Quaternary ammonium compounds, bis(C16-18 and C18-unsatd. alkyl)dimethyl, chlorides

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:

supporting study

Study period:

From 2002-10-11 to 2003-12-02

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Cross-reference

Reason / purpose for cross-reference:

read-across source

Reference

Endpoint:

sub-chronic toxicity: oral

Type of information:

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

Adequacy of study:

supporting study

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

assessment report

Sex:

female

Route of administration:

oral: gavage

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

no mortality throughout study period

Mortality:

mortality observed, treatment-related

Description (incidence):

no mortality throughout study period

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

mid and high dose

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

mid and high dose

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

no effects observed

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

high dose

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

mid and high dose

Urinalysis findings:

not examined

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

high dose

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

mid and high dose

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

mid and high dose

Histopathological findings: neoplastic:

not examined

Details on results:

CLINICAL SIGNS AND MORTALITY:
General clinical signs: 6000 ppm: Emaciated appearance in 1/10 male and 2/10 female; Soft feces in 1/10 males and 8/10 females. Further in females: Area of hair loss or scattered hair (6/10) and one female showed a round back. Although 1/10 males given 1500 ppm showed piloerection, no noteworthy clinical signs were observed in any other group over the considered period.
Detailed clinical signs - Except soft feces noted in females at 6000 ppm, the detailed clinical observation had shown no perturbation of the autonomic or physiological functions in any treated group.
- No mortality occurred during the study period.

BODY WEIGHT AND WEIGHT GAIN: Decreased food consumption and body weight gain which were noted in treated males and females at 1500 ppm during the first week of the study, were considered to be related to palatability of the compound in the diet, resulting in a lower food consumption and lower body weight gain. The body weight gain and food consumption of treated animals at 3000 and 6000 ppm was affected by the oral administration (dietary admixture) of DDAC.


FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): When compared to controls, food consumption of animals treated at 3000 and 6000 ppm was affected during the whole treatment period, with a more marked effect at the highest concentration.
Also in the 1500 ppm group the food intake is decreased during the first treatment week.

OPHTHALMOSCOPIC EXAMINATION: No relevant ophthalmological findings were observed in any group.

HAEMATOLOGY: When compared with control values, a decrease in WBC count was recorded in all treated males and females treated at 6000 ppm. A tendency for lower WBC was observed in treated females at 3000 ppm.

CLINICAL CHEMISTRY: - increase in sodium, chloride and inorganic phosphorus plasma levels at 6000 ppm,
- a tendency for decrease in glucose levels in treated groups at 6000 ppm and in females treated at 3000 ppm,
- increase in urea and creatinine plasma levels in all treated male groups, and only a tendency for higher urea values in females treated at 6000 ppm,
- decrease in protein concentration in the 6000 ppm group,
- increase in A/G ratio in males treated at 3000 and 6000 ppm and in females treated at 6000 ppm. This difference could be explained by a decrease in globulins in males. In females, decrease in albumin was noted at 6000 ppm,
- dose-related decreased triglyceride in treated males, a tendency for lower cholesterol values in treated females at 6000 ppm,
- increase in transaminase activities in females treated at 6000 ppm.

ORGAN WEIGHTS: Relative and absolute organ weight differences to control are mostly related to low body weights in 6000 ppm group. Absolute kidney weights are also depressed in mid-dose groups.

GROSS AND HISTOPATHOLOGY: Distended coecum with feces in 9/10 males and all females of the high dose group, and in 4/10 males and 3/10 females of the mid-dose group. Similarly distended colon with faeces was observed in 4/10 males and 2/10 females of the high-dose group.
A brownish colour of the mesenteric lymphnodes was seen in 2/10 males and 4/10 females of the high-dose group, and in one male of the mid-dose group. Histiocytosis and mastocytosis in the mesenteric lymph node were seen at histopathology at and above 3000 ppm.

OTHER FINDINGS
Functional Observation Battery (FOB) - There were no perturbation of either autonomic or physiological functions at any dose-levels.

