Fatty acids, rape-oil, hydrogenated, reaction products with diethylenetriamine, di-Me sulfate-quaternized

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

No studies available.

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

the study does not need to be conducted because a pre-natal developmental toxicity study is available

Reason / purpose for cross-reference:

data waiving: supporting information

Effect on fertility: via oral route

Endpoint conclusion:

no study available

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Effects on developmental toxicity

Description of key information

Based on the results of the read across pre-natal study, the test substance, 'di-C18-22 AAEMIM-MS', is not expected to be a development toxicant.

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Type of information:

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

Adequacy of study:

key study

Study period:

From May 13, 1991 to April 23, 1992

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

KL2 due to RA study

Justification for type of information:

Refer to section 13 of IUCLID for details on the read-across justification. The study with the read across substance is considered sufficient to fulfil the information requirements as further explained in the provided endpoint summary.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

EPA OPP 83-3 (Prenatal Developmental Toxicity Study)

Deviations:

no

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

Test animals
- Age: Males approx. 63 d; Females: approx. 56 d
- Weight: Males 265 – 320 g upon arrival; Females 172 – 200 g upon arrival
- Fasting period before study: no data given
- Housing: Before mating males were housed singly, females were housed two to a cage.
- During mating 1 male and 1 female were housed individually.
- After mating copulation plug positive females and therefore considered successfully mated
- were housed singly for the duration of the study in stainless steel wire mesh cages.
- Diet: Access ad libitum; Ground Certified Rodent Chow ® (#5002, Ralston Purina Co., St. Louis, MO).
- Water: ad libitum, municipal tap water
- Acclimation period: approx. 2 wks
Environmental Conditions
- Temperature (°C): 19 – 25
- Humidity (%): relative 40 – 70
- Photoperiod: 12 h dark/12 h light; light period 5 – 17 h

In-Life dates: From May 27-30, 1991 to June 14 (gd 15); May 28 – 30, 1991 was gd0.
- Treatment began on gd6 (June 3-5, 1991). Dams were sacrificed on gd21 (June 18 – 20, 1991)

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on exposure:

Preparation of dosing solutions:
Control: Milli-Q ® water, volume according to highest test substance volume selected (1,42 mlL/ kg bw/day)
Dose levels: The dose levels referring to the active ingredient were 100, 300, and 1000 mg/kg bw/day administered undiluted once a day, based on the dam's body weight on gestational day 6, the percent active ingredient (76,6%) and the specific gravity of the test substance (0,92 g/cm3).
Dosage volumes were not changed to account for increases in gestational body weight after gd6. Accordingly the applied test substance volumes were 0,142, 0,426, and 1,42 mg/kg bw/day

Analytical verification of doses or concentrations:

no

Details on analytical verification of doses or concentrations:

Not applicable

Details on mating procedure:

Proof of pregnancy: Females were checked once daily for vaginal copulation plugs in the morning and the paperboard beneath the cages was checked twice daily for dropped copulation plugs. The observation of a vaginal or dropped copulation plug was considered evidence of successful mating. Each male was paired only once in this study.

Duration of treatment / exposure:

10 d, from gd6 through gd15

Frequency of treatment:

Once daily

Duration of test:

22 days (till gd21)

Dose / conc.:

0 mg/kg bw/day

Remarks:

based on active ingredient (a.i.)

Dose / conc.:

100 mg/kg bw/day

Remarks:

based on active ingredient (a.i)

Dose / conc.:

300 mg/kg bw/day

Remarks:

based on active ingredient (a.i)

Dose / conc.:

1 000 mg/kg bw/day

Remarks:

based on active ingredient (a.i)

No. of animals per sex per dose:

25

Evaluated pregnant rats: group 1-4: 21/23/22/25 (females with viable fetuses)

Animals evaluated for maternal toxicity: group 1-4: 22/23/24/25

Control animals:

yes, concurrent vehicle

Details on study design:

