Registration Dossier

Diss Factsheets

Administrative data

Key value for chemical safety assessment

Toxic effect type:
dose-dependent

Effects on fertility

Description of key information

Screening study: No adverse effects were observed up to the limit dose of 1000 mg/kg bw/day.


 


EOGRTS: No adverse effects were observed in an EOGRTS with basic test design up to the limit dose of 1000 mg/kg bw/day (read across from MDEA-Esterquat C16-18 and C18 unsatd.).

Link to relevant study records

Referenceopen allclose all

Endpoint:
extended one-generation reproductive toxicity - basic test design (Cohorts 1A, and 1B without extension)
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
This read-across hypothesis corresponds to Scenario 2 of the Read-Across Assessment Framework (RAAF), ECHA, March 2017.

This scenario covers the analogue approach for which the hypothesis is based on different compounds with the same type of effect(s). For the REACH information requirement under consideration, the effects obtained in a study conducted with one source substance are used to predict the effects that would be observed in a study with the target substance if it were to be conducted. The same type of effect(s) or absence of effect is predicted. The predicted strength of the effects may be similar or based on a worst-case approach.

Namely, the absence of adverse effects observed in two studies with the structurally similar source substance MDEA-Esterquat C16-18 and C18 unsatd. is used to predict the toxicological properties of the target substance Partially unsaturated TEA-Esterquat. In this case, the absence of adverse effects is predicted for the endpoints toxicity to reproduction and for prenatal developmental toxicity.

The read-across from the existing toxicological studies conducted with the source substances are considered as an appropriate adaptation to the standard information requirements of the REACH Regulation for the target substance, in accordance with the provisions of Annex XI, 1.5 of the REACH Regulation. The justification of the proposed read-across approach is elaborated in the next chapters.

Property-specific read-across hypothesis
The read-across approach will be applied for the endpoints toxicity to reproduction and developmental toxicity.

The property specific read-across hypothesis is that both source and target substances have similar properties (absence of effects) because:
• they share structural similarities with common functional groups: One quaternised ethanolamine moiety, one to three ethanol ester groups but mainly two ester groups, each with typical UVCB distribution with similar long-chain fatty acids of natural origin.
• they are manufactured from comparable (triethanolamine or diethanolamine) or nearly identical precursors (long-chain fatty acids), methyl-quaternization agents dimethyl sulfate or methyl chloride) under similar conditions. Therefore, comparable breakdown products via physical and biological processes (hydrolysis and biological degradation resp. metabolization), which result in partially structural similar chemicals are evident.

The following supporting evidence is provided:
Anchor studies for the target substance:
• To predict the absence of effects of a prenatal developmental toxicity study, a reproductive toxicity screening study conducted with the target and source substance is provided, and a prenatal developmental toxicity study in rats with the source substance is provided.
• To predict the absence of effects of an EOGRTS study, a reproductive toxicity screening study conducted with the target and source substance is provided, and an EOGRTS study with the source substance is provided.
• To further strengthen the evidence, repeated dose toxicity studies are provided for both, source and target substances.
Consistency is demonstrated by the absence of quantitative variations in the effects (or absence of effect(s)) for both substances and for all relevant properties.


2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)

see attacted justification for read-across

3. ANALOGUE APPROACH JUSTIFICATION

see attacted justification for read-across

4. DATA MATRIX

see attacted justification for read-across
Reason / purpose for cross-reference:
read-across: supporting information
Reason / purpose for cross-reference:
read-across source
Species:
rat
Sex:
male/female
Route of administration:
oral: gavage
Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: no toxicologically relevant adverse effects observed
Key result
Critical effects observed:
no
Dose descriptor:
NOAEL
Generation:
F1 (cohort 1A)
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no toxicologically relevant adverse effects observed
Dose descriptor:
NOAEL
Generation:
F1 (cohort 1B)
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no toxicologically relevant adverse effects observed
Key result
Reproductive effects observed:
no
Conclusions:
In conclusion, the dosage of 1000 mg/kg/day was considered the NOAEL for general and reproductive toxicity and pups development in Parental generation and for general and reproductive toxicity in Cohort 1A and 1B.
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
07 January 2020 - 08 March 2020
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Italia S.p.A., Calco (Lecco), Italy
- Females (if applicable) nulliparous and non-pregnant: yes
- Age and weight at study initiation: 7 to 8 weeks old and
weighing 200 to 225 g for males and 175 to 200 g for females
- Fasting period before study: no
- Housing:
from arrival to mating: up to 5 of one sex to a cage, in polysulfone
solid bottomed cages measuring 59.5×38×20cm (Tecniplast Gazzada S.a.r.l., Buguggiate, Varese). Nesting material was provided inside suitable bedding bags and changed at least twice a week.
During mating: one male to one female in clear polysulfone cages measuring 42.5×26.6×18.5cm with a stainless steel mesh lid and floor (Tecniplast Gazzada
S.a.r.l., Buguggiate, Varese)
After mating: males were re-caged as they were before mating; females were transferred to individual solid bottomed cages for the gestation period, birth and lactation (measuring 42.5×26.6×18.5 cm). Nesting material was provided inside suitable bedding bags. In addition, suitable nesting material (Scobis 0Mucedola) was provided as necessary. Nesting material was changed at least two times a week.
- Diet (e.g. ad libitum): laboratory rodent diet (4 RF 21,Mucedola S.r.l., Via G. Galilei, 4, 20019 SettimoMilanese (MI), Italy), ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 32 days