Dose descriptor:

NOAEL

Effect level:

1 500 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Body weight; food consumption; gross pathology; histopathology

Dose descriptor:

LOAEL

Effect level:

3 000 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Body weight; food consumption; gross pathology; histopathology

Critical effects observed:

not specified

At 6000 ppm, main treatment-related findings were: soft feces, body weight loss (week 1), lower body weight gain and food consumption, perturbation of hematological and blood biochemical parameters, distension of the cecum/colon with feces in all animals, histiocytosis, mastocytosis and sinusal hemorrhage in the mesenteric lymph node, consistent with a continued action of a mild irritant. At 3000 ppm, body weight gain and food consumption were affected, changes at clinical pathology were noted, the cecum was distended with feces in about third of the animals, histiocytosis and mastocytosis in the mesenteric lymph node were seen at histopathology. At 1500 ppm, minor changes in hematological and blood biochemical parameters were recorded which were not considered to be of toxicological relevance. The most obvious effect is the lowering of the food intake caused by the palatability of the compound, which at lower levels improves after some time. Some effects seen in haematology and blood chemistry at the higher dose levels could very well result from a poor alimentary status as opposed to a specific toxic mechanism. At the highest dose level, the finding of distended cecum and/or colon becomes noteworthy, and possibly a higher severity of histiocytosis at microscopic examination, although this aspect is also already observed in controls.

Conclusions:

The NOAEL of the test material was determined to be 1500 ppm (corresponding to 61.5 mg test item/kg/d for the males and 71.7 mg test item/kg/d for the females).



Under the test conditions, the NOAEL of the test material was determined to be 1500 ppm(corresponding to 42 mg DDAC/kg/d for the males and 49 mg DDAC/kg/d for the females) in rats

Executive summary:

The study used as source investigated repeated oral toxicity. The study results of the source compound were considered applicable to the target compound. Justification and applicability of the read-across approach (structural analogue) is outlined in the read-across report in section 13 or find a link in cross reference “assessment report”.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2004

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

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:

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

Details on test material:

- Name of test material (as cited in study report): Arquad 2.10-40
- Composition of test material, percentage of components: ca. 40% Didecyldimethylammonium chloride (CAS no.: 7173-51-5) in water only.
- Batch No.: FP019980246
- Physical state: Colourless to pale yellowish liquid
- Analytical purity: 42.9% active ingredient in water
- Stability: Stable

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Cesarian Obtained, Barrier Sustained-Virus Antibody Free (COBS-VAF®).
- Age at study initiation: 6 wk at Day 1 of treatment
- Weight at study initiation: Males: 185 g; females: 153 g

Administration / exposure

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

- Type: Oral via diet in feed
- Vehicle: Plain diet

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

No data

Duration of treatment / exposure:

93 days

Frequency of treatment:

Daily

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 1500, 3000 and 6000 ppm of test material in diet corresponds to active substance DDAC (males/females): 1500 ppm: 42 mg/kg/d /49 mg/kg/d; 3000 ppm : 84 / 96 mg/kg/d; 6000 ppm : 175 / 201 mg/kg/d (males / females respectively)
Basis:
actual ingested

No. of animals per sex per dose:

10 animals/sex/dose

Control animals:

yes, plain diet

Details on study design:

None

Positive control:

None

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes

DETAILED CLINICAL OBSERVATIONS: Yes

BODY WEIGHT: Yes
- Time schedule for examinations: Once a wk

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: at beginning and end of study

HAEMATOLOGY: Yes
- Number of animals: all animals
- Time points: end of study
- Parameters: Haematocrit, haemoglobin concentration, erythrocyte count, total and differential leukocyte count, platelet count, prothrombin time

CLINICAL CHEMISTRY: Yes
- Number of animals: all animals
- Time points: end of study
- Parameters: sodium, potassium, chloride, calcium, Inorg. Phosphorous, glucose, urea, total bilirubin, creatinine, total protein, albumin, total cholesterol, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, lipids.

