Quaternary ammonium compounds, bis(hydrogenated tallow alkyl)dimethyl, acetates

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

Oral: NOAEL; OECD 421, rat. NOAEL for reproductive toxicity was125mg/kg/day. Reliability = 2 (read-across substance).

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

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

Adequacy of study:

key study

Study period:

From 1999-06-16 to 1999-12-14

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Although this study is guideline and GLP compliant and would normally be assigned a reliability of 1 (reliable without restrictions), this study is used for read-across and therefore has been assigned a reliability of 2 (reliable with restrictions).

Justification for type of information:

The primary component of all substances provide complete coverage of 68334-33-8. 68334-33-8 shares high structural similarity with 61789-80-8, 68783-78-8, 107-64-2, and 112-02-7. As 68334-33-8 is a UVCB its components encapsulate the other substances except for the counter ion (Cl-). In solution, the counter ions will dissociate from the parent structures. Therefore, we are comparing substances of equivalent nature. CAS 107-64-2 represents the C18 boundary of the 61789-80-8. Ignoring the salt component CAS 61789-80-8 is equivalent to CAS 68334-33-8. 68783-78-8 is a worst case of both 68334-33-8 and 61789-80-8 since it is unsaturated and the SP2 carbon-carbon bonds are of higher reactivity and a more likely site of metabolic activation. The primary component of CAS 112-02-7 is a substructure of all the other substances. Additional documentation, provided within the IUCLID Assessment Reports section, supports the read-across approach.

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

according to guideline

Guideline:

OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Crj: CD(SD)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Italia S.p.A Calco (Italy)
- Age at study initiation: 8 weeks old
- Weight at study initiation: males: 225-250 g, Females: 200-225 g
- Fasting period before study: Not documented
- Housing: The rats were divided by sex and kept in Makrolon cages measuring 425 x 266 x 150mm, type 3D-Tecniplast Gazzada s.r.l., Buguggiate - Varese, each fitted with stainless steel cover-feed rack. During pre-mating, 2 males and 2 females were housed together. During the mating period, 1 male and female were housed together while during pregnancy and lactation, the females were housed individually. Dust free poplar/fir wood chips, heat processed for resin removal were used for bedding.
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±2°C
- Humidity (%): 55±15%
- Air changes: 15-20 air changes/hour
- Photoperiod: 12hrs dark / 12 hrs light

IN-LIFE DATES: From: June 16th 1999 To: July 25th 1999

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: daily

VEHICLE
- Justification for use and choice of vehicle (if other than water): corn oil for solubility reasons
- Concentration in vehicle: 0, 6.25, 12.5 and 50 mg/ml
- Amount of vehicle: the volume administered was 10 ml/kg bw/day
Every day, appropriate amounts of the test substance were weighed , mixed in corn oil, magnetically stirred for few minutes and warmed up to 50-60°c for 15-20 minutes. Then the formulates were magnetically stirred again to obtain final suspensions at the final concentrations of 6.25, 12.5 and 50 mg/ml. Formulates were kept under magnetic stirring until the end of daily administration. All formulates were administered within 4 hours of the preparation.

Details on mating procedure:

- M/F ratio per cage: one male with one female
- Length of cohabitation: 7 evenings/week for a maximum of 14 evenings for 16 hours at a time
- Proof of pregnancy: sperm in vaginal smear] referred to as day 0 of pregnancy
- Further matings after two unsuccessful attempts: no
- After successful mating each pregnant female was caged individually

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The concentration of the test substance were checked on two occasions with a two phase titration method according to ISO 2271.
Actual concentrations of 6.2, 13.2, 55.1 mg/mL for the first assay and 6.4, 13.4, 53.8 mg/mL for the second assay were found for the respective nominal concentrations of 6.25, 12.5 and 50 mg/mL.