Dose selection rationale: Based on a former dose range finding study
Rationale for animal assignment (if not random): The rat is a commonly used rodent species for embryotoxic studies.
Group size per dose level: 22/23/23/25 pregnant animals per dose group

Maternal examinations:

Cage side observations: yes
Time schedule: daily
Detailed clinical observations: Yes
Time schedule: daily
Clinical signs: Yes
daily observations for behavior, external appearance and general condition
Viability: Yes
daily checks twice daily for morbidity and mortality
Body weight: yes
Time schedule for examinations: Maternal body weights were taken on gd0, gd6 (prior to the onset of dosing), gd9, 12, 15, 18, and 21
Food consumption: yes
Food consumption was measured at three-day intervals throughout the study (gd0 through 21).
Water consumption: no
Post-mortem examinations: Yes
Organs examined: Macroscopic examination of uteri, ovaries, cervix, vagina, peritoneal and thoracic cavities.
Maternal liver and Gravid Uteri weights, Number of Corpora lutea, Number and status of Implantation sites.

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
Gravid Uterus weight: yes
Number of corpora lutea: yes
Number of implantations: yes
Number of early resorptions: yes
Number of late resorptions: yes

Fetal examinations:

Fetal examinations
External examinations: yes: all litter
Soft tissue examinations: yes; approx. 50%
Skeletal examinations: yes, 50%
Head examinations: yes
other: Number of fetuses (live and dead), sex and viability of fetuses, variations and malformations, including cleft palate

Statistics:

Levene's test for equality of variances, analysis of variance (ANOVA) and t-tests. The t-tests were used when the F value from the ANOVA was significant. When Levene's test indicated equal variances, and the ANOVA was significant, a pooled t-test was used for pair wise comparisons. When Levene's test indicated heterogeneous variances, all groups were compared by an ANOVA for unequal variances followed, when necessary, by a separate variance t-test for pair wise comparisons. Nonparametric data were statistically evaluated using the Kruskal-Wallis test, followed by the Mann-Whiney U test when appropriate. Incidence data were compared using the Fisher's Exact Test. With the exception of analyses for fetal malformation and variation data, all statistical evaluations were performed using BMDP Statistical Software (Dixon, 1990). For all statistical tests, the probability value of <0,05 (two-tailed) was used as the critical level of significance.

Details on maternal toxic effects:

Maternal toxic effects: no effects
Details on maternal toxic effects: No substance related maternal toxicity detected up to the highest dose tested.

Key result

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day

Basis for effect level:

other: maternal toxicity

Details on embryotoxic / teratogenic effects:

No embryotoxicity or teratogenicity detected up to the highest dose tested

Key result

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day

Based on:

other:

Basis for effect level:

other: embryotoxicity

Key result

Abnormalities:

no effects observed

Key result

Developmental effects observed:

no

TOXIC RESPONSE/EFFECTS BY DOSE LEVEL:

Mortality:

300 mg/kg bw/day dose group: One female became moribund and was sacrificed on gd10.

Clinical signs:

300 mg/kg bw/day dose group: 2 dams exhibited audible respiration during or subsequent to the treatment period.

One dam exhibited swelling in the face, urogenital area wetness, gasping and perinasal and perioral encrustation. This animal was sacrificed due to her moribund condition on gd10. None of these signs were considered to be test substance related due to their absence in the dose range finding study up to doses of 1875 mg/kg bw/day and the lack of any dose relation in the current study.

1000 mg/kg bw/day dose group: 3 dams exhibited audible respiration during or subsequent to the treatment period.

No treatment-related differences in gestational parameters including total number of implantations, number of viable and nonviable implants in any dose group.

Body weight: No treatment-related effects on gestational body weights and body weight gain, corrected body weight, corrected body weight gain, absolute and relative liver weight, and gravid uterine weight . Fetal body weights per litter were not affected by treatment.

Increased food consumption in the 100 mg/kg bw/day dose group was not considered to be related to treatment due to the lack of a dose-relationship.