URINALYSIS: No

Sacrifice and pathology:

-Organ weights: Yes, all animals
-Organs: adrenals, liver, kidneys, adrenals, testes, epididymides, uterus, ovaries, thymus, thyroid, spleen, brain, heart

-Gross and histopathology: Yes, all animals of control and high dose group, animals that prematurely died, and all macroscopic lesions.
Animals of intermediate dose group also investigated in full when possible effects are seen in highest dose group.
-Organs: Macroscopic lesions, Adrenals, Aorta, Brain (including medulla/pons, cerebellar and cerebral cortex), Cecum, Colon, Duodenum, Epididymides, Esophagus, Eyes with Harderian glands, Femoral bone with articulation, Heart, Ileum, Jejunum, Kidneys, Liver, Lungs with bronchi, Lymph nodes
(mandibular and mesenteric), Mammary glands/area, Ovaries (with oviducts), Pancreas, Pituitary gland, Prostate (dorso-lateral and ventral),
Rectum, Salivary glands (sublingual and submandibular), Sciatic nerve, Seminal vesicles (including coagulation gland), Skeletal muscle, Skin,
Spinal cord (cervical, thoracic and lumbar), Spleen, Sternum with bone marrow, Stomach with forestomach, Testes, Thymus, Thyroids with
parathyroids, Tongue, Trachea, Urinary bladder, Uterus (horns and cervix), Vagina.
Additionally, mesenteric lymph nodes of the low- and intermediate-dose groups were examined.

Other examinations:

-Other examinations: Functional observation battery (FOB)

Statistics:

Depending on normal distribution (Kolmogorov–Lilliefors test, possibly after a Logarithmic transformation of the values – except
for organ weights) and following testing of homogeneity of variances between groups: Bartlett test (3 or more groups) or Fisher test (2 groups).
If Homogeneous: Student test (2 groups) or Dunnett test (3 or more groups),
Else: Dunn test (3 or more groups) or Mann–Whitney/Wilcoxon test (2 groups)
If not normal distribution: Dunn test

Results and discussion

Results of examinations

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

no mortality throughout study period

Mortality:

mortality observed, treatment-related

Description (incidence):

no mortality throughout study period

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

mid and high dose

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

mid and high dose

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

no effects observed

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

high dose

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

mid and high dose

Urinalysis findings:

not examined

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

high dose

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

mid and high dose

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

mid and high dose

Histopathological findings: neoplastic:

not examined

Details on results:

CLINICAL SIGNS AND MORTALITY:
General clinical signs: 6000 ppm: Emaciated appearance in 1/10 male and 2/10 female; Soft feces in 1/10 males and 8/10 females. Further in females: Area of hair loss or scattered hair (6/10) and one female showed a round back. Although 1/10 males given 1500 ppm showed piloerection, no noteworthy clinical signs were observed in any other group over the considered period.
Detailed clinical signs - Except soft feces noted in females at 6000 ppm, the detailed clinical observation had shown no perturbation of the autonomic or physiological functions in any treated group.
- No mortality occurred during the study period.

BODY WEIGHT AND WEIGHT GAIN: Decreased food consumption and body weight gain which were noted in treated males and females at 1500 ppm during the first week of the study, were considered to be related to palatability of the compound in the diet, resulting in a lower food consumption and lower body weight gain. The body weight gain and food consumption of treated animals at 3000 and 6000 ppm was affected by the oral administration (dietary admixture) of DDAC.


FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): When compared to controls, food consumption of animals treated at 3000 and 6000 ppm was affected during the whole treatment period, with a more marked effect at the highest concentration.
Also in the 1500 ppm group the food intake is decreased during the first treatment week.

OPHTHALMOSCOPIC EXAMINATION: No relevant ophthalmological findings were observed in any group.

HAEMATOLOGY: When compared with control values, a decrease in WBC count was recorded in all treated males and females treated at 6000 ppm. A tendency for lower WBC was observed in treated females at 3000 ppm.

CLINICAL CHEMISTRY: - increase in sodium, chloride and inorganic phosphorus plasma levels at 6000 ppm,
- a tendency for decrease in glucose levels in treated groups at 6000 ppm and in females treated at 3000 ppm,
- increase in urea and creatinine plasma levels in all treated male groups, and only a tendency for higher urea values in females treated at 6000 ppm,
- decrease in protein concentration in the 6000 ppm group,
- increase in A/G ratio in males treated at 3000 and 6000 ppm and in females treated at 6000 ppm. This difference could be explained by a decrease in globulins in males. In females, decrease in albumin was noted at 6000 ppm,
- dose-related decreased triglyceride in treated males, a tendency for lower cholesterol values in treated females at 6000 ppm,
- increase in transaminase activities in females treated at 6000 ppm.

