Reaction mass of bis(N-decyl-N,N-dimethyldecan-1-aminium) carbonate and N-decyl-N,N-dimethyldecan-1-aminium hydrogen carbonate

Repeated dose toxicity: dermal

Administrative data

Endpoint:

sub-chronic toxicity: dermal

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

- Reliability: GLP study according to international guidelines - Read-across justification: As both source (Didecyldimethylammonium chloride, DDAC) and target chemicals (Didecyldimethylammonium carbonate, DDA carbonate) have identical organic cations with hydrophobic side chains - the only difference is the inorganic anion carbonate or chloride with negligible contribution to the hazard properties- both substances can be predicted to have similar movement and fate characteristics.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Report date:

1988

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Test material form:

liquid: viscous

Details on test material:

- Name of test material: Didecyldimethylammonium chloride (DDAC)

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 8 weeks
- Weight at study initiation: males: 221.3-264.5 g; females: 166.7-219.11 g
- Fasting period before study: none
- Housing: individually housed in stainless steel cages
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 3 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°F): 66 to 75
- Humidity (%): 40 and 70
- Photoperiod (hrs dark / hrs light): 12

IN-LIFE DATES:
From: 11 January 1987
To: 12 April 1987

Administration / exposure

Type of coverage:

occlusive

Vehicle:

water

Details on exposure:

TEST SITE
- Area of exposure: 4" x 4"
- % coverage: 100 %
- Type of wrap if used: Vetrap® Bandaging Tape
- Time intervals for shavings or clippings: Weekly and as needed

REMOVAL OF TEST SUBSTANCE
- Washing: After removal of the wraps, the application site was rinsed. Water was liberally applied to the treatment area and the area was blotted using a 4" x 4" (8-ply) gauze pad.
- Time after start of exposure: 6 hours

TEST MATERIAL
- Amount(s) applied: 2.0 ml/kg bw
- Concentration (if solution): 0, 2, 6 and 12 mg/kg bw
- Constant volume or concentration used: Yes

VEHICLE
- Justification for use and choice of vehicle: Water
- Amount(s) applied: 2 mg/kg bw
- Concentration: 0, 0.1, 0.3 and 0.6% (w/w)

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Before initiation of the study, a trial batch of the dose solutions was prepared to assess the homogeneity and stability of the test material in water. For homogeneity, 3 samples each from the top, middle, and bottom of the mixing vessel were analyzed. Stability was evaluated 7 and 14 days after preparation by determining the test material concentration in triplicate samples in solutions prepared at the high and low concentrations and stored at room temperature.
Samples of the actual dosing solutions were taken and analyzed for test material concentration (prior to being used for dosing) during study weeks 1 through 4. In subsequent weeks, the samples were stored refrigerated, and a sample from the preparations made in weeks a and 13 were
analyzed.
Homogeneity studies performed on samples from all dosing solutions indicated that the test material was uniformly distributed. Stability studies were conducted on solutions (0.1 % and 0.6 %) stored at room temperature. These analyses indicated that the test material was stable in solution for at least 14 days. The values of analyses for the stability tests for both solutions ranged from 95.0 to 102.2 percent of nominal when assayed at 0, 7, and 14 days.
Concentration verification analyses (mean of duplicate assays) for the solutions prepared for the study ranged from 94.2 to 108.0 percent of nominal for all 3 concentrations.

Duration of treatment / exposure:

All animals were dosed for 13 weeks.

Frequency of treatment:

5 days/week, Monday through Friday. The females were also dosed on Saturday of the 13th week so that only 2 days elapsed between the administration of the last dose and the final sacrifice. This was done so that the elapsed time from dosing to sacrifice was the same as for the males.

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 2, 6 and 12 mg/kg bw/day
Basis:
nominal per unit body weight

No. of animals per sex per dose:

15/sex

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Three graduated dosage levels of the test material were evaluated in three groups of rats, anticipating that at the higher dosage level(s) some toxicological or pharmacological effect(s) will be observed and that at the lower dosage level(s) no treatment-related effects will be seen.
- Rationale for animal assignment (if not random): random
- Rationale for selecting satellite groups: no satellite groups selected
- Post-exposure recovery period in satellite groups: none
- Section schedule rationale (if not random): none

Positive control:

Not applicable

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily

DETAILED CLINICAL OBSERVATIONS: Yes / No / No data
- Time schedule:

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

BODY WEIGHT: Yes
- Time schedule for examinations: Body weight data were collected weekly for all animals.