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

Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
-dosing solutions were prepared in concentrations of 10, 30 and 100 mg/mL

- dose volume: 10mL/kg body weight



Details on mating procedure:
- M/F ratio per cage: 1:1
- Length of cohabitation: until positive identification occurred
- Proof of pregnancy: vaginal plug / sperm in vaginal smear referred to as day 0 of pregnancy
- After successful mating each pregnant female was caged individually
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Results of the analyses were within the acceptability limits for suspensions
(85-115% for concentration and CV < 10% for homogeneity)
Duration of treatment / exposure:
males: for 14 days prior to pairing and thereafter until the day before necropsy, for a total of 30 days
females: 14 days prior to pairing and thereafter during pairing, post coitum and post partum periods until Day 13 post partum, for a total of 50 to 55 days
Frequency of treatment:
daily
Details on study schedule:
- F1 parental animals not mated (screenign study)
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Dose / conc.:
300 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Parental animals: Observations and examinations:
MORTALITY: Yes
twice daily

CLINICAL SIGNS: Yes
Once before commencement of treatment and at least once daily during the study


BODY WEIGHT: Yes
- Time schedule for examinations:
Males: weekly from allocation to termination
Females: weekly from allocation to positive identification of mating and on
gestation Days 0, 7, 14 and 20; Dams: Days 1, 4, 7 and 13 post partum and on the day of necropsy


FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes


WATER CONSUMPTION: No

OTHER:
thyroid hormone determination (T3, T4 and TSH)
Males at termination
Females at termination
Pups on Days 4 and 13 or 14 post partum

Blood collection for detection of test item - Dams and pups
Dams
Blood samples (approximately 0.3 mL) were taken from the tail vein of 5 females/group on Day 18 post coitum and Day 13 post partum at 1 time point (4 hours post-dose).
Pups
On Day 13 post partum blood samples (approximately 0.3 mL) were taken from 1 pup/sex/litter from 5 selected dams of each group (5 pups/sex/group). Blood samples were withdrawn at necropsy under light ether anaesthesia from the heart (intracardiac puncture).
Oestrous cyclicity (parental animals):
Before allocation (stock females)
Oestrous cycle was monitored by vaginal smears in all females ordered for the study for 2 weeks before allocation, in order to exclude females showing irregular oestrus cyclicity.

After allocation (females allocated to groups)
Vaginal smears were taken in the morning from Day 1 of dosing up to positive identification of copulation. The vaginal smear data were examined to determine the following:
1. anomalies of the oestrous cycle;
2. pre-coital interval (i.e., the number of nights paired prior to the detection of mating).

Before despatch to necropsy
Vaginal smears were also taken from all females before despatch to necropsy and the oestrous cycle phase recorded.
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: yes
- size of each litter was adjusted by eliminating extra pups by random
selection to yield, as nearly as possible, four pups per sex per litter. Partial adjustment was acceptable. At least one culled male and one
culled female was selected for hormone determination with the exception of the following females where the selection was performed on the basis of the pups available:
- Group 1 : female no. X12500005: 2 female pups selected
female no. X12500009: 1 male pups selected
- Group 3 : female no. X12500041: 1 female pup selected
female no. X12500047: 1 female pup selected
- Group 4 : female no. X12500077: 2 female pups selected

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, anogenital distance (AGD), pup weight on the day of AGD, presence of nipples/areolae in male pups

GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities

thyroid hormone determination (T3, T4 and TSH): Pups on Days 4 and 13 or 14 post partum
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals 30 days of treatment.
- Maternal animals: All surviving animals on Day 14 post partum

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

HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in Table 1 were prepared for microscopic examination and weighed, respectively.
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring were sacrificed at 13 days of age.
- These animals were subjected to postmortem examinations as follows:

GROSS NECROPSY
- Gross necropsy consisted of external examinations

– Internal examination - Gonads were inspected from all pups in order to confirmthe
sex previously determined by external examination.
– Nipples retention at Day 14 post partum - No nipples were found in pups to be
retained on Day 14 post partum. Data were not tabulated, but will be archived with
all raw data.

HISTOPATHOLOGY / ORGAN WEIGTHS
thyroid weights
Statistics:
Standard deviationswere calculated as appropriate. For variables such as bodyweight, food consumption, hormone determination and organ weight the significance of the differences amongst group means was assessed by Dunnett’s test or a modified t test, depending on the homogeneity of data. Statistical analysis of histopathological findings was carried out by means of the non-parametric Kolmogorov-Smirnov test if n was more than 5. The non-parametric Kruskal-Wallis analysis of variance was used for the other parameters.
Intergroup differences between the control and treated groups were assessed by the nonparametric version of theWilliams test. The criterion for statistical significance was p<0.05.
Reproductive indices:
Copulation Index
Fertility Index
Pre-implantation loss
Pre-natal loss