Duration of treatment / exposure:

From 14 days prior to mating until a maximum dosing period of 28 days for males.
From 14 days prior to mating, during mating, pregnancy and up to day 3 of lactation for females. Females with positive smear who did not give birth were treated until killing (Day 25 of presumed pregnancy). Females without positive smear were treated up until 25 days after the end of the mating period.

Frequency of treatment:

once daily

Dose / conc.:

62.5 mg/kg bw/day (nominal)

Dose / conc.:

125 mg/kg bw/day (nominal)

Dose / conc.:

500 mg/kg bw/day (nominal)

No. of animals per sex per dose:

10/sex/dose

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale::
Dosage were selected on a basis of a maximum tolerated dose study in male and female rats (no further details available)

Positive control:

no

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: recorded daily.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: recorded daily

BODY WEIGHT: Yes
- Time schedule for examinations: All rats were weighed on a weekly basis during the pre-mating and mating periods. During the pregnancy period, the females were weighed on days 0, 7, 14 and 20 and during the lactation on days 0 (parturition day), and day 4.

FOOD CONSUMPTION :
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
Durign the pre-mating period of the male and female F0 generation, food was distributed in weighed amounts. The leftover amounts of the weighed food allocated to each cage were recorded once a week in order to calculate food consumption in g/rat/day. Food consumption was then recorded on days 7, 14 and 20 during pregnancy and on day 4 of lactation.

WATER CONSUMPTION: No data

Oestrous cyclicity (parental animals):

Not examined

Sperm parameters (parental animals):

Parameters examined in male parental generations:
In all males: testis weight, epididymis weight,
In the control and high dose-groups: histology with special emphasis on stages of spermatogenesis and histology on intersticial cell structure.

Litter observations:

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities

GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities; possible cause of death was determined for pups born or found dead.

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving animals were killed the day after the end of treatment.
- Maternal animals: All surviving animals were killed on day 4 of lactation with their pups.

GROSS NECROPSY
Gross pathology examination was performed.

HISTOPATHOLOGY / ORGAN WEIGHTS
- The ovaries and uteri (including horns, cerfix and vagina) testes, epididymides, accessory sex organs were weighed and then fixed in formalin (ovaries and uteri) or in Bouins fluid for histology

- The uteri of apparently non-pregnant females were stained using the method of Salewski and examined for the presence of implantation sites.

- Additional sections of testes were stained with PAS- hematoxylin to allow spermatogenesis to be classified into 14 stages, each stage based primarily upon the changes of the acrosome and head morphology of the younger generation of spermatids.

Postmortem examinations (offspring):

SACRIFICE
- The F1 offspring were sacrificed at 4 days of age.

GROSS NECROPSY
- Gross necropsy consisted of external examinations

Statistics:

- The probablility of survival per group was determined using the product-limit procedure of Kaplan-Meyer. The Log-Rank test was applied in order to detect differences in survival among groups. A trend test was also applied.
- The following tests were used for the purpose of comparing treated and control groups:
To compare frequencies, the heterogeneity test (CHI square 2xN) and Fischers exact test were applied. The Trend test was also applied.
To compare the mating distribution during the time allowed to the F0 generation for mating, the Log-rank test was applied.
All the other data were compared with ANOVA and Dunnett's test (when necessary) for homogeneous variance and kruskal-Wallis non parametric ANOVA for not homogeneous variance.

Reproductive indices:

Mating index (F0): percent ratio of animals with positive smears plus females found to be pregnant but without positive smears to the animals mated.

Fertility index (F0): percent ratio of females having evident signs of pregnancy to females having positive vaginal smears plus females found to be pregnant but without positive vaginal smears.

Pregnancy index (F0): the precent ratio of females with live births to the pregnant females.

Pre-coital interval (F0): calculated on the dams which proved pregnant and was expressed for each group as the mean time lapse (in days) between the beginning of the mating period and the ascertainment that copulation had occurred.

Pregnancy period (F0): the duration of pregnancy was determined for all those dams that reached term of pregnancy as being the time that elapsed between the day vaginal smear proved positive and the day of parturition.

