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Key value for chemical safety assessment

Effects on fertility

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Referenceopen allclose all

Endpoint:
two-generation reproductive toxicity
Type of information:
read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
2008
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
RA study
Justification for type of information:
Refer to the section 13 for details on the read across justification. The toxicity to reproduction study with the read across substance is considered sufficient to fulfil the information requirements as further explained in the provided endpoint summary.
Reference:
Composition 1
Qualifier:
according to
Guideline:
OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Test material information:
Composition 1
Specific details on test material used for the study:
- Purity: 49.9%
- C12-16-benzyldimethylammonium chloride (CAS no.: 68424-85-1) in water only.
- Specification: C12-16 BKC (C12: 72.0%; C14: 27.7%; C16: 0.3%)
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Route of administration:
oral: feed
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
- Exposure period: about 18 wk.
P0 and P1:
Premating exposure period (males): 10 wk.
Premating exposure period (females): 10 wk.
- Duration of test: P0 pre-mating 10 wk, until F2 weaning.
Frequency of treatment:
Continuously
Details on study schedule:
Groups of 25 male and 25 female Sprague-Dawley rats were administered the test substance (purity 49.9%) at levels of 0, 500, 2000 or 4000 ppm in their diet over a period of 10 wk before mating, 2 wk during mating, and until after weaning of the pups (total period 18 wk). From every litter one or two pups were selected to again obtain 25 animals per sex and dose group for the F1 generation. These animals were similarly treated as the parent (P1) animals throughout premating, mating, pregnancy until sacrifice, after weaning of F2 progeny.
Remarks:
Doses / Concentrations:
1000, 2000 and 4000 ppm of test substance
No. of animals per sex per dose:
25
Control animals:
yes, concurrent vehicle
Parental animals: Observations and examinations:
- Examination of P0 and P1 generation:
Clinical signs and mortality were checked daily. Food consumption and body weight were recorded at designated intervals. Males and females were paired for a 2-wk period, until mating was obtained. The P1 females were allowed to deliver normally, and rear their progeny. Pregnancy and litter parameters were recorded.
- During lactation, the pups (F1 generation) were observed daily for survival and clinical signs; body weight was recorded at designated intervals; the sex-ratio was recorded. On Day 4 post-partum, the size of each litter was adjusted to obtain eight pups per litter (four males and four females).
- Reflex development was assessed at designated time-points.
Sperm parameters (parental animals):
Epididymal and testicular sperm parameters were evaluated for both P0 and P1 males.
Litter observations:
Examination of F1/P1 generation:
- On Day 22 post-partum, one male and one female pup per litter were selected to constitute the F1/P1 generation, which comprised 25 males and 25 females per group. The F1/P1 animals were observed daily for clinical signs and mortality. Body weight and food consumption were recorded once a week. Sexual development of both males and females was assessed.
- Neurobehavioural tests were conducted at designated intervals to assess auditory and visual functions. Spontaneous locomotor activity was also evaluated when the animals were between 7 and 8 wk old.
- After sexual maturity, F1 male and F1 female animals were paired. The F1 females were allowed to deliver normally, and rear their progeny. Pregnancy and litter parameters were recorded. During lactation, the pups (F2 generation) were observed daily for survival and clinical signs; body weight was recorded at designated intervals; the sex-ratio was recorded. On day 4 post-partum, the size of each litter was adjusted to obtain eight pups per litter (four males and four females).
- Reflex development was assessed at designated time-points.
Postmortem examinations (parental animals):
Terminal examination of P0 and P1 animals:
- After weaning of their respective progeny, P0 and P1 parent males and females were sacrificed. Designated organs were weighed for P0 and P1 parents, as well as brain, spleen and thymus of one pup per sex per litter of each generation.
- Epididymal and testicular sperm parameters were evaluated for both P0 and P1 males.
- A macroscopic post-mortem examination was performed on all P0 and P1 parent males and females. Any pups which died or were killed prematurely during the lactation period were also submitted for a macroscopic post-mortem examination. Macroscopic lesions, reproductive organs, adrenals and pituitary glands were sampled in all parent animals. A microscopic examination was performed on macroscopic lesions, reproductive organs, adrenals, and pituitary glands of all P0 and P1 parents of the control and high dose groups.
- Particularly detailed histopathological examinations were performed for the ovaries and the testes.
Postmortem examinations (offspring):
- A macroscopic post-mortem examination was performed on three pups per sex and per litter of each P0 and P1 (F1 and F2 generation) females killed at weaning. Any pups which died or were killed prematurely during the lactation period were also submitted for a macroscopic post-mortem examination. In all pups, the macroscopic lesions were preserved.
Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
- At 4000 ppm, a slightly lower mean body weight gain was recorded during most of the dosing period in both parental males and females (and was associated with reduced liver weights).
- At 2000 and 500 ppm, a marginally to slightly lower body weight gain was noted over all the dosing period for the males.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
- At 4000 ppm, a slightly to moderately lower mean food consumption was recorded during most of the dosing period in both parental males and females.
- At 2000 and 500 ppm, a marginally to slightly lower mean food consumption was noted over all the dosing period for the males.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
- At 4000, ppm, reduced liver weights were observed.
- At 2000 ppm, lower liver weights in parental animals were recorded.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
- At 4000 ppm, necropsy of these animals revealed dilation of the cecum, colon or ileum in some animals (more marked in P0 parents).
- At 2000 ppm, necropsy of parents revealed dilatation of the cecum in some animals.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
- At 4000 and 2000 ppm, no effects were noted at histopathological examination of sexual organs.

Histopathological findings: neoplastic:
not examined
Reproductive function: estrous cycle:
not examined
Reproductive function: sperm measures:
no effects observed
Description (incidence and severity):
- At 4000 and 2000 ppm, no effects were noted on sperm parameters.

