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: oral

Administrative data

Endpoint:

sub-chronic toxicity: oral

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/OECD 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

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPP 82-1 (90-Day Oral Toxicity)

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: 241.2-308.7 g; females: 166.6-211.1 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: 15 September 1987
To: 16 December 1987

Administration / exposure

Route of administration:

oral: feed

Vehicle:

ethanol

Remarks:

approximately 10 % by weight

Details on oral exposure:

DIET PREPARATION
- Rate of preparation of diet: fresh diet was prepared and offered to the animals each week
- Mixing appropriate amounts with: ground rodent feed
- Storage temperature of food: room temperature

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Experimental diets were analyzed using a gas chromatographic procedure. Homogeneity of the test material at each diet concentration was established prior to the start of the study. Stability of the test material in diets at the 3000 and 100 ppm concentrations was determined in open glass feed jars and in the polyethylene storage containers prior to dosing. Diet concentrations were verified for all dose levels for the first four weeks of the study prior to administration of the diets to the animals. In subsequent weeks, samples of each test diet were retained from each preparation, and all samples from weeks 8 and 13 were analyzed with one control sample.
These analyses indicated that the test material was stable in the diets for at least 20 days under both storage conditions for all dose levels. The value of analyses for the stability tests for both diets ranged from 91.5 to 101.3 % of nominal when assayed at 13 and 20 days. Concentration verification analyses (mean of duplicate assays) for the diets prepared for the study ranged from 93.0 to 106.6 % of nominal for all 5 concentrations.

Duration of treatment / exposure:

90 to 91 days

Frequency of treatment:

7 days/week

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 100, 300, 600, 1000, or 3000 ppm AI
Basis:
nominal in diet

No. of animals per sex per dose:

15 males/15 females

Control animals:

yes, plain diet

Details on study design:

- Dose selection rationale: Five graduated dosage levels of the test material were evaluated in five 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
- Time schedule: weekly

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption and body weights were recorded weekly for each animal: Yes

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: prior to the start of dosing and prior to final sacrifice
- Dose groups that were examined: all dose groups

HAEMATOLOGY: Yes
- Time schedule for collection of blood: just prior to sacrifice
- Animals fasted: Yes
- Anaesthetic used for blood collection: Yes (methoxyflurane)
- How many animals: from 10 randomly selected animals/sex/group
- 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: just prior to sacrifice
- Animals fasted: Yes
- Anaesthetic used for blood collection: Yes (methoxyflurane)
- How many animals: from 10 randomly selected animals/sex/group
- Parameters checked:
AST (SCPT)
ALT (SCOT)
creatinine
alkaline phosphatase
gamma glutamyl transpeptidase
glucose
urea nitrogen
total protein
albumin
globulin (calculated)
A/C ratio (calculated)
total bilirubin
direct bilirubin
indirect bilirubin (calculated)
calcium
phosphorus
sodium
potassium
chloride

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
Gross lesions
Brain
Cerebral cortex
Cerebellar cortex
Medulla/pons
Eyes
Pituitary
Salivary gland
Heart
Aorta
Thymic region
Thyroid - parathyroid complex
Spinal cord
Cervical, Thoracic, Lumbar
Pancreas
Liver (3 lobes)
Kidneys
Urinary bladder
Testes
Prostate
Epididymis
Ovaries
Vagina
Uterus (corpus and cervix)
Lungs with mainstem bronchi
Adrenals
Trachea
Esophagus
Stomach
Duodenum, Jejunum, Ileum,
Cecum, Colon, Rectum (entire organs)
Exorbital lacrymal glands
Spleen
Lymph nodes (mesenteric & non-mesenteric)
Skeletal muscle
Sciatic nerve
Mammary gland (female)
Skin
Sternum (including marrow)
Femur

HISTOPATHOLOGY: Yes
Histologic examinations were performed on following tissues:
Gross lesions
Brain, incl. cerebra1 cortex, cerebellar cortex, medulla/pons
Pituitary
Salivary gland
Heart
Aorta
Pancreas
Liver (3 lobes)
Gall bladder
Kidneys
Testes
Epididymis
Thymic region
Thyroid - parathyroid complex
Spinal cord
Urinary bladder
Lungs with mainstem bronchi
Adrenals
Trachea
Esophagus
Stomach
Duodenum, Jejunum, Ileum, Cecum, Colon
Testes
Prostate
Epididymis
Ovaries
Uterus (corpus and cervix)
Spleen
Lymph nodes (mesenteric & mediastinal)
Sciatic nerve
Sternum (including marrow)
A total of 10 randomly selected animals/sex/group sacrificed in the control and 1000 ppm dose groups. The 1000 ppm dose level was selected for complete histologic examination since it was the highest dose level without significant mortality. The lungs, liver, kidneys, testes (males), stomach, duodenum, and gross lesions were examined for 10 animals/sex/group (including all survivors) from the other 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:

effects observed, treatment-related

Description (incidence and severity):