NECROPSY FINDINGS IN DAMS AT TERMINATION:

One female in the 300 mg/kg bw/day dose group which was sacrificed due to her moribund condition, had gas-filled intestines, mucoid fluid or material in the nose turbinates, abnormal material in the trachea and discolored and consolidated lungs. One further animal in this dose group had no implants in one uterine horn.

Two females in the 1000 mg/kg bw/day dose group contained blood in the uterus (detected with Hemastix reagent strips) One further female had discolored lungs (dark red) in all lobes.

REPRODUCTION DATA OF DAMS (0, 100, 300, 1000 mg/kg bw):

-         Corpora lutea:338 (16.1 per dam), 380 (16.5 per dam), 364 (15.7 per dam), 387 (15.5 perdam)

-         Implantation sites (Total implants): 340 (15.5 per dam), 379 (16.5 per dam)*, 363 (15.7 perdam)**, 382 (15.3 per dam)

-         Resorptions: 10 (0.5 per dam), 7 (0.4 per dam), 12 (0.5 per dam), 53 (2.7 per dam)**

-         Early resorptions:13 (0.6 per dam), 21 (0.9 per dam), 17 (0.7 per dam), 14 (0.6 per dam)

-         Late resorptions:0 (0 per dam), 1 (0 per dam), 2 (0.1 per dam), 0 (0 per dam)

-         Live (Viable) fetuses:327 (14.9 per dam), 356 (15.5 per dam), 343 (14.3 per dam),367 (14.7 per dam)

-         Malformations (external, skeletal, soft tissue) (fetal incidence): none significantly different from Control

-         Skeletal variations (fetal incidence):

-         Findings: Anterior arch of atlas poorly ossified; Statistical significance of affected litters in 1000mg/kg bw/day dose group;

% affected litters (71.4, 87.0, 86.4 96.0*)

-         - Skeletal variations (fetal incidence):

-         Findings: Majority of forelimbs unossified; Statistical significance of affected litters in 300mg/kg bw/day dose group;

% affected litters (38.1, 26.1, 4.5**, 24.0)

-         - Soft Tissue variations (fetal incidence):

-         Findings: Excessive bleeding at umbilicus; Statistical significance of affected litters in100 mg/kg bw/day dose group;

% affected litters (23.8, 0*, 27.3, 40.0)

-         Other External variations (fetal incidence): none significantly different from control

Weight of fetuses: No treatment related effects on fetal body weights (all fetuses, male or female) observed in any group. The statistically significant* differences in mean male and female body weights in litters at 100 mg/kg bw/day were not considered to be treatment-related due to the lack of a dose-response relationship.

* Significantly different from control, p= 0.05 using two-tailed Fisher's exact test

** Significantly different from control, p= 0.01 using two-tailed Fisher's exact test

Conclusions:

Based on the results of the read across study, the test substance maternal, emryotoxic and teratogenic toxicity NOAELs of the test substance were considered to be ≥1000 mg a.i./kg bw/day.

Executive summary:

