N,N,N-trimethylanilinium chloride

Toxicity to reproduction

Administrative data

Endpoint:

two-generation reproductive toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Justification for type of information:

Data from secondary source

Data source

Reference

Reference Type:

secondary source

Title:

Robust Summaries & Test Plans for test material Joint Venture HPV Chemicals Challenge

Author:

Consumer Specialty Products Association (CSPA)

Year:

2011

Bibliographic source:

Robust Summaries & Test Plans for test material, Joint Venture HPV Chemicals Challenge 2011

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)

Principles of method if other than guideline:

Reproductive toxicity study of test material was performed on Sprague Dawley rats

GLP compliance:

not specified

Limit test:

no

Justification for study design:

No data available

Test material

Test material information

Constituent 1

Reference substance name:

Quaternary ammonium compounds, benzyl-C12-16-alkyldimethyl, chlorides

EC Number:

270-325-2

EC Name:

Quaternary ammonium compounds, benzyl-C12-16-alkyldimethyl, chlorides

Cas Number:

68424-85-1

Molecular formula:

Unspecified

IUPAC Name:

Alkyl dimethyl benzyl ammonium chloride

Details on test material:

- Name of test material (as cited in study report):Alkyl dimethyl benzyl ammonium chloride
- Molecular formula:unspecified
- Molecular weight :368.0448g/mol
- Substance type: Organic

Test animals

Species:

rat

Strain:

Sprague-Dawley

Details on species / strain selection:

No data available

Sex:

male/female

Details on test animals or test system and environmental conditions:

No data available

Administration / exposure

Route of administration:

oral: feed

Vehicle:

not specified

Details on exposure:

Details on exposure
PREPARATION OF DOSING SOLUTIONS: test material mixed with feed

DIET PREPARATION
- Rate of preparation of diet (frequency):No data available
- Mixing appropriate amounts with (Type of food)
- Storage temperature of food: No data available
VEHICLE
- Justification for use and choice of vehicle (if other than water):
- Concentration in vehicle:
- Amount of vehicle (if gavage): No data available

- Lot/batch no. (if required): No data available
- Purity: No data available

Details on mating procedure:

- M/F ratio per cage:1:1
- Length of cohabitation: three weeks
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: [no / yes (explain)]
- After successful mating each pregnant female was caged (how):
- Any other deviations from standard protocol:

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

F0: 19 weeks (from 1st prebreed dose to last F0
sacrifice)
F1: 24 weeks (from 1st F1 wean to last F1 sacrifice)
F2: to weaning
Premating exposure period for males: F0: 10 weeks; F1: 10 to 14 weeks (beginning with
1st weaning)
Premating exposure period for females: F0: 10 weeks; F1: 10 to 14 weeks (beginning with
1st weaning)

Frequency of treatment:

7 days/week

Details on study schedule:

After a 10-week pre-breed period rats were mated (one male to one female) for three weeks to produce the F1 generation. At weaning, 28 F1 weanlings/sex/group were selected to produce the F2 generation,After their pre-breed exposure, F1 animals were paired as described above. All procedures during mating, gestation, and lactation of the F1 parents and selected F2 weanlings were performed as described above.

Doses / concentrations

Remarks:

0,300, 1000 and 2000 ppm ( Mean consumption for the males and females throughout the study was 22.5, 72.6 and 145.5 mg/kg/day for the 300, 1000 and 2000 ppm groups, respectively.

No. of animals per sex per dose:

Total:224
0 mg/kg/day:28 male and 28 female
23 mg/kg/day:28 male and 28 female
73 mg/kg/day:28 male and 28 female
145 mg/kg/day:28 male and 28 female

Control animals:

yes, concurrent no treatment

Details on study design:

No data available

Positive control:

No data available

Examinations

Parental animals: Observations and examinations:

Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: yes

DETAILED CLINICAL OBSERVATIONS: Yes

Time schedule: daily


BODY WEIGHT: Yes
Time schedule for examinations: daily
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes Food consumption was determined weekly.

Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data: No data available


WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
Time schedule for examinations:

OTHER:

Oestrous cyclicity (parental animals):

No data available

Sperm parameters (parental animals):

No data available

Litter observations:

number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies were observed

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving animals [describe when, e.g. as soon as possible after the last litters in each generation were produced.]
- Maternal animals: All surviving animals [describe when, e.g. after the last litter of each generation was weaned.]

GROSS NECROPSY: yes
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]

HISTOPATHOLOGY / ORGAN WEIGHTS: yes
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.

Postmortem examinations (offspring):

SACRIFICE
- The F1 offspring not selected as parental animals and all F2 offspring were sacrificed at [#?] days of age.
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows:

GROSS NECROPSY: yes
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]

HISTOPATHOLOGY / ORGAN WEIGTHS: yes
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.

Statistics:

The results of the quantitative continuous variables (e.g., body weights, food consumption, etc.) were compared for the three treatment groups and one control group by use of Levene’s test for equal variances, analysis of variance and (pooled or separate variance) t-test. Non-parametric data were statistically evaluated using the Kruskall-Wallis test followed by the Mann-Whitney U test for pairwise comparisons when appropriate. Frequency data were compared using the Fisher’s exact test.

