Didecyldimethylammonium chloride

Endpoint summary

Key value for chemical safety assessment

Toxic effect type:

concentration-driven

Effects on fertility

Link to relevant study records

Reference

Reference 1

Endpoint:

two-generation reproductive toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

30 Oct, 2003 - 21 May, 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Deficiencies: Yes F0 animals: the day 11 samples taken for stability in closed bags and open feeders, were actually sampled on day 12 by error, males D26143 and D26154 given 1500 ppm were weighed again on day 17 due to aberrant values on day 15, for females D26364, pup No. 5 and D26393, pup No. 10 (given respectively 500 and 1500 ppm), macroscopic examination was performed although not scheduled in the study plan (data not presented in the study report), Male D26146 (given 1500 ppm): left testis and epididymis were mislaid before microscopic examination. F1 animals: according to the Study plan, the dosing period was 20 weeks. However, the study was finished in week 18, the dietary admixtures were analyzed in week 18 rather than week 20, as specified in the Study plan, because the study finished early, female D26509 given 4000 ppm and females D26439 and D26446 given 0 ppm were mated one extra day by error (the females had been positive for mating the previous day), during the mating periods (F0 and F1 animals), the food consumption was not recorded (typing error in the final study plan), organ weights with aberrant values have been excluded from mean calculation: thyroid glands of male D26210 (given 500 ppm), adrenal glands of male D26232 (given 1500 ppm) and pituitary gland of male D26234 (given 1500 ppm), male D26226 (given 500 ppm): dilated renal pelvis was observed for left kidney at macroscopic examination but no microscopic examination was performed, male D26262 (given 4000 ppm): adrenal glands were not found for the preparation of slides, female D26457 (given 500 ppm): pituitary gland was mislaid after weighing, female D26493, pup No. 4 (given 1500 ppm) found dead on day 2 post-partum was mislaid before macroscopic examination. These deviations were considered not to have compromised the validity or integrity of the study

Qualifier:

according to guideline

Guideline:

OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.3800 (Reproduction and Fertility Effects)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

Species: Rat
Strain: Sprague-Dawley, Crl CD® (SD) IGS BR
Source: Charles River Laboratories France, L’Arbresle, France.
Sex: 100 males and 100 females
Age/weight at study initiation: At the beginning of the treatment period, the animals were approximately 5 to 6 weeks old and had a mean body
weight of 137 g (range: 104 g to 168 g) for the males and 118 g (range: 96 g to 139 g) for the females.
Number of animals per group: 25 males and 25 females
Duration of mating: 2 weeks
Deviations from standard protocol: Female D26509 given 4000 ppm and females D26439 and D26446 given 0 ppm were mated one extra day
by error (the females had been positive for mating the previous day).
Control animal: Yes

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on exposure:

Administration/ Exposure: Oral
Duration of exposure before mating: 10 weeks
Duration of exposure in general P, F1, F2 males, females From beginning of the study until sacrifice of Parent, F1, F2-generation
Type: oral In food
Concentration Food consumption per day: 0, 500, 1500 and 4000 ppm ad libitum, for groups 1, 2, 3 and 4, respectively.
Vehicle: The test item was mixed in the powdered maintenance diet UAR A04C P2.5 (UAR, Villemoisson, Epinay-sur-Orge, France).
Controls: Vehicle

Details on mating procedure:

Males and females were paired for, at most, a 2-week period, until mating was obtained.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Doses were verified for test substance concentrations in regular intervals during the study.

Duration of treatment / exposure:

Exposure: Premating exposure period (males): 10 weeks
Premating exposure period (females): 10 weeks
Duration of test: F0 pre-mating 10 weeks, until F2 weaning. (Continuously)

Frequency of treatment:

Continuously

Details on study schedule:

F0 generation: 10 weeks before mating, during the mating period until sacrifice (weaning of pups) for the males, 10 weeks before mating, during the mating, pregnancy and lactation periods (until day 21 post-partum) for the females.

A group of 25 males and 25 females received untreated diet alone, under the same experimental conditions, and acted as a reference control group. 

F1 generation: At weaning of the F1 generation, on day 22 post-partum, three groups of 25 male and 25 female Sprague-Dawley rats received the same test item, under the same experimental conditions as above, during their growth, adulthood, mating, pregnancy and lactation, until weaning of the pups (F2 generation).

A group of 25 males and 25 females received untreated diet alone under the same experimental conditions and acted as a reference control group. 

Remarks:

Doses / Concentrations:
500, 1500, and 4000 ppm of test substance (40% active DDAC) (i.e., corresponding to 203, 608 and 1620 ppm of DDAC)
Basis:
nominal in diet

Remarks:

Doses / Concentrations:
14, 39 and 109 mg a.i./kg bw/day for males and 18, 51 and 137 mg a.i./kg bw/day in females
Basis:
nominal in diet

No. of animals per sex per dose:

25 per sex per group

Control animals:

yes, concurrent vehicle

Details on study design:

Refer to study design above

Positive control:

Not applicable

Parental animals: Observations and examinations:

Examination of F0 generation: Clinical signs and mortality were checked daily. Food consumption and body weight were recorded at designated intervals.  Males and females were paired for, at most, a 2-week period, until mating was obtained.  The F0 females were allowed to deliver normally, and rear their progeny. Pregnancy and litter parameters were recorded. The F0 parent males and females were sacrificed after weaning of their progeny. During lactation, the pups from the F1 generation were observed daily for survival and clinical signs. Body weight was measured at designated intervals and 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 endpoints.