Offspring viability indices:
Post-natal loss at Day 0 post partum
Post-natal loss at Day 4 post partum (before culling)
Post natal loss at Day 13 post partum (after culling)
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Salivation was the treatment-related clinical sign recorded during the study. This sign was occasionally noticed in all males and females treated at 1000mg/kg/day, starting from the pre-mating phase until sacrifice (end of mating for males and post partum phase for females). Males treated at 300mg/kg/day also showed salivation, but this sign appeared later in the study, mostly at the end of the mating was considered not adverse.
Other signs, such as scabs, damaged eye, hairloss and isolation in cage, were sporadically recorded during the study. These findings were considered as incidental or related to in-life procedures and were, thus, deemed as minor clinical signs, not related to treatment with the test item.
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
No differences in body weight were observed in treated animals of both sexes, when compared to controls.
Statistically significant differences in body weight gain were observed:
– decrease in low and high dose groups of males on Day 8 of treatment
– decrease in low dose group of females on Day 1 of treatment
– increase in high dose females on Day 7 post partum
These differences were occasional and with no dose relationship, therefore considered unrelated to treatment.
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Endocrine findings:
no effects observed
Description (incidence and severity):
Thyroid hormones: No changes were recorded between control and treated groups.
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
No treatment-related changes were noted during the microscopic organ exminations.
The sporadic lesions, reported in control and treated animals, were considered to be an expression of spontaneous/or incidental pathology, seen in this species and age in this kind of studies and under the test site's experimental conditions.

Seminiferous tubules were evaluated with respect to their stage in the spermatogenic cycle and to the integrity of the various cell types within the different stages; regular layering in the germinal epithelium was noted.
Reproductive function: oestrous cycle:
no effects observed
Reproductive function: sperm measures:
no effects observed
Reproductive performance:
no effects observed
Fate of females
All females mated.
The number of females with live pups on Day 13/14 post partum was 10 in the control group and 10 in all treated groups, dosed at 100, 300 and 1000 mg/kg/day.

Oestrous cycle, reproductive parameters, pairing combination and mating
performance
No anomalies were noted in the oestrous cycle and pre-coital interval of the treated females, when compared to controls.
All females mated, all gave birth and all conceivings were within 5 days of mating.

The copulatory indices were 100% for controls and treated animals. The fertility indices were 100% for control animals and 100% for all treated groups.

Implantation sites, pre-implantation loss data, pre-natal loss dataandgestation
length of females
Implantations and pre-implantation loss, total litter size and pre-natal loss (as percentage) did not show dose-related or treatment-related differences.
Gestation periods were comparable between control and treated groups. All dams gave birth between Days 21 and 23 post coitum.

Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no signs of treatment-related toxicity
Key result
Critical effects observed:
no
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
Small size, cold to touch, pallor, apparently no food intake (milk) and small appearance were in general the clinical signs noted in the pups of control and treated groups.
Dermal irritation (if dermal study):
not examined
Mortality / viability:
mortality observed, non-treatment-related
Description (incidence and severity):
No significant differences in total litter size, live litter size, mean pup loss (percentage), and sex ratio of pups were observed among the treated and the control dams at birth and on Days 1, 4, 13 and 14 post partum.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No significant differences in mean pup weights (percentage) were observed among the treated and the control dams at birth and on Days 1, 4, 13 and 14 post partum.
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Sexual maturation:
not examined
Anogenital distance (AGD):
effects observed, non-treatment-related
Description (incidence and severity):
Statistically significant decreases in the anogenital distances were noted in male pups (up to 14%) and female pups (up to 32%) of treated groups in respect to the controls. However, the above results were considered incidental and unrelated to treatment, considering the following:

- all values were within the range of ERBC historical control data (0.55 - 3.14mm/g1⁄3 for males and 0.48 - 2.09mm/g1⁄3 for females). What are the control group means of the historical data? Compare mean litter values with mean litter values of historical control data. To might be able to show that It Is some individual litter causing the decrease, and not a decrease in all litters.
- there was no relation to the dose. (Compound identified as endocrine disruptors show marked effects on AGD in a dose-dependent manner. Camilla Lindgren Schwartz, et al., Archives of Toxicology volume 93, pages253–272(2019)) – Absence of other correlated findings considered androgen-mediated endpoints (such as areola/nipple retention, cryptorchidism, decreased reproductive organ weights, and malformation incidence)
- the testing was performed at high doses, not representing real live exposure ("For these evaluations, there should be stronger emphasis on effect doses and to what extend supra-high test reliably recapitulate what occurs at more human relevant doses." Camilla Lindgren Schwartz, et al., Archives of Toxicology volume 93, pages253–272(2019)).

Nipple retention in male pups:
no effects observed
Description (incidence and severity):
No nipples were observed in male pups on Day 13/14 post partum.
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
No significant differences were noted in thyroid weight between controls and pups of treated groups (1 pup/sex/litter).
Gross pathological findings:
no effects observed
Description (incidence and severity):
The majority of decedent pups from all groups were found autolysed or cannibalised without any possibility for a macroscopic examination of internal organs. However, at external examination, performed in pups not extremely cannibalised, no findings were noted.
Culled pups sacrificed on Day 4 post partum and those sacrificed onDays 13/14 post partum did not show any abnormalities.
Histopathological findings:
not examined
Other effects:
no effects observed
Description (incidence and severity):
Thyroid hormones in Pups - Day 13/14 post partum
No changes were recorded between control and treated groups.
Behaviour (functional findings):
not examined
Developmental immunotoxicity:
not examined
Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no signs of treatment-related toxicity
Key result
Reproductive effects observed:
no

REPRODUCTIVE PARAMETERS OF ANIMALS – SUMMARY

 

 

Group

1

2

3

4

Copulatory Index% (males)

 

100.0

100.0

100.0

100.0

Fertility Index% (males)

 

100.0

100.0

100.0

100.0

Copulatory Index% (females)

 

