Amines, polyethylenepoly-, tetraethylenepentamine fraction

Repeated dose toxicity: dermal

Administrative data

Endpoint:

short-term repeated dose toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

April-May 1986

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

Purity of a.i. has been indicated; complete composition of the test substance is not available (available in departmental study file).

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1986

Report date:

1986

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 410 (Repeated Dose Dermal Toxicity: 21/28-Day Study)

Version / remarks:

1981

Deviations:

yes

Remarks:

20 instead of 21 days of exposure, occlusive dressing

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Amines, polyethylenepoly-, tetraethylenepentamine fraction

EC Number:

292-587-7

EC Name:

Amines, polyethylenepoly-, tetraethylenepentamine fraction

Cas Number:

90640-66-7

Molecular formula:

C8H23N5, C10H25N5

IUPAC Name:

(2-aminoethyl)[2-({2-[(2-aminoethyl)amino]ethyl}amino)ethyl]amine; bis(2-aminoethyl)({2-[(2-aminoethyl)amino]ethyl})amine

Test material form:

liquid

Test animals

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Ray Nichols Rabbitry, Lumberton, TX, USA
- Age at study initiation: ca. 5 months
- Weight at study initiation: ca. 3.1 - 4.0 kg
- Fasting period before study: not applicable
- Housing: individually in stainless steel cages
- Diet: restricted to 4 oz per animal per day
- Water: ad libitum
- Acclimation period: at least 14 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 72 ± 5 degr F (22 ± 1.5 degr C)
- Humidity (%): 50
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Type of coverage:

occlusive

Vehicle:

water

Details on exposure:

TEST SITE
- Area of exposure: 10 x 15 cm
- % coverage: 100
- Type of wrap if used: an occlusive bandage of absorbent gauze and non-absorbent cotton; the bandage was held in place using a lycra/spandex jacket
- Time intervals for shavings or clipplings: periodically as required

REMOVAL OF TEST SUBSTANCE
- Washing (if done): not reported
- Time after start of exposure: ca. 6 h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): a 10% solution was applied with the volume adjusted to provide the required dose
- Concentration (if solution): 10%
- Constant volume or concentration used: yes, constant concentration

VEHICLE
- Justification for use and choice of vehicle (if other than water): water
- Amount(s) applied (volume or weight with unit): 1 mL/kg BW

USE OF RESTRAINERS FOR PREVENTING INGESTION: no (but animals wore jackets)

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Test material concentration (100 mg/g) was measured 3 times during the study in triplo. % of target concentrations were 87.0, 95.3 and 90.9%, respectively. Stability was measured 2, 4 and 7 days after preparation. % of Day 0 was, 92.1, 59.4 and 59.9%, respectively.

Duration of treatment / exposure:

20 days

Frequency of treatment:

5 days/week, ca. 6h/day

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

50 mg/kg bw/day (nominal)

Doses / concentrations 2

Dose / conc.:

100 mg/kg bw/day (nominal)

Doses / concentrations 3

Dose / conc.:

200 mg/kg bw/day (nominal)

No. of animals per sex per dose:

5

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: based on a probe study using 2 female rabbits per group, at levels of 0, 50, 100 or 200 mg/kg bw/day for 4 days
- Rationale for animal assignment (if not random): computer-generated tables of random numbers
- Rationale for selecting satellite groups: not used
- Post-exposure recovery period in satellite groups: not used
- Section schedule rationale (if not random): no info

Positive control:

not used

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily on working days (one additional routine monitoring on these days and on weekend and holidays for dead animals and check for water and food availability)

DETAILED CLINICAL OBSERVATIONS: No

DERMAL IRRITATION (if dermal study): Yes
- Time schedule for examinations: daily upon removal when bandage and jacket were removed.

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION: Since the animals were maintained on a restricted diet and normally ate all their ration, measurement of food consumption was not doen. Qualitative changes in consumption were monitored by a periodic visual check.