A study was conducted to determine the teratogenicity of the read across substance, ‘di-C16-18 and C18-unsatd. AAEMIM-MS' (active: 75%), according to EPA OPP 83 -3 Guideline, in compliance with GLP. During Gestation Day 6-15, four groups of 25 female Sprague CD rats/dose were dosed gavage at dose levels of 0, 100, 300, and 1000 mg/kg bw/day referring to 100% active substance, based on range finding study results conducted at doses up to 1875 mg/kg bw/day. Animals were sacrificed on Day 22. Examinations included daily observations of behaviour and clinical symptoms, daily food consumption, and body weight every three days. Upon sacrifice, number of corpora lutea, implantations and resorptions were examined. In the main study, no signs of maternal toxicity were observed, except one female in the 300 mg/kg bw/day group become moribund and was sacrificed on gestation Day 10. Also no foetal mortality was observed. Numbers of corpora lutea and resorptions were comparable. At 300 mg/kg bw/day and 1000 mg/kg bw/day dose groups 2 and 3 dams exhibited audible respiration during or subsequent to the treatment period. No treatment-related differences in gestational parameters including total number of implantations, number of viable and nonviable implants in any dose group. No treatment-related effects on gestational body weights and body weight gain, corrected body weight, corrected body weight gain, absolute and relative liver weight and gravid uterine weight were observed. Foetal body weights per litter were also not affected by treatment. Increased food consumption in the 100 mg/kg bw/day dose group was not considered to be related to treatment due to the lack of a dose-relationship. The animal in the 300 mg/kg bw/day dose group which was sacrificed due to moribund condition, had gas-filled intestines, mucoid fluid or material in the nose turbinates, abnormal material in the trachea and discoloured and consolidated lungs. One further animal in this dose group had no implants in one uterine horn. Two females in the 1000 mg/kg bw/day dose group contained blood in the uterus (detected with Hemastix reagent strips) another female had discoloured lungs (dark red) in all lobes. However, no statistically significant maternal toxic or embryotoxic/teratogenic effects were seen up to highest tested dose of 1000 mg/kg bw/day. Under the study conditions, the NOAEL for maternal and embryotoxic effects/teratogenicty in rats was established at 1000 mg/kg bw/day (Neeper-Bradley, 1992). Based on the results of the read across study, similar absence of toxicity and NOAELs can be expected for the test substance, ‘di-C18-22 AAEMIM-MS’.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

Guideline compliant study

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 to determine the teratogenicity of the read across substance, ‘di-C16-18 and C18-unsatd. AAEMIM-MS' (active: 75%), according to EPA OPP 83 -3 Guideline, in compliance with GLP. During Gestation Day 6-15, four groups of 25 female Sprague CD rats/dose were dosed gavage at dose levels of 0, 100, 300, and 1000 mg/kg bw/day referring to 100% active substance, based on range finding study results conducted at doses up to 1875 mg/kg bw/day. Animals were sacrificed on Day 22. Examinations included daily observations of behaviour and clinical symptoms, daily food consumption, and body weight every three days. Upon sacrifice, number of corpora lutea, implantations and resorptions were examined. In the main study, no signs of maternal toxicity were observed, except one female in the 300 mg/kg bw/day group become moribund and was sacrificed on gestation Day 10. Also no foetal mortality was observed. Numbers of corpora lutea and resorptions were comparable. At 300 mg/kg bw/day and 1000 mg/kg bw/day dose groups 2 and 3 dams exhibited audible respiration during or subsequent to the treatment period. No treatment-related differences in gestational parameters including total number of implantations, number of viable and nonviable implants in any dose group. No treatment-related effects on gestational body weights and body weight gain, corrected body weight, corrected body weight gain, absolute and relative liver weight and gravid uterine weight were observed. Foetal body weights per litter were also not affected by treatment. Increased food consumption in the 100 mg/kg bw/day dose group was not considered to be related to treatment due to the lack of a dose-relationship. The animal in the 300 mg/kg bw/day dose group which was sacrificed due to moribund condition, had gas-filled intestines, mucoid fluid or material in the nose turbinates, abnormal material in the trachea and discoloured and consolidated lungs. One further animal in this dose group had no implants in one uterine horn. Two females in the 1000 mg/kg bw/day dose group contained blood in the uterus (detected with Hemastix reagent strips) another female had discoloured lungs (dark red) in all lobes. However, no statistically significant maternal toxic or embryotoxic/teratogenic effects were seen up to highest tested dose of 1000 mg/kg bw/day. Under the study conditions, the NOAEL for maternal and embryotoxic effects/teratogenicty in rats was established at 1000 mg/kg bw/day (Neeper Bradley, 1992). Based on the results of the read across study, similar absence of toxicity and NOAELs can be expected for the test substance, ‘di-C18-22 AAEMIM-MS’.

Justification for classification or non-classification

Based on the results of the read across study, the test substance, 'di-C18-22 AAEMIM-MS', is not expected to be a development toxicant. Therefore, no classification is warranted according to the EU CLP criteria (Regulation 1272/2008/EC).​