Treatment-related signs were observed in the high dose group only and consisted of signs consistent with general cachexia (emaciation, pallor, etc.). 12/15 male and 13/15 female rats died at 3000 ppm in the diet.

Mortality:

mortality observed, treatment-related

Description (incidence):

Treatment-related signs were observed in the high dose group only and consisted of signs consistent with general cachexia (emaciation, pallor, etc.). 12/15 male and 13/15 female rats died at 3000 ppm in the diet.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

No treatment-related effects on body weight were observed at 1000 ppm or lower. Body weights were significantly lower for survivors at 3000 ppm.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

No treatment-related effects on food consumption were observed at 1000 ppm or lower. Food consumption was significantly lower for survivors at 3000 ppm.

Ophthalmological findings:

no effects observed

Description (incidence and severity):

No treatment-related findings were observed at any treatment level.

Haematological findings:

no effects observed

Description (incidence and severity):

No treatment-related findings were observed at any treatment level except for in the survivors from the 3000 ppm group. Alterations in blood parameters for this group were considered of little toxicological significance.

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

No treatment-related findings were observed at any treatment level except for in the survivors from the 3000 ppm group. Alterations in blood parameters for this group were considered of little toxicological significance.

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

No treatment-related findings were observed at any treatment level except for in the survivors from the 3000 ppm group. Alterations in blood parameters for this group were considered of little toxicological significance.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

Treatment-related findings were limited to animals from the 3000 ppm group, the principle finding was intestinal ileus consisting of distended fluid and gas-filled viscera affecting the cecum and colon

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Histologic findings were consistent with the moribund condition of the animals.

Details on results:

MORTALITY DATA
Twelve male rats from the 3000 ppm group were found dead or sacrificed in a moribund condition from Day 8 to Day 17, and twelve of the 15 females from this group died from Day 5 to Day 16. All of these deaths were attributed to treatment with the test material. One female from the 300 ppm (22 mg/kg bw/day) died on Day 81 of the study. Death was preceded with observations related to general cachexia and the death was not considered to be treatment related.

CLINICAL OBSERVATIONS
Treatment-related clinical signs were observed in both sexes from the high dose group only. In general, the signs indicated general cachexia (emaciation, unkemp appearance, pallor, hunched posture etc.) and were observed in animals that died or were sacrificed in a moribund condition and, to a lesser extent, in the 3 animals/sex that survived to sacrifice. In addition, loose faeces was observed in all animals from the high dose group and associated perianal redness was observed in the animals that survived. All other clinical signs recorded during the study for all groups were sporadic and not related to treatment with the test material.

BODYWEIGHT
No effect on body weight or weight gain was observed for either sex in any group other than in the high dose (3000 ppm) group. Animals of both sexes from the high dose group showed marked weight loss during the initial stages of the study, and the weights and weight gains for the surviving rats in this group remained reduced from the controls throughout the dosing period.

FOOD CONSUMPTION
The data for the animals with significant food spillage are removed from each test period. Food consumption was markedly reduced in males and females from the 3000 ppm dose group in the first three weeks of the study. The food consumption for the three males and three females that survived through the study was also reduced, but to a lesser extent than observed in the first three weeks. No effects on food consumption were observed in either sex from any other dose group. Statistically significant differences from controls in males from other dose groups were inconsistent and not dose related and were, therefore, considered spurious.
The mean test material intake over the entire study was 6.2, 18.5, 36.8 and 60.7 mg/kg bw/day for the males and 7.5, 22.3, 44.4 and 74.3 mg/kg bw/day for the females from the 100, 300, 600 and 1000 ppm groups, respectively. Due to the extensive mortality and food spillage in the 3000 ppm group, an accurate daily dosage could not be calculated for these animals.