Post implantation losses: No. implantations-No. live pups / No. implantations x 100.

Offspring viability indices:

Birth Index: No. of live newborns at birth/No. of implantations x 100.

Viabililty index on Day 4: No. of live pups on day 4 after birth / No. of live births x 100.

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

* At 500mg/kg/day:
- All females treated with 500mg/kg/day showed dyspnea, loss of weight and soft stools after a few treatments. One female also displayed chromadacryorrhea and 5 females also showed slight to moderate dilation of the abdomen. Seven of the ten males at 500 mg/kg/day showed dyspnoea and soft stools.
* At 125 and 62.5 mg/kg/day, there were no clinical signs.

Mortality:

mortality observed, treatment-related

Description (incidence):

* At 500mg/kg/day:
- One female and one male died after 12 and 10 treatments respectively.

* At 125 and 62.5 mg/kg/day, there were no deaths.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

- Pre-mating period:
* At 500mg/kg/day:
The body weight of the treated males and females decreased significantly in the first week of treatment. In the second week of treatment the body weight remained significantly lower than controls in the case of males and lower without reaching significance level in females. Consequently, body weight gain during the whole pre-mating period was lower in the case of females and significantly decreased in the case of males.
The mean daily food consumption of the treated males was significantly lower during the pre-mating period. The mean daily food consumption of the treated females was slightly lower than that of the control females, but without reaching significance.
* At 125 and 62.5 mg/kg/day, there were no effects on the body weight and food consumption during the pre-mating period.

- Pregnancy period:
* At 500mg/kg/day:
The mean body weight of the treated female was still lower than that of the control females during the pregnancy period, reaching significance level on days 14 and 20.
The mean daily food consumption of the 500mg/kg/day treated group was significantly lower only in the first week of pregnancy.
* At 125 and 62.5 mg/kg/day, there were no effects on the body weight and food consumption during the pre-mating period.

- Lactation period:
* At 500mg/kg/day:
The mean body weight of the treated females was still significantly lower than that of the control group on day 0 of lactation.
The mean daily food consumption of the treated female was slightly lower than that of the control group without reaching significance.
* At 125 and 62.5 mg/kg/day, there were no effects on the body weight and food consumption during the lactation period.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

only high dose

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

Histology of F0 testes, epididymides and ovaries did not show any compound-related changes. In particular, no changes were seen in the testicular stages performed in the PAS-hematoxylin stained sections.

Other effects:

not examined

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

effects observed, treatment-related

Description (incidence and severity):

only high dose* At 500mg/kg/day:
The mating and fertility indices were slightly lower than those of the control group. From the 9 paired females 7 revealed to be sperm positive (77 %) after mating, 6 out of 9 (67 %) revealed to be pregnant, and 5 out of 6 (83 %) delivered live litters. One animal revealed to have fully resorbed. Mean pre-coital time was longer in this group (about 6.1 days) when compared to the controls and the lower dosage groups (1.5 to 2.1 days).No effects on parturition were found at any dose. The pregnancy length was similar in all experimental groups. The mean value per litter of live born was slightly lower in the 500 mg/kg/day treated group when compared to controls.
* At 125 and 62.5 mg/kg/day, there were no effects on reproductive performance.

Dose descriptor:

NOAEL

Effect level:

125 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: See remarks

Critical effects observed:

no

Clinical signs:

no effects observed

Mortality / viability:

mortality observed, treatment-related

Description (incidence and severity):

* At 500mg/kg/day:
The birth index was statistically decreased compared to controls. The percentage of post implantation losses was increased to 19% per litter in comparison to about 6% per litter in the controls thus resulting in a lower rate of live borns of 83% in comparison to 94% in the controls.
Viability index on postnatal day 4 was in the range of those of the controls.
* At 125 and 62.5 mg/kg/day, there were no effects on the viability.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

No statistically significant differences were found between the body weight of the treated pups and the control pups during the 4 days of lactation.