Oestrous cyclicity (parental animals):

Refer to study design

Sperm parameters (parental animals):

Refer to study design

Litter observations:

Examination of F1 generation; On day 22 post-partum, one male and one female pup per litter were selected to constitute the F1 generation, which comprised 25 males and 25 females per group. The F1 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. Neurobehavioral tests were conducted at designated intervals to assess auditory and visual function. Spontaneous locomotor activity was also evaluated twice over a 60-min interval when the animals were 7 and 8 weeks old. When the animals were 12 weeks old, F1 males and F1 females were paired.
The F1 females were allowed to deliver normally, and rear their progeny. Pregnancy and litter parameters were recorded. During lactation, the pups from 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 endpoints.

Postmortem examinations (parental animals):

Terminal examination of F0 and F1 animals: After weaning of their respective progeny, F0 and F1 parent males and females were sacrificed. Designated organs were weighed for F0 and F1 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 in both F0 and F1 males.
A macroscopic post-mortem examination was performed on all F0 and F1 parent males and females and on three pups per sex and per litter of each F0 and F1 female killed at weaning.  Any pups which died or were killed prematurely during the lactation period were also submitted for macroscopic post-mortem examination.  Macroscopic lesions, reproductive organs, adrenals and pituitary glands were sampled in all parent animals. In all pups, the macroscopic lesions were preserved. A microscopic examination was performed on macroscopic lesions, reproductive organs, adrenals, and pituitary glands of all F0 and F1 parents of the control and high dose groups. A detailed histopathological examination was performed on the ovaries and the testes.

Postmortem examinations (offspring):

Terminal examination of F0 and F1 animals: After weaning of their respective progeny, F0 and F1 parent males and females were sacrificed. Designated organs were weighed for F0 and F1 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 in both F0 and F1 males.
A macroscopic post-mortem examination was performed on all F0 and F1 parent males and females and on three pups per sex and per litter of each F0 and F1 female killed at weaning.  Any pups which died or were killed prematurely during the lactation period were also submitted for macroscopic post-mortem examination.  Macroscopic lesions, reproductive organs, adrenals and pituitary glands were sampled in all parent animals. In all pups, the macroscopic lesions were preserved. A microscopic examination was performed on macroscopic lesions, reproductive organs, adrenals, and pituitary glands of all F0 and F1 parents of the control and high dose groups. A detailed histopathological examination was performed on the ovaries and the testes.

Statistics:

No data

Reproductive indices:

Please refer to study design for F0 animals

Offspring viability indices:

Please refer to study design for F1 animals

Clinical signs:

no effects observed

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Other effects:

no effects observed

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

no effects observed

F0 generation
Test group 4 (4000 ppm): - no treatment-related deaths and no remarkable clinical signs,  - for the males, a lower body weight gain during the premating period (days 1-15), and food consumption was affected throughout the study,  - for the females, a lower body weight gain was noted during the first week of the premating period, during the gestation and the lactation, and food consumption was affected from week 2 of the study, but no effect was detected during the lactation, - there were no apparent effects on mating, fertility, fecundity or delivery,  - the pup body weight gain was significantly lower than controls from day 4 p.n.,  - no disturbances of the reflex development in the pups,  - none of the seminology parameters evaluated in F0 parents males were affected,  - distension of cecum with feces was seen at the autopsy in 13/25 F0 males, with no histopathological findings in this organ,  - higher adrenal weights (p<0.01) in F0 females correlated with enlargement of the gland and cortical cell hypertrophy seen at histopathology, - lower spleen weights in male and female pups sacrificed at weaning (p<0.01),  - no treatment-related findings were observed in male and female genital organs, and the accessory organs.          
Test group 3 (1500 ppm): - no treatment-related deaths, - no remarkable clinical signs, - neither body weight gain nor food consumption of parent animals were affected, - there were no apparent effects on mating, fertility, fecundity or delivery, - no effect on the pup body weight gain, - no disturbances of the reflex development in the pups, - none of the seminology parameters evaluated in F0 parents males were affected, - no treatment-related differences in organ weights were recorded, - neither macroscopic nor microscopic findings were observed.
Test group 2 (500 ppm): - no treatment-related deaths, - no remarkable clinical signs, - neither body weight gain nor food consumption of parent animals were affected, - there were no apparent effects on mating, fertility, fecundity or delivery, - no effect on the pup body weight gain, - no disturbances of the reflex development in the pups, - none of the seminology parameters evaluated in F0 parents males were affected, - no treatment-related differences in organ weights were recorded, - neither macroscopic nor microscopic findings were observed.

