Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Link to relevant study records
Reference
Endpoint:
toxicity to reproduction
Remarks:
other: Repeated Dose 90-Day Oral Toxicity in Rodents
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Study period:
1992-10-30 to 1993-02-05
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
other: OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
other: Crl CD BR
Sex:
male/female
Route of administration:
oral: gavage
Vehicle:
water
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
Daily, 7 days/week, for 13 weeks
Remarks:
Doses / Concentrations:
1, 10, 500 mg/kg bw/d
Basis:
actual ingested
No. of animals per sex per dose:
15
Control animals:
yes, concurrent vehicle
Dose descriptor:
NOAEL
Effect level:
500 mg/kg bw/day (actual dose received)
Sex:
male/female
Basis for effect level:
other: There were no significant adverse test substance-related effects on reproductive organs (organ weights of ovary and testis and histopathology of gonads).
Reproductive effects observed:
not specified
Conclusions:
The results from the evaluation of reproductive organs, especially organ weights of ovary and testis and histopathology of gonads from this 91-day repeated dose study revealed no indications of any substance-related effects up to and including the highest test dose of 500 mg/kg bw/day.
Executive summary:

In a subchronic toxicity study comparable to OECD guideline 408, MDEA-Esterquat C16-18 and C18 unsatd.  (10 % a.i.) was administered to 15 Charles River CD rats / sex/ dose by gavage at dose levels of 1, 10 and 500 mg/kg bw/ day for a period of 13- weeks. One control group received the vehicle, deionized water, and a second control group received pH-adjusted, deionized water (pH 2.5). The regimen for both control groups was identical to treatment groups.

The following parameters were monitored during the study: clinical observations (detailed, weekly; mortality, morbidity, and overt signs of toxicity, twice a day); body weights (weekly); food consumption (weekly); clinical pathology (haematology, blood chemistry, and urinalysis; at termination); opothalmoscopic examinations (once pre-test and prior to sacrifice); macroscopic pathologic examination; absolute and relative organ weights; microscopic pathology.

The following reproductive organs were included in the examinations: absolute and relative organ weigths of ovary and testis and histopathology of gonads, mammary gland (females only), prostate and seminal vesicle, uterus with cervix and vagina.

There were no changes in any of these parameters that were considered to be toxicologically significant or test-substance related.

The no effect level (NOEL) for this study is the high dose level of 500 mg/kg bw/day of the test article.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
500 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

For the assessment of effects of MDEA Esterquat C18 satd. to fertility, a 28 day oral toxicity study with the target substance as well as a subacute toxicity study and a subchronic toxicity study conducted with the structurally similar source substance MDEA-Esterquat C16-18 and C18 unsatd. are available.

In a Repeated Dose, 28-day Oral Toxicity Study performed according to the First Addendum to OECD Guidelines for Testing of Chemicals, Section 4, No. 407, "Repeated Dose 28-day Oral Toxicity Study in Rodents" adopted 03 October 2008 and Directive 96/54 EEC B.7., the test item MDEA Esterquat C18 satd.suspended in Aqua ad injectionem (10 % w/w) as vehicle was orally administered in graduated doses to three groups of male and female rats (HsdRccHan: WIST) by gavage.

The main study included 4 groups (control, 62.5, 250, 1000 mg/kg bw) with each 5 male and 5 female animals and the recovery study included 2 groups (control, 1000 mg/kg bw) each with 5 male and 5 female animals. No test substance-related findings were detected or observed in clinical examinations, body weights, food consumption values, haematology, urinalysis, neurobehavior, clinical biochemistry or gross pathology evaluations, including organ weights of ovary, uterus with cervix, testis, epididymides, prostate with seminal vesicles and coagulating glands and the gross pathology and the histopathology of the reproductive organs (gonads, prostate and seminal vesicle, epididymides, uterus and vagina). The NOEL fertility is therefore ≥ 1000 mg/kg bw/day in this study.

In the repeated dose toxicity studies with the source substance MDEA-Esterquat C16-18 and C18 unsatd. , Charles River CD rats (Sprague-Dawley) were treated with the test substance by oral gavage for 28 days (25 animals/sex/dose) or for 91 days (15 animals/sex/dose) at dose levels of 0, 1, 10, or 500 mg/kg bw/day. In both studies, no adverse test substance-related effects were found for any parameter measured, especially regarding organ weights of ovary and testes and histopathology of gonads, uterus, mammary gland, prostate and seminal vesicle.

Further testing for effects concerning fertility is considered not to be necessary because up to 1000 mg/kg bw/day there were no indications for substance-related effects in general and especially regarding organ weights (ovary and testis) and histopathology of gonads from the repeated dose toxicity studies. In addition this is substantiated by the absence of effects on maternal reproduction, embryo lethality or embryotoxicity in the developmental toxicity study in rats performed with the source substance MDEA-Esterquat C16-18 and C18 unsatd. with doses of up to 1000 mg/kg bw/day.