FOOD EFFICIENCY: No

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: not applicable

HAEMATOLOGY: Yes
- Time schedule for collection of blood: immediately prior to necropsy
- Anaesthetic used for blood collection: Yes (no info which was used)
- Animals fasted: No
- How many animals: all
- Parameters examined: RBC, WPC, platelet count, haemoglobin, packed cell volume

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: immediately prior to necropsy
- Animals fasted: No
- How many animals: all
- Parameters examined: blood urea nitrogen, ALP, ALAT, glucose, total protein, albumin, Cl, Na, K, globulin (calculated)

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

OTHER:

Sacrifice and pathology:

GROSS PATHOLOGY: Yes (including eyes)
HISTOPATHOLOGY: Yes (complete set) of control and high dose animals; skin (application site and a non-treated section), liver kidneys and gross pathologic lesions were aslo examined from low and mid dose animals
Organ weights: brain, liver, kidneys, adrenals, ovaries, testes

Other examinations:

None

Statistics:

Yes, for body weights, organ weights, clinical chemistry, haematology.

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Dermal irritation:

effects observed, treatment-related

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

effects observed, treatment-related

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Histopathological findings: neoplastic:

no effects observed

Other effects:

not examined

Details on results:

CLINICAL SIGNS AND MORTALITY: one control female was removed from the study on day 14 because of a broken back.
There were no signs other than local dermal effects. These were dose-related and consisted of red dermal test sites on days 1-2. By study Day 6, animals tretaed with 200 mg/kg bw showed very red and slightly swollen test sites; on Day 16 the skin of some of these females was severely irritated with some crusting and bleeding. Other animals of the 200 mg/kg bw group and those of the 100 mg/kg group had irritated skin that did not progress further.

BODY WEIGHT AND WEIGHT GAIN: no effects

FOOD CONSUMPTION: no effects

FOOD EFFICIENCY: not measured

WATER CONSUMPTION: not measured

OPHTHALMOSCOPIC EXAMINATION: not applicable

HAEMATOLOGY: no effects

CLINICAL CHEMISTRY: no effects

URINALYSIS: not measured

NEUROBEHAVIOUR: not measured

ORGAN WEIGHTS: no effects

GROSS PATHOLOGY: confined to the skin of the 100 and 200 mg/kg groups, characterised by multifocal areas of epidermal and dermal necrosis often covered with crusts comprised of dry serum and necrotic debris.

HISTOPATHOLOGY: NON-NEOPLASTIC
Only dose-related changes in the skin consisting of: multifocal necrosis of the epidermis with extension into the dermis, acanthotic and hyperkeratotic areas, underlying dermis had a subacute inflammatory response consisting of lymphocytes, macrophages and heterophils. Dermal arterioles contained marginated leukocytes, hair folicles showed keratinization. Grading of the skin lesions reflected the extent if the lesion and the degree of functional damage.

HISTOPATHOLOGY: NEOPLASTIC (not applicable)

HISTORICAL CONTROL DATA (not applicable)

OTHER FINDINGS: none

Effect levels

1

Dose descriptor:

NOAEL

Remarks:

local

Effect level:

1.25 mg/cm² per day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

dermal irritation

2

Dose descriptor:

NOEL

Remarks:

systemic

Effect level:

>= 200 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

clinical signs

gross pathology

haematology

mortality

Target system / organ toxicity

Critical effects observed:

no

Any other information on results incl. tables

The NOAEL for the local effects is 50 mg/kg bw/day. Taking into account a mean body weight of this dose group of 3745 g and an exposed are of 150 cm² 50 mg/kg bw/day corresponds to about 1.25 mg/cm²/day.

Applicant's summary and conclusion

Conclusions:

At 100 and 200 mg/kg bw/day, the only lesions noted were skin lesions; the NOEL was 50 mg/kg bw/day.

Executive summary:

Groups of 5 male and 5 female rabbits were treated on their skin with 50, 100 or 200 mg Amines, polyethylenepoly-, tetraethylenepentamine fraction per kg/bw ca. 6 h/day day, 5 days/week for a period of 31 days (under occlusive conditions). These levels were based on a 4 -day RF study using the same levels using 2 females per group. Controls were treated with the vehicle, distilled water. No changes were observed in body weights, food intake, haematology, clinical chemistry, and organ weights. The only lesions observed were local skin lesions; the degree of irritation was dose-related; effects in the 200 mg/kg group were generally more severe than in the 100 mg/kg group. There were no indications of systemic toxicity. Because no changes were seen in the 50 mg/kg group, the NOEL was 50 mg/kg bw/day.