Key result

Dose descriptor:

NOAEL

Effect level:

ca. 1 500 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

food consumption and compound intake

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Sexual maturation:

no effects observed

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Gross pathological findings:

no effects observed

Histopathological findings:

no effects observed

F1 generation
Test group 4 (4000 ppm): - no treatment-related deaths, - no remarkable clinical signs, - for the males, there was a lower body weight gain until day 43 of the premating period and food consumption was affected, mainly during the same period of the study, - for the females, a lower body weight gain was noted during the first 2 weeks of the premating period, from GD 7 (GD: gestation day) and during the lactation. The food consumption was affected during the premating period, during GD 0-14 and during days 7 14 p.p., - there were no apparent effects on mating, fertility, fecundity or the progress of delivery,  - the pup body weight gain was significantly lower than controls during the lactation, - no disturbances of the reflex development in the pups, - none of the seminology parameters evaluated in F1 parents males were affected, - none of the organ weight differences recorded in F1 parents were directly treatment-related, - lower spleen weights were recorded in F1 pups, - no relevant findings were seen at necropsy of parents or pups, - neither macroscopic nor microscopic findings were observed.
Test group 3 (1500 ppm): - no treatment-related deaths, - no remarkable clinical signs, - neither body weight gain nor food consumption of parent animals were affected, - there were no apparent effects on mating, fertility, fecundity or the progress of delivery, - increase in the male ratio, - no effect on the pup body weight gain, - no disturbances of the reflex development in the pups, - none of the seminology parameters evaluated in F1 parents males were affected. - no treatment-related differences in organ weights were recorded, - neither macroscopic nor microscopic findings were observed.
Test group 2 (500 ppm): - no treatment-related deaths, - no remarkable clinical signs, - neither body weight gain nor food consumption of parent animals were affected, - there were no apparent effects on mating, fertility, fecundity or the progress of delivery, - increase in the male ratio, - no effect on the pup body weight gain, - no disturbances of the reflex development in the pups, - none of the seminology parameters evaluated in F1 parents males were affected. - no treatment-related differences in organ weights were recorded, - neither macroscopic nor microscopic findings were observed.

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

ca. 1 500 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

food consumption and compound intake

Key result

Dose descriptor:

NOAEL

Generation:

F2

Effect level:

ca. 4 000 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No treatment related effects on mating behaviour, fertility and gestation, of each generation and for development, growth and survival of each progeny

Reproductive effects observed:

not specified

Result: Not toxic to reproduction
concentrations of 500, 1500 or 4000 ppm test substance,
corresponding to 203, 608 or 1620 ppm of DDAC.

The estimation of the achieved dosages of test item
(expressed as mg of DDAC/kg/day) are summarized in the
following table:
Mean achieved dosages (in mg DDAC/kg/day)
Concentration (ppm)         500         1500         4000
F0 generation
Males
. premating (days 1-71)         17         49         137
. post-mating (days 85-120)     10         30         84
Females
. premating (days 1-71)         20         58         157
. gestation (GD 0-20)          15         45         52
. lactation (days 1-21 p.n.)    31         93         261
F1 generation
Males
. premating (days 1-64)         22         65         183
. post-mating (days 85-120)     10         31         94
Females
. premating (days 1-64)         23         69         198
. gestation (GD 0-20)          17         48         57
. lactation (days 1-21 p.n.)    36         93         263

Conclusions:

Under the study conditions, the NOAEL for parental toxicity was 1500 ppm (39 mg a.i./kg bw/day) for the male and female animals. The NOAEL for mating behaviour, fertility and gestation of each generation and for development, growth and survival of each progeny was 4000 ppm (109 mg a.i./kg bw/day).

Executive summary:

The test substance was administered, in accordance with OECD Guideline 416, by dietary admixture to male and female Sprague-Dawley rats at 500, 1500 or 4000 ppm (equivalent to an average of 14, 39 and 109 mg a.i./kg bw/day for males and 18, 51 and 137 mg a.i./kg bw/day in females, calculated from the mean achieved dose levels in the parental generation) before and through mating and in gestation until the end of the lactation period in both F0 and F1 generations. At 4000 ppm, F0 and F1 parents showed significantly lower body weight gains and reduced food consumption. At 1500 or 500 ppm, no relevant changes were noted. Treatment with the test substance had no effect on the reproductive parameters in F0 and F1 parental rats at treatment levels up to 4000 ppm. No effect was observed on mating, fertility, gestation, fecundity or delivery at any concentration for either generation. No effect was recorded on litter parameters and on pre- and post-natal development of either generation at any concentration. Apart from cortical cell hypertrophy found in the adrenal glands of F0 females treated at 4000 ppm and lower spleen weight in F1 pups of the same group, no other treatment-related findings were seen upon microscopic examination of the concerned tissues. Consequently, under the study conditions, the NOAEL for parental toxicity was 1500 ppm (39 mg a.i./kg bw/day) for the male and female animals. The NOAEL for mating behaviour, fertility and gestation of each generation and for development, growth and survival of each progeny was 4000 ppm (109 mg a.i./kg bw/day) (CIT, 2008).   

Reference 2

Endpoint:

two-generation reproductive toxicity

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Report peer-reviewed by governmental competent authority.