The DNELs fertility have been derived from results of the sub-chronic repeated dose toxicity study, taking full account of the potential increased uncertainty resulting fromlower sensitivity and the limited scope of the repeated dose toxicity studies for detecting effects on reproductive organs by applying an additional assessment factor. These derived DNELs are therefore relevant and appropriate for risk assessment purposes. In addition the results of exposure and risk assessments covering all relevant exposures throughout the life cycle of the substance demonstrate a low exposure and a RCR value below 1 in all scenarios of manufacture, formulation, professional use and indirect exposure of humans via the environment (see Chapter 9 and 10 of the CSR). 

There are no data gaps for effects on fertility. No human data are available. However, there is no reason to believe that these results from rat would not be applicable to humans.

Justification for read-across

For details on substance identity, toxicokinetics and detailed toxicological profiles, please refer also to the general justification for read-across given in chapter 5 of the CSR and attached as pdf document to section 7 of the IUCLID file.

 

a. Structural similarity and functional groups

The target substance, MDEA Esterquat C18 satd., consists of an amine backbone (MDEA = Methyldiethanol amine) esterified with the long chain fatty acid stearic acid (C18 satd.; IV (iodine value) < 1). The main reaction product is the dialkylester compound, next to that small amounts of the monoalkylester may be formed. The amine function is quaternised with two methyl groups. The counter ion is Chloride.

The source substance, MDEA EsterquatC16-18 and C18 unsatd., consists of the same amine backbone (MDEA = Methyldiethanol amine) but esterified with a mixture of the long chain fatty acids C16, C18 and C18 unsaturated (IV < 25). The main reaction product is the dialkylester compound, next to that small amounts of the monoalkylester may be formed. The amine function is quaternised with two methyl groups. The counter ion is Chloride.

The source and the target substance share structural similarities with common functional groups (quaternary amines, esters, and fatty acid chains with comparable length and degree of saturation).

b. Common breakdown products

The metabolism expected to occur is hydrolysis of the ester-bond by esterases. However, the rate of hydrolysis is assumed to be low. The fraction of metabolised molecules would result in free fatty acids and Dimethyl-DEA (DEA = Diethanolamine). The carboxylic acids are further degraded by the mitochondrial beta-oxidation process (for details see common text books on biochemistry). The fatty acids enter normal metabolic pathways and are therefore indistinguishable from fatty acids from other sources including diet. The quaternary ammonium ions are not expected to be further metabolised, but excreted unchanged via the urine. 

c. Differences

The differences in fatty acid chain length (higher percentage of C16 in the source substance MDEA Esterquat C16 -18 and C18 unsatd.) and degree of saturation (higher degree of unsaturation of the C18 fatty acid chains in the source substance substance MDEA Esterquat C16-18 and C18 unsatd.) could be relevant for local toxicity (skin and eye irritation) but arebut are not considered to be of relevance for reproductive toxicity.

Comparison of molecular structures and toxicity data of MDEA Esterquat C18 satd. and MDEA-Esterquat C16-18 and C18 unsatd.

 

 

Source substances

Target substance

 

Endpoints

 

MDEA-Esterquat C16-18 and C18 unsatd.

 

MDEA Esterquat C18 satd.

Repeated Dose Toxicity/Fertility

equivalent to OECD 407 (oral 4 week study)

 

25 rats/dose/sex in the groups treated with 1,10 and 500 mg/kg/d

 

 

The test substance did not produce any test related toxicological responses up to levels of 500 mg/kg bw/day. The results from the evaluation of reproductive organs, especially organ weights of ovary and testis and histopathology of gonads revealed no indications of any substance-related effects up to and including the highest test dose of 500 mg/kg bw/day.

 

 

 

NOAEL = 500 mg/kg bw/day(general systemic toxicity/fertility)

according to OECD 407 (oral 4 week study, including 14-day recovery period)

 

5 rats/dose/sex in the groups treated with 62.5, 250 and 1000 mg/kg/d

 

No test substance-related findings up to the highest tested dose. The results from the evaluation of reproductive organs (Organ weights of ovary, uterus with cervix, testis, epididymides, prostate with seminal vesicles and coagulating glands and the gross pathology and the histopathology of the reproductive organs (gonads, prostate and seminal vesicle, epididymides, uterus and vagina)

 

NOAEL = 1000 mg/kg bw/day(general systemic toxicity/fertility)

equivalent to OECD 408 (oral 13 week study)

 

15 rats/dose/sex in the groups treated with 1,10 and 500 mg/kg/d

 

No biologically significant adverse effects were observed in any test group. The results from the evaluation of reproductive organs, especially organ weights of ovary and testis and histopathology of gonads from this 91-day repeated dose study revealed no indications of any substance-related effects up to and including the highest test dose of 500 mg/kg bw/day.

 

NOAEL = 500 mg/kg bw/day(general systemic toxicity/fertility)

 

 

Developmental Toxicity

according to OECD guideline 414(Prenatal Developmental Toxicity Study;
RL 1, GLP)

25 female Wistar rats/dose exposedby gavageat dose levels of 0, 50, 250 and 1000 mg/kg bw/d.