Reproductive performance:
no effects observed
Description (incidence and severity):
- At 4000 ppm, slightly lower pup body weight was observed (with lower spleen weights).
- At 2000 ppm, no effects were observed on parental fertility as assessed by normal mating, gestation and delivery.
- At 500 ppm, no effects were noted on mating, fertility, gestation, fecundity or delivery of either generation or on development of their progeny.
As lower food consumption is known to occur due to palatability of compound. As no other substance related effects were seen than can be attributed to lower food intake, the level of 500 ppm test substance in the diet, corresponding to 16-25 mg/kg bw/day, should be regarded as NOAEL for P0 and P1generation.
Key result
Dose descriptor:
NOAEL
Remarks:
systemic toxicity
Effect level:
500 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
food consumption and compound intake
organ weights and organ / body weight ratios
gross pathology
Remarks on result:
other: the effect level ranged from 16 to 25 mg/kg bw/day
Key result
Dose descriptor:
NOAEL
Remarks:
reproductive toxicity
Effect level:
2 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
reproductive performance
Remarks on result:
other: dose range from 61 to 101 mg/kg bw/day
Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
4 000 ppm
System:
other: reproductive performance
Treatment related:
yes
Dose response relationship:
yes
Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
- At 4000 ppm, a slightly to moderately lower mean body weight gain was recorded during most of the dosing period in both parental males and females (and was associated with reduced liver weights).
- At 2000 ppm, a marginally to slightly lower mean body weight gain was noted.
- At 500 ppm, a marginally to slightly lower mean body weight gain was observed in both sexes. This was associated with lower liver weights of parental males and females.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
- At 4000 ppm, a slightly to moderately lower mean food consumption was recorded during most of the dosing period in both parental males and females.
- At 2000 and 500 ppm, a marginally to slightly lower mean food consumption was noted over all the dosing period in both sexes.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
- At 4000 ppm, reduced liver weights were observed.
- At 2000 and 500 ppm, lower liver weights in parental animals were recorded.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
- At 4000 ppm, necropsy of these animals revealed dilation of the cecum, colon or ileum in some animals (more marked in F0 parents).
- At 2000 ppm, necropsy of parents revealed dilatation of the cecum in a single animal. 
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
- At 4000 and 2000 ppm, no effects were noted at histopathological examination of sexual organs.
Histopathological findings: neoplastic:
not examined
Reproductive function: sperm measures:
no effects observed
Description (incidence and severity):
- At 4000 and 2000 ppm, no effects were noted on sperm parameters.

Description (incidence and severity):
- At 4000 ppm, slightly lower pup body weight was observed for each progeny and was associated, for the F2 generation pups, with a reduction in litter size (as a consequence of lower number of implantation sites of P1 females) and a delay in sexual development. Lower spleen weights were also noted for each progeny.
- At 2000 ppm, no effects were noted on parental fertility as assessed by normal mating, gestation and delivery.
- At 500 ppm, no effects were noted on mating, fertility, gestation, fecundity or delivery and development of their progeny.
As lower food consumption is known to occur due to palatability of compound. As no other substance related effects were seen than can be attributed to lower food intake, the level of 500 ppm test substance in the diet, corresponding to 16-25 mg/kg bw/day, should be regarded as NOAEL for P0 and P1generation.
Key result
Dose descriptor:
NOAEL
Remarks:
systemic toxicity
Effect level:
500 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
food consumption and compound intake
organ weights and organ / body weight ratios
gross pathology
Remarks on result:
other: the effect level ranged from 16 to 25 mg/kg bw/day
Key result
Dose descriptor:
NOAEL
Remarks:
reproductive toxicity
Effect level:
2 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
food consumption and compound intake
organ weights and organ / body weight ratios
gross pathology
Remarks on result:
other: dose range from 61 to 101 mg/kg bw/day
Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
4 000 ppm
System:
other: reproductive performance
Treatment related:
yes
Dose response relationship:
yes
Clinical signs:
not examined
Dermal irritation (if dermal study):
not examined
Mortality / viability:
mortality observed, treatment-related
Description (incidence and severity):
- At 4000 ppm, the litter size at birth was reduced. 

Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
- At 4000 ppm, slightly reduced pup body weight was observed. Pup weight gain was also slightly lower during lactation.
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Sexual maturation:
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
- At 4000 ppm, lower spleen weights were noted.
- At 2000 ppm, except for a marginally lower spleen weight of the progeny, no other effects were noted on their development.
Gross pathological findings:
no effects observed
Histopathological findings:
no effects observed
Behaviour (functional findings):
not examined
Developmental immunotoxicity:
not examined
Key result
Dose descriptor:
NOAEL
Remarks:
general toxicity
Generation:
F1
Effect level:
2 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
organ weights and organ / body weight ratios
Key result
Dose descriptor:
NOAEL
Remarks:
developmental toxicity
Generation:
F1
Effect level:
2 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
organ weights and organ / body weight ratios
Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
4 000 ppm
System:
other: body weight and organs weight
Organ:
spleen
Treatment related:
yes
Dose response relationship:
yes
Clinical signs:
not examined
Dermal irritation (if dermal study):
not examined
Mortality / viability:
mortality observed, treatment-related
Description (incidence and severity):
- At 4000 ppm, the litter size at birth were reduced (as a consequence of lower number of implantation sites of F1 parent females).
- At 2000 ppm, no effects were seen in F2 offspring regarding pup survival until weaning.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
- At 4000 ppm, slightly lower pup body weight was observed. The pup weight gain was slightly reduced during lactation, 
- At 2000 ppm, no effects were seen regarding pup development.
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Sexual maturation:
effects observed, treatment-related
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
- At 4000 ppm, lower spleen weights were noted.
- At 2000 ppm, except for a marginally lower spleen weight, no other effects were noted on their development.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
At 4000 ppm, dilatation of the caecum with faeces was observed in 4/25 males and 2/25 females.
At 2000 ppm, no effects were seen regarding pup development and after sacrifice at weaning.
Histopathological findings:
not examined
Behaviour (functional findings):
not examined
Developmental immunotoxicity:
not examined
Key result
Dose descriptor:
NOAEL
Remarks:
general / developmental toxicity
Generation:
F2
Effect level:
2 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
sexual maturation
mortality
body weight and weight gain
organ weights and organ / body weight ratios
gross pathology
Key result
Dose descriptor:
LOAEL
Remarks:
general / developmental toxicity
Generation:
F2
Effect level:
4 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
sexual maturation
mortality
body weight and weight gain
organ weights and organ / body weight ratios
gross pathology
Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
4 000 ppm
System:
other: spleen weight
Treatment related:
yes
Dose response relationship:
yes
Key result
Reproductive effects observed:
yes
Lowest effective dose / conc.:
2 000 ppm
Treatment related:
yes
Relation to other toxic effects:
reproductive effects as a secondary non-specific consequence of other toxic effects
Dose response relationship:
yes