Qualifier:

no guideline followed

GLP compliance:

not specified

Limit test:

no

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: Exposure period: Continuous
Premating exposure period (males): 10 weeks F1A, and approx. 19 weeks F1B
Premating exposure period (females): 10 weeks F1A, and approx. 19 weeks F1B
Duration of test: Weaning second generation (Continuous)

Frequency of treatment:

Continuous

Details on study schedule:

A total of 28 males and 28 females were evaluated at each dose level. Animals were exposed to DDAC for 10 weeks prior to mating, and each of the two generations produced two litters. The original rats, called the FO generation, were randomly paired within dose groups and mated over a three-week period to produce the F1A generation Exposures continued through mating, gestation, parturition, and lactation. at least 10 days after weaning the F1A litters, FO parents were mated in different male-female pairings, within dose groups, to produce the F1B generation. Exposures to DDAC again occurred from mating to lactation. After the F1B animals were weaned, F0 parents were necropsied and high dose and control animals were examined for histopathologic lesions.

Selected F1 parents wene exposed to the same concentrations of DDAC as their parents for at least 10 weeks, and were then paired as described above to produce F2A and F2B generations. Mating, gestation, lactation, and necropsy of the F1 parents and selected F2A and F2B pups were performed as outlined above, except that no F2 animals were selected as parents.

Remarks:

Doses / Concentrations:
0, 300, 750, or 1500 ppm
Basis:
nominal in diet

No. of animals per sex per dose:

28 per sex per dose

Control animals:

yes

Details on study design:

Refer to the study schedule above

Positive control:

Not applicable

Parental animals: Observations and examinations:

Details not available

Oestrous cyclicity (parental animals):

Details not available

Sperm parameters (parental animals):

Details not available

Litter observations:

Details not available

Postmortem examinations (parental animals):

Details not available

Postmortem examinations (offspring):

Details not available

Statistics:

Details not available

Reproductive indices:

Details not available

Offspring viability indices:

Details not available

Clinical signs:

no effects observed

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Other effects:

no effects observed

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

no effects observed

Parental toxicity was observed at 1500 ppm (112.6 mg/kg/day), limited to body weight reduction, weight gain depression, and decreased food consumption.

Key result

Dose descriptor:

NOAEL

Effect level:

ca. 750 ppm

Based on:

act. ingr.

Sex:

not specified

Basis for effect level:

body weight and weight gain

food consumption and compound intake

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Sexual maturation:

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings:

no effects observed

Postnatal toxicity at 1500 ppm was indicated by reduced pup body weights.

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

ca. 750 ppm

Based on:

act. ingr.

Sex:

not specified

Basis for effect level:

body weight and weight gain

Key result

Dose descriptor:

NOAEL

Generation:

F2

Effect level:

ca. 750 ppm

Based on:

act. ingr.

Sex:

not specified

Basis for effect level:

body weight and weight gain

Reproductive effects observed:

not specified

Result: No adverse reproductive effects. Only effects reduced body weights in parents and pups at 1500 ppm.

Conclusions:

The NOEL for both adults and offspring was 750 ppm, indicating no increased risk to offspring in the absence of adult toxicity.

Executive summary:

A reproduction study using Sprague-Dawley rats involved dietary administration of DDAC at target dosage levels of 0, 300, 750 or 1500 ppm for two generations. Dose levels in terms of mg/kg/day decreased throughout the study as animal body weight increased, and varied within and between dose groups and generations, so are not reported here. Results indicate that continuous exposure to DDAC in the diet for two generations resulted in no adverse reproductive effects. Parental toxicity was observed at 1500 ppm (112.6 mg/kg bw/day), limited to body weight reduction, weight gain depression, and decreased food consumption. Postnatal toxicity at 1500 ppm was indicated by reduced pup body weights. The NOEL for both adults and offspring was 750 ppm, indicating no increased risk to offspring in the absence of indications of adult toxicity (Henderson, 1992).

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

30 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

Good quality. In line with the data presented in the DDAC assessment report for Product Type 8 conducted under Directive 98/8/EC (evaluating Competent Authority: Italy, June 2015

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

The test substance was administered, in accordance with OECD Guideline 416, by dietary admixture to male and female Sprague-Dawley rats at 500, 1500 or 4000 ppm (equivalent to an average of 14, 39 and 109 mg a.i./kg bw/day for males and 18, 51 and 137 mg a.i./kg bw/day in females, calculated from the mean achieved dose levels in the parental generation) before and through mating and in gestation until the end of the lactation period in both F0 and F1 generations. At 4000 ppm, F0 and F1 parents showed significantly lower body weight gains and reduced food consumption. At 1500 or 500 ppm, no relevant changes were noted. Treatment with the test substance had no effect on the reproductive parameters in F0 and F1 parental rats at treatment levels up to 4000 ppm. No effect was observed on mating, fertility, gestation, fecundity or delivery at any concentration for either generation. No effect was recorded on litter parameters and on pre- and post-natal development of either generation at any concentration. Apart from cortical cell hypertrophy found in the adrenal glands of F0 females treated at 4000 ppm and lower spleen weight in F1 pups of the same group, no other treatment-related findings were seen upon microscopic examination of the concerned tissues. Consequently, under the study conditions, the NOAEL for parental toxicity was 1500 ppm (39 mg a.i./kg bw/day) for the male and female animals. The NOAEL for mating behaviour, fertility and gestation of each generation and for development, growth and survival of each progeny was 4000 ppm (109 mg a.i./kg bw/day) (CIT, 2008).   