Sexual maturation:

not examined

Description (incidence and severity):

not applicable in this type of test

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

not examined

Histopathological findings:

not examined

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

125 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: At the higher dose-level of 500 mg/kg bw (which is also the highest dose of the study), the live birth index was statistically decreased compared to controls (83% versus 94%).

Critical effects observed:

no

Reproductive effects observed:

no

Conclusions:

Under these experimental conditions, , the NOAEL for the dams and their offspring was determined to be 125 mg/kg/day.

Executive summary:

The objective of this study was to evaluate the potential reproductive/developmental toxicity of dioctadecyldimethylammonium chloride (96.8% active) according to a GLP conform Reproduction/Developmental Toxicity Screening Test (OECD Guideline 421).

Groups of 10 rats (CRL:CD (SD) BR) per sex were treated with dosages of 0, 62.5, 125, and 500 mg/kg bw/day by gavage (administration volume 10 ml/kg bw/day) using corn oil as a vehicle for the control group. Males were treated daily from two weeks before mating, during mating and until a dosing period of a total of 28 days had been completed. Females were treated daily from two weeks before mating until the 4 th day of lactation. Subsequently these females weresacrificedwith their pups.

At daily doses of 500 mg/kg bw one male and one female died after 12 and 10 treatments respectively. Clinical observations revealed dyspnea, soft stools in all females and almost all males. Half of the females also showed slight to moderate dilation of the abdomen. Body weight loss of about 14 to 15 g was observed in both sexes during the first week of treatment. Further, statistically significantly lower mean daily food consumption was observed in the males during the premating period and in the dams during the first week of pregnancy. Statistically significantly lower mean dam body weights were observed after 14 and 20 days of gestation and at the day of birth after delivery.

No toxicologically relevant effects were observed at dosages of 125 and 62.5 mg/kg bw/day.

At sacrifice of the parental animals no significant differences were found in the organ weights of uterus, ovaries, testes and epididymides. Histopathology of testes, epididymides and of the ovaries of the animals of the 500 mg/kg dose groups did not show any compound related changes. No substance related changes were reported for the evaluation of testicular stages of spermatogenesis performed in the PAS-hematoxylin stained sections.

At the dosages of 62.5 and 125 mg/kg/day all of the 10 paired females revealed to be sperm positive after mating, all revealed to be pregnant and all delivered live litters. The numbers of corpora lutea had not been evaluated during this study.

At 500 mg/kg/day, from the 9 paired females 7 revealed to be sperm positive (77 %) after mating, 6 out of 9 (67 %) revealed to be pregnant, and 5 out of 6 (83 %) delivered live litters. One animal revealed to have fully resorbed. Mean pre-coital time was longer in this group (about 6.1 days) when compared to the controls and the lower dosage groups (1.5 to 2.1 days).

After birth, for the animals treated with 62.5 or 125 mg/kg/day there were no substance related biological differences in their pregnancy outcome in comparison to the control group. At 500 mg/kg/day, the percentage of post implantation losses was increased by 19% per litter in comparison to about 6% per litter in the controls and in the lower treatment groups, thus resulting in a statistically significantly lower rate of live borns of 83% in comparison to 94% in the controls and in the lower treatment groups. Viability index on postnatal day 4 was in the range of the controls and the lower treatment groups.

For all dose groups under investigation no statistically significant differences were found for the body weights of male and female pups at birth and on postnatal day 4. External abnormalities have not been reported.

Based on the findings of this study, it was concluded that the No-observed-adverse-effect level (NOAEL) regarding reproductive toxicity was 125 mg/kg bw/day.

