Poly(oxy-1,2-ethanediyl), α-[2-(dide- cylmethylammonio)ethyl]- .omega.- hydroxy-, propanoate (salt) (Bardap 26)

Repeated dose toxicity: dermal

Administrative data

Endpoint:

sub-chronic toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

January to April 1988

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1988

Report date:

1988

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EPA OPP 82-3 (Subchronic Dermal Toxicity 90 Days)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 411 (Subchronic Dermal Toxicity: 90-Day Study)

Deviations:

no

Principles of method if other than guideline:

Not applicable.

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

DDAC

IUPAC Name:

DDAC

Constituent 2

Reference substance name:

1-Decanaminium, N-decyl-N,N-dimethyl-, chloride

IUPAC Name:

1-Decanaminium, N-decyl-N,N-dimethyl-, chloride

Constituent 3

Reference substance name:

Didecyldimethylammonium chloride

EC Number:

230-525-2

EC Name:

Didecyldimethylammonium chloride

Cas Number:

7173-51-5

Molecular formula:

C22 H48 N.Cl

IUPAC Name:

N-decyl-N,N-dimethyldecan-1-aminium chloride

Constituent 4

Reference substance name:

N,N-Didecyl-N,N-dimethylammonium Chloride

IUPAC Name:

N,N-Didecyl-N,N-dimethylammonium Chloride

Constituent 5

Reference substance name:

Bardac 2280

IUPAC Name:

Bardac 2280

Details on test material:

The test material was Bardac 2280 (didecyldimethylammonium chloride in aqueous/alcohol solution: DDAC; CAS RN 7173-51-5), batch number B-1889, obtained from Lonza, Inc. The substance was described as a very viscous honey-coloured liquid, supplied as 80.8% a.s. in aqueous/ethanol solution. DDAC, is hydrolytically and photolytically stable under the conditions of this study and has been shown to be stable in aqueous, alcohol and alcohol/aqueous solutions for extended periods, e.g. at least seven years under standard laboratory conditions.

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

The test animals were male and female Sprague-Dawley rats obtained from Charles River Breeding Laboratories, MI, USA. They were acclimatised for approximately 3 weeks prior to study initiation. The rats were 8 weeks old at study initiation and weighed 221.3 - 264.5 g (males) and 166.7 - 219.11 g (females). Pretest health screens and viral screens were performed. Upon arrival the rats were housed in pairs in suspended stainless steel cages with mesh floors. After 1 week they were housed individually. Ground Purina Certified Rodent Chow #5002 and tap water were provided ad libitum. Temperature was maintained from 66-75°F and relative humidity was maintained from 40 to 70%. Lighting was provided on a 12 hour light/dark cycle, and there were at least 8 air changes per hour.
In life dates: 11 January 1988 to 12 April 1988.

Administration / exposure

Type of coverage:

occlusive

Vehicle:

water

Details on exposure:

The concentration of the test substance (expressed in terms of didecyldimethylammonium chloride; 80.8%) in the vehicle was 0, 0.1, 0.3 and 0.6% (w/w). Solutions were prepared weekly and stored at room temperature. Controls were dosed with water (the vehicle) at a constant dosing volume of 2.0 ml/kg bw.

Eight days prior to the first dose, the hair was clipped from the dorsal area of the trunk. All of the animals were wrapped in the manner used during the study for 6 hours/day for 4 days during the week prior to dosing. Animals that did not adapt to the pre-treatment wrapping procedure were not used in the study. Prior to the first dose, the fur was reclipped in preparation for dosing. Animals were reclipped in the afternoons as needed throughout the study, and on each Friday afternoon. The test material was applied directly to the back at each application. The entire application site was covered with a sterile 8-ply gauze pad and the rats were wrapped using Vetrap bandaging tape and secured with elastic tape. Dressings were left in place for approximately 6 hours. After dressing removal, the application site was rinsed with water and blotted using an 8-ply gauze pad. All animals were dosed 5 days/week, for 13 weeks. The females were also dosed on Saturday of the 13th week so that only 2 days elapsed between administration of the last dose and the final sacrifice.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Homogeneity studies were performed on samples from all dosing solutions, and indicated that the test material was uniformly distributed. Stability studies indicated that the test material was stable in solution for at least 14 days. Concentration analyses ranged from 94.2 to 108.0% of nominal for all 3 concentrations.

Duration of treatment / exposure:

90 days

Frequency of treatment:

5 days/week; 6 hours/day (Monday through Friday)

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

2 mg/kg bw/day (nominal)

Doses / concentrations 2

Dose / conc.:

6 mg/kg bw/day (nominal)

Doses / concentrations 3

Dose / conc.:

12 mg/kg bw/day (nominal)

No. of animals per sex per dose:

15/sex/dose

Control animals:

yes, concurrent vehicle

Details on study design:

Controls were exposed to the vehicle only (water). All solutions of the test substance were corrected for percentage active ingredient. The highest dose administered (0.6%) was chosen as the maximum dose that could be applied dermally without causing significant skin irritation. Only animals with an intact and normal epidermis were used in the study.

Positive control:

Not applicable.