Exposure day 6 through 15 post coitum

 

NOAEL = 1000 mg/kg bw/day (general tolerability in the females and for the fetal organism).

NOAEL = 1000 mg/kg bw/day (maternal reproduction).

NOAEL = 1000 mg/kg/day (teratologic effects

Read Across from source substance

 

Fatty acids

<C16      <7%                                            

C16, 16‘  26-35%

C18         42-52%

C18‘        15-20%

C18‘‘,18‘‘‘ ≤ 1.5%

>C18       ≤ 2%

C16 8%

C18 92%

Headgroup

MDEA

MDEA

 

 Quality of the experimental data of the analogues

The target substance MDEA Esterquat C18 satd. and the source substance MDEA Esterquat C16-18 and C18 unsatd. have both been tested in reliable studies equivalent to OECD 407 including evaluation of reproductive organs . In addition, a reliable subchronic study according to OECD 408. with assessment of reproductive organs and a reliable prenatal developmental toxicity study according to OECD 414 with the source substance MDEA-Esterquat C16-18 and C18 unsatd is available. All tests have been conducted according to GLP criteria.

Therefore this data have no uncertainties and can be used in an analogue approach.

The available data from the target and the source chemical are sufficiently reliable to justify the read-across approach.

 

Classification and labelling (Human Health)

The source substance MDEA-Esterquat C16-18 and C18 unsatd. is not classified for any human health effects similar to the target substance MDEA Esterquat C18 satd. which is also not classified regarding human health hazards.

 

Conclusion for read-across

 

Adequate and reliable scientific information indicates that the source and target substances and their subsequent degradation products have similar toxicity profiles under the experimental conditions in the considered studies for the endpoint reproductive toxicity.The results of the repeated dose toxicity tests and the prenatal developmental toxicity study show a comparable low systemic toxicity with NOAEL ≥ 500 mg/kg bw/day for both the target and the source substance. Neither the target substance MDEA Esterquat C18 satd. nor the source substance MDEA-Esterquat C16-18 and C18 unsatd. showed a substance-related effect on reproduction (fertility) up to the highest dose of 1000 and 500 mg/kg bw/day, respectively.

The structural similarities between the source and the target substances and the similarities in their breakdown products presented above further support the read-across hypothesis.Details are given in the general justification for read-across.

 


Short description of key information:
- Subacute (28 day) study; oral (gavage), rat (Wistar); 5/sex/dose), according to OECD guideline 407, GLP; NOAEL = 1000 mg/kg bw/day (highest dose level tested)
- Subacute (28 day) study; oral (gavage), rat (Charles River CD; 25/sex/dose), equivalent to OECD guideline 407, GLP; NOAEL = 500 mg/kg bw/d (highest dose level tested); read across from MDEA-Esterquat C16-18 and C18 unsatd.
- Subchronic (91 day) study; oral (gavage), rat (Crl CD BR; 15/sex/dose), equivalent to OECD guideline 408, GLP; NOAEL = 500 mg/kg bw/d (highest dose level tested); read across from MDEA-Esterquat C16-18 and C18 unsatd.

Justification for selection of Effect on fertility via oral route:
OECD guideline study, no deviations, GLP

Effects on developmental toxicity

Description of key information
- Prenatal developmental toxicity study; oral (gavage); rat (Wistar, 25/group, dosed from day 6 through 15 post coitum); OECD Guideline 414; GLP; NOEL = 1000 mg/kg bw/d, read across from MDEA-Esterquat C16-18 and C18 unsatd.
Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1992-10-09 to 1992-11-21
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Limit test:
no
Species:
rat
Strain:
other: Rat WIST HanIbm: WIST (SPF)
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd.
- Age at study initiation: No data. (Age at pairing: 12 weeks, minimum).
- Weight at study initiation: No data. (189-228 g at day 0 post coitum).
- Fasting period before study: N/A.
- Housing: individually in Makrolon cages (type-3) with wire mesh tops and standarized granulated softwood bedding (Lignocel, Schill AG, CH 4132 Muttenz/Switzerland).
- Diet (e.g. ad libitum): pelleted standard Kilba 343 rat/mouse maintenance diet ("Kilba", Klingentalmuehle AG, CH 4303 Kaiseraugust/Switzerland) ad libitum. (Batch no. 65-92).
- Water (e.g. ad libitum): tap water in bottles ad libitum.
- Acclimation period: 10 days (minimum).


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3 (air-conditioned).
- Humidity (%): 40-70.
- Air changes (per hr): 10-15.
- Photoperiod (hrs dark / hrs light): 12-hrs artificial fluorescent light/12 hours dark.


IN-LIFE DATES: From: 1992-10-09 To: 1992-11-21
Route of administration:
oral: gavage
Vehicle:
other: NOAEL emBe-distelled water, previously adjusted with hydrochloric acid to pH 2.5-2.6
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: The mixtures of the test substance and vehicle were prepared daily before administration. The test substance was weighed into a glass beaker on a tared precision balance (Mettler PE 360) and the vehicle added (w/v). The mixtures were prepared using a homogenizer. During the daily administration period, homogeneity was maintained using a magnetic stirrer. All animals received a dose volume of 10 ml/kg bw with a daily adjustment of the individual volume to the actual body weight.