The mean achieved dosages of the test substance for the dose-levels of 500, 2000 and 4000 ppm of test substance were as follows:
F0 generation
- males (Days 1 to 106): 16, 61 and 123 mg/kg bw/day, respectively,
- females:

during premating period (Days 1 to 71): 19, 74 and 154 mg/kg bw/day, respectively,
    during pregnancy period (Days 0 to 20 p.c.): 18, 69 and 145 mg/kg bw/day, respectively,
    during lactation period (Days 1 to 21 p.p.): 37, 159 and 326 mg/kg bw/day, respectively.
F1 generation
- males (Days 1 to 120): 24, 96 and 202 mg/kg bw/day, respectively,
- females:
    during premating period (Days 1 to 64): 32, 127 and 269 mg/kg bw/day, respectively,
    during pregnancy period (Days 0 to 20 p.c.): 21, 83 and 164 mg/kg bw/day, respectively,
    during lactation period (Days 1 to 21 p.p.): 41, 162 and 323 mg/kg bw/day, respectively.

The actual intake of test substance for both males and females given 500, 2000 and 4000 ppm throughout the study is approximately 16-25, 61-101 and 123-208 mg/kg bw/day, respectively  for the F0 generation and 24-31, 96-123 and 202-252 mg/kg bw/day for the F1 generation.

Conclusions:
Based on the results of the read across study, the rat NOEL for parental toxicity was established at 500 ppm for the male and the female animals. The rat NOEL for mating behaviour, fertility and gestation of each generation and for development, growth and survival of each progeny was determined to be 2000 ppm (i.e., 61 to 101 mg/kg bw/day (nominal) (i.e., equivalent to 30.5 to 50.5 mg a.i./kg bw/day) and 96 to 123 mg/kg bw/day (nominal) (i.e., equivalent to 48 to 61.5 mg a.i./kg bw/day for the F0 and F1 generation respectively)).
Executive summary:

A study was conducted to determine the toxicity to reproduction of the read across substance, quaternary ammonium compounds, benzyl C12-C16 (even numbered)-alkyldimethyl chlorides (C12-16 ADBAC), according to OECD Guideline 416, in compliance with GLP. In this two-generation study, the test substance was administered in the diet to male and female Sprague Dawley rats at dose levels of 0, 500, 2,000 and 4,000 ppm (purity 49.9%) (corresponding to 0 mg (a.i.)/kg bw/day, 16-25 and 24-31 mg/kg bw/day or 8-12.5 and 12-15.5 mg a.i./kg bw/day in males and females, 61-101 and 96-123 mg/kg bw/day or 30.5-50 and 48 to 61.5 mg a.i./kg bw/day in males and females and 123-208 and 202-252 mg/kg bw/day or 61-104 and 101-126 mg a.i./kg bw/day in males and females, respectively). Doses were administered before and throughout mating and gestation until the end of the lactation period in both P0 and P1 generations. At 2,000 ppm, P0 (males) and P1 (both sexes) showed marginally to slightly lower body weight gains and reduced food consumption. Necropsy of parents of both generations revealed dilatation of the caecum in some animals. This was associated with lower liver weights in parental animals of both generations. At 4,000 ppm, in P0 and P1 generations, number of implantation sites and litter size at birth were reduced. The progenies (F1 and F2) also showed lower pup weights. Pup weight gain was slightly lower during lactation. The weight of the spleen was also reduced. Upon necropsy, dilatation of the caecum with faeces was observed in 4/25 males and 2/25 females in F2. Treatment with the test substance had no effect on mating, fertility and behavioural parameters in P0 and P1 parental Sprague-Dawley rats at treatment levels up to 2,000 ppm. No effect was recorded on litter parameters and on pre- and post-natal development of either generation at 2,000 ppm. Based on the results of the read across, the rat NOEL for parental toxicity was determined to be 500 ppm for the male and the female animals. The rat NOEL for mating behaviour, fertility and gestation of each generation and for development, growth and survival of each progeny was established at 2,000 ppm (61 to 101 mg/kg bw/day (nominal) (equivalent to 30.5 to 50.5 mg a.i./kg bw/day and 96 to 123 mg/kg bw/day (nominal) (equivalent to 48 to 61.5 mg a.i./kg bw/day for the F0 and F1 generation respectively)) (Foulon, 2008)

