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Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
two-generation reproductive toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data from secondary literature

Data source

Referenceopen allclose all

Reference Type:
secondary source
Title:
Reasoned opinion on the setting of an import tolerance for test material
Author:
European Food Safety Authority (EFSA)
Year:
2013
Bibliographic source:
EFSA Journal 2013
Reference Type:
secondary source
Title:
Draft Assessment Report (DAR): Didecyldimethylammonium Chloride
Author:
European Food Safety Authority (EFSA)
Year:
2008
Bibliographic source:
Draft Assessment Report (DAR): Didecyldimethylammonium Chloride, Volume 3, Annex B, part 2, B.6 April 2008

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)
Principles of method if other than guideline:
Two generation reproduction study of test material was performed on Sprague Dawley rats orally by gavage.
GLP compliance:
not specified
Limit test:
no
Justification for study design:
No data available

Test material

Constituent 1
Chemical structure
Reference substance name:
Didecyldimethylammonium chloride
EC Number:
230-525-2
EC Name:
Didecyldimethylammonium chloride
Cas Number:
7173-51-5
Molecular formula:
C22H48N.Cl
IUPAC Name:
1-Decanaminium, N-decyl-N,N-dimethyl-, chloride
Details on test material:
- Name of test material (as cited in study report):Didecyldimethylammonium Chloride
- Molecular formula : C22H48N.Cl
- Molecular weight : 362.081g/mol
- Substance type: Organic
- Physical state: Solid

Test animals

Species:
rat
Strain:
Sprague-Dawley
Remarks:
CD®
Details on species / strain selection:
No data available
Sex:
male/female
Details on test animals or test system and environmental conditions:
No data available

Administration / exposure

Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on exposure:
Details on exposure
PREPARATION OF DOSING SOLUTIONS:
Undiluted test material given by oral route in food
DIET PREPARATION
- Rate of preparation of diet (frequency):No data available
- Mixing appropriate amounts with (Type of food )
- Storage temperature of food: No data available
VEHICLE
- Justification for use and choice of vehicle (if other than water): none
- Concentration in vehicle: 0, 20, 50 and 100 mg /kg bw/day
- Amount of vehicle (if gavage): No data available

- Lot/batch no. (if required): No data available
- Purity: No data available
Details on mating procedure:
- M/F ratio per cage: No data available
- Length of cohabitation: 21 days
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy No data available
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: [no / yes (explain)] No data available
- After successful mating each pregnant female was caged (how): No data available
- Any other deviations from standard protocol: No data available
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
starting 10 weeks before mating (F0) or at weaning (F1), throughout mating, gestation and lactation periods until scheduled necropsy
Frequency of treatment:
daily
Details on study schedule:
F0 rats were mated twice to produce F1a and F1b litters. Twenty-eight parents/sex of the F2 generation were selected from the F1b litters. They were mated twice to produce F2a and F2b litters. Mating was allowed for up to 21 days. Necropsy was performed on 10 pups/sex/dose from the F1a and F1b generation
Doses / concentrations
Remarks:
0, 20, 50 and 100 mg /kg bw/day
No. of animals per sex per dose:
Total:448
For F0 generation
0 mg/kg bw/day:28 male and 28 female
20mg/kg bw/day:28 male and 28 female
50mg/kg bw/day:28 male and 28 female
100 mg/kg bw/day:28 male and 28 female
For F1 generation
0 mg/kg bw/day:28 male and 28 female
20mg/kg bw/day:28 male and 28 female
50mg/kg bw/day:28 male and 28 female
100 mg/kg bw/day:28 male and 28 female
Control animals:
yes
Details on study design:
No data available
Positive control:
No data available

Examinations

Parental animals: Observations and examinations:
Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: yes

DETAILED CLINICAL OBSERVATIONS: Yes

Time schedule:


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

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


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

OTHER:
Oestrous cyclicity (parental animals):
No data available
Sperm parameters (parental animals):
No data available
Litter observations:
No data available
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals [describe when, e.g. as soon as possible after the last litters in each generation were produced.]
- Maternal animals: All surviving animals [describe when, e.g. after the last litter of each generation was weaned.]

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

HISTOPATHOLOGY / ORGAN WEIGHTS: yes
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring not selected as parental animals and all F2 offspring were sacrificed at [#?] days of age.
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows:

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

HISTOPATHOLOGY / ORGAN WEIGTHS
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.
Statistics:
No data available
Reproductive indices:
No data available
Offspring viability indices:
No data available

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
no effects observed
Description (incidence and severity):
No treatment-related clinical signs were observed
Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Description (incidence):
There were no treatment related deaths during the study. One male at each dosing level was killed due to trauma or caging accident or sacrificed moribund.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Body weight gain was reduced in males and females at 100 mg/kg/day food during the premating period.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Food consumption was reduced at 100 mg/kg /day during the premating period in males and females.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
No treatment-related abnormalities were observed, including histopathological examinations
Histopathological findings: neoplastic:
not specified
Other effects:
not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed
Description (incidence and severity):
There were no changes detected between parental animals of the treated and control groups in mating indices and pregnancy rates. No treatment-related changes were detected in litter size, live birth index, viability index, lactation index and sex ratio of the F0 offspring

Effect levels (P0)

Dose descriptor:
NOAEL
Effect level:
100 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
reproductive performance
Remarks on result:
other: No effects on reproductive parameters

Target system / organ toxicity (P0)

Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not specified
Mortality / viability:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
F1 pup weight gains were initially similar to those in the control group, but were significantly decreased on day 21 and 28 post-partum at 100 mg/kg /day (82-87% and 83-86% of control for F1a males and females, and 76-85% and 78-83% of control for F1b males and females, resp.).
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
no effects observed
Description (incidence and severity):
F1a and F1b pups showed no treatment-related macroscopic findings.
Histopathological findings:
no effects observed
Other effects:
not specified

Developmental neurotoxicity (F1)

Behaviour (functional findings):
not specified

Developmental immunotoxicity (F1)

Developmental immunotoxicity:
not specified

Details on results (F1)

No treatment-related changes were observed in litter size, live birth index, viability index, lactation index or sex ratio

Effect levels (F1)

open allclose all
Dose descriptor:
NOAEL
Generation:
F1a
Effect level:
50 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
viability
clinical signs
mortality
body weight and weight gain
gross pathology
Remarks on result:
other: overall no effects on developmental parameters
Dose descriptor:
NOAEL
Generation:
F1b
Effect level:
50 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
viability
mortality
body weight and weight gain
gross pathology
Remarks on result:
other: No effects on developmental parameters

Target system / organ toxicity (F1)

Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Results: F2 generation

General toxicity (F2)

Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality / viability:
not specified
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
F2a pup weight gains were decreased at 100 mg/kg /day on day 21 and 28 post-partum (85-89% and 83-90% of control for males and female resp.). F2b pup weight gains were decreased at 100 mg/kg/day on day 21 and 28 post-partum (85-89% and 84-86% of control).
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
no effects observed
Description (incidence and severity):
F2a and F2b pups showed no treatment-related macroscopic finding
Histopathological findings:
not specified
Other effects:
not specified

Developmental neurotoxicity (F2)

Behaviour (functional findings):
not specified

Developmental immunotoxicity (F2)

Developmental immunotoxicity:
not specified

Effect levels (F2)

open allclose all
Dose descriptor:
NOAEL
Generation:
F2a
Effect level:
50 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
gross pathology
Remarks on result:
other: overall no effects on developmental parameters
Dose descriptor:
NOAEL
Generation:
F2b
Effect level:
50 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
gross pathology
Remarks on result:
other: overall no effects on developmental parameters

Target system / organ toxicity (F2)

Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Overall reproductive toxicity

Reproductive effects observed:
not specified
Treatment related:
not specified
Relation to other toxic effects:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Any other information on results incl. tables

The results of the study are summarized in table

Dose

(mg/kg food)

0

300

750

1500

dr

m

f

m

f

m

f

m

f

         F0animals:

 

 

Mortality

no treatment-related findings

 

Clinical signs

no treatment-related findings

 

Body weight gain

 

 

 

dc

dc

 

Food consumption

 

 

 

dc

dc

 

Mating/fertility/gestation

no treatment-related findings

 

Oestrus cycle

not evaluated

 

Sperm evaluation

not evaluated

 

Organ weight

not determined

 

Pathology

 

 

Macroscopy

no treatment-related findings

 

Microscopy

no treatment-related findings

 

F1 pups:

 

 

Litter size

no treatment-related findings

 

Post implantation loss (%)

not evaluated

 

Live birth index

no treatment-related findings

 

Viability index

no treatment-related findings

 

Lactation index

no treatment-related findings

 

Sex ratio

no treatment-related findings

 

Clinical signs

not determined

 

Body weight gain (F1a + F1b)

day 21+28 post-partum

 

 

 

dc

dc

 

Sexual maturation

not determined

 

Organ weights

not determined

 

Pathology

 

 

Macroscopy

no treatment-related findings

 

F1 animals:

 

 

Mortality

no treatment-related findings

 

Clinical signs

no treatment-related findings

 

Body weight gain

no treatment-related findings

 

Food consumption

- premating period

 

 

 

 

dc

 

dc

 

Mating/fertility/gestation

no treatment-related findings

 

Oestrus cycle

not evaluated

 

Sperm evaluation

not evaluated

 

Organ weight

not determined

 

Pathology

 

 

Macroscopy

no treatment-related findings

 

Microscopy

no treatment-related findings

 

       F2 pups:

 

 

Litter size

no treatment-related findings

 

Post implantation loss (%)

not determined

 

Live birth index

no treatment-related findings

 

Viability index

no treatment-related findings

 

Lactation index

no treatment-related findings

 

Sex ratio

no treatment-related findings

 

Clinical signs

not evaluated

 

Body weight

F2a pups

day 21 post-partum

day 28 post-partum

F2b pups

day 21+28 post-partum

 

 

 

 

 

 

 

 

dc

dc

 

dc

 

 

dc

dc

 

dc

 

Organ weights

not determined

 

Pathology

 

 

Macroscopy

no treatment-related findings

 

dr :dose related

dc/ic :statistically significantly decreased/increased compared to the controls

d/i :decreased/increased, but not statistically significantly compared to the controls

a/r :absolute/relative organ weight

Applicant's summary and conclusion

Conclusions:
No Observed Adverse Effect Level (NOAEL) for F0 generation was considered to be 100mg/kg/day, As No treatment related changes were noted in mating behaviour, conception and gestation and the NOAEL for developmental toxicity was considered to be 50 mg/kg /day, based on decreased body weight. When female Sprague Dawley rats were treated with test material orally.
Executive summary:

Two generation reproduction study of test material was performed on male and female Sprague Dawley CD® rats in accordance with OECD guideline 416 (1983). Although oestrus cycle, sperm parameters and sexual maturation of pups was not investigated, no reproductive effects were observed indicating indirectly no effect on these parameters. The study is considered acceptable. The test material mixed with food in dose concentration0, 20, 50 and 100 mg /kg bw/day (0, 300, 750 and 1500 mg/kg food ).28 male and 28 female placed in each dose group. Diet starting 10 weeks before mating (F0) or at weaning (F1), throughout mating, gestation and lactation periods until scheduled necropsy. F0 rats were mated twice to produce F1a and F1b litters. Twenty-eight parents/sex of the F2 generation were selected from the F1b litters. They were mated twice to produce F2a and F2b litters. Mating was allowed for up to 21 days. Necropsy was performed on 10 pups/sex/dose from the F1a and F1b generation. Clinical signs, mortality, body weight and Food consumption were noted in each generation. 

 No treatment-related clinical signs were observed. There were no treatment related deaths during the study. One male at each dosing level was killed due to trauma or caging accident or sacrificed moribund. Body weight gain was reduced in males and females at 100 mg/kg/day food during the premating period. Food consumption was reduced at 100 mg/kg /day during the premating period in males and females. At necropsy no treatment-related abnormalities were observed, including histopathological examinations there were no changes detected between parental animals of the treated and control groups in mating indices and pregnancy rates. No treatment-related changes were detected in litter size, live birth index, viability index, lactation index and sex ratio of the F0 offspring.

 F1 pup weight gains were initially similar to those in the control group, but were significantly decreased on day 21 and 28 post-partum at 100 mg/kg /day (82-87% and 83-86% of control for F1a males and females, and 76-85% and 78-83% of control for F1b males and females, resp.). F1a and F1b pups showed no treatment-related macroscopic findings. F1 parental animals showed no treatment-related mortality or clinical signs. Body weight gain was only incidentally reduced for F1 parents. Body weights of males and females at 100 mg/kg /day were already significantly reduced at the beginning of mating due to the reduced body weight gain during their lactation period. Body weights of females were still decreased during gestation and were equal to controls at the end of lactation due 10 increased body weight gains relative to controls during gestation and lactation (F28 and F2b). Food consumption was significantly reduced at 100mg/kg /day in F1 males until week 7 and F1 females during the whole pre-mating period. No changes were noted between parental animals of the treated and control groups in mating indices, pregnancy rates and male fertility. At necropsy no treatment-related abnormalities were observed, including histopathological examination. No treatment-related changes were observed in litter size, live birth index, viability index,l actation index or sex ratio of the F, offspring (F2pups). F2apup weight gains were decreased at 100 mg/kg /day on day 21 and 28 post-partum (85-89% and 83-90% of control for males and female resp.). F2bpup weight gains were decreased at 100 mg/kg/day on day 21 and 28post-partum (85-89%and 84-86% of control).F2a andF2b pups showed no treatment-related macroscopic finding. Hence No Observed Adverse Effect Level (NOAEL) for F0 generation was considered to be 100mg/kg/day, As No treatment related changes were noted in mating behaviour, conception and gestation and the NOAEL for developmental toxicity was considered to be 50 mg/kg /day, based on decreased body weight. When female Sprague Dawley rats were treated with test material orally.