DIET PREPARATION
- Rate of preparation of diet (frequency): N/A.
- Mixing appropriate amounts with (Type of food): N/A.
- Storage temperature of food: N/A.


VEHICLE: Bi-distilled water (previously adjusted with hydrochloric acid to pH 2.5-2.6).
- Justification for use and choice of vehicle (if other than water): N/A.
- Concentration in vehicle: N/A.
- Amount of vehicle (if gavage): 10 ml/kg.
- Lot/batch no. (if required): N/A.
- Purity: N/A.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Samples of the test substance/vehicle dilutions were taken on two occasions during the dosing period of the study and sent to the Sponsor for analysis (concentration, homogeneity and stability over 2-hrs).
Details on mating procedure:
- Impregnation procedure: cohabitation.
- If cohoused:
- M/F ratio per cage: 1:1.
- Length of cohabitation: overnight
- Further matings after two unsuccessful attempts: no data.
- Verification of same strain and source of both sexes: no data. (The male rats used for mating were in the possession of the testing laboratory. The fertility of these males was proved and was continuously controlled).
- Proof of pregnancy: sperm in vaginal smear/vaginal plug referred to as day 0 post coitum.
- Any other deviations from standard protocol: After mating, female rats were removed and allocated to the test groups.
Duration of treatment / exposure:
Day 6 through 15 post coitum.
Frequency of treatment:
Once daily in the morning.
Duration of test:
Females were sacrificed on day 21 post coitum.
No. of animals per sex per dose:
25/group.
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Dosages were based on the results of the dose range-finding sudy (RCC Project 326158--see cross reference section).
- Rationale for animal assignment (if not random): Mated rats were assigned to the different groups using a computer-generated random algorithm.
- Rationale for method of administration: International guidelines recognize the efficacy of oral administration.
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes.
- Time schedule: The animals were checked at least twice daily for any mortalities. The animals were observed at least twice daily for signs of reaction to treatment and/or symptoms of ill health.


DETAILED CLINICAL OBSERVATIONS: No


BODY WEIGHT: Yes
- Time schedule for examinations: Body weights were recordered daily, from day 0 until day 21 post coitum.
- Body weight gains from days 0-6 p.c., 6-9 p.c., 9-12 p.c., 12-16 p.c., 6-16 p.c., 16-21 p.c., and 6-21 p.c. were calculated.


FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes. Food consumption was recorded for the following periods: days 0-6, 6-9, 9-12, 12-16, and 16-21 post coitum.
- Food consumption for each animal determined and mean daily diet consumption calculated as g food consumed per period/days per period: Yes.
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No


WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No


POST-MORTEM EXAMINATIONS: Yes. Any female sacrificed or found dead during the study was subjected to gross macroscopic examination of all internal organs.
- Sacrifice on day # 21 post coitum by CO2 asphyxiation. Fetuses were removed by Caesarean section.
- Organs examined: Emphasis on the uterus and its contents, position of the fetuses in the uterus, and number of corpora lutea.
- Specimens of abnormal tissue were fixed in neutral phosphate buffered 4% formaldehyde solution.


Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes.
- Gravid uterus weight: Yes. (The uteri (and contents) of all females with live fetuses were weighed at necropsy on day 21 post coitum to enable the calculation of the corrected body weight gain).
- Number of corpora lutea: Yes.
- Number of implantations: Yes. If no implantation sites were evident, the uterus was placed in an aqueous solution of ammonium sulfide to accentuate possible haemorrhagic areas of implantation sites.
- Number of early resorptions: Yes.
- Number of late resorptions: Yes.
- Other: position of the fetuses was recordered.
Fetal examinations:
The fetuses were removed from the uterus, sexed, weighed individually, examnined for gross external abnormalities and allocated to one of the following procedures: Wilson's slicing technique or skeletal examination.

- External examinations: Yes: all per litter.
- Soft tissue examinations: Yes: half per litter. Wilson's slicing technique for examination of the viscera and brain. One half of the live fetuses from each litter were fixed in a mixture of ethyl alcohol, formol and acetic acid. After examination, the sections were preserved in a solution of ethyl alcohol and glycerine (one fetus/container).
- Skeletal examinations: Yes: half per litter. Fetuses were placed in a solution of potassium hydroxide for clearing and stained with alizarin red S (modified technique). The skeletons were examined and all abnormalities were recordered. The specimens were preserved individually in plastic bags.
- Head examinations: No data.
Statistics:
The following statistical methods were used to analyze body weights, food consumption, reproduction and skeletal examination data: means and standard deviations, univariate one-way analysis of variance, Dunnett-test, Steel-test, Fisher's Exact test for 2x2 tables. Individual values, means, standard deviations and t-statistics were rounded off before printing.
Indices:
N/A.
Historical control data:
N/A.
Details on maternal toxic effects:
Maternal toxic effects:no effects

Details on maternal toxic effects:
MORTALITY: No test substance related deaths were noted. One female in group 2 (50 mg/kg), was found dead in the evening of day 10 post coitum. At first daily inspection in the morning, this female had ruffled fur, ataxia, tremor and bleeding from the vagina. This single finding in the low dose
group was considered to be incidental.