Endpoint:
two-generation reproductive toxicity
Type of information:
read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
1990
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
RA study
Justification for type of information:
Refer to the section 13 for details on the read across justification. The toxicity to reproduction study with the read across substance is considered sufficient to fulfil the information requirements as further explained in the provided endpoint summary.
Reference:
Composition 1
Qualifier:
according to
Guideline:
EPA OPP 83-4 (Reproduction and Fertility Effects)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Test material information:
Composition 1
Specific details on test material used for the study:
- Name of test material (as cited in study report): Alkyldimethylbenzylammonium chloride (ADBAC)
- Physical state: Pale yellow viscous liquid
- Analytical purity: 81.09% active substance in aqueous/ethanol solution.
- Lot/batch No.: 7293K
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
Test animals
- Source: Sprague-Dawley CD rats were obtained from Charles River Breeding Laboratories, Kingston, NY, USA
- Age at study initiation: Six weeks
- Weight at study initiation: 212.2-213.4 g (males); 148.3-150.2 g (females)
- Housing: Individually in stainless steel, wire mesh cages (22.5x15.5x18.0 cm; mating cages 22.5x31.0x18.0 cm)
- Diet: Certified Ground Rodent Chow # 5002 (Ralstor Purina Co., St. Louis, MO), ad libitum
- Water: Tap water, ad libitum. Water was provided by an automatic watering system with demand control valves mounted on each cage rack.
- Acclimation period: Two weeks

Environmental conditions
- Temperature: 66-73°F
- Humidity: 40-60%
Route of administration:
oral: feed
Vehicle:
other: Certified Ground Rodent Chow # 5002
Details on exposure:
Diet preparation: A concentrated premix was prepared by direct addition of the test substance to ground chow and mixing for approximately an hour. Test diets were prepared by appropriate dilutions of the concentrated premix or higher diet concentrations.
- Rate of preparation of diet (frequency): Fresh diet was prepared weekly.
- Mixing appropriate amounts with (Type of food): Certified Ground Rodent Chow # 5002
- Storage temperature of food: Diets were stored in polyethylene containers at room temperature.
Details on mating procedure:
- M/F ratio per cage: 1:1
- Mating procedure: After pre-breeding exposure the animals were mated on the basis of one male to one female selected randomly within each dose group for a period of 21d to produce F1 generation.
- Proof of pregnancy: Copulation plug and/or vaginal sperm as Day 0 of gestation.
- After the first 7d of the mating period females of unsuccessfully mated pairs were placed with males of other unmated pairs within the same dose group; after an additional 7d, unsuccessfully mated pairs were similarly exchanged again for a period of 7d or until successful mating had occurred, whichever came first, allowing for a total of 21d to mate.
- After successful mating each pregnant female was caged: Pregnant females were housed individually. On Day 20 of gestation each pregnant female was transferred to a shoe-box cage.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Experimental diets were analyzed for stability, homogeneity and concentration of test substance using high pressure liquid chromatography.
- Homogeneity study indicated that the distribution of the test substance in the test diet was uniform.
- Stability study indicated that the dosed feed was stable for at least 14d when stored in open glass feed jars at room temperature. Dosed feed was stable for at least 21d when stored in closed polyethylene containers at room temperature.
- Concentration verification analyses of the dosed feed indicated that the mean concentrations of the test substance in the diet for the 300, 1000 and 2000 ppm dosage levels were 95.3-109.0% of nominal for 300 ppm, 95.6-107.9% of nominal for 1000 ppm and 94.7-108.0% of nominal for 2000 ppm.
Duration of treatment / exposure:
P0 generation: 19 weeks (from 1st prebreed dose to last F0 sacrifice)
F1 generation: 25 weeks (from 1st F1 wean to last F1 sacrifice)
F2 generation: Until weaning
Frequency of treatment:
Daily
Details on study schedule:
- P1 parental animals not mated until 17-18 weeks after selection from the F1 litters.
- Selection of parents from F1 generation when pups were 28d old.
- Number of P1 generation animals selected: 28 males and 28 females

Remarks:
Doses / Concentrations:
0, 300, 1000 or 2000 ppm test substance (i.e., equivalent to 0, 16-31, 51-102, and 100-188 mg/kg bw/day (males) and 0, 21-32, 67-106 and 139-198 mg/kg bw/day (females)).
Basis: nominal in diet
No. of animals per sex per dose:
28
Control animals:
yes, plain diet
Details on study design:
- Rationale for animal assignment: Animals were randomly distributed based on body weight
- Animal identification: By ear tags
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Twice daily

CLINICAL SIGNS: Yes
- Time schedule: Once daily for overt clinical signs.

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly during prebreed and mating for both sexes. For females on gestational Day 0, 6, 15, and 20 and on postnatal Day 0, 7, 14, and 21.

FOOD CONSUMPTION AND COMPOUND INTAKE: Yes
- Time schedule: Weekly during prebreed for both sexes. For females in 3- or 4-day intervals throughout gestation and to postnatal Day 14.
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes

WATER CONSUMPTION AND COMPOUND INTAKE: No

Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: Yes, maximum of 8 pups/litter (4/sex/litter as nearly as possible). Excess pups were subjected to detailed external examination and then sacrificed.

PARAMETERS EXAMINED
The following parameters were examined in F1 and F2 offspring: Live/Still births on the day of birth (postnatal Day 0), survival indices at Days 0, 4, 7 and 14 after birth and weaning, weight on postnatal Day 1, 4, 7, 14, and at weaning (Day 21), and physical abnormalities for all pups at birth and throughout the pre-weaning period.

GROSS EXAMINATION OF DEAD PUPS: The thoracic and abdominal organs from pups which died after Day 4 were preserved for subsequent histopathological examination.
Postmortem examinations (parental animals):
SACRIFICE: Yes
- How many animals: All animals
- Necropsy method: Animals were anesthetized with methoxyflurane and exsanguinated by severing the brachial vessel.

GROSS NECROPSY: Yes
- How many animals: All animals sacrificed in P0 and P1 parental animals
- Gross necropsy consisted of: Vagina, uterus, ovaries, gross lesions, testes, epididymis, seminal vesicles and prostrate.