100.0

100.0

100.0

100.0

Fertility Index% (females)

 

100.0

100.0

100.0

100.0

 

 

IMPLANTATION, PRE-IMPLANTATION LOSS DATA, PRE-NATAL LOSS DATA AND GESTATION LENGTH OF FEMALES - GROUP MEAN DATA

 

Group

 

Corpora Lutea

Implantations

Pre-implantation loss %

Live Litter size at birth

Pre-natal loss %

Gestation

length (days)

 

1

Mean

SD

(n)

18.20

1.62

10

18.10

1.66

10

0.56

1.77

10

16.80

2.25

10

7.23

8.75

10

22.10

0.32

10

2

Mean

SD

(n)

16.80

3.19

10

16.80

3.19

10

0.00

0.00

10

15.00

3.02

10

10.11

10.74

10

21.90

0.32

10

3

Mean

SD

(n)

16.20

2.98

10

16.00

3.53

10

2.22

7.02

10

14.00

4.16

10

13.26

14.25

10

22.10

0.32

10

4

Mean

SD

(n)

17.00

2.58

10

16.70

2.67

10

1.82

2.94

10

15.50

2.51

10

7.08

8.55

10

22.10

0.32

10

Statistical analysis: Kruskall Wallis test

William’s test if group mean differences are different from control at p < 0.05

* = mean value of group is significantly different from control

Conclusions:
The toxic effects on Sprague Dawley rats of both sexes were investigated after repeated dosing with the test item. Furthermore, effects of the test item on male and female reproductive performance were examined, i.e. gonadal function, mating behaviour, conception, development of conceptus, parturition and early lactation of the offspring.
Groups of 10 males and 10 females received the test item, by gavage, at dosages of 0, 100, 300 and 1000 mg/kg body weight/day. An additional group of 10 males and 10 females received the vehicle alone (softened water) at the dose volume of 10 mL/kg and acted as control.
No effects were observed in the in vivo parameters of parental animals: no toxicologically relevant clinical signs and no effects on body weight, body weight gain and food consumption were noted at any dose level investigated. Thyroid hormones (T3, T4 and TSH) evaluated in parental males did not show any changes of toxicological relevance. No remarkable differences were noted at post mortem examination including organ weights and no treatment-related macroscopic and microscopic changes were observed in treated animals, when compared to controls. Futhermore, no effects on the spermatogenic cycle were described. No intergroup differences were seen in oestrous cycle, pre-coital intervals, copulatory and fertility indices.
No significant differences were observed in the number of corpora lutea, implantations, total litter size, pre-implantation loss, pre-birth loss and gestation length between control and treated females. Litter data at birth and on Days 1, 4 and 13 post partum and sex ratios were also comparable between treated and control females. There were neither treatment-related signs at clinical observation of pups, nor at necropsy of deceased pups or those sacrificed after culling or at term. The statistically significant decrease noted in the ano-genital distance of male and female pups on Day 1 of age in all treated groups was not considered of toxicological relevance, as there was no clear relation to the dose and as all values were within ERBC historical control data. No nipples were observed on Day 13 of age, in male pups at all dose levels. Thyroid hormones levels and pups thyroid weight were unaffected by treatment.
Analysis of plasma samples collected at 4 hours post-dose, showed that after oral administration of PC-2019-848, parental females were systemically exposed to the test item (detected as the physiological degradation product TEA-Core) both on Day 18 post coitum and Day 13 post partum. The systemic exposure increased with the dose but not with the duration of treatment. Systemic exposure was also considered demonstrated in not dosed pups on Day 13 post partum, although with values below the quantification limits (BLOQ).
Based on the results obtained in this study, the NOAEL (No Observed Adverse Effect Level) for both general toxicity and reproduction/developmental toxicity was considered to be 1000 mg/kg/day for both males and females.
Executive summary:

 The toxic effects on Sprague Dawley rats of both sexes were investigated after repeated dosing with the test item partially unsaturated TEA-Esterquat in accordance with OECD Guideline 421 (adopted 29  July 2016).


Furthermore, effects of the test item on male and female reproductive performance were examined, i.e. gonadal function, mating behaviour, conception, parturition and early lactation of the offspring.


The vehicle was water (was water softened by reverse osmosis. All doses (0, 100, 300 and 1000 mg/kg/day) were administered orally, by gavage at a dose volume of 10mL/kg body weight.


 


Males were treated for 14 days prior to pairing and during pairing with females until the day before necropsy, for a total of 30 days. Females were treated for 14 days prior to pairing, during pairing and throughout the gestation and lactation periods until Day 13 post partum.


The following investigations were performed: body weight, body weight gain, clinical signs, food consumption, macroscopic observations, organ weights and histopathological examination.


In addition he following assessments performed: oestrous cycle evaluation for parental females (2 weeks before start of dosing, during pre-mating and mating phases, prior to necropsy), mating performance, thyroid hormone determination (parental males and pups at Day 13/14 post partum) and collection of litter data.


Clinical signs, anogenital distance, external and/or internal examination were recorded for pups. Thyroid hormone levels and thyroid weight were also determined in 1 pup/sex/group randomly selected at Day 14 post partum.


Routine histopathological examination was performed only in control and high dose groups and it included identification of the stages of the spermatogenic cycle in male animals.