CLINICAL SIGNS OF REACTION TO TREATMENT: In group 4 (1000 mg/kg), one female had bleeding from the vagina on days 15-16 post coitum. At scheduled Caesarean section, this female had one empty implantation site, only. No abnormal findings were noted in the dams of groups 1 (0 mg/kg), 2 (50 mg/kg), or 3 (250 mg/kg) with the exception of the dam in group 2 (50 mg/kg) mentioned above.

FOOD CONSUMPTION: Up to and including the highest dose level of 1000 mg/kg, food consumption of the dams was similar in all groups. No test substance related effects were noted.

BODY WEIGHTS: The differences in mean body weights between the control group and any dose group did not reveal any test substance related effect. In all groups the body weight gain and the corrected body weight gain (corrected for uterus weight) were similar. The dams of group 4 (1000 mg/kg), had statistically significantly increased mean body weights on day 5, between days 7-13 and on day 15 post coitum. These findings were considered to be a consequence of the slightly increased initial mean body weight: 213 g compared to 207 g in the control group, on day 0 post coitum.

REPRODUCTION DATA: Values for post-implantation loss were: 4.8 and 8.7 in groups 1 (0 mg/kg) and 4 (1000 mg/kg), respectively. The latter value attained statistical significance. Although the post-implantation losses were within the normal range of the historical control data of this strain of animals, two additional females (which were not included in the statistical analysis) had total post-implantation losses (i.e. implantation sites, only). Therefore, the increased post-implantation loss in group 4 (1000 mg/kg) was considered to be a slight effect of treatment with the test substance. In groups 2 (50 mg/kg) and 3 (250 mg/kg), none of the reproduction parameters as assessed by the mean number per dam of corpora lutea and implantation sites, pre- or post-implantation losses, and mean number of fetuses per dam were adversely affected by treatment with the test substance.

NECROPSY FINDINGS: No test substance related abnormal findings were noted at terminal necropsy of the dams in any group. One female in group 2 (50 mg/kg) which was found dead on day 10 post coitum had reddish lungs with dark red foci, reduced right kidney and enlarged left kidney with nodules and calcus in pelvis (both kidneys with pelvic dilation) and in the urinary badder dark red discoloration of the mucosa and urine with hemorrhagic contents were noted. No abnormal findings were noted in the dams of groups 1 (0 mg/kg), 3 (250 mg/kg), or 4 (1000 mg/kg).
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Basis for effect level:
other: maternal toxicity
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Basis for effect level:
other: developmental toxicity
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
EXTERNAL EXAMINATION: No test substance related abnormal findings were noted. Neither the type nor the incidences of the abnormal findings listed below indicated test substance related effects: In each group 1 (0 mg/kg) and 3 (250 mg/kg), one fetus with reduced mouth and shortened and tapering lower jaw was noted. In group 2 (50 mg/kg), one runt (fetal weight < 2.5 g) was noted. In group 4 (1000 mg/kg), one fetus with several abnormal findings--generally oedematous, cranioschisis with excencephaly, protusion of the tongue, eventration and both eyes partially missing--was noted.

SEX RATIOS: In all groups, the sex ratios of the fetuses were not affected by the treatment with the test substance.

BODY WEIGHTS: The mean fetal body weights were similar in all groups. No test substance related effects were evident.

VISCERAL EXAMINATION BY WILSON TECHNIQUE: No test substance related abnormal findings were noted at visceral examination of the fetuses. In group 3 (250 mg/kg), in the fetus which had reduced mouth, shortened and tapering lower jaw at external examination a small palatoschisis was additionally observed at visceral examination. No abnormal findings were noted in groups 1 (0 mg/kg), 2 (50 mg/kg), or 4 (1000 mg/kg).

SKELETAL EXAMINATION: The incidences of abnormal findings at skeletal examination were as following:
2/145 (=1%) fetuses in 2/25 litters in group 1 (0 mg/kg);
10/141 (=7%) fetuses in 6/24 litters in group 2 (50 mg/kg);
4/136 (=3%) fetuses in 4/25 litters in group 3 (250 mg/kg) and
4/116 (=3%) fetuses in 4/21 litters in group 4 (1000 mg/kg).
Mainly: wavy or fused ribs, abnormally or incompletely ossified sternebrae, dumbbell shaped or hemicentric thoracic vertebral body and in two fetuses in group 2 (50 mg/kg) fused os interparietale and occipitale were noted. Neither the type nor the incidences of these abnormal findings indicated test substance related effects. In all groups, the stage of the skeletal development of the fetuses was similar. The few statistically significant differences (on individual basis, only) between the control group and the dose group 4 (1000 mg/kg) in the ossification grade of the phalangeal nuclei did not indicate any test substance related effect. All values evaluated in this study were in the normal range of the historical control data of this strain of animals.
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Basis for effect level:
other: embryotoxicity
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Basis for effect level:
other: fetotoxicity
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Basis for effect level:
other: teratogenicity
Abnormalities:
not specified
Developmental effects observed:
not specified

N/A.