Effect on fertility: via oral route

Dose descriptor:

NOAEL

125 mg/kg bw/day

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

No reproductive studies with the test substance are available. Therefore, reproductive/developmental screening study with DODMAC (CAS 107-64-2) was used as read across to fulfil the data gap for repeated dose toxicity. The primary component of all substances provide complete coverage of 68334-33-8.  68334-33-8 shares high structural similarity with 61789-80-8, 68783-78-8, 107-64-2, and 112-02-7. As 68334-33-8 is a UVCB its components encapsulate the other substances except for the counter ion (Cl-). In solution, the counter ions will dissociate from the parent structures.  Therefore, we are comparing substances of equivalent nature. CAS 107-64-2 represents the C18 boundary of the 61789-80-8. Ignoring the salt component CAS 61789-80-8 is equivalent to CAS 68334-33-8. 68783-78-8 is a worst case of both 68334-33-8 and 61789-80-8 since it is unsaturated and the SP2 carbon-carbon bonds are of higher reactivity and a more likely site of metabolic activation. The primary component of CAS 112-02-7 is a substructure of all the other substances. Additional documentation, provided within the IUCLID Assessment Reports section, supports the read-across approach.

In a reproductive/developmental toxicity screening study according to OECD Guideline 421 following the principles of Good Laboratory practices, groups of 10 males and 10 female rats received daily doses of 0, 62.5, 125 and 500 mg/kg bw/day by gavage. Males were treated daily from two weeks before mating, during mating and until a dosing period of a total of 28 days had been completed. Females were treated daily from two weeks before mating until the 4th day of lactation.

 

At daily doses of 500 mg/kg bw one male and one female died after 12 and 10 treatments respectively. Clinical observations revealed dyspnoea, soft stools in all females and almost all males. Half of the females also showed slight to moderate dilation of the abdomen. Body weight loss was observed in both sexes during the first week of treatment. Further, statistically significantly lower mean daily food consumption was observed in the males during the premating period and in the dams during the first week of pregnancy. Statistically significantly lower mean dam body weights were observed after 14 and 20 days of gestation and at the day of birth after delivery. No toxicologically relevant effects were observed at dosages of 125 and 62.5 mg/kg bw/day.

 

At sacrifice of the parental animals, no significant differences were found in the organ weights of uterus, ovaries, testes and epididymides. Histopathology of testes, epididymides and of the ovaries of the animals of the 500 mg/kg dose groups did not show any compound related changes. No substance related changes were reported for the evaluation of testicular stages of spermatogenesis performed in the PAS-haematoxylin stained sections.

 

At the dosages of 62.5 and 125 mg/kg/day all of the 10 paired females revealed to be sperm positive after mating, all revealed to be pregnant and all delivered live litters. The numbers of corpora lutea had not been evaluated during this study. At 500 mg/kg/day, from the 9 paired females 7 revealed to be sperm positive (77 %) after mating, 6 out of 9 (67 %) revealed to be pregnant, and 5 out of 6 (83%) delivered live litters. One animal revealed to have fully resorbed. Mean pre-coital time was longer in this group (about 6.1 days) when compared to the controls and the lower dosage groups (1.5 to 2.1 days).

 

After birth, for the animals treated with 62.5 or 125 mg/kg/day there were no substance related biological differences in their pregnancy outcome in comparison to the control group. At 500 mg/kg/day, the percentage of post implantation losses was increased by 19% per litter in comparison to about 6% per litter in the controls and in the lower treatment groups, thus resulting in a statistically significantly lower rate of live born of 83% in comparison to 94% in the controls and in the lower treatment groups. However, the viability index on postnatal day 4 was in the range of the controls and the lower treatment groups.

 

For all dose groups under investigation no statistically significant differences were found for the body weights of male and female pups at birth and on postnatal day 4. External abnormalities have not been reported. The No-observed-adverse-effect level (NOAEL) regarding reproductive toxicity was established at 125 mg/kg bw/day and adopted for an existing EU risk assessment on this compound as well. Additionally, from oral repeated dose toxicity studies performed with dioctadecyldimethylammonium chloride or dihydrogenatedtallowdimethylammonium chloride, no indications with regard to histopathological changes of reproductive organs exist.

