Didecyldimethylammonium chloride

Repeated dose toxicity: oral

Administrative data

Endpoint:

chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

14 Nov, 2003 - 13 Mar, 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The study was performed in accordance with the OECD and OPPTS protocols, although there were few subsequent amendments, with the following deviations from the agreed Study plan: - the temperature and relative humidity recorded in the animal room were sometimes out of the target ranges specified in the Study plan, - on day 47, animals were checked for mortality or signs of morbidity only once, instead of at least twice as stated in the Study plan, - in order to decrease cross-contamination risks, the racks were no longer rotated around the room from day 280, - I. Gaou took on the responsibility of this study from 19 October 2004, instead of 18 November 2003 as specified by error in her statement, - further to the receipt of a new certificate of analysis, the test substance contained 39.6% DDAC instead of 40.5%, - the concentration of preparations was additionally checked in week 104. These deviations were not considered to have compromised the validity or integrity of the study.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 453 (Combined Chronic Toxicity / Carcinogenicity Studies)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: US EPA guideline OPPTS 870.3700, August 1998

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Didecyldimethylammonium chloride

EC Number:

230-525-2

EC Name:

Didecyldimethylammonium chloride

Cas Number:

7173-51-5

Molecular formula:

C22H48N Cl

IUPAC Name:

didecyldimethylammonium chloride

Details on test material:

Test substance: as prescribed by 1.2 as typical marketed substance (act: 40%, water: 60%)

Composition: Contains 39.6% DDAC (CASno.: 7173-51-5) in water only.
Batchno: WIR03048

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Administration / exposure

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

Dietary admixture

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The analysis of mean achieved dose levels were performed at regular intervals.

Duration of treatment / exposure:

52 weeks for toxicology group and 104 weeks for carcinogenicity group.

Frequency of treatment:

Daily

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
15.6, 32.6 and 66.1 mg a.i./kg bw/day for males and 18.2, 39.2 and 77.2 mg a.i./kg bw/day for females (52 weeks)
Basis:
nominal in diet

Doses / concentrations 2

Remarks:

Doses / Concentrations:
12.6, 27.3 and 55.4 mg a.i./kg bw/day for males and 15.7, 33.8 and 69.5 mg a.i./kg bw/day for females (104 weeks)
Basis:
nominal in diet

No. of animals per sex per dose:

Three groups of 70 male and 70 female rats in total. Twenty males and 20 females in each group were used for toxicological investigations and treated for 52 weeks. Fifty males and 50 females in each group were used to investigate the carcinogenic potential. One additional group of 60 male and 60 female rats received untreated diet under the same experimental conditions and acted as a toxicology control group (10 males and 10 females) and as a carcinogenicity control group (50 males and 50 females).

Control animals:

yes, concurrent vehicle

Details on study design:

Three groups of 70 male and 70 female rats were treated with the test substance containing 39.6% of the active substance DDAC (didecyldimethylammonium chloride) by daily oral administration (dietary admixtures) at the constant concentrations of 700, 1500 or 3000 ppm (corresponding to 277.2, 594 or 1188 ppm of DDAC). The dietary admixtures were supplied ad libitum. Twenty males and 20 females in each group were used for toxicological investigations and treated for 52 weeks. Fifty males and 50 females in each group were used to investigate the carcinogenic potential and were treated for 104 weeks. One additional group of 60 male and 60 female rats received untreated diet under the same experimental conditions and acted as a toxicology control group (10 males and 10 females) and as a carcinogenicity control group (50 males and 50 females).

Positive control:

Not applicable

Examinations

Observations and examinations performed and frequency:

The animals were checked at least once a day for mortality and twice daily for clinical signs. In addition, detailed clinical examinations were made once a week. After 6 months of treatment, all animals were palpated every 2 weeks in order to record the time of onset, location, size, appearance and progression of palpable masses. Body weights were recorded once during the pre-treatment period, on the first day of treatment, once a week during the first 13 weeks of the treatment period and then once every 4 weeks until the end of the study. Food consumption was recorded once a week during the first 13 weeks of the study, once every 3 months between weeks 14 and 26, once a month between weeks 27 and 39 and then every 4 weeks until the end of the study. Hematology, blood biochemistry and urinalysis investigations were performed on all surviving animals allocated to toxicology sub-groups in weeks 13, 26 and 51. The differential white cell count was determined for all surviving control animals and those treated at 3000 ppm in weeks 52, 78 and 104.