HISTOPATHOLOGY : Yes
- How many animals: All animals of control and high dose groups sacrificed in P0 and P1 parental animals
- Tissues evaluated: Vagina, uterus, ovaries, gross lesions, testes, epididymis, seminal vesicles and prostrate and any tissues or organs showing gross alterations from the low and mid dose groups
Postmortem examinations (offspring):
SACRIFICE
- The F1 offsprings not selected as parental animals and all F2 offsprings were sacrificed at 7d of age.

GROSS NECROPSY
- Examination for gross lesions was performed on any pup appearing abnormal or dying on test and for ten randomly selected F1 and F2 weanlings/sex/group.

Statistics:
- The results of the quantitative continuous variables (e.g., body weights, food consumption, etc.) were compared between the three treatment groups and one control group using 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:
Mating index, fertility index and gestational index were determined.
Offspring viability indices:
Live birth index, 4-d survival index, 7-d survival index, 14-d survival index, 21-d survival index and lactation index

Clinical signs:
no effects observed
Description (incidence and severity):
No significant signs of toxicity during the pre-breed, mating, gestation or lactation periods at any dose for either generation were observed.



Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
- During the 10-week pre-breed exposure, P0 males exhibited no reduction in body weight. During the same period, P0 females at 2000 ppm exhibited reduction in body weight during Weeks 5, 6, 9 and 10 of pre-breed treatment. Body weight gain was also reduced at 2000 ppm for one week (Week 8-9) during the pre-breed period.
- On lactation Day 21 mean body weight of P0 dams at 2000 ppm exhibited a significant increase. Increased lactation body weight gain was observed at 2000 ppm throughout lactation.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
- Food consumption in the P0 females at 2000 ppm was reduced for the first four exposure weeks. Food consumption in P0 males was significantly reduced at 2000 ppm for the first week of treatment only.
- At P0 breed to produce F1 litters, food consumption during gestation and lactation was unaltered by treatment.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
not examined
Other effects:
effects observed, treatment-related
Reproductive function: estrous cycle:
not examined
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed
Description (incidence and severity):
Mating index, fertility index and gestational index
Key result
Dose descriptor:
NOAEL
Remarks:
systemic toxicity
Effect level:
1 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
food consumption and compound intake
Remarks on result:
other: equivalent to 51-102 mg/kg bw/day in males and 67-106 mg/kg bw/day in females
Key result
Dose descriptor:
NOEL
Remarks:
reproductive toxicity
Effect level:
2 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no reproductive toxicity observed
Remarks on result:
not determinable due to absence of adverse toxic effects
Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
1 000 ppm
System:
other: body weight and food consumption
Treatment related:
yes
Dose response relationship:
yes
Clinical signs:
no effects observed
Description (incidence and severity):
No significant signs of toxicity during the pre-breed, mating, gestation or lactation periods at any dose for either generation were observed.
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
At 2000 ppm, only slight reduction were observed in males and females
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
not examined
Reproductive function: estrous cycle:
not examined
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed
Description (incidence and severity):
Mating index, fertility index and gestational index
Key result
Dose descriptor:
NOAEL
Remarks:
systemic toxicity
Effect level:
1 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
Remarks on result:
other: equivalent to 51-102 mg/kg bw/day in males and 67-106 mg/kg bw/day in females
Key result
Dose descriptor:
NOEL
Remarks:
reproductive toxicity
Effect level:
2 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no reproductive toxicity observed
Remarks on result:
not determinable due to absence of adverse toxic effects
Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
2 000 ppm
System:
other: body weight
Treatment related:
yes
Dose response relationship:
yes
Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not examined
Mortality / viability:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
The F1 litters exhibited reduced body weight per litter on postnatal Days 21 and 28 at 2000 ppm. F1 pup body weight gain was reduced during lactation Days 14-21 and 21-28.
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings:
no effects observed
Behaviour (functional findings):
not examined
Developmental immunotoxicity:
not examined
Key result
Dose descriptor:
NOAEL
Remarks:
general toxicity
Generation:
F1
Effect level:
1 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
Key result
Dose descriptor:
NOEL
Remarks:
developmental toxicity
Generation:
F1
Effect level:
2 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects observed
Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
2 000 ppm
System:
other: body weight
Treatment related:
yes
Dose response relationship:
yes
Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not examined
Mortality / viability:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
F2 pup weights per litter were reduced at 2000 ppm on postnatal Day 28. Pup weight gain was also reduced at 2000 ppm during lactation Days 14-21 and for Days 21-28.
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings:
no effects observed
Behaviour (functional findings):
not examined
Developmental immunotoxicity:
not examined
Key result
Dose descriptor:
NOAEL
Remarks:
general toxicity
Generation:
F2
Effect level:
1 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
Key result
Dose descriptor:
NOEL
Remarks:
developmental toxicity
Generation:
F2
Effect level:
2 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects observed
Remarks on result:
not determinable due to absence of adverse toxic effects
Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
1 000 ppm
System:
other: body weight
Treatment related:
yes
Dose response relationship:
yes
Key result
Reproductive effects observed:
no
Conclusions:
Based on the results of the read across study, dietary administration of the test substance at dose levels of 0, 300, 1000 and 2000 ppm for two generations to Sprague-Dawley rats was well tolerated and no signs of reproductive toxicity were observed at any dose level. The rat NOAEL (systemic toxicity) for both parental generation (P0 and P1) and offsprings (F1 and F2) was considered to be 1000 ppm (in diet), equivalent to 51-102 and 67-106 mg/kg bw/d for male and female respectively. The rat NOEL (reproductive toxicity) was established at 2000 ppm (the highest dose tested).
Executive summary:

A study was conducted to determine the toxicity to reproduction of the read across substance, quaternary ammonium compounds, benzyl C12-C16 (even numbered)-alkyldimethyl chlorides (C12-16 ADBAC), according to US EPA OPP 83 -4, in compliance with GLP. The test substance was orally administered to Sprague-Dawley CD rats (28/sex/group) at dose levels of 0, 300, 1,000 or 2,000 ppm test substance (equivalent to 0, 16-31, 51-102, and 100-188 mg/kg bw/day (males) and 0, 21-32, 67-106 and 139-198 mg/kg bw/day (females)) in the diet. There was one control group of 28 animals/sex. Following a 10 week pre-breed exposure period, the P0 rats were randomly paired within dose groups for a 3 week mating period to produce the F1 generation. Exposure continued through mating, gestation, parturition and lactation. At weaning, 28 F1 weanling/sex/group were randomly selected and exposed to the same dietary concentration of the test substance as their parents for 10 weeks. After their pre-breed exposure, F1/P1 animals were paired to produce the F2 generation. The animals were observed twice daily and clinical findings, body weights and food consumption were recorded periodically. All P0 and P1 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/dose were randomly selected and necropsied and examined for gross lesions. P0 female body weights and food consumption were reduced at 2,000 ppm during the pre-breeding exposure period. For the pre-breeding period of the P1 animals, only slight reductions in body weight gain were observed for males in the high dose group. No other treatment-related effects were observed on any of the reproductive parameters. Based on the results of the study, it can be stated that, dietary administration of the test substance at dose levels of 0, 300, 1,000 and 2,000 ppm for two generations to Sprague-Dawley rats was well tolerated and no sign of reproductive toxicity was observed at any dose level. Based on the results of the read across study, the rat NOAEL (systemic toxicity) for both parental generation (P and F1) and offsprings (F1 and F2) was considered to be 1,000 ppm (in diet), equivalent to 51-102 or 41.3-83 mg a.i./kg bw/day in males and 67-106 mg/kg bw/day or 54-86 mg a.i./kg bw/day for female, respectively. The rat NOEL (reproductive toxicity) was established at 2000 ppm (the highest dose tested) (Neeper-Bradley, 1990).

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
30.5 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
Two 2-generation reproductive toxicity studies conducted with the read-across substance C12-C16 ADBAC are available. Hence, the information requirement for this tonnage band is sufficiently met with the available data.
Additional information

Study 1. A study was conducted to determine the toxicity to reproduction of the read across substance according to OECD Guideline 416, in compliance with GLP. In this two-generation study, the test substance was administered in the diet to male and female Sprague Dawley rats at dose levels of 0, 500, 2,000 and 4,000 ppm (purity 49.9%) (corresponding to 0 mg (a.i.)/kg bw/day, 16-25 and 24-31 mg/kg bw/day or 8-12.5 and 12-15.5 mg a.i./kg bw/day in males and females, 61-101 and 96-123 mg/kg bw/day or 30.5-50 and 48 to 61.5 mg a.i./kg bw/day in males and females and 123-208 and 202-252 mg/kg bw/day or 61-104 and 101-126 mg a.i./kg bw/day in males and females, respectively). Doses were administered before and throughout mating and gestation until the end of the lactation period in both P0 and P1 generations. At 2,000 ppm, P0 (males) and P1 (both sexes) showed marginally to slightly lower body weight gains and reduced food consumption. Necropsy of parents of both generations revealed dilatation of the caecum in some animals. This was associated with lower liver weights in parental animals of both generations. At 4,000 ppm, in P0 and P1 generations, number of implantation sites and litter size at birth were reduced. The progenies (F1 and F2) also showed lower pup weights. Pup weight gain was slightly lower during lactation. The weight of the spleen was also reduced. Upon necropsy, dilatation of the caecum with faeces was observed in 4/25 males and 2/25 females in F2. Treatment with the test substance had no effect on mating, fertility and behavioural parameters in P0 and P1 parental Sprague-Dawley rats at treatment levels up to 2,000 ppm. No effect was recorded on litter parameters and on pre- and post-natal development of either generation at 2,000 ppm. Based on the results of the read across, the rat NOEL for parental toxicity was determined to be 500 ppm for the male and the female animals. The rat NOEL for mating behaviour, fertility and gestation of each generation and for development, growth and survival of each progeny was established at 2,000 ppm (61 to 101 mg/kg bw/day (nominal) (equivalent to 30.5 to 50.5 mg a.i./kg bw/day and 96 to 123 mg/kg bw/day (nominal) (equivalent to 48 to 61.5 mg a.i./kg bw/day for the F0 and F1 generation respectively)) (Foulon, 2008).

Study 2. A study was conducted to determine the toxicity to reproduction f the read across substance according to US EPA OPP 83 -4, in compliance with GLP. The test substance was orally administered to Sprague-Dawley CD rats (28/sex/group) at dose levels of 0, 300, 1,000 or 2,000 ppm test substance (equivalent to 0, 16-31, 51-102, and 100-188 mg/kg bw/day (males) and 0, 21-32, 67-106 and 139-198 mg/kg bw/day (females)) in the diet. There was one control group of 28 animals/sex. Following a 10 week pre-breed exposure period, the P0 rats were randomly paired within dose groups for a 3 week mating period to produce the F1 generation. Exposure continued through mating, gestation, parturition and lactation. At weaning, 28 F1 weanling/sex/group were randomly selected and exposed to the same dietary concentration of the test substance as their parents for 10 weeks. After their pre-breed exposure, F1/P1 animals were paired to produce the F2 generation. The animals were observed twice daily and clinical findings, body weights and food consumption were recorded periodically. All P0 and P1 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/dose were randomly selected and necropsied and examined for gross lesions. P0 female body weights and food consumption were reduced at 2,000 ppm during the pre-breeding exposure period. For the pre-breeding period of the P1 animals, only slight reductions in body weight gain were observed for males in the high dose group. No other treatment-related effects were observed on any of the reproductive parameters. Based on the results of the study, it can be stated that, dietary administration of the test substance at dose levels of 0, 300, 1,000 and 2,000 ppm for two generations to Sprague-Dawley rats was well tolerated and no sign of reproductive toxicity was observed at any dose level. Based on the results of the read across study, the rat NOAEL (systemic toxicity) for both parental generation (P and F1) and offsprings (F1 and F2) was considered to be 1,000 ppm (in diet), equivalent to 51-102 or 41.3-83 mg a.i./kg bw/day in males and 67-106 mg/kg bw/day or 54-86 mg a.i./kg bw/day for female, respectively. The rat NOEL (reproductive toxicity) was established at 2000 ppm (the highest dose tested) (Neeper-Bradley, 1990).

Effects on developmental toxicity

Link to relevant study records

Referenceopen allclose all

Endpoint:
developmental toxicity
Type of information:
read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
1985
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Remarks:
RA study
Justification for type of information:
Refer to the section 13 for details on the read across justification. The developmental toxicity / teratogenicity study with the read across substance is considered sufficient to fulfil the information requirements as further explained in the provided endpoint summary.
Reference:
Composition 1
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Version / remarks:
see 'Principles of method if other than guideline'
Deviations:
yes
Remarks:
short exposure time i.e., on gestation Days 7-18 only
Principles of method if other than guideline:
- 20 mated female rabbits per group were exposed for Days 7 - 18 of gestation to 2.0 mL/kg bw/day of the test substance topically (2h per day) at concentrations of 0, 0.5, 1.0, or 2.0% (i.e., equivalent to 0, 10, 20 and 40 mg/kg bw/day, respectively). The control group was treated with deionised water.
- Animals were observed twice daily for signs of toxicity, including skin irritation from Days 7 to 29. Body weights and food consumption were recorded. A gross necropsy was conducted on animals that died. Foetuses less than 28d old were fixed in buffered neutral formalin and those 28d or older were cleared and stained. All surviving dams were sacrificed at study termination on gestation Day 29. An examination of the uterus and ovaries was conducted. Following removal of the foetuses the abdominal and thoracic cavities and organs of the dams were examined. At sacrifice foetuses were identified, weighed and examined externally for defects. Gross dissection and examination of viscera, and internal sex determination also were conducted on each foetus. Finally, an examination of the skeleton for anomalies and ossification variations was conducted after clearing and alizarin red staining of the foetuses.
GLP compliance:
yes
Limit test:
no
Test material information:
Composition 1
Specific details on test material used for the study:
- Name of test material (as cited in study report): 1-Hexadecanaminium, N,N,N-trimethyl-, chloride
- Physical state: Liquid
- Analytical purity: 25% aqueous solution
Species:
rabbit
Strain:
New Zealand White
Route of administration:
dermal
Vehicle:
water
Details on exposure:
TEST SITE:
Shaved dorsal area.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Rinsed with water and dried.
- Time after start of exposure: 2 h.

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2 mL/kg
- Concentration (if solution): 0, 0.5, 1.0 and 2.0%
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
Days 7 - 18 of gestation.
Frequency of treatment:
Once daily (2 hours).
Duration of test:
Days 0 - 29 of gestation.
Remarks:
Concentrations: 0, 0.5, 1.0, or 2.0% (i.e., equivalent to 0, 10, 20 and 40 mg/kg bw/day, respectively)
No. of animals per sex per dose:
20 pregnant females per dose.
Control animals:
yes, concurrent vehicle
Maternal examinations:
Dams were observed twice daily for signs of toxicity, including skin irritation from Days 7 through 29. Body weights were taken on gestation Days 0, 3, 6, 9, 12, 15, 18, 21, 24, 27 and 29. Individual food consumption was measured daily. A gross necropsy was conducted on dams that died in an attempt to determine the cause of death. All surviving dams were sacrificed at study termination on gestation Day 29. An examination of the uterus (including the number and location of live and dead foetuses, early and late resorptions, and implantation sites) and ovaries (including the number of corpora lutea) was conducted. Following removal of the foetuses the abdominal and thoracic cavities and organs of the dams were examined. Uteri from females that appeared non-gravid were placed in 10% ammonium sulphide solution for confirmation of pregnancy.
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
Fetal examinations:
- External examinations: Yes
- Soft tissue examinations: Yes
- Skeletal examinations: Yes
Statistics:
Body weight changes and food consumption and number of early and late resorptions, dead foetuses, total implantations, corpora lutea, skeletal abnormalities, and mean fetal body weight were compared by analysis of variance (Bartlette's). If variance was not significant, then treatment-control comparisons were made using the least significant difference (LSD) criterion. If variance was significant, then comparison was made using the t-test for unequal variances and the Wilcoxon, Mann-Whitney rank sum test. Additionally, a regression and lack of fit were performed on each of these parameters. The number of pregnancies per group, the percentage of skeletal abnormalities and soft tissue malformations were analysed by Fisher's exact test.
Clinical signs:
no effects observed
Dermal irritation (if dermal study):
effects observed, treatment-related
Description (incidence and severity):
Skin irritation was observed at all doses with the severity and duration of erythema, oedema, desquamation, atonia and coriaceousness increased in a dose-dependent manner.
Mortality:
mortality observed, non-treatment-related
Description (incidence):
Two control, one intermediate and one high dose doe died during the study. The cause of death could not be determined. Two of the does that died aborted prior to death (one control and one intermediate dose group animal).
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
A slight increase in congested lungs was observed for the high dose group at necropsy.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
not examined
Number of abortions:
effects observed, non-treatment-related
Description (incidence and severity):
Two additional abortions occurred, one each in the intermediate and high dose groups. None of these deaths or abortions were considered related to test substance toxicity.
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
Effects on pregnancy duration:
not examined
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
not examined
Changes in number of pregnant:
not examined
Details on maternal toxic effects:
Maternal toxic effects: no test substance related significant effects.
Key result
Dose descriptor:
NOAEL
Remarks:
systemic toxicity
Effect level:
40 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Basis for effect level:
other: no significant treatment-related maternal toxic effects were observed up to the highest tested dose.
Key result
Dose descriptor:
NOAEL
Remarks:
developmental toxicity
Effect level:
40 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Basis for effect level:
other: no significant treatment-related foetal development effects were observed up to the highest tested dose.
Key result
Abnormalities:
no effects observed
Fetal body weight changes:
not examined
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
not examined
Fetal/pup body weight changes:
not examined
Changes in litter size and weights:
no effects observed
Changes in postnatal survival:
not examined
External malformations:
no effects observed
Skeletal malformations:
no effects observed
Visceral malformations:
no effects observed
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects: no effects

Details on embryotoxic / teratogenic effects:
The incidence of foetal malformations, as well as genetic and developmental variations in the treated groups were comparable to that of the control group.
Key result
Dose descriptor:
NOAEL
Remarks:
general toxicity
Effect level:
40 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: no effects observed up to the highest dose tested
Key result
Dose descriptor:
NOAEL
Remarks:
developmental toxicity
Effect level:
40 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: no developmental effects observed up to the highest dose tested
Key result
Abnormalities:
no effects observed
Key result
Developmental effects observed:
no
Conclusions:
Based on the results of the read across study, the NOAEL of the test substance for maternal as well as developmental toxicity was found to be 40 mg/kg bw/day in rabbits.
Executive summary:

A study was conducted to determine the developmental toxicity / teratogenicity of the read across substance, cetrimonium chloride (C16 TMAC), according to a method similar to OECD Guideline 414, in compliance with GLP. This experiment was performed in New Zealand White rabbits. Twenty mated female rabbits per group were exposed topically (daily for 2 hours) from Days 7 to 18 of gestation at concentrations of 0, 0.5, 1.0, or 2.0% (equivalent to 0, 10, 20 and 40 mg a.i./kg bw/day, respectively). The control group was treated with deionised water only. Clinical condition and reactions to treatment were recorded at least once daily. Body weights were recorded on Days 0, 3, 6, 9, 12, 15, 18, 21, 24, 27 and 29 of gestation. All surviving females were sacrificed on Day 29 of gestation and the foetuses were removed by caesarean section. At necropsy the females were examined macroscopically. Live foetuses were weighed, sexed and were examined for visceral and skeletal abnormalities. Two control animals, one intermediate and one high dose died during the study. Two of the rabbits that died, aborted prior to death (one control and one intermediate dose). Two additional abortions occurred, one each in the intermediate and high dose groups. Deaths or abortions were not considered to be related to the test substance. No treatment-related maternal body weight or food intake effects were noted. The incidence of foetal malformations, as well as genetic and developmental variations in the treated groups were comparable to that of the control group. No other treatment-related effects were noted.Based on the results of the read across study, the NOAEL of the test substance for maternal as well as developmental toxicity was found to be 40 mg/kg bw/day in rabbits (Albridge, 1985).

Endpoint:
developmental toxicity
Data waiving:
other justification
Justification for data waiving:
other:
Reason / purpose:
data waiving: supporting information
Related information:
Composition 1
Reason / purpose:
data waiving: supporting information
Related information:
Composition 1
Reason / purpose:
data waiving: supporting information
Related information:
Composition 1
Reason / purpose:
data waiving: supporting information
Related information:
Composition 1
Reason / purpose:
data waiving: supporting information
Related information:
Composition 1
Abnormalities:
not specified
Developmental effects observed:
not specified
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
40 mg/kg bw/day
Study duration:
subacute
Species:
rabbit
Quality of whole database:
Well-documented study which meets basic scientific principles.
Additional information

A study was conducted to determine the developmental toxicity / teratogenicity of the read across substance acoording to a method similar to OECD Guideline 414, in compliance with GLP. This experiment was performed in New Zealand White rabbits. Twenty mated female rabbits per group were exposed topically (daily for 2 hours) from Days 7 to 18 of gestation at concentrations of 0, 0.5, 1.0, or 2.0% (equivalent to 0, 10, 20 and 40 mg a.i./kg bw/day, respectively). The control group was treated with deionised water only. Clinical condition and reactions to treatment were recorded at least once daily. Body weights were recorded on Days 0, 3, 6, 9, 12, 15, 18, 21, 24, 27 and 29 of gestation. All surviving females were sacrificed on Day 29 of gestation and the foetuses were removed by caesarean section. At necropsy the females were examined macroscopically. Live foetuses were weighed, sexed and were examined for visceral and skeletal abnormalities. Two control animals, one intermediate and one high dose died during the study. Two of the rabbits that died, aborted prior to death (one control and one intermediate dose). Two additional abortions occurred, one each in the intermediate and high dose groups. Deaths or abortions were not considered to be related to the test substance. No treatment-related maternal body weight or food intake effects were noted. The incidence of foetal malformations, as well as genetic and developmental variations in the treated groups were comparable to that of the control group. No other treatment-related effects were noted.Based on the results of the read across study, the NOAEL of the test substance for maternal as well as developmental toxicity was found to be 40 mg/kg bw/day in rabbits (Albridge, 1985).

Justification for classification or non-classification

Based on the available negative data on structurally similar C16 TMAC and C12-16 ADBAC, the test substance is not expected to be a reproductive toxicant with regards to fertility or developmental effects. Therefore, no classification is required according to Directive 67/548/EEC and Regulation (EC) 1272/2008.