FOOD CONSUMPTION: Yes
- Time schedule for examinations: Food consumption data were collected weekly for all animals.

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: Prior to initiation and prior to sacrifice.
- Dose groups that were examined:

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Blood was obtained just prior to sacrifice from all surviving animals via the retroorbital sinus for clinical chemistry and hematology analyses.
- Anaesthetic used for blood collection: Yes (methoxyflurane)
- Animals fasted: Yes (overnight)
- How many animals: All animals
- Parameters checked:
total leukocyte count
erythrocyte count
hemoglobin
hematocrit
erythrocyte indices
platelet count
differential leukocyte count
reticulocyte count

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Blood was obtained just prior to sacrifice from all surviving animals via the retroorbital sinus for clinical chemistry and hematology analyses.
- Animals fasted: Yes (overnight)
- How many animals: All animals
- Parameters checked:
AST (SGPT)
ALT (SGOT)
creatinine
alkaline phosphatase
gamma glutamyl transpeptidase
glucose
urea nitrogen
total protein
albumin
globulin (calculated)
A/G ratio (calculated)
total bilirubin
direct bilirubin
indirect bilirubin (calculated)
calcium
phosphorus
sodium
potassium
chloride

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
Gross lesions
Brain, incl. cerebral cortex, cerebellar cortex, medulla/ pons
Eyes
Pituitary
Salivary gland (mandibular with submandibular lymph node)
Heart
Aorta
Thymic region
Thyroid - parathyroid complex
Lungs with mainstem bronchi
Adrenals
Spinal cord
Cervical, Thoracic, Lumbar
Pancreas
Liver (3 lobes)
Kidneys
Urinary bladder
Testes
Prostate
Epididymis
Ovaries
Vagina
Uterus (corpus and cervix)
Spleen
Lymph nodes (mesenteric & mediastinal)Trachea
Esophagus
Stomach
Duodenum, Jejunum, Ileum,
Cecum, Colon, Rectum (entire organs)
Exorbital lacrymal gland
Femur (including articular surface)
Skeletal muscle
Sciatic nerve
Mammary gland (female)
Skin (application site & non-application site)
Sternum (including marrow)
Feet and ears were saved for identification purposes.

HISTOPATHOLOGY: Yes
All harvested tissues (see list above) from all male and female rats in the control and high dose groups were processed histologically and examined microscopically. 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.

Other examinations:

None.

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

Description (incidence and severity):

Skin irritation (erythema, oedema) was observed at 6 and 12 mg/kg bw/day primarily early in the study (Days 5-8).

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Ophthalmological findings:

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Details on results:

CLINICAL OBSERVATIONS AND MORTALITY
No treatment-related clinical signs of toxicity were observed in either sex during the 90-day treatment period. Gradable skin irritation (erythema and/or edema) was observed principally on Days 5 and 8 for several males and during the first 5 weeks for several females from some groups. Therefore, the skin of the treatment area was evaluated using the Draize scoring system for erythema and edema for all animals after dosing on study Days 5, 12, 19, 26,and 33, and prior to dosing on study Days 8, 15, 22, 29, and 36.
Signs of skin irritation included erythema (scored as barely perceptible or well defined) at the application site of animals from the low dose group (1 female), mid dose group (2 males and 4 females), and high dose group (6 males and 14 females). Edema (scored as barely perceptible or well defined) was also observed in animals from the mid dose group (1 male and 2 females) and high dose group (1 male and 9 females). Erythema and edema were observed only for females in the high dose treatment group after study Week 2 with the exception of isolated observations of erythema for one male in the high dose group and one female in each of the low and mid dose groups. Erythema and edema were observed for females in the high dose group through study Weeks 5 and 4, respectively. Mild exfoliation in the treatment area was also observed for approximately half of the males and most of the females in the high dose groups throughout the dosing period.
No male rats died during the study. One female from the low dose group and one female from the high dose group were found dead on study Days 77 and 63, respectively, and one female from the mid dose group was sacrificed in a moribund condition on Day 75. None of the deaths were attributed to treatment with the test substance.