LABORATORY FINDINGS
No treatment related effects on haematology or serum chemistry measurements occurred in any group of either sex except for the remaining three animals/sex in the 3000 ppm dose group. Included in the changes in the animals from the high dose group were decreased glucose and protein concentrations in both sexes, decreased albumin and gobulin concentrations in females, and increased erythrocyte count, haemoglobin concentration, and hematocrit concentrations in males. These changes were probably related to the debilitated state of the animals and/or the intestinal alterations described below.

GROSS NECROPSY FINDINGS AND HISTOPATHOLOGY
No treatment-related effect on organ weights (absolute or relative) was observed for male or female rats administered the teat material at 1000 ppm or lower. Decreased absolute organ weights and weights relative to brain weight, and increased organ to body weight ratios in the 3 surviving males and the 3 surviving females from the 3000 ppm dose group were consistent with the reduced body weights and were not considered related to direct organ toxicity of the test material. Statistically significant differences compared to the controls in females from the dose groups other than the 3000 ppm group were not dose-related, were of minor magnitude, and were considered spurious.

ORGAN WEIGHTS AND FINAL BODY WEIGHTS
Treatment-related changes in gross and histological pathology are restricted to the animals that died or were sacrificed in a moribund condition and the survivors of the 3000 ppm dose group. The principal observations at necropsy for the animals from the 3000 ppm dose group were intestinal ileus consisting of distended fluid and gas-filled viscera affecting the cecum and colon. These lesions tended to be more extensive in the animals that were found dead or sacrificed moribund than in those that survived to final sacrifice. Histological findings in the animals that died or were sacrificed moribund included contracted spleens, indicative of terminal dehydration and shock, and glycogen depletion of the liver assumed to be secondary to inanition.
In addition, stomach irritation may have been evident in some animals that died although this finding was difficult to confirm due to autolysis. Typhlitis, seen as a minimal to mild lymphoplasmacytic infiltrate in the cecal lamina propria, was observed in all animals from the 3000 ppm dose group that survived the 13-week exposure period. No other treatment-related gross or microscopic findings were recorded for males or females from any dose group.

Effect levels

1

Dose descriptor:

NOEL

Effect level:

1 000 ppm

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

other: Treatment of rats with 1000 ppm or less of test material in the diet for 13 weeks resulted in no treatment-related differences in any measurements.

2

Dose descriptor:

NOEL

Effect level:

61 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Sex:

male

3

Dose descriptor:

NOEL

Effect level:

74 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Sex:

female

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

The no observed adverse effect level was determined to be 1000 ppm (61 mg/kg/d for males, 74 mg/kg/d for females). The adverse effects observed are likely to be secondary to G.I. tract irritation therefore the NOAEL is for local toxicity.

Executive summary:

A study was carried out according to OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents) and EPA OPP 82-1 (90-Day Oral Toxicity) 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. Sprague-Dawley rats (15/sex/group) were exposed to test material in the diet at concentrations of 0, 100, 300, 600, 1000, or 3000 ppm for 90 (males) or 91 (females) days. The doses corresponded to approximate mean intake levels of 6, 18, 37, and 61 mg/kg bw/day for the males and 8, 22, 44, and 74 mg/kg bw/day for the females of the 100, 300, 600, and 1000 ppm groups, respectively. Due to high mortality in the 3000 ppm group, daily intakes could not be calculated.

Mortality (80 %) for both sexes resulted from dietary exposure to test material at a concentration of 3000 ppm. The three rats of each sex from this group that survived to the end of treatment were observed to have markedly reduced body weights, fluid- or gas-filled intestines at necropsy, and typhlitis. Clinical pathologic changes including decreased serum glucose and protein concentrations in both sexes, decreased albumin and globulin concentrations in females, and increased erythrocyte count, and hemoglobin and hematocrit concentrations in males were considered to be related to the debilitated condition of the animals and/or the gastro-intestinal lesions. Gross pathologic findings and microscopic lesions in the animals that died or were sacrificed in a moribund condition supported ileus and shock as the probable cause of death or the moribund condition. In addition, 3000 ppm test material may have been irritating to the gastric mucosa.

Treatment of rats with 1000 ppm or less of test material in the diet for 13 weeks resulted in no treatment-related differences in any measurements. The no observable effect level of test material for this strain of rats is considered to be 1000 ppm (61 mg/kg bw/day males, 74 mg/kg bw/day females).