Examinations

Observations and examinations performed and frequency:

Animals were observed once a week for detailed clinical observations, and six days a week for overt clinical signs. Skin reactions were scored according to a modified Draize procedure when signs were observed on Friday after dosing and on Monday before dosing. Observations for mortality and moribundity were made twice daily. Bodyweight data and food consumption data were collected weekly for all animals. Ophthalmoscopic examinations took place prior to study initiation and prior to sacrifice. Blood samples were collected from all animals prior to sacrifice for haematology and clinical chemistry analyses. The following haematology parameters were evaluated: erythrocyte count, haemoglobin, haematocrit, erythrocyte indices, platelet count, total leukocyte count, differential leukocyte count, reticulocyte count. The following clinical chemistry parameters were determined: glucose, urea nitrogen, creatinine, AST (SGPT), ALT (SGOT), creatine kinase, gamma glutamyl transpeptidase, alkaline phosphatase, total protein, total cholesterol, albumin, globulin, A/G ratio, total bilirubin, direct bilirubin, indirect bilirubin, calcium, phosphorous, sodium, potassium, chloride.

Sacrifice and pathology:

All surviving rats were necropsied and examined for gross pathologic changes. The following tissues were processed for histopathology: gross lesions, spinal cord, brain, pituitary, thyroid, thymic region, trachea, lungs, heart, salivary gland, liver, spleen, kidneys, adrenals, pancreas, testes, epididymides, prostate, seminal vesicles, ovaries, uterus, vagina, mammary gland, skin, oesophagus, stomach, duodenum, jejunum, ileum, caecum, colon, rectum, urinary bladder, lymph nodes, sciatic nerve, sternum, femur, skeletal muscle, eyes, aorta, exorbital lacrymal gland. Histopathologic examinations were performed on the harvested tissues from all animals in the control and high dose groups. In addition, the skin, lungs, liver, kidneys and all gross lesions were processed and examined histologically from all animals in the mid and low dose groups. Organ weights were obtained for the liver, kidneys, spleen, heart, brain with stem, adrenals, testes, ovaries.

Other examinations:

No other examinations reported.

Statistics:

Parametric variables were intercompared for the dose and control groups using Levene’s test for homogeneity of variances, by analysis of variance and by pooled variance t-tests. Non-parametric data were analyzed by the Kruskal-Wallis test or by the Wilcoxon rank sum test as modified by Mann-Whitney. Frequency data were compared using Fisher’s exact tests.

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:

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

effects observed, treatment-related

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Histopathological findings: neoplastic:

no effects observed

Other effects:

not specified

Details on results:

MORTALITY
One female each from the low and high dose groups died during the study on Days 77 and 63 respectively, and one female from the mid dose groups was sacrificed moribund on Day 75. These deaths were not considered related to treatment.

CLINICAL SIGNS
No treatment-related signs were observed in either sex at any dose throughout the 90-day dosing period. Skin irritation (erythema, oedema) was observed at 6 and 12 mg a.s./kg/d primarily early in the study (Days 5-8).

BODY WEIGHT
No treatment-related effects in either sex at any dose.

FOOD CONSUMPTION
No treatment-related effects in either sex at any dose.

OPHTHALMOSCOPIC EXAMINATION
No treatment-related effects in either sex at any dose.

HAEMATOLOGY
No treatment-related effects in either sex at any dose.

CLINICAL CHEMISTRY
No treatment-related effects in either sex at any dose.

ORGAN WEIGHTS
No treatment-related effects in either sex at any dose. Statistically significant differences (adrenal glands relative to body weight in males from the mid dose group and spleen weight relative to body weight in females from the mid dose group) were considered spurious.

GROSS PATHOLOGY
Treatment-related gross findings indicative of minimal to mild skin irritation were observed at the two highest doses.

HISTOPATHOLOGY: NON-NEOPLASTIC
Micropscopically, lesions wer eobserved in the treated skin for some animals from each treatment level. The greatest incidence of animals with clear histologic evidence of dermal and/or epidermal inflammation (epidermitis, dermatitis, vacuolar degeneration, haemorrhage, ulceration) occurred for females at the high dose. No other treatment-related observations were recorded.

Effect levels

Key result

Dose descriptor:

NOAEL

Effect level:

12 mg/kg bw/day (nominal)

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

other: Equivalent to 14.9 mg test substance/kg bw/d. No treatment-related effects were observed at any dose.

Target system / organ toxicity

Key result

Critical effects observed:

not specified

Any other information on results incl. tables

No further information available.

Applicant's summary and conclusion

Conclusions:

The NOAEL was considered to be 12 mg a.s./kg bw/d (equivalent to 14.9 mg test substance/kg bw/d), the highest dose tested.

Executive summary:

Male and female Sprague-Dawley rats were exposed dermally to Bardac 2280 (Didecyldimethylammonium Chloride aqueous/alcohol solution) for 90 days, according to USEPA OPP 82 -3 and OECD 411. The test substance was applied at 0, 2, 6 and 12 mg a.s./kg bw/d under occlusive dressings for 6 hours/day, 5 days/week for 90 days. The rats were observed for clinical signs and mortality and signs of dermal irritation. Bodyweights and food consumption were recorded weekly. At study termination ophthalmoscopic examinations, haematology and clinical chemistry analyses, gross necropsy and histopathology were carried out.

Other than a brief period of skin irritation at the two highest doses observed early in the study (Day 5 -8), and gross and microscopic indication of skin irritation after 90-days of treatment, no effects from repeated dermal exposure to the test substance were observed. The NOAEL was considered to be 12 mg a.s./kg bw/d, the highest dose tested (equivalent to 14.9 mg test substance/kg bw/d) .

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