg/kg bw based on well-defined erythema, very slight edema, fissuring of skin, sloughing of skin and scattered necrosis of the dose area. The LOAEL for systemic effects was 50 mg/kg bw based on effects on necropsy.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
Study duration:
subchronic
Species:
rat

Additional information

Repeated dose toxicity- oral:

A repeated dose (31 day) oral toxicity study via feed in rats was conducted with the test substance (equivalent to OECD 407, Klimisch 4; American Cyanamid Company, 1968). The substance was incorporated into the diet to give final concentrations of 0.083% and 0.208% and fed to groups of 5 males and 5 females for 31 days. There were no deaths and no relevant gross pathology noted in either sex at autopsy. The NOAEL was deemed to be 239 mg/kg/day based on the lack of effects reported at this dose level.

A subacute, preliminary dose range finding study was performed in rabbits with the test substance administered via oral gavage at a single occasion at 300 and 600 mg/kg, or daily for 11 days at 300 mg/kg (Renaut, 2015; equivalent to OECD 407, Klimisch 2). Dose levels of 300 mg/kg were unsuitable for repeat dosing in rabbit and lead to marked findings in stomach and GI tract. A high dose of 150 mg/kg/day was recommended in a subsequent embryo-fetal development study.

Repeated dose toxicity - dermal:

A 28 -day dermal toxicity study was conducted in methods comparable to OECD guideline 410 (supporting, Klimisch 2; Pharmakon Research International Inc., 1989). Groups of 5 male and 5 female rats were exposed to the test substance 6 hours/day, 5 days/week for 4 weeks at 50, 100, 250, 500, and 750 mg/kg. Clinical signs observed in the 250, 500, and 750 mg/kg dose groups were very slight to well-defined erythema, very slight to severe edema, fissuring of skin, sloughing of skin and necrosis of the dose area. Terminal necropsy revealed mottled lungs, red foci throughout the lungs, yellow discoloration of the left lateral lobe of the liver, tan foci on the medial lobe of the liver and mottled kidneys in the test substance treated groups.

Based on the results of the 28 -day study, 50, 80, and 250 mg/kg were used in the subsequent 90 -day dermal study.

In a key, subchronic repeated dose toxicity study, the substance was applied dermally to rats 6 hours/day, 5 days per week, for 90 days with a 28-day recovery period at doses of 0, 50, 80, and 250 mg/kg/day (Pharmakon research International Inc., 1990; key). No systemic effects were reported at any dose. Therefore, the NOAEL for systemic effects was deemed to be 250 mg/kg/day (ie the highest dose tested). However, significant irritation was observed in the high dose group. Therefore, the NOAEL for local, dermal effects was deemed to be 80 mg/kg/day.

Justification for classification or non-classification

The NOAEL for systemic toxicity was equal to or greater than 250 mg/kg in the 90-day dermal toxicity study. Because the cut off value for category 2 specific target organ toxicity repeated exposure - dermal is 20-200 mg/kg/day, no classification is justified for the test substance based on the criteria laid down in the CLP regulation (EC) 1272/2008.