Reproductive indices:

No data available

Offspring viability indices:

No data available

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality:

not specified

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Males exhibited no reduction in body weight. Females at 2000 ppm had reductions in body weight for weeks 5, 6, 9 and 10 of treatment. Body weight gain was reduced at 2000 ppm for one week during the prebreed treatment. Food consumption in F0 females at 2000 ppm was reduced for the first four exposure weeks.

Females in the 2000 ppm group showed significant reductions in body weights on day 0 of gestation, but no body weight gain reductions. Body weight gains throughout lactation and body weights on lactation day 21 were increased in the females in the 2000 ppm group.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

During gestation and lactation was unaffected by test substance treatment.

Food efficiency:

not specified

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

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

No treatment-related lesions were observed in the necropsy of F0 males and females. No treatment-related lesions were observed in the
histopathologic examination of selected organs from F0 males and females at 2000 ppm.

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

no effects observed

Description (incidence and severity):

Reproductive parameters were unaffected by treatment.

Effect levels (P0)

Dose descriptor:

NOAEL

Effect level:

145 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

food consumption and compound intake

histopathology: non-neoplastic

reproductive performance

Remarks on result:

other: No effects on reproductive parameters were observed

Target system / organ toxicity (P0)

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Treatment related:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Results: P1 (second parental generation)

General toxicity (P1)

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality:

no mortality observed

Description (incidence):

There were no apparent treatment-related deaths of adult animals on study.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

F1 males at 2000 ppm exhibited no reduction in body weights but did have reduced weight gain in the second treatment week.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

Food consumption was reduced for F1 males at 2000 ppm for two of the 10 treatment weeks. There were no significant effects on F1 females.

Food efficiency:

not specified

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

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

No treatment-related lesions were observed in the necropsy of F1 males and females.
There were no treatment-related lesions observed in the histopathologic examination of selected organs from the 2000 ppm group.

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Reproductive function / performance (P1)

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

no effects observed

Description (incidence and severity):

Reproductive parameters were unaffected by treatment. Maternal body weights at 2000 ppm were unaffected during the gestational and lactational periods. Gestational food consumption was reduced for days 7 - 11 and 14 - 17 at 2000 ppm.

Effect levels (P1)

Dose descriptor:

NOAEL

Effect level:

145 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

mortality

body weight and weight gain

food consumption and compound intake

histopathology: non-neoplastic

reproductive performance

Remarks on result:

other: Reproductive parameters were unaffected by treatment.

Target system / organ toxicity (P1)

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Treatment related:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Results: F1 generation

General toxicity (F1)

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality / viability:

no mortality observed

Description (incidence and severity):

No effects of treatment on postnatal deaths (postnatal days 0 - 28) were observed.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Pups exhibited reduced body weights per litter on days 21 (weaning) and 28 (postweaning) at 2000 ppm. F1 pup body weight gains were reduced for lactation days 14 - 21 and 21 - 28 (postweaning).

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

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

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Histopathological findings:

no effects observed

Description (incidence and severity):

No treatment-related lesions were observed in the necropsy of F1 pups that died during lactation or of randomly selected F1 pups (10/sex/dose)

Other effects:

not specified

Developmental neurotoxicity (F1)

Behaviour (functional findings):

not specified

Developmental immunotoxicity (F1)

Developmental immunotoxicity:

not specified

Effect levels (F1)

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

73 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

not specified

Basis for effect level:

viability

mortality

body weight and weight gain

histopathology: non-neoplastic

Remarks on result:

other: Pups exhibited reduced body weights per litter on days 21 (weaning) and 28 (postweaning) at 2000 ppm. F1 pup body weight gains were reduced for lactation days 14 - 21 and 21 - 28 (postweaning).

Target system / organ toxicity (F1)

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Treatment related:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Results: F2 generation

General toxicity (F2)

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality / viability:

no mortality observed

Description (incidence and severity):

Perinatal deaths and lactational survival were unaffected by treatment.

Description (incidence and severity):

Pup body weights per litter were reduced at 2000 ppm at postnatal day 28 (postweaning). Pup weight gains per litter also were reduced at 2000 ppm for lactational days 14 - 21 (preweaning) and for days 21 - 28 (postweaning).

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

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

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Histopathological findings:

no effects observed

Description (incidence and severity):

There were no treatment-related lesions observed in the necropsy of F2 pups that died during lactation or of randomly selected F2 pups
(10/sex/group)

Other effects:

not specified

Developmental neurotoxicity (F2)

Behaviour (functional findings):

not specified

Developmental immunotoxicity (F2)

Developmental immunotoxicity:

not specified

Effect levels (F2)

Dose descriptor:

NOAEL

Generation:

F2

Effect level:

73 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

not specified

Basis for effect level:

viability

mortality

body weight and weight gain

histopathology: non-neoplastic

Remarks on result:

other: No developmental toxic effects observed

Target system / organ toxicity (F2)

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Treatment related:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Overall reproductive toxicity

Reproductive effects observed:

not specified

Treatment related:

not specified

Relation to other toxic effects:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Applicant's summary and conclusion

Conclusions:

No Observed Adverse Effect Level (NOAEL) for reproductive toxicity was considered to beapproximately 145 mg/kg/day (2000ppm)and for developmental toxicity considered to be approximately 73mg/kg /day(1000ppm). When male and female SpragueDawley rats were treated with test material orally.

