Propylidynetrimethanol, ethoxylated, esters with acrylic acid, reaction products with diethylamine

Endpoint summary

Key value for chemical safety assessment

Toxic effect type:

dose-dependent

Effects on fertility

Description of key information

A screening study is available to evaluate the reprotoxic potential of Diethylamine modified ethoxylated trimethylolpropane triacrylate (OECD 421).
The test item was administered daily by oral gavage to male and female Sprague-Dawley rats, for 2 weeks before pairing, during pairing, and for females, during gestation and until day 4p.p., at dose-levels of100, 300 or 1000/600 mg/kg/day. There were adverse findings in animals treated with 1000/600 mg/kg/day: three test item-related deaths, marked clinical signs (piloerection, hypoactivity, emaciated appearance, breathing difficulties), lower body weight gain, body weight losses and lower food consumption. There were no toxicologically important effects on reproductive, developmental and pathology parameters.
Based on the experimental conditions of this study: the NOAEL for parental toxicity was considered to be 300 mg/kg/day, and the NOAEL for reproductive performance (mating and fertility) and the NOAEL for toxic effects on progeny were considered to be 600 mg/kg/day (the highest dose).

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

12 October 2012 - 25 January 2013

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)

Version / remarks:

1995

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: breeder: Charles River Laboratories Italia, Calco, Italy
- Age/mean body weight at study initiation: on the first day of treatment, the males were approximately 10 weeks old and had a mean body weight of 370 g (range: 339 g to 407 g) and the females were approximately 9 weeks old and had a mean body weight of 208 g (range: 174 g to 239 g)
- Fasting period before study: no
- Housing: the females were individually housed, except during pairing and lactation, in polycarbonate cages. The males were individually housed, except during pairing, in wire-mesh cages.
- Diet: SSNIFF R/M-H pelleted diet (free access)
- Water: tap water filtered with a 0.22 µm filter (free access)
- Acclimation period: 7 days before the beginning of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C except once up to +25.2°C
- Humidity (%): 50 ± 20%
- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air
- Photoperiod (hrs dark / hrs light): 12 h/12 h.

IN-LIFE DATES: 23 October 2012 to 16 December 2012.

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
The test item was administered as a solution in the vehicle. It was mixed with the required quantity of vehicle. The dose formulations were kept under magnetic stirring at least 30 minutes before delivery to the study room.
No correction factor was applied to the calculation of dose levels.
The frequency of dose formulation preparation was on a daily basis. The dose formulations were delivered to the study room at room temperature and protected from light.

VEHICLE
- Vehicle choice: test item soluble in corn oil which is commonly used in this type of study
- Concentration in vehicle: 20, 60, 200 (from days 1 to 15) and 120 mg/mL (from day 16)
- Amount of vehicle (if gavage): 5 mL/kg/day.

Details on mating procedure:

- M/F ratio per cage: 1/1
- Length of cohabitation (mating period): until mating occured or 14 days has elapsed then with another male until mating occured within 7 days
- Proof of pregnancy: vaginal plug or sperm in the morning vaginal lavage referred to as day 0 post-coitum
- After successful mating each pregnant female was caged individually

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Type of method: GC-FID
Test item concentrations: remained within an acceptable range of variation compared to nominal values.
Homogeneity: not assessed, dose formulation is a solution
Stability: not assessed, dose formulation prepared daily

Duration of treatment / exposure:

In the males:
- 2 weeks before mating,
- during the mating period,
- until sacrifice (i.e. at least 5 weeks in total),

In the females:
- 2 weeks before mating,
- during the mating period,
- during pregnancy,
- during lactation until day 4 post-partum inclusive,
- until sacrifice for females which had not delivered.

Frequency of treatment:

Daily (except for a few animals which were not treated for one day during a premating period because of poor health condition).

Details on study schedule:

- No F1 parents (only one generation mated)
- Age at mating of the mated animals in the study: 11-12 weeks

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)

Remarks:

1000 (from days 1 to 15) and 600 mg/kg/day (from day 16)

No. of animals per sex per dose:

10 animals per sex per dose.

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale:
The dose-levels were selected in agreement with the Sponsor, following the results of a previous 4-week toxicity study performed in the same species and strain. In this study, three groups of 5 males and 5 females received the test item by gavage at 100, 300 or 1000 mg/kg/day as a solution in corn oil. Another group of five males and five females received the vehicle only. All high-dose animals and half of the mid-dose animals had ptyalism. A few animals had hunched posture, loud breathing and/or thin appearance towards the end of the treatment period, particularly the males. Males had a mean body weight slightly lower than controls, together with slightly reduced food consumption at the end of the treatment period. There were no obvious effects in mean hematology, blood biochemistry, urinalysis, organ weight and macroscopic examination results.
As there were no adverse effects in the 4-week toxicity study, the same dose-levels were selected for the study (100, 300 or 1000 mg/kg/day).

- Rationale for animal assignment: computerized stratification procedure.

Positive control:

no (not required)

Parental animals: Observations and examinations:

MORTALITY / MORBIDITY
- Time schedule: at least twice a day during the treatment period.

CLINICAL OBSERVATIONS:
- Time schedule: once a day during the treatment period.

BODY WEIGHT:
- Time schedule: Males: on the first day of treatment, then once a week until sacrifice. Females: on the first day of treatment, then once a week until mating, on Days 0, 7, 14 and 20 post-coitum and days 1 and 5 post-partum.

FOOD CONSUMPTION:
- Time schedule: Males: on the first day of treatment, then once a week until pairing. Females: on the first day of treatment, then once a week until pairing, on Days 0, 7, 14 and 20 post-coitum and days 1 and 5 post-partum.