 

 A reproduction study using Sprague-Dawley rats involved dietary administration of test substance at target dosage levels of 0, 300, 750 or 1500 ppm for two generations. Dose levels in terms of mg/kg/day decreased throughout the study as animal body weight increased, and varied within and between dose groups and generations, so are not reported here. Results indicate that continuous exposure in the diet for two generations resulted in no adverse reproductive effects. Parental toxicity was observed at 1500 ppm (112.6 mg/kg bw/day), limited to body weight reduction, weight gain depression, and decreased food consumption. Postnatal toxicity at 1500 ppm was indicated by reduced pup body weights. The NOEL for both adults and offspring was 750 ppm, indicating no increased risk to offspring in the absence of indications of adult toxicity (Henderson, 1992).  

Further, the DDAC assessment report for Product Type 8 conducted under Directive 98/8/EC (evaluating Competent Authority: Italy, June 2015, attached in Section 13 of the IUCLID dataset) reported one additional 2-generation study in rats. In this study, based on reduced body weight and food consumption in F0 and F1 animals, the maternal NOAEL was established at 750 ppm (≥31 mg/kg bw/day). In the offsprings, except for reduced body weight there were no other adverse effects leading to establishment of NOAEL at 750 ppm (≥31 mg/kg bw/day). There were also no treatment related effects on any reproductive parameters were observed at any dose level. Further, the biocides assessment report, cited slightly different converted doses in mg/kg bw/day for the EQC (Chevalier 2008) study. The parental, fertility and offspring NOAELs were all established at 608 ppm (>30 mg/kg bw/day). The RMS further stated that:“Available studies do not indicate any specific potential for reproductive toxicity. Observed effects concern solely general toxicity”. The overall NOAEL (fertility/parental effects) was concluded to be 608 ppm or 30 mg/kg bw/day, but it was not considered relevant for systemic toxicity. 

Therefore, in line with the biocides assessment report, the NOAELs from the 2-generation reproductive toxicity studies in rats have not been considered further for systemic risk assessment.  

Effects on developmental toxicity

Link to relevant study records

Reference

Reference 1

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.3700 (Prenatal Developmental Toxicity Study)

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Species: Rabbit
Strain: KBL New Zealand White, Specific Pathogen Free (S.P.F.).
Source: Charles River Laboratories (Elevage Scientifique des Dombes, Châtillon-sur-Chalaronne, France).
Sex: Female
Age/weight at study initiation: 18-20 weeks; mean body weight of 3494 g (range: 3105 g to 3940 g).
Number of animals per group: 22
The animals were individually housed in stainless steel cages, in a barriered rabbit unit, under specific pathogen free (SPF) standard laboratory
conditions: temperature : 18 ± 3°C; relative humidity : 50 ± 20%; light/dark cycle : 8h dark/16h light (5:00 - 21:00);
ventilation : about 8 to 10 cycles/hour of filtered, non-recycled air.
Control animals: Yes
Mating period: Were mated at the breeder's facilities. The day of confirmed mating (visual assessment) was designated as day 0 post-coitum (p.c.) or day 0 of gestation (GD 0). The animals were supplied to CIT on day 1 post-coitum; consequently they were acclimated for a period of 5 days before
the beginning of the treatment period.

Route of administration:

oral: gavage

Vehicle:

water

Details on exposure:

Administration/ Exposure: Oral
Duration of exposure Rabbit: day 6-28 of gestation
Postexposure period: None, as exposure period was until day prior to term.
Type: oral gavage
Concentration Gavage; 0, 4, 12 or 32 mg DDAC/kg bw . The choice of the dose-levels was based on a preliminary study of effects on embryo-fetal
development in rabbits (CIT/Study No. 26153 RSL), in which the test item, DDAC, was administered to pregnant female rabbits, from day 6 to day
28 of pregnancy at the dose-levels of 4, 16 or 32 mg DDAC/kg/day.
Vehicle: Aqueous solution with purified water.
Concentration in vehicle: 0, 1.33, 4, 10.66 mg/mL. The test item dosage forms were prepared daily and stored at room temperature prior to use.
Chemical analysis of DDAC in dosage form: The concentration of samples taken from each control and test item dosage form prepared for use on
the first day of treatment and on the last day of treatment was determined. Values were found to be in agreement with nominal concentration ± 10%.
Total volume applied: 3 mL/kg BW/day
Controls: Vehicle

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

At the start and at end of the study.

Details on mating procedure:

Females were mated at breeder's facilities and the day of confirmed mating was designated as Day 0.

Duration of treatment / exposure:

Day 6 to 28 post coitum

Frequency of treatment:

Daily

Duration of test:

24 days

Remarks:

Doses / Concentrations:
Gavage 0, 4, 12 or 32 mg a.i./kg bw/day
Basis:
actual ingested

No. of animals per sex per dose:

22

Control animals:

yes, concurrent no treatment

Details on study design:

Groups of 22 mated female KBL New Zealan White rabbits were gavaged the test substance at 4, 12 and 32 mg/kg/day from day 6 to day 28 post coitum. One group of 22 mated females received the vehicle alone. Clinical sign and mortality were checked daily. Bosy weight and food consumption were recorded at designated intervals. On day 29 post coitum, the does were sacrificed and subjected to macroscopic examination of the major organs. The gravid uterus was weighed. The fetuses removed and litter parameters were examined. The fetuses were then weighed and subjected to external and internal tissue, organs and bone examination.