 


1.2 Fate of females


Details of the pregnancy status were as follows:


Groups                                                                         1          2          3          4


Non-pregnant females                                                   0          0          0          0


Conceiving 1 - 5 days                                                    10        10        10        10


No. of females with live pups on Day 13/14 post partum 10         10        10        10


 


1.3 Mortality and clinical signs


No mortality occurred during the study.


Salivation was the most relevant and treatment-related clinical sign recorded during the


study. This sign was occasionally noticed in all males and females treated at 1000mg/kg/day,


starting from the pre-mating phase until sacrifice (end of mating for males and post partum


phase for females). Males treated at 300mg/kg/day also showed salivation, but this sign


appeared later in the study, mostly at the end of the mating phase.


 


1.4 Body weight and body weight gain


No differences in body weight were observed in treated animals of both sexes, when compared


to controls and no differences considered treatment-related were observed in body


weight gain of treated animals.


 


1.5 Food consumption


Food consumption was unaffected by treatment.


 


1.6 Blood detection of test item


Systemic exposure of pups through the mother milk was also demonstrated by the presence of the physiological degradation product TEA-Core in pups on Day 13 post partum, i.e. prior to weaning.  However, quantification was not possible since the detected values of TEA-Core were below the limit of quantification (BLOQ).


 


1.7 Oestrous cycle, reproductive parameters, pairing combination and mating


performance


Oestrous cycles, pre-coital interval, copulatory index and fertility index did not show any


treatment-related intergroup differences.


 


1.8 Implantation sites, pre-implantation loss data, pre-natal loss data And gestation


length of females


Implantation sites, pre-implantation and pre-natal loss and gestation length did not show treatment-related differences.


 


1.9 Litter data and sex ratio of pups


No significant differences in total and live litter size, pup loss, litter weights and mean pup weight were observed among treated and control females at birth and on Days 1, 4 and 13 post partum. Sex ratio did not show any significant differences between groups.


 


1.10 Clinical signs of pups


No signs considered treatment related were seen in pups of treated groups.


 


1.11 Anogenital distance


The statistically significant decrease noted in the ano-genital distance of male and female pups on Day 1 of age in all treated groups was considered incidental and unrelated to treatment, considering that: in both sexes all values were within the range of ERBC historical control data and there was no relation to the dose.


In addition, in females, the decrease observed represents a trend towards feminisation and is not considered an adverse effect (i.e. an increase in AGD of females, suggesting masculinisation); in males, there was no correlations with other findings considered androgen-mediated endpoints (such as areola/nipple retention, cryptorchidism, decreased reproductive organ weights, and malformation incidence).


 


1.12 Thyroid hormones


Parental animals


No differences were recorded between treated and control groups.


Pups - Day 14 post partum


No differences were recorded between treated and control groups.


 


1.13 Necropsy findings in decedent pups, inpupssacrificedonDays 4and13/14


post partum and nipple count


No findings were seen at necropsy in decedent pups or in those sacrificed on Days 4 and 13 post partum.


No nipples were observed in male pups on Day 13 post partum.


 


1.14 Pups thyroid weight on Day 13/14 post partum


No significant differences were noted in thyroid weight between controls and pups of treated groups.


 


1.15 Terminal body weight and organ weights


No changes attributable to the treatment were observed in terminal body, absolute and relative organ weight of treated animals, when compared to the controls.


 


1.16 Macroscopic observations


No significant differences were noted at post mortem examination in treated animals, when compared to the controls.


 


1.17 Microscopic observations


No treatment-related changes were observed in treated animals, when compared to the controls.


Seminiferous tubules were evaluated with respect to their stage in the spermatogenic cycle and to the integrity of the various cell types within the different stages; regular layering in the germinal epithelium was noted.


 


1.18 Conclusion


In conclusion, no signs of treatment-related toxicity were observed at any of the dose levels investigated following treatment with the test item, when administered to rats by oral route at dose levels of 100, 300 and 1000 mg/kg/day. Based on the results of the present study, the NOAEL (No Observed Adverse Effect Level) for general, reproductive and developmental toxicity was considered to be 1000 mg/kg/day for males and females.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 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

A Reproduction / Developmental Toxicity Screening Test is available for the target substance partially unsatd. TEA-Esterquat as well as for the source substance MDEA-Esterquat C16-18 and C18 unsatd. An Extended One Generation Reproduction Toxicity Study is available for the source substance MDEA-Esterquat C16-18 and C18 unsatd. A justification for read-across is attached to Iuclid section 13.

 

Screening study

Target substance partially unsatd. TEA-Esterquat

The toxic effects on Sprague Dawley rats of both sexes were investigated after repeated dosing with the test item partially unsaturated TEA-Esterquat in accordance with OECD Guideline 421 (adopted 29  July 2016).

Furthermore, effects of the test item on male and female reproductive performance were examined, i.e. gonadal function, mating behaviour, conception, parturition and early lactation of the offspring.

The vehicle was water (was water softened by reverse osmosis. All doses (0, 100, 300 and 1000 mg/kg/day) were administered orally, by gavage at a dose volume of 10mL/kg body weight.

Males were treated for 14 days prior to pairing and during pairing with females until the day before necropsy, for a total of 30 days. Females were treated for 14 days prior to pairing, during pairing and throughout the gestation and lactation periods until Day 13 post partum.

The following investigations were performed: body weight, body weight gain, clinical signs, food consumption, macroscopic observations, organ weights and histopathological examination.