Conclusions:
NOAEL = 1000 mg/kg bw/day (general tolerability in the females and for the fetal organism).
NOAEL = 1000 mg/kg bw/day (maternal reproduction).
NOAEL = 1000 mg/kg/day (teratologic effects)
Executive summary:

In the developmental toxicity study (OECD guideline 414), groups of 25 mated female Wistar rats were treated with the test substance MDEA-Esterquat C16-18 and C18 unsatd. orally by gavage once daily from day 6 through 15 post coitum, at dose levels of 0, 50, 250 and 1000 mg/kg bw/day. Females were sacrificed on day 21 post coitum and the fetuses were removed by Caesarean section.

At 50, 250, and 1000 mg/kg, for the dams no test substance-related deaths or clinical signs as were noted as reaction to treatment. Up to and including the highest dose level of 1000 mg/kg, food consumption and body weight development of the dams were not affected by treatment with the test substance. At necropsy, no test substance-related abnormal findings in the dams were noted in any group.

A slight (statistically non significant) decrease in the number of corpora lutea and a slight (statistically non significant) increase in pre-implantation losses were observed in the high dose group (17.4 %) as compared to the controls (12.8 %); however, this is not a substance-related effect, since exposure started at gestation day 6, the day of implantation.

The slightly but statistically significant increased post-implantation losses at the high dose of (8.7 %) as compared to the controls (4.8 %) resulted in a slightly reduced portion of total fetuses per implantation site (91.3%) and in a reduced mean litter size (10.5 fetuses/litter) as compared to the controls (95.2 %; 11.2 fetuses/litter). The values were within the range of historical control values (3.9 % to 11.6 % for post-implantation losses and 10.2 to 12.2 fetuses/litter).

At 1000 mg/kg, two females with total post-implantation losses were noted. One of the two females had bleeding from the vagina on days 15-16 post coitum. However these two animals had only two and one corpora lutea, respectively, and were obviously not fit for reproduction. A small but significant increase in post-implantation losses was noted for the remaining females; however, the increase was within the historical data of the laboratory. These findings were considered by the authors to be a potentially effect of the test substance.

As the values were well within the range of the historical control values recorded at the same laboratory, it is likely that the effects observed are incidental and therefore not treatment related.

At 50 and 250 mg/kg, no test substance-related effects were noted on the maternal reproductive parameters, assessed by the mean number per dam of corpora lutea and implantation sites, pre- or post-implantation losses, and by the mean number of fetuses per dam.

Up to and including the highest dose level of 1000 mg/kg, no adverse effects on the fetal parameters were recorded. No external, skeletal or soft tissue malformations and no external variations were found. Mean fetal body weights and the sex ratios of the fetuses were comparable in all groups.

The maternal NOAEL is 1000 mg/kg bw/day. 

The developmental NOAEL is 1000 mg/kg bw/day.

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
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

Prenatal developmental toxicity data on the target substance MDEA Esterquat C18 satd. is not available. However, a prenatal developmental toxicity study with the source substance MDEA-Esterquat C16-18 and C18 unsatd. is available.

In a study according to OECD guideline 414, groups of 25 mated female Wistar rats were treated with MDEA-Esterquat C16-18 and C18 unsatd. orally by gavage once daily from day 6 through 15 post coitum, at dose levels of 0, 50, 250 and 1000 mg/kg bw/day. Females were sacrificed on day 21 post coitum and the fetuses were removed by Caesarean section.

At 50, 250, and 1000 mg/kg bw/d, for the dams no test substance-related deaths or clinical signs as were noted as reaction to treatment. Up to and including the highest dose level of 1000 mg/kg bw/d, food consumption and body weight development of the dams were not affected by treatment with the test substance. At necropsy, no test substance-related abnormal findings in the dams were noted in any group.

A slight (statistically non significant) decrease in the number of corpora lutea and a slight (statistically non significant) increase in pre-implantation losses were observed in the high dose group (17.4 %) as compared to the controls (12.8 %); however, this is not a substance-related effect, since exposure started at gestation day 6, the day of implantation.

The slightly but statistically significant increased post-implantation losses at the high dose of (8.7 %) as compared to the controls (4.8 %) resulted in a slightly reduced portion of total fetuses per implantation site (91.3%) and in a reduced mean litter size (10.5 fetuses/litter) as compared to the controls (95.2 %; 11.2 fetuses/litter). The values were within the range of historical control values (3.9 % to 11.6 % for post-implantation losses and 10.2 to 12.2 fetuses/litter).