FOOD CONSUMPTION
The data for the animals with significant food spillage are removed from each test period. No treatment-related differences were observed for either sex at any dose level.

BODY WEIGHTS
No treatment-related effects on body weight or weight gain were observed for either sex from any dose group. The occasional statistically significant increases in weight gain observed for females from the low and mid dose groups compared to controls were considered a result of random variation.

OPHTHALMIC EXAMINATIONS
No treatment-related findings were made for either sex at any dose level.

CLINICAL PATHOLOGY
No treatment-related effects on hematology or serum chemistry measurements occurred in any group of either sex.

ORGAN WEIGHTS AND FINAL BODY WEIGHTS
No treatment-related effect on organ weights (absolute or relative) was observed for male or female rats administered test material.
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) in treated animals compared to controls were considered spurious.

GROSS PATHOLOGY AND HISTOPATHOLOGY
Treatment-related gross findings indicative of minimal to mild skin irritation were observed at the two highest dosage levels used in this study. Microscopically, lesions were observed 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, hemorrhage, ulceration) occurred for females at the high dose. No other gross or microscopic pathologic findings were attributed to treatment with the test material.

Effect levels

Dose descriptor:

NOAEL

Effect level:

12 mg/kg bw/day (nominal)

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

other: see 'Remark'

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

Other than mild skin irritation, the occluded cutaneous treatment of male and female Sprague-Dawley rats with test material five days per week for 13 weeks at dosage levels of 2.0, 6.0, or 12.0 mg/kg bw/day resulted in no systemic toxicity as measured by clinical signs, food consumption, body weights or weight gain, ophthalmic changes, hematology or clinical chemistry measurements, gross pathology or histopathology. Since the 0.6 % concentration represented the maximum concentration which could be applied without producing more than slight skin irritation, and the 2.0 mL/kg bw volume represented the maximum volume which could be applied without run-off, the 12 mg/kg bw/day dosage level represented the maximum dosage level that could be evaluated in this test system. In addition, a local effect LOAEL of 2 mg/kg bw/day can be identified from this study.

Executive summary:

A study was carried out according to EPA OPP 82-3 (Subchronic Dermal Toxicity 90 Days) using the structural analog Didecyldimethylammonium chloride (DDAC). In view of the chemical and structural similarities (the relevant chemical part of both, DDAC and DDACarbonate, under the conditions of this test is the common quaternary ammonium cation Didecyldimethylammonium+), it is considered that the data are adequate for DDACarbonate. Solutions of test material were applied to the clipped backs of Sprague-Dawley CD® rats. The solutions were applied daily, five days per week, for 13 weeks. The dosing sites were occluded for at least 6 hours each day of dosing after which time the dressings were removed and the application site rinsed with tap water and blotted dry. Concentrations of 0, 0.1, 0.3 and 0.6 % (w/w) were evaluated in groups consisting of 15 male and 15 female rats. All solutions were administered at a constant volume of 2.0 mL/kg bw/day. The three concentrations, therefore, corresponded to dosage levels of 2, 6, and 12 mg/kg bw/day. Since the 0.6 % concentration represented the maximum concentration which could be applied without producing more than slight skin irritation, and the 2.0 mL/kg bw volume represented the maximum volume which could be applied without run-off, the 12 mg/kg bw/day dosage level represented the maximum dosage level that could be evaluated in this test system.

No treatment-related systemic toxicity as measured by clinical signs, food consumption, body weights or weight gain, ophthalmic changes, hematology or clinical chemistry measurements, gross pathology or histopathology was observed. Mild skin irritation at the treatment site was observed grossly in most animals treated with test material at a concentration of 0.6 %. Mild skin irritation was also observed grossly in a few animals at the 0.3 % concentration and in one female at the lowest concentration (0.1 %).

Changes in skin at the application site, indicative of irritation, were also observed microscopically for some animals at all dose levels. These microscopic changes were most evident in the female rats from the mid and high dose groups.

The systemic NOAEL was determined to be 12 mg/kg bw/day for males and females. In addition, a local effect NOAEL of 2 mg/kg bw/day could be identified from this study.