In addition he following assessments performed: oestrous cycle evaluation for parental females (2 weeks before start of dosing, during pre-mating and mating phases, prior to necropsy), mating performance, thyroid hormone determination (parental males and pups at Day 13/14 post partum) and collection of litter data.

Clinical signs, anogenital distance, external and/or internal examination were recorded for pups. Thyroid hormone levels and thyroid weight were also determined in 1 pup/sex/group randomly selected at Day 14 post partum.

Routine histopathological examination was performed only in control and high dose groups and it included identification of the stages of the spermatogenic cycle in male animals.

 

Fate of females

Details of the pregnancy status were as follows:

Groups                                                                            1         2          3          4

Non-pregnant females                                                     0         0          0          0

Conceiving 1 - 5 days                                                      10       10        10        10

No. of females with live pups on Day 13/14 post partum10       10        10        10

 

No mortality occurred during the study.

Salivation was the most relevant and treatment-related clinical sign recorded during the study. This sign was occasionally noticed in all males and females treated at 1000mg/kg/day, starting from the pre-mating phase until sacrifice (end of mating for males and post partum phase for females). Males treated at 300mg/kg/day also showed salivation, but this sign appeared later in the study, mostly at the end of the mating phase.

No differences in body weight were observed in treated animals of both sexes, when compared to controls and no differences considered treatment-related were observed in body weight gain of treated animals.

Food consumption was unaffected by treatment.

Oestrous cycles, pre-coital interval, copulatory index and fertility index did not show any

treatment-related intergroup differences. Implantation sites, pre-implantation and pre-natal loss and gestation length did not show treatment-related differences.  

No significant differences in total and live litter size, pup loss, litter weights and mean pup weight were observed among treated and control females at birth and on Days 1, 4 and 13 post partum. Sex ratio did not show any significant differences between groups.

No changes attributable to the treatment were observed in terminal body weight, absolute and relative organ weight of treated animals, when compared to the controls.

No significant differences were noted at post mortem macroscopic examination in treated animals, when compared to the controls.

No treatment-related microscopic changes were observed in treated animals, when compared to the controls.

Seminiferous tubules were evaluated with respect to their stage in the spermatogenic cycle and to the integrity of the various cell types within the different stages; regular layering in the germinal epithelium was noted. No differences in thyroid hormones were recorded between treated and control groups in parental animals.

Systemic exposure of pups through the mother milk was also demonstrated by the presence of the physiological degradation product TEA-Core in pups on Day 13 post partum, i.e. prior to weaning.  However, quantification was not possible since the detected values of TEA-Core were below the limit of quantification (BLOQ).

 No clinical signs considered treatment related were seen in pups of treated groups.

The decrease in the anogenital distance noted in males and females pup of treated groups was not considered attributed to treatment. No findings were seen at necropsy in decedent pups or in those sacrificed on Days 4 and 13 post partum. No nipples were observed in male pups on Day 13 post partum. No differences in thyroid hormones were recorded between treated and control groups in pups on Day 14 post partum. No significant differences were noted in thyroid weight between controls and pups of treated groups.

 

In conclusion, no signs of treatment-related toxicity were observed at any of the dose levels investigated following treatment with the test item, when administered to rats by oral route at dose levels of 100, 300 and 1000 mg/kg/day. Based on the results of the present study, the NOAEL (No Observed Adverse Effect Level) for general, reproductive and developmental toxicity was considered to be 1000 mg/kg/day for males and females.

 

 

Source substance MDEA-Esterquat C16-18 and C18 unsatd.

The toxic effects on Sprague Dawley rats of both sexes were investigated after repeated dosing with MDEA-Esterquat C16-18 and C18 unsatd. in accordance with OECD TG 421 (adopted on 29 July 2016). Furthermore, effects of the test item on male and female reproductive performance were examined, i.e. gonadal function, mating behaviour, conception, development of conceptus, parturition and early lactation of the offspring.

Groups of 10 males and 10 females received the test item, by gavage, at dosages of 0, 100, 300 and 1000 mg/kg body weight/day. An additional group of 10 males and 10 females received the vehicle alone (softened water) at the dose volume of 10 mL/kg and acted as control.

No effects were observed in the in vivo parameters of parental animals: no relevant clinical signs and no effects on body weight, body weight gain and food consumption were noted at any dose level investigated. Thyroid hormones (T3, T4 and TSH) evaluated in parental males did not show any changes of toxicological relevance. No remarkable differences were noted at post mortem examination including organ weights and no treatment-related macroscopic and microscopic changes were observed in treated animals, when compared to controls. Futhermore, no effects on the spermatogenic cycle were described. No intergroup differences were seen in oestrous cycle, pre-coital intervals, copulatory and fertility indices.

No significant differences were observed in the number of corpora lutea, implantations, total litter size, pre-implantation loss, pre-birth loss and gestation length between control and treated females. Litter data at birth and on Days 1, 4 and 13 post partum and sex ratios were also comparable between treated and control females.

There were neither treatment-related signs at clinical observation of pups, nor at necropsy of deceased pups or those sacrificed after culling or at term. The statistically significant decrease noted in the ano-genital distance of female pups on Day 1 of age in all treated groups cannot be considered as adverse, since it had a positive trend (feminisation effect).

No nipples were observed on Day 13 of age, in male pups at all dose levels. Increases of circulating T3 levels were found in mid- and high dose male pups sacrificed at Day 14 of age. However, no clear consistency of hormones variations, dose and/or sex relation were noted to finally address an adverse effect of the test compound on thyroid function.

Analysis of plasma samples showed that after oral administration of the test item, females were exposed to the test item (detected as Core 134) both on Day 18 post coitum and Day 13 post partum, as well as, via milk, in pups on Day 13 post partum. The exposure increased with the dose and the duration of treatment (Day 18 post coitum corresponds to approximately 33 days of treatment; Day 13 post partum corresponds to approximately 50 days of treatment.

Based on the results obtained in this study, the NOAEL (No Observed Adverse Effect Level) for both general toxicity and reproduction/developmental toxicity was considered to be 1000 mg/kg/day for both males and females.

These results were used to set the dose levels for the subsequent EOGRTS.

 

Extended One Generation Reproduction Toxicity Study

In an extended one generation reproduction toxicity study in accordance with OECD TG 443 (adopted on 25 June 2018) the pre- and post-natal effects of MDEA-Esterquat C16-18 and C18 unsatd. on development, as well as a thorough evaluation of systemic toxicity were investigated in male and female rats of the parental generation treated for 10 weeks before pairing and during gestation and lactation period.

40 male and 40 female pups/group, from different litters, were selected and assigned to Cohort 1 A and Cohort 1 B in order to follow the toxicity effect in the second generation administered from weaning (Day 21 of age) up to 13 weeks.

In addition, the study provided and/or confirmed information about the effects of the test item on the integrity and performance of the adult male and female reproductive systems. The following parameters were considered: gonadal function, oestrous cycle, epididymal sperm maturation, mating behaviour conception, pregnancy, parturition and lactation.

 

Parental generation (F0)

The dose levels of 100, 300 and 1000 mg/kg/day were selected for parental animals and were administered orally by gavage. The control group received softened water.

Males were treated for 10 weeks prior to pairing, through the mating period and thereafter until the day before necropsy, for a total of 196-100 days.

Females were treated for 10 weeks prior to pairing, during mating, gestation and post partum periods until Day 21 post partum, for at least 85 days.

The number of females with live pups on Days 21/22 post partum was: 21 in the control, 22 in the low dose (100 mg/kg/day), 24 in the mid-dose (300 mg/kg/day) and 22 in the high dose (1000 mg/kg/day) groups.

At the daily and weekly clinical observation, no signs considered adverse and no effect in the neurotoxicity assessment were observed.

Body weight, body weight gain and food consumption were unaffected by treatment.

No changes that could be considered adverse were seen in haematology, coagulation and clinical chemistry parameters of treated animals compared to controls.

Changes noted in thyroid hormone levels of treated males and high dose females were considered to be unrelated to treatment since the simultaneous changes of two or three hormones (conditions which could represent a pathological significance) were sporadic and not dose-related, and no histopathological changes were recorded.

Exposure to Core 134 (metabolite used to monitor the presence of the test item in plasma) was demonstrated in all treated dams on Day 18 post coitum and Day 21 post partum at 1 and 4 hours after treatment and in pups of Groups 3 and 4 on Day 21 post partum, after approximately 24 hours of treatment of the respective dams. No exposure to Core 134 was detected in pups of Group 2.

No treatment-related anomalies were noted in the oestrous cycle of the treated females, when compared to controls. Copulatory and fertility indices did not show any treatment-related differences among treated and control groups. Implantation, pre-natal loss, litter data and sex ratio did not show any changes of toxicological relevance. No significant differences in the anogenital distance were seen between control and treated groups both for male and female pups. No nipples were observed in male pups. Clinical signs and findings at necropsy and organs weight did not reveal any treatment-related or adverse effect.

Sperm analysis performed in all treated males was comparable to controls. Enumeration of ovarian follicles performed in control and high dose females did not show any treatment-related effects. No relevant changes were seen in bone marrow evaluation.

No treatment-related changes were noted in animals sacrificed at the end of treatment at organ weight, macroscopic and microscopic examination.

 

Cohorts 1A and 1B

No mortality occurred in Cohort 1A and Cohort 1B animals.

At the daily and weekly clinical observation, no signs of toxicological relevance and no effect in the neurotoxicity assessment were observed in treated males and females of both Cohorts.

Body weight, body weight gain and food consumption of both sexes of both Cohorts were unaffected by treatment.

Oestrous cycle, vaginal opening and balano-preputial skin folds separation did not eveal

differences considered adverse in Cohort 1A and Cohort 1B animals.

No changes were recorded in the haematology, coagulation, clinical chemistry and urinalysis parameters between treated and control animals of Cohort 1A.

The increase in Triiodothyronine level noted in low dose males was considered unrelated to treatment, in the absence of a of dose-relation.

No sign of alteration in the immune cell distribution was observed in splenocytes of treated animals of Cohort 1A.

No treatment-related changes were noted at sperm analysis (in Cohort 1 A), organ weights, gross pathology and the histopathological examination (in Cohort 1 A). Enumeration of ovarian follicles enumeration and the staging of spermatogenic cycle performed did not show any treatment-related effects in Cohort 1A and Cohort 1B.

 

In conclusion, the dosage of 1000 mg/kg/day was considered the NOAEL for general and reproductive toxicity and pups development in Parental generation and for general toxicity in Cohort 1A and 1B.

 

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.

Effects on developmental toxicity

Description of key information
NOEL=1000 mg/kg bw/d, prenatal developmental toxicity study; oral (gavage); rat (Wistar, 25/group, dosed from day 6 through 15 post coitum); OECD Guideline 414; GLP; read across from MDEA-Esterquat C16-18 and C18 unsatd. 
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
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
This read-across hypothesis corresponds to Scenario 2 of the Read-Across Assessment Framework (RAAF), ECHA, March 2017.

This scenario covers the analogue approach for which the hypothesis is based on different compounds with the same type of effect(s). For the REACH information requirement under consideration, the effects obtained in a study conducted with one source substance are used to predict the effects that would be observed in a study with the target substance if it were to be conducted. The same type of effect(s) or absence of effect is predicted. The predicted strength of the effects may be similar or based on a worst-case approach.

Namely, the absence of adverse effects observed in two studies with the structurally similar source substance MDEA-Esterquat C16-18 and C18 unsatd. is used to predict the toxicological properties of the target substance Partially unsaturated TEA-Esterquat. In this case, the absence of adverse effects is predicted for the endpoints toxicity to reproduction and for prenatal developmental toxicity.

The read-across from the existing toxicological studies conducted with the source substances are considered as an appropriate adaptation to the standard information requirements of the REACH Regulation for the target substance, in accordance with the provisions of Annex XI, 1.5 of the REACH Regulation. The justification of the proposed read-across approach is elaborated in the next chapters.

Property-specific read-across hypothesis
The read-across approach will be applied for the endpoints toxicity to reproduction and developmental toxicity.

The property specific read-across hypothesis is that both source and target substances have similar properties (absence of effects) because:
• they share structural similarities with common functional groups: One quaternised ethanolamine moiety, one to three ethanol ester groups but mainly two ester groups, each with typical UVCB distribution with similar long-chain fatty acids of natural origin.
• they are manufactured from comparable (triethanolamine or diethanolamine) or nearly identical precursors (long-chain fatty acids), methyl-quaternization agents dimethyl sulfate or methyl chloride) under similar conditions. Therefore, comparable breakdown products via physical and biological processes (hydrolysis and biological degradation resp. metabolization), which result in partially structural similar chemicals are evident.

The following supporting evidence is provided:
Anchor studies for the target substance:
• To predict the absence of effects of a prenatal developmental toxicity study, a reproductive toxicity screening study conducted with the target and source substance is provided, and a prenatal developmental toxicity study in rats with the source substance is provided.
• To predict the absence of effects of an EOGRTS study, a reproductive toxicity screening study conducted with the target and source substance is provided, and an EOGRTS study with the source substance is provided.
• To further strengthen the evidence, repeated dose toxicity studies are provided for both, source and target substances.
Consistency is demonstrated by the absence of quantitative variations in the effects (or absence of effect(s)) for both substances and for all relevant properties.


2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)

see attacted justification for read-across

3. ANALOGUE APPROACH JUSTIFICATION

see attacted justification for read-across

4. DATA MATRIX

see attacted justification for read-across
Reason / purpose for cross-reference:
read-across: supporting information
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Species:
rat
Route of administration:
oral: gavage
Dose / conc.:
50 mg/kg bw/day (actual dose received)
Dose / conc.:
250 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
25/group.
Control animals:
yes, concurrent vehicle
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Basis for effect level:
other: maternal toxicity
Abnormalities:
no effects observed
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:
no effects observed
Developmental effects observed:
no
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)
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

No experimental data are available for the target substance partially unsatd. TEA-Esterquat. However, a prenatal develomental toxicity study is available for the closely related source substance MDEA-Esterquat C16-18 and C18 unsatd. A justification for read-across is attached to iuclid section 13.

 

In the developmental toxicity study, 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, 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. 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.

There are indications that the slightly increased incidences of post-implantation losses at 1000 mg/kg bw/day are due to some maternal (toxic) effects, which could not be further evaluated because this dose level has not been tested in the repeated dose toxicity studies.

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. Other long-term studies with this high dose level are not available to further evaluate substance-related toxicity. Therefore, the impact of the maternal effects cannot be evaluated in depth. 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/day. Therefore, the aspect of prenatal developmental toxicity is sufficiently covered.

Additional data from another substance of the esterquat family (HEQ-based esterquat, CAS-no 19467-38-0) are described in the HERA RAR Esterquats (2009). There was no increased embryolethality, foetotoxicity, nor any specific defect in the foetuses which could be attributable to maternal exposure to the HEQ-based esterquat up to 1000 mg/kg bw/day in the rat.

 

Performance of a prenatal developmental toxicity study in a second species is not necessary in accordance with Annex XI (3.2 (a)) of REACH Regulation (EC) No 1907/2006 based on a quantitative assessment. 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, consumer use and indirect exposure of humans via the environment. The very small spectrum of identified uses as referred to in Annex VI section 3.5 is documented in chapters 9 and 10 in this CSR. The exposure assessment and risk characterisation according to Article 14(4) and Annex I, 5 of REACH Regulation (EC) No 1907/2006 have been conducted solely for the purpose of justifying this waiver, given that the substance is not classified as dangerous or as PBT/vPvB according to any of the Regulation’s criteria.

Strictly conservative DNELs have been derived from results of the available test data which include a pre-natal developmental study and subacute and subchronic 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 the omission of the information requirement by 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 relevant and appropriate both for the information requirement to be omitted and 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 classification or non-classification

Based on the available data, partially unsatd. TEA-Esterquat does not need to be classified for reproductive toxicity (fertility and development) according to regulation (EC) 1272/2008. Thus, no labelling is required.

Additional information