ORGAN WEIGHTS: Relative and absolute organ weight differences to control are mostly related to low body weights in 6000 ppm group. Absolute kidney weights are also depressed in mid-dose groups.

GROSS AND HISTOPATHOLOGY: Distended coecum with feces in 9/10 males and all females of the high dose group, and in 4/10 males and 3/10 females of the mid-dose group. Similarly distended colon with faeces was observed in 4/10 males and 2/10 females of the high-dose group.
A brownish colour of the mesenteric lymphnodes was seen in 2/10 males and 4/10 females of the high-dose group, and in one male of the mid-dose group. Histiocytosis and mastocytosis in the mesenteric lymph node were seen at histopathology at and above 3000 ppm.

OTHER FINDINGS
Functional Observation Battery (FOB) - There were no perturbation of either autonomic or physiological functions at any dose-levels.

Effect levels

1

Dose descriptor:

NOAEL

Effect level:

1 500 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Body weight; food consumption; gross pathology; histopathology

2

Dose descriptor:

LOAEL

Effect level:

3 000 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Body weight; food consumption; gross pathology; histopathology

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

At 6000 ppm, main treatment-related findings were: soft feces, body weight loss (week 1), lower body weight gain and food consumption, perturbation of hematological and blood biochemical parameters, distension of the cecum/colon with feces in all animals, histiocytosis, mastocytosis and sinusal hemorrhage in the mesenteric lymph node, consistent with a continued action of a mild irritant. At 3000 ppm, body weight gain and food consumption were affected, changes at clinical pathology were noted, the cecum was distended with feces in about third of the animals, histiocytosis and mastocytosis in the mesenteric lymph node were seen at histopathology. At 1500 ppm, minor changes in hematological and blood biochemical parameters were recorded which were not considered to be of toxicological relevance. The most obvious effect is the lowering of the food intake caused by the palatability of the compound, which at lower levels improves after some time. Some effects seen in haematology and blood chemistry at the higher dose levels could very well result from a poor alimentary status as opposed to a specific toxic mechanism. At the highest dose level, the finding of distended cecum and/or colon becomes noteworthy, and possibly a higher severity of histiocytosis at microscopic examination, although this aspect is also already observed in controls.

Applicant's summary and conclusion

Conclusions:

The NOAEL of the test material was determined to be 1500 ppm (corresponding to 42 mg DDAC/kg/d for the males and 49 mg DDAC/kg/d for the females).



Under the test conditions, the NOAEL of the test material was determined to be 1500 ppm(corresponding to 42 mg DDAC/kg/d for the males and 49 mg DDAC/kg/d for the females) in rats

Executive summary:

The potential systemic toxicity of the test material (40% didecyldimethylammonium chloride in water) was evaluated following repeated oral administration to rats according to OECD Guideline 408. The study was conducted in compliance with the principles of Good Laboratory Practice regulations. The test material was given by dietary admixture to Sprague-Dawley rats for 13 weeks at the concentration of 1500, 3000 or 6000 ppm (corresponding to 42, 84 or 175 mg of DDAC/kg bw/day for the males and 49, 96 or 201 mg of DDAC/kg bw/day for the females). All animals were monitored for toxic effects including clinical observations, body and organ weights, hematology, serum chemistry, macroscopic and microscopic evaluations. At 6000 ppm, main treatment-related findings were: soft feces, body weight loss (at week 1), lower body weight gain and food consumption, perturbation of hematological and blood biochemical parameters, distension of the cecum/colon with feces in all animals, histiocytosis, mastocytosis and sinusal hemorrhage in the mesenteric lymph node, consistent with a continued action of an irritant. At 3000 ppm, body weight gain and food consumption were affected, changes at clinical pathology were noted, the cecum was distended with feces in about third of the animals, histiocytosis and mastocytosis in the mesenteric lymph node were seen at histopathology. At 1500 ppm, minor changes in hematological and blood biochemical parameters were recorded which were not considered to be of biological significance and therefore considered of no toxicological relevance. Based on these findings it was concluded that the NOAEL of didecyldimethylammonium chloride following repeated dietary administration of 90 days was 1500 ppm which corresponds to 42 mg DDAC/kg bw /day for the males and 49 mg DDAC/kg bw/day for the females. The overall NOAEL for both males and females is set at 46 mg/kg bw/day.