Executive summary:

The two generation reproductive toxicity of test material was performed on male and female SpragueDawley rats. The test material mixed with feed in dose concentration 300, 1000 and 2000 ppm (approximately 22.5, 72.6 and 145.5 mg/kg/day base on Mean consumption for the males and females throughout the study). A total of 28 male and 28 female rats were evaluated at each dose level. After a 10-week pre-breed period rats were mated (one male to one female) for three weeks to produce the F1 generation. Exposures continued through mating, gestation, parturition and lactation. At weaning, 28 F1 weanlings/sex/group were selected to produce the F2 generation, and were then exposed to the same dietary concentrations of test material as their parents for 10 weeks. After their pre-breed exposure, F1 animals were paired as described above. All procedures during mating, gestation, and lactation of the F1 parents and selected F2 weanlings were performed as described above. All F0 and F1 parental animals were necropsied and examined for gross lesions; selected reproductive tissues from the high dose and control groups were examined histologically as were other tissues with gross lesions. Ten F1 and F2 weanlings/sex/group were randomly selected and examined for gross lesions. Remaining nonselected F1 and F2 pups at weaning were euthanized and discarded after the necropsy of the selected pups.

After 10 weeks Premating Exposure for paraents males exhibited no reduction in body weight. Females at 2000 ppm had reductions in body weight for weeks 5, 6, 9 and 10 of treatment. Body weight gain was reduced at 2000 ppm for one week during the prebreed treatment. Food consumption in F0 females at 2000 ppm was reduced for the first four exposure weeks. While Reproductive parameters were unaffected by treatment. Females in the 2000 ppm group showedsignificant reductions in body weights on day 0 of gestation, but no body weight gain reductions.Body weight gains throughout lactation and body weights on lactation day 21 were increased in thefemales in the 2000 ppm group. Food consumption during gestation and lactation was unaffected bytest substance treatment during gestation and lactation period in F0 Female. After postmortem .No treatment-related lesions were observed in the necropsy of F0 males and females. No treatment-related lesions were observed in the histopathologic examination of selected organs from F0 males and females at 2000 ppm.

F1 males at 2000 ppm exhibited no reduction in body weights but did have reduced weight gain in the second treatment week. Food consumption was reduced for F1 males at 2000 ppm for two of the 10 treatment weeks. There were no significant effects on F1 females. Reproductive parameters were unaffected by treatment. Maternal body weights at 2000 ppm were unaffected during the gestational and lactational periods. Gestational food consumption was reduced for days 7 - 11 and 14- 17 at2000 ppm. After postmortem in F1 male and female .No treatment-related lesions were observed in the necropsy of F1 males and females. There were no treatment-related lesions observed in the histopathologic examination of selected organs from the 2000 ppm group. There were no apparenttreatment-related deaths of adult animals on study.

In F1 litter,Pups exhibited reduced body weights per litter on days 21 (weaning) and 28 (postweaning) at 2000 ppm. F1 pup body weight gains were reduced for lactation days 14 - 21 and 21 – 28 (postweaning). No effects of treatment on postnatal deaths (postnatal days 0 - 28) were observed. No treatment-related lesions were observed in the necropsy of F1 pups that died during lactation or of randomly selected F1 pups (10/sex/dose).While in F2 Litters,Pup body weights per litter were reduced at 2000 ppm at postnatal day 28 (postweaning). Pup weight gains per litter also were reduced at 2000 ppm for lactational days 14 - 21 (preweaning) and for days 21 - 28 (postweaning). Perinatal deaths and lactational survival were unaffected by treatment. There were no treatment-related lesions observed in the necropsy of F2 pups that died during lactation or of randomly selected F2 pups (10/sex/group).

Exposure of rats to the test substance in the diet for two generations resulted in parental toxicityat the target dose level of 145mg/kg bw/day (2000 ppm): perinatal toxicity was concomitant with parental toxicity, being well-defined at 145mg/kg bw/day (2000 ppm.There were notreatment-related reproductive effects observed in this study. The A/D ratio (the dose level at which there were no observable effects in adults/the dose level at which there were no observable effects on offspring) is 1 (1000 ppm/1000 ppm) indicating no increased risk to the offspring in the absence of indications of adult toxicity. HenceNo Observed Adverse Effect Level (NOAEL) for reproductive toxicity was considered to beapproximately 145 mg/kg/day (2000ppm)and for developmental toxicity considered to be approximately 73mg/kg /day(1000ppm). When male and femaleSpragueDawley rats were treated withtest materialorally.