At 1000 mg/kg bw/d, two females with total post-implantation losses were noted. One of the two females had bleeding from the vagina on days 15-16 post coitum. However these two animals had only two and one corpora lutea, respectively, and were obviously not fit for reproduction. A small but significant increase in post-implantation losses was noted for the remaining females; however, the increase was within the historical data of the laboratory. These findings were considered by the authors to be a potentially effect of the test substance. At 50 and 250 mg/kg bw/d, no test substance-related effects were noted on the maternal reproductive parameters, assessed by the mean number per dam of corpora lutea and implantation sites, pre- or post-implantation losses, and by the mean number of fetuses per dam.

Up to and including the highest dose level of 1000 mg/kg bw/d, no adverse effects on the fetal parameters were recorded. No external, skeletal or soft tissue malformations and no external variations were found. Mean fetal body weights and the sex ratios of the fetuses were comparable in all groups.

The slightly increased post-implantation losses at the high dose compared to the controls could be caused by some maternal toxicity, incidentally and therefore not treatment related or through a direct toxic effect on the fetus. Since two females of the high dose had total post-implantation losses (implantation-sites only) and one of these females showed vaginal bleeding, this may indicate that the post-implantation losses are due to (some) maternal toxicity effects although there were no adverse effects observed in the subacute study with the target substance MDEA Esterquat C18 satd. at doses up to 1000 mg/kg bw/day. As the values were well within the range of the historical control values recorded at the same laboratory, it is likely that the effects observed are incidental and therefore not treatment related. Since there was no effect on fetal body weight and no increased incidences of abnormal fetuses, it is assumed that the post-implantation losses are not due to a direct effect on the fetus.

The results of the prenatal developmental toxicity study do not indicate a substance-related effect on the fetus up to the limit dose of 1000 mg/kg bw/d. Therefore, the aspect of prenatal developmental toxicity is sufficiently covered.  

Strictly conservative DNELs have been derived from results of the available test data which include a pre-natal developmental study and sub-acute and sub-chronic studies with evaluation of reproductive endpoints. No substance-related adverse effects were found in any of the tests conducted and the NOAELs used to derive the DNELs correspond to the maximum doses tested. The DNELs fertility have been derived from results of the sub-chronic repeated dose toxicity study, taking full account of the potential increased uncertainty resulting from thelower sensitivity and the limited scope of the repeated dose toxicity studies for detecting effects on reproductive organsby applying an additional assessment factor. DNELs for developmental toxicity derived from the developmental toxicity study were higher than the DNELs for fertility. Thus, the fertility DNELs are also protective for development. These derived DNELs are therefore relevant and appropriate for risk assessment purposes.

The exposure and risk assessments are included in chapters 9 and 10 of this CSR. The final conclusion is based on the risk characterisation ratio (RCR). Comparison of all the derived DNELs with the results of the exposure assessment shows that exposures in all life cycle stages of the substance are well below the derived DNELs even under the precautionary assumptions applied.

There are no data gaps for effects on development. No human data are available. However, there is no reason to believe that these results from rat would not be applicable to humans.

 

Justification for read-across

For details on substance identity, toxicokinetics and detailed toxicological profiles, please refer also to the general justification for read-across given in chapter 5 of the CSR and attached as pdf document to section 7 of the IUCLID file.

 

a. Structural similarity and functional groups

The target substance, MDEA Esterquat C18 satd., consists of an amine backbone (MDEA = Methyldiethanol amine) esterified with long chain fatty acid stearic acid (C18 satd.; IV (iodine value) < 1). The main reaction product is the dialkylester compound, next to that small amounts of the monoalkylester may be formed. The amine function is quaternised with two methyl groups. The counter ion is Chloride.

The source substance, MDEA EsterquatC16-18 and C18 unsatd., consists of the same amine backbone (MDEA = Methyldiethanol amine) but esterified with a mixture of the long chain fatty acids C16, C18 and C18 unsaturated (IV < 25). The main reaction product is the dialkylester compound, next to that small amounts of the monoalkylester may be formed. The amine function is quaternised with two methyl groups. The counter ion is Chloride.

The source and the target substance share structural similarities with common functional groups (quaternary amines, esters, and fatty acid chains with comparable length and degree of saturation).

b. Common breakdown products

The metabolism expected to occur is hydrolysis of the ester-bond by esterases. However, the rate of hydrolysis is assumed to be low. The fraction of metabolised molecules would result in free fatty acids and Dimethyl-DEA (DEA = Diethanolamine). The carboxylic acids are further degraded by the mitochondrial beta-oxidation process (for details see common text books on biochemistry). The fatty acids enter normal metabolic pathways and are therefore indistinguishable from fatty acids from other sources including diet. The quaternary ammonium ions are not expected to be further metabolised, but excreted unchanged via the urine. 

c. Differences

The differences in fatty acid chain length (higher percentage of C16 in the source substance MDEA Esterquat C16 -18 and C18 unsatd.) and degree of saturation (higher degree of unsaturation of the C18 fatty acid chains in the source substance substance MDEA Esterquat C16-18 and C18 unsatd.) could be relevant for local toxicity (skin and eye irritation) but arebut are not considered to be of relevance for developmental toxicity.