Effects on developmental toxicity

Description of key information

Oral: NOAEL; OECD 414, rat. NOAEL for teratogenicity was≥500 mg/kg/day, the highest dose tested. Reliability = 2 (read-across substance).

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Type of information:

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

Adequacy of study:

supporting study

Study period:

1975

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

Study predated formal guideline requirements and GLP but performed according to existing scientific standards. Reliabilty check by Expert Panel (American Chemistry Council Fatty Nitrogen Derivatives) passed.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

yes

Remarks:

modifications in exposure period

GLP compliance:

no

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

no data

Route of administration:

other: oral gavage; oral diet

Vehicle:

not specified

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

days 6 - 13 or 6 - 18 of gestation

Frequency of treatment:

daily

Duration of test:

days 0 - 21 of gestation

Dose / conc.:

100 mg/kg bw/day (actual dose received)

Remarks:

oral gavage

Dose / conc.:

500 mg/kg bw/day (actual dose received)

Remarks:

oral gavage

Dose / conc.:

508 mg/kg bw/day (actual dose received)

Remarks:

0.65% active in diet; oral diet

No. of animals per sex per dose:

25 rats per group (10 animals sacrified after 13 days, 15 animals after 21 days of gestation)

Control animals:

yes, concurrent vehicle

yes, plain diet

Details on study design:

Two concurrent control groups: One receiving the gavage vehicle isopropanol, one receiving control feed only

Dose descriptor:

NOAEL

Remarks:

via gavage

Effect level:

> 500 mg/kg bw/day (nominal)

Basis for effect level:

other: highest dose tested

Dose descriptor:

NOAEL

Remarks:

via diet

Remarks on result:

not determinable

Remarks:

slight effects on body weight gain

Abnormalities:

no effects observed

Dose descriptor:

NOAEL

Remarks:

via gavage

Effect level:

> 500 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: highest dose tested

Dose descriptor:

NOAEL

Remarks:

via diet

Effect level:

> 508 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: highest dose tested

Abnormalities:

no effects observed

Developmental effects observed:

no

Conclusions:

Based on the results from this study ditallowdimethylammonium chloride is not considered to be a developmental toxicant.

Executive summary:

Ditallowdimethylammonium chlorid (DTDMAC)was investigated for developmental toxicity in the rat. 25 female rats per group were administered the test substance either by oral gavage at dose levels of 100 and 500 mg active ingredient per kg body weight per day (vehicle was 15% isopropanol) or in the diet at a dose level of 0.65% active ingredient beginning on day 6 of gestation. Two control groups were run concurrently. One received the gavage vehicle and the other received control feed only.

Ten rats per group were sacrificed after the day 13 treatment and 15 rats per group were treated through day 18 and sacrificed on day 21 of gestation. Body weights were taken on days 0, 6, 9, 12, 15, 18 and 21 of gestation. Food consumption was evaluated for days 12 and 18 of gestation. Clinical observations were made daily for signs of pharmacologic or toxicologic effect and mortality. Gross necropsies were conducted on all surviving rats, moribund rats and rats that died spontaneously. At necropsy, for rats sacrificed on day 13 of gestation, the uterus (number and location for each horn of resoptions, embryos and implantation sites) and ovaries (number of corpora lutea of pregnancy per ovary) were observed. At necropsy for rats sacrificed on day 21 of gestation, the uterus (number and location for each horn of live fetuses, dead fetuses, early and late resorptions and impantation sites) and ovaries ( number of corpora lutea of pregnancy per ovary) were observed. The necropsy for all maternal rats also included observations for obvious abnormalities and the following tissues were examined: heart, lung, stomach, liver, pancreas, spleen, mesenteric lymph nodes, jejunum, kidney, adrenal, bladder and ovary.