Maternal examinations:

Refer to study design

Ovaries and uterine content:

Refer to study design

Fetal examinations:

Refer to study design

Statistics:

Refer to study design

Indices:

Refer to study design

Historical control data:

Refer to study design

Details on maternal toxic effects:

Maternal toxic effects:yes

Details on maternal toxic effects:
In absence of a dose-related increase of death/abortion at 4 and 12 mg DDAC/kg/day and with the abortion of one female in the control group, the mortality at 4 and 12 mg DDAC/kg/day were considered to be most probably incidental and not related to treatment with the test item. 32 mg DDAC mg/kg/day, death/abortion in 3 females were ascribed to the treatment with the test item. When given 12 mg DDAC/kg/day, colored urine and loud breathing was observed in one female from GD 22 each. Both animals however, showed no findings at autopsy.  At 32 mg DDAC/kg/day, the number of animals showing clinical signs was greater than in the control group, and most of these clinical signs (colored urine, soft feces, soiled urogenital area) were recorded at the end of the gestation period, along with an absence of feces observed for three animals. Presence of blood in the bedding was noted in two females on GD 16 or 22 and one female showed an emaciated appearance on GD 29. No other clinical signs were considered to be treatment-related; absence of feces is commonly recorded at the end of pregnancy and was therefore not considered to be treatment-related at 4 mg DDAC/kg/day. Overall, there seems to be a dose related effect, tending to lower body weights at higher dose levels which corresponding to lower food consumption related to palatability of the compound in the diet.

Key result

Dose descriptor:

NOAEL

Effect level:

ca. 4 mg/kg bw/day

Based on:

act. ingr.

Basis for effect level:

other: maternal toxicity

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:yes

Details on embryotoxic / teratogenic effects:
There is a big variability in litter data. The low dose group shows higher number of dead fetuses, post implantation loss and lower fetal weight compared to control. On the other hand, the number of live fetuses is even higher than of control. At the mid-dose group comparable to control again. The high-dose group of 32 mg DDAC/kg/d resulted to an increased post-implantation loss, decrease in live fetuses with a decreased fetal body weight, and high number of dead fetuses. Upon examination of the soft tissue, presence of whitish fluid in the abdomens of 5/6 fetuses (one litter) was observed. No relevant malformations or variations were observed during the fetal evaluation. At lower concentrations, litter parameters were not affected by the test treatment and no evidence of treatment-related findings were noted at skeletal examinations of the fetuses.

Key result

Dose descriptor:

NOAEL

Effect level:

ca. 12 mg/kg bw/day

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

other: teratogenicity

Key result

Abnormalities:

not specified

Key result

Developmental effects observed:

not specified

Result: no specific concerns for developmental toxicity by DDAC.

In absence of a dose-related increase of death/abortion at 4 and 12 mg DDAC/kg/day and with the abortion of one female in
the control group, the mortality at 4 and 12 mg DDAC/kg/day were considered to be most probably incidental and not
related to treatment with the test item. 32 mg DDAC mg/kg/day, death/abortion in 3 females were ascribed to the
treatment with the test item.
When given 12 mg DDAC/kg/day, colored urine and loud breathing was observed in one female from GD 22 each. Both
animals however, showed no findings at autopsy.  At 32 mg DDAC/kg/day, the number of animals showing clinical
signs was greater than in the control group, and most of these clinical signs (colored urine, soft feces, soiled
urogenital area) were recorded at the end of the gestation period, along with an absence of feces observed for three
animals. Presence of blood in the bedding was noted in two females on GD 16 or 22 and one female showed an emaciated
appearance on GD 29. No other clinical signs were considered to be treatment-related; absence of feces is commonly
recorded at the end of pregnancy and was therefore not considered to be treatment-related at 4 mg DDAC/kg/day.
Overall, there seems to be a dose related effect, tending to lower body weights at higher dose levels which corresponding
to lower food consumption related to palatability of the compound in the diet.

There is a big variability in litter data. The low dose group shows higher number of dead fetuses, post implantation
loss and lower fetal weight compared to control. On the other hand, the number of live fetuses is even higher than
of control. At the mid-dose group comparable to control again. The high-dose group of 32 mg DDAC/kg/d resulted to an
increased post-implantation loss, decrease in live fetuses with a decreased fetal body weight, and high number of dead
fetuses.
Upon examination of the soft tissue, presence of whitish fluid in the abdomens of 5/6 fetuses (one litter) was
observed. No relevant malformations or variations were observed during the fetal evaluation.
At lower concentrations, litter parameters were not affected by the test treatment and no evidence of treatment-related
findings were noted at skeletal examinations of the fetuses.

Conclusions:

Based on the results of this study, there are no specific concerns for developmental toxicity by DDAC.