Comparison of molecular structures and toxicity data ofMDEA Esterquat C18 satd. and MDEA-Esterquat C16-18 and C18 unsatd.

 

 

Source substances

Target substance

 

Endpoints

 

MDEA-Esterquat C16-18 and C18 unsatd.

 

MDEA Esterquat C18 satd.

Developmental Toxicity

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

according to OECD guideline 414(Prenatal Developmental Toxicity Study;
RL 1, GLP)

25 female Wistar rats/dose exposedby gavageat dose levels of 0, 50, 250 and 1000 mg/kg bw/d.

Exposure day 6 through 15 post coitum

 

NOAEL = 1000 mg/kg bw/day (general tolerability in the females and for the fetal organism).

NOAEL = 1000 mg/kg bw/day (maternal reproduction).

NOAEL = 1000 mg/kg/day (teratologic effects

Read Across from source substance

 

Repeated Dose Toxicity/Fertility

according to OECD 407 (oral 4 week study)

 

25 rats/dose/sex in the groups treated with 1,10 and 500 mg/kg/d

 

 

The test substance did not produce any test related toxicological responses up to levels of 500 mg/kg bw/day. The results from the evaluation of reproductive organs, especially organ weights of ovary and testis and histopathology of gonads revealed no indications of any substance-related effects up to and including the highest test dose of 500 mg/kg bw/day.

 

 

 

NOAEL = 500 mg/kg bw/day(general systemic toxicity/fertility)

according to OECD 407 (oral 4 week study, including 14-day recovery period)

 

5 rats/dose/sex in the groups treated with 62.5, 250 and 1000 mg/kg/d

 

'No test substance-related findings up to the highest tested dose. The results from the evaluation of reproductive organs (Organ weights of ovary, uterus with cervix, testis, epididymides, prostate with seminal vesicles and coagulating glands and the gross pathology and the histopathology of the reproductive organs (gonads, prostate and seminal vesicle, epididymides, uterus and vagina)

 

NOAEL = 1000 mg/kg bw/day(general systemic toxicity/fertility)

according to OECD 408 (oral 13 week study)

 

15 rats/dose/sex in the groups treated with 1,10 and 500 mg/kg/d

 

No biologically significant adverse effects were observed in any test group. The results from the evaluation of reproductive organs, especially organ weights of ovary and testis and histopathology of gonads from this 91-day repeated dose study revealed no indications of any substance-related effects up to and including the highest test dose of 500 mg/kg bw/day.

 

NOAEL = 500 mg/kg bw/day(general systemic toxicity/fertility)

 

 

Fatty acids

<C16      <7%                                            

C16, 16‘  26-35%

C18         42-52%

C18‘        15-20%

C18‘‘,18‘‘‘ ≤ 1.5%

>C18       ≤ 2%

C16 8%

C18 92%

Headgroup

MDEA

MDEA

 

 Quality of the experimental data of the analogues

The source substance MDEA-Esterquat C16-18 and C18 unsatd. has been tested in a reliable prenatal developmental toxicity study according to OECD 414. The target substance MDEA Esterquat C18 satd. and the source substance MDEA Esterquat C16-18 and C18 unsatd. have both been tested in reliable studies equivalent to OECD 407 including evaluation of reproductive organs . An additional reliable subchronic study according to OECD 408 the source substance MDEA-Esterquat C16-18 and C18 unsatd. with assessment of reproductive organs is available. All tests have been conducted according to GLP criteria.

Therefore this data have no uncertainties and can be used in an analogue approach.

The available data from the target and the source chemical are sufficiently reliable to justify the read-across approach.

 

Classification and labelling (Human Health)

The source substance MDEA-Esterquat C16-18 and C18 unsatd. is not classified for any human health effects similar to the target substance MDEA Esterquat C18 satd. which is also not classified regarding human health hazards.

 

Conclusion for read-across

Adequate and reliable scientific information indicates that the source and target substances and their subsequent degradation products have similar toxicity profiles under the experimental conditions in the considered studies for the endpoint developmental toxicity.The results of the repeated dose toxicity tests and the prenatal developmental toxicity study show a comparable low systemic toxicity with NOAEL ≥ 500 mg/kg bw/day for both the target and the source substance. Neither the target substance MDEA Esterquat C18 satd. nor the source substance MDEA-Esterquat C16-18 and C18 unsatd. showed a substance-related effect on reproduction (fertility) up to the highest dose of 1000 and 500 mg/kg bw/day, respectively.

The structural similarities between the source and the target substances and the similarities in their breakdown products presented above further support the read-across hypothesis. Details are given in the general justification for read-across.


Justification for selection of Effect on developmental toxicity: via oral route:
OECD guideline study, no deiviatons, GLP

Justification for classification or non-classification

Results of existing studies with the target substance and a structurally very similar source substance indicate that MDEA Esterquat C18 satd. does not need to be classified for toxicity to reproduction, developmental toxicity and teratogenicity according to the criteria given in regulation (EC) 1272/2008 or the former European directive on classification and labelling 67/548/EEC. Therefore labelling is not necessary.

 

Additional information