REPRODUCTION (apart from indices):
- Pre-coital time and duration of gestation were recorded

Oestrous cyclicity (parental animals):

fresh vaginal lavage (stained with methylene blue), each morning during the pairing period, until females are mated.

Sperm parameters (parental animals):

Parameters examined in males of parental generation:
- testis weight (all groups) + microscopic evaluation (control and high-dose groups)
- epididymis weight (all groups) + microscopic evaluation (control and high-dose groups)
- microscopic evaluation of stages of the spermatogenic cycle and testicular interstitial cells (control and high-dose groups).

Litter observations:

STANDARDISATION OF LITTERS: No

PARAMETERS EXAMINED:
- number and sex of pups,
- number of live, dead and cannibalized pups,
- presence of gross anomalies, weight gain, clinical signs.

GROSS EXAMINATION OF DEAD AND SURVIVING PUPS:
- external and internal abnormalities.

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: all surviving animals after the end of the mating period after at least 5 weeks of treatment
- Female animals: all surviving animals = day 5 post-partum or, for females which had not delivered yet, day 26 post-coitum

GROSS NECROPSY
Macroscopic post-mortem examination of principal thoracic and abdominal organs

HISTOPATHOLOGY
A microscopic examination was performed on:
- epididymides, testes, ovaries from all animals of the control and high-dose groups (groups 1 and 4) sacrificed at the end of the treatment period (at the end of the pairing period for males and on day 5 p.p. for females) and from prematurely dead animals,
- all macroscopic lesions of all groups.

Special emphasis was paid to the stages of spermatogenesis in the male gonads and histopathology of interstitial testicular cell structure.


ORGAN WEIGHTS: epididymides, testes, ovaries.

Postmortem examinations (offspring):

SACRIFICE: on day 5 post-partum

GROSS NECROPSY: on all pups (surviving and found dead)

HISTOPATHOLOGY: No

ORGAN WEIGTHS: No

Statistics:

Yes

Reproductive indices:

Pre-implantation loss = 100 * (Number of corpora lutea - Number of implantation sites) / Number of corpora lutea
Post-implantation loss = 100 * (Number of implantation sites - Number of live pups / Number of implantation sites
Mating index = 100 * (Number of mated animals / Number of paired animals)
Fertility index = 100 * (Number of pregnant female partners / Number of mated pairs)
Gestation index = 100 * (Number of females with live born pups / Number of pregnant females)

Offspring viability indices:

Live birth index = 100 * (Number of live born pups / Number of delivered pups)
Viability index on day 4 p.p. = 100 * (Number of surviving pups on day 4 p.p. / Number of live born pups).

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Piloerection, hypoactivity, emaciated appearance and breathing difficulties noted at 1000 mg/kg/day in surviving animals were similar to those observed in prematurely dead animals and were considered to be test item-related and adverse. Breathing difficulties were observed within the first treatment days and were still noted in some females during gestation and lactation even after the reduction of the dose-level to 600 mg/kg/day. These breathing difficulties were sometimes, for a few animals, present before the next treatment. Piloerection, hypoactivity and emaciated appearance appeared in the second half of treatment at 1000 mg/kg/day and lasted a few days only.
Ptyalism was observed with a dose-relationship in incidence and duration from 100 mg/kg/day in males and females. This finding was considered not to be adverse.
One female at 1000/600 mg/kg/day had piloerection, pallor of extremities and emaciated appearance throughout lactation. A relationship between its clinical signs and the test item treatment was considered to be unlikely and more related to gestation and parturition problems and incidental. Indeed, it was the only animal with these clinical signs appearing at the end of the treatment period. In addition, this female had the longest duration of gestation (23 days), the highest post-implantation loss (36%) and the lowest litter weight (mean pup body weight of 5.3 g vs. mean of 7.2 in controls), with only seven live pups on day 1 p.p. which were dehydrated and thin until scheduled death on day 5 p.p.

Dermal irritation (if dermal study):

not examined

Mortality:

mortality observed, treatment-related

Description (incidence):

At 1000 mg/kg/day, there were three premature deaths which were ascribed to the test item treatment: one male prematurely sacrificed on day 2, and one male and one female on day 11. Poor health condition preceding the premature sacrifice included piloerection, pallor of extremities, cold to the touch, hypoactivity, breathing difficulties and/or emaciated appearance. Marked body weight loss was recorded for these animals before sacrifice.
The cause of moribundity was not evident at macroscopic and microscopic examinations.
There was at 1000 mg/kg/day one found dead female on day 15. Only loud breathing and ptyalism were noted on the day before and the female was gaining weight from day 8. There were no relevant findings at pathology. A relationship with the test item treatment was unclear.
There were no other unscheduled deaths during the study, in particular after the decrease of the high dose-level.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Males treated at 1000 mg/kg/day had a mean body weight gain severely lower than controls in the first week of treatment. In the second week of the treatment period, mean body gain became comparable to that of the controls (the slightly lower value was due to one male with body weight loss). Thereafter, there was a dose related higher mean body weight gain than controls in test item groups in males. These changes of body weight gain in males did not toxicologically impact the terminal mean body weight but were considered to be adverse in view of the severity of the differences from controls.

There were no toxicologically important effects on female mean body weight and mean body weight gain during premating and gestation. In lactation, females given 1000/600 mg/kg/day had a lower mean body weight gain than controls.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

Males treated at 1000 mg/kg/day had a mean food consumption lower than controls (-19%) in the first