Executive summary:

A study was conducted in rabbits according to OECD Guideline and 414 and EPA OPPTS guideline to determine the prenatal developmental toxicity of the test substance. Groups of 22 mated female KBL New Zealand White rabbits were gavaged the test substance at 4, 12 and 32 mg/kg/day from day 6 to day 28post coitum. One group of 22 mated females received the vehicle alone. Clinical sign and mortality were checked daily. Body weight and food consumption were recorded at designated intervals. On day 29 post coitum, the does were sacrificed and subjected to macroscopic examination of the major organs. The gravid uterus was weighed. The foetuses removed and litter parameters were examined. The foetuses were then weighed and subjected to external and internal tissue, organs and bone examination. Based on the results of this study, there are no specific concerns for developmental toxicity by DDAC. LO(A)EL maternal toxic effects: 32 mg a.i./kg bw/day resulted from clear signs of maternal toxicity (mortality, abortion, clinical signs, body weight loss, increased incidence autopsy findings). At 12 mg/kg bw/day, one female displayed coloured urine and another female loud breathing, each from GD 22. There is no real critical effect, but more a general display of toxicity, possibly related to reduced food intake due to palatability. Based on this, the LOAEL could be set at 12 mg a.i./kg bw/day. NO(A)EL maternal = 4 mg/kg bw/day. LO(A)EL embryotoxic/teratogenic effects: 32 mg DDAC/kg/day. Effects observed are increased number of dead fetuses, lower number of live foetuses, increased post-implantation loss and a decreased foetal body weight. 5/6 foetuses had whitish fluid in the abdomens upon soft tissue examination. NO(A)EL embryotoxic / teratogenic effects = 12 mg a.i./kg bw/day

Reference 2

Endpoint:

developmental toxicity

Type of information:

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

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

no guideline followed

GLP compliance:

no

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Route of administration:

dermal

Vehicle:

ethanol

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

Gestation Day 6-15

Frequency of treatment:

Daily

Duration of test:

16 days

Remarks:

Doses / Concentrations:
0.5 mL of 4.4, 6.6 or 9.9%
Basis:
nominal conc.

Remarks:

Doses / Concentrations:
8, 16.5 or 33 mg per animal
Basis:
nominal conc.

No. of animals per sex per dose:

Approx 20 dams/group

Control animals:

yes, concurrent vehicle

Details on study design:

Sex: female

Details on maternal toxic effects:

Maternal toxic effects:yes

Details on maternal toxic effects:
Only dermal irritation observed

Key result

Dose descriptor:

NOAEL

Effect level:

ca. 200 mg/kg bw/day (nominal)

Based on:

act. ingr.

Basis for effect level:

other: maternal toxicity

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Key result

Dose descriptor:

NOAEL

Effect level:

ca. 200 mg/kg bw/day (nominal)

Based on:

act. ingr.

Sex:

not specified

Basis for effect level:

other: teratogenicity

Key result

Abnormalities:

not specified

Key result

Developmental effects observed:

not specified

Conclusions:

No embryotoxicity was observed up to the highest dose of 200 mg/kg/day dermal application. Maternal toxicity was observed in terms of local irritation only.

Executive summary:

The study involved dermal application, of 0.5 mL of 0, 4.4, 6.6, 9.9% dimethyldistearyl ammonium chloride to the uncovered shaved skin of 20 mated Sprague-Dawley rats/group upon GD 6 -15 (9.9% corresponds to approximately 200 mg/kg/day). The doses induced local adverse maternal reaction (skin reactions), but no systemic toxicity. No effects on litter size, post implantation loss, litter weights, mean foetal weights were seen. No signs of foetal abnormalities were also observed.

Reference 3

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

abstract

Remarks:

Only brief study summary available

GLP compliance:

not specified

Species:

rabbit

Strain:

New Zealand White

Route of administration:

oral: gavage

Duration of treatment / exposure:

Gestation Day 6-18

Frequency of treatment:

Daily

Remarks:

Doses / Concentrations:
0.0, 1.0, 3.0, or 10.0 mg/kg bw/day
Basis:

Control animals:

yes, concurrent vehicle

Details on study design:

Sex: female

Key result

Dose descriptor:

NOAEL

Effect level:

ca. 1 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: maternal toxicity

Key result

Dose descriptor:

NOAEL

Effect level:

ca. 3 mg/kg bw/day

Based on:

test mat.

Sex:

not specified

Basis for effect level:

other: teratogenicity

Key result

Abnormalities:

not specified

Key result

Developmental effects observed:

not specified

Result: Non teratogenic

Conclusions:

The NOEL for maternaltoxicity was set at 1.0 mg/kg bw/day, and the NOEL for developmental toxicity was set at 3.0 mg/kg bw/day

Executive summary:

In a developmental toxicity study, sixteen mated New Zealandfemale white rabbits per dosage group were administered 0.0,1.0, 3.0, or 10.0 mg/kg/day of DDAC by gavage at gestational days 6 through 18. The vehicle was deionized water. 4 out of 16 does in the highest dose died prior to GD 13. One doe at 10 mg/kg bw/day and two does at 1.0 mg/kg bw/day delivered early and were removed from the study. No females aborted. Maternal toxicity, including mortality, was evident at 10 mg/kg bw/day. Non-lethal indications of maternal toxicity were evident at 3 and 10 mg/kg bw/day, as evidenced by reduced weight gain and clinical signs during the treatment period. Developmental toxicity including increased incidence of foetal mortality and reduced foetal body weight per litter, were observed only at the highest dose. No teratogenic effects were observed at any dose employed. The NOEL for maternal toxicity was set at 1.0 mg/kg bw/day, and the NOEL for developmental toxicity was set at 3.0 mg/kg bw/day.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

0.8 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

Good quality. Effects secondary to local effects; in line with the data presented in the DDAC assessment report for Product Type 8 conducted under Directive 98/8/EC (evaluating Competent Authority: Italy, June 2015

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Additional information

A study was conducted according to OECD Guideline and 414 and EPA OPPTS guideline to determine the prenatal developmental toxicity of the test substance in rabbits. Groups of 22 mated female KBL New Zealand White rabbits were gavaged the test substance at 4, 12 and 32 mg/kg/day from Day 6 to Day 28 post coitum. One group of 22 mated females received the vehicle alone. Clinical sign and mortality were checked daily. Body weight and food consumption were recorded at designated intervals. On Day 29 post coitum, the does were sacrificed and subjected to macroscopic examination of the major organs. The gravid uterus was weighed. The foetuses removed and litter parameters were examined. The foetuses were then weighed and subjected to external and internal tissue, organs and bone examination. Based on the results of this study, there are no specific concerns for developmental toxicity. LO(A)EL maternal toxic effects: 32 mg a.i./kg bw/day resulted from clear signs of maternal toxicity (mortality, abortion, clinical signs, body weight loss, increased incidence autopsy findings). At 12 mg a.i./kg bw/day, one female displayed coloured urine and another female loud breathing, each from Gestation Day (GD) 22. There is no real critical effect, but more a general display of toxicity, possibly related to reduced food intake due to palatability. Based on this, the LOAEL could be set at 12 mg a.i./kg bw/day. NO(A)EL maternal = 4 mg a.i./kg bw/day. LO(A)EL embryotoxic/teratogenic effects: 32 mg a.i./kg bw/day. Effects observed are increased number of dead fetuses, lower number of live foetuses, increased post-implantation loss and a decreased foetal body weight. 5/6 foetuses had whitish fluid in the abdomens upon soft tissue examination. NO(A)EL embryotoxic / teratogenic effects = 12 mg a.i./kg bw/day (CIT, 2003 and 2005 a and b).   

Another developmental study involved dermal application of 0.5 mL of 0, 4.4, 6.6 and 9.9% test substance to the uncovered shaved skin of 20 mated Sprague-Dawley rats/group on GD 6 to 15 (9.9% corresponds to approximately 200 mg/kg/day). The doses induced local adverse maternal reaction (skin reactions), but no systemic toxicity. No effects on litter size, post implantation loss, litter weights, mean foetal weights were seen. No signs of foetal abnormalities were observed (HRC, 1983).  

 In a developmental toxicity study, 16 mated New Zealand female white rabbits per dosage group were administered 0.0,1.0, 3.0, or 10.0 mg/kg/day of DDAC by gavage at GD 6 to 18. The vehicle was deionized water. Six out of 16 does in the highest dose died prior to GD 13. One doe at 10 mg/kg bw/day and two does at 1.0 mg/kg bw/day delivered early and were removed from the study. No females aborted. Maternal toxicity, including mortality, was evident at 10 mg/kg bw/day. Non-lethal indications of maternal toxicity were evident at 3 and 10 mg/kg bw/day, as evidenced by reduced weight gain and clinical signs during the treatment period. Developmental toxicity including increased incidence of foetal mortality and reduced foetal body weight per litter, were observed only at the highest dose. No teratogenic effects were observed at any dose. The NOEL for maternal toxicity was set at 1.0 mg a.i./kg bw/day, and the NOEL for developmental toxicity was set at 3.0 mg/kg bw/day (Henderson, 1992). 

Further, the DDAC assessment report for Product Type 8 conducted under Directive 98/8/EC (evaluating Competent Authority: Italy, June 2015, attached in Section 13 of the IUCLID dataset) reported two additional studies in rats and rabbits. In these studies, based on local effects, reduced body weight and/or food consumption, which was considered secondary, the maternal NOELs in rats and rabbits were established at 0.8 and 1 mg/kg bw/day respectively. Except for reduced fetal weight in the rabbit study, there no other development toxicity including teratogenicity observed at any dose leading to establishment of respective NOELs at >16.2 mg/kg bw/day in rats and at 3 mg a.i./kg bw/day in rabbits. The lowest NOAEL for maternal toxicity was concluded to be 0.8 mg/kg bw/day, which was not considered relevant for systemic toxicity.  

Therefore, in line with the biocides assessment report, the NOAELs from the development toxicity studies have not been considered further for systemic risk assessment.  

Justification for classification or non-classification

Based on the results of the read across two-generation reproductive toxicity and pre-natal development toxicity studies in rats and rabbits, the test substance does not warrant a classification according to EU CLP (Regulation EC/1272/2008) criteria.