Sacrifice and pathology:

Surviving toxicology sub-group animals were killed at the end of the 52-week treatment period and carcinogenicity sub-group animals were killed at the end of the 104-week treatment period. A full macroscopic post-mortem examination was performed on all the animals. Designated organs were weighed and selected tissue specimens preserved. A microscopic examination was performed on all masses, and on designated tissues from control animals and those treated at 3000 ppm of the test substance sacrificed at the end of the 52 or 104-week treatment periods, and from animals that died prematurely.

Other examinations:

None

Statistics:

Analysis of survival data - Kaplan Meier technique and Peto's method.
Analysis of tumor incidence: Peto's method, one-tailed exact test and Armitage's test.

Results and discussion

Results of examinations

Clinical signs:

no effects observed

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:

no effects observed

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:

no effects observed

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Histopathological findings: neoplastic:

no effects observed

Details on results:

Mortality/Survival rate: No effects.
Clinical signs: No effects 
Macroscopic investigations: No effects
Haematology: No effects 
Clinical Chemistry: No effects 
Urinalysis: No effects 
Pathology: Toxicity groups (52 weeks) There was an increase in incidence of reduced size in the thymus of females in all treated groups, which correlated with an (non-significant) increase in lymphoid atrophy at microscopic examination. The organ weights were not significantly affected. This may to be due to an unusually low incidence in the control group, however a treatment-related effect cannot be excluded. There were no other treatment-related findings. All other necropsy findings were typical of animals of this strain and age.
Carcinogenicity groups (104 weeks): No effects
Organ Weights: No effects 
Histopathology: Neoplastic findings: No effects;
Non-neoplastic findings for toxicity groups (52 weeks) The following statistically significant treatment-related findings were present:
- mesenteric lymph node: increase in hemorrhage in high-dose males
- peyer's patches: decrease in germinal centers in high-dose females. There were no other treatment-related findings
Non-neoplastic findings for carcinogenicity groups (104 weeks) The following statistically significant treatment-related findings were present:
Mesenteric lymph node: 
- increase in histiocytosis in high-dose females
- increase in mastocytosis in high-dose males and females
Peyer's patches: 
- decrease in germinal centres in high-dose males and females. There were no other treatment-related findings.

Effect levels

1

Key result

Dose descriptor:

LOAEL

Effect level:

ca. 55.4 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Sex:

male

Basis for effect level:

body weight and weight gain

2

Key result

Dose descriptor:

NOAEL

Effect level:

ca. 27.3 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Sex:

male

Basis for effect level:

body weight and weight gain

3

Key result

Dose descriptor:

LOAEL

Effect level:

ca. 69.5 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Sex:

female

Basis for effect level:

body weight and weight gain

4

Key result

Dose descriptor:

NOAEL

Effect level:

ca. 33.8 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Sex:

female

Basis for effect level:

body weight and weight gain

5

Key result

Dose descriptor:

LOAEL

Effect level:

ca. 62 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

body weight and weight gain

6

Key result

Dose descriptor:

NOAEL

Effect level:

ca. 31 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

body weight and weight gain

Target system / organ toxicity

Key result

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

Based on the results, the LOAEL for toxicity was considered to be 3000 ppm and the NOAEL 1500 ppm (equivalent to 31 mg a.i./kg bw/day, combined males/females). The test substance was not found to be carcinogenic under the conditions of this study.

Executive summary:

A study was conducted in accordance with OECD Guideline 453 and OPPTS Guideline 870.3700 to evaluate the chronic toxicity of the test substance (40% a.i.). The test substance was administered daily to Sprague-Dawley rats by dietary admixture at the concentrations of 700, 1500, and 3000 ppm of for 52 weeks (toxicology sub-group) or for 104 weeks. The test substance did not induce any treatment-related mortality or clinical signs when administered daily for 52 or 104 weeks. At 3000 ppm, the mean body weight and mean body weight gain of the carcinogenicity subgroup animals were slightly lower than in controls (-26%), correlating in females with slightly lower mean food consumption during the first 13 weeks. There were no significant differences in hematological, biochemical and/or urinalysis parameters at any treated dose-level for animals of either sub-group, compared with controls. There were no macroscopic findings attributable to the test item at any of the tested dose-levels. The non-neoplastic histopathological findings confined to the mesenteric lymph nodes and Peyer’s patches were consistent with the continued action of a mild irritant and are considered to be of limited toxicological significance. There were no treatment-related neoplastic findings at histologic al examination. Consequently, the LOAEL for toxicity was considered to be 3000 ppm (equivalent to 62 mg/kg bw/day combined male/female) and the NOAEL 1500 ppm (equivalent to 31 mg a.i./kg bw/day, combined male/female). The test substance was not found to be carcinogenic under the conditions of this study (CIT, 2008).