With regard to maternal data, no adverse effects attributable to DTDMAC administration were noted in comparison of pregnancy and mortality rates. Depressed body weight gains during gestation were noted in the group that received the test substance in the diet together with less food consumption values compared to the group that received control feed. Early deliveries and abortions, necropsy findings, and reproduction data were considered not to be affected by treatment with the test substance. An increase in resorptions was observed for the 100 mg/kg/day gavage group compared to the isopropanol control group (7.1% vs 2.1%). This difference was not considered a treatment-related effect due to the lack of dose response and a low value for the control group compared to historical data from the laboratory.

With regard to fetal data, no differences considered to be related to the adminsitration of ditallowdimethylammonium chloride were noted in fetal size and sex, variations in degree of ossification or malformations.

Based on the results of this study, the NOAEL for maternal toxicity was considered to be greater 500 mg/kg body weight per day by gavage. No NOAEL for maternal toxicity was established for dietary treatment because of the slight effects on body weight gain. With regard to developmental toxicity the NOAEL was established to be greater 500 mg/kg body weight per day by gavage and greater 508 mg/kg body weight per day via the diet.

Effect on developmental toxicity: via oral route

Dose descriptor:

NOAEL

500 mg/kg bw/day

Species:

rat

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

No developmental toxicity studies with the test substance are available. Therefore, a developmental screening study with DTDMAC was used for classification purposes.

Ditallowdimethylammonium chloride (DTDMAC) was investigated for developmental toxicity in the rat. 25 female rats per group were administered the test substance either by oral gavage at dose levels of 100 and 500 mg active ingredient per kg body weight per day (vehicle was 15% isopropanol) or in the diet at a dose level of 0.65% active ingredient beginning on day 6 of gestation. Two control groups were run concurrently. One received the gavage vehicle and the other received control feed only.

Ten rats per group were sacrificed after the day 13 treatment and 15 rats per group were treated through day 18 and sacrificed on day 21 of gestation. Body weights were taken on days 0, 6, 9, 12, 15, 18 and 21 of gestation. Food consumption was evaluated for days 12 and 18 of gestation. Clinical observations were made daily for signs of pharmacologic or toxicologic effect and mortality. Gross necropsies were conducted on all surviving rats, moribund rats and rats that died spontaneously. At necropsy, for rats sacrificed on day 13 of gestation, the uterus (number and location for each horn of resorptions, embryos and implantation sites) and ovaries (number of corpora lutea of pregnancy per ovary) were observed. At necropsy for rats sacrificed on day 21 of gestation, the uterus (number and location for each horn of live foetuses, dead foetuses, early and late resorptions and implantation sites) and ovaries (number of corpora lutea of pregnancy per ovary) were observed. The necropsy for all maternal rats also included observations for obvious abnormalities and the following tissues were examined: heart, lung, stomach, liver, pancreas, spleen, mesenteric lymph nodes, jejunum, kidney, adrenal, bladder and ovary.

 

With regard to maternal data, no adverse effects attributable to DTDMAC administration were noted in comparison of pregnancy and mortality rates. Depressed body weight gains during gestation were noted in the group that received the test substance in the diet together with less food consumption values compared to the group that received control feed. Early deliveries and abortions, necropsy findings, and reproduction data were considered not to be affected by treatment with the test substance. An increase in resorptions was observed for the 100 mg/kg/day gavage group compared to the isopropanol control group (7.1% vs 2.1%). This difference was not considered a treatment-related effect due to the lack of dose response and a low value for the control group compared to historical data from the laboratory.

 

With regard to foetal data, no differences considered to be related to the administration of ditallowdimethylammonium chloride were noted in foetal size and sex, variations in degree of ossification or malformations.

 

Based on the results of this study, the NOAEL for maternal toxicity was considered to be greater 500 mg/kg body weight per day by gavage. No NOAEL for maternal toxicity was established for dietary treatment because of the slight effects on body weight gain. With regard to developmental toxicity the NOAEL was established to be greater 500 mg/kg body weight per day by gavage and greater 508 mg/kg body weight.

Justification for classification or non-classification

Based on the available data for the read-across substance, the test substance does not need to be classified for reproductive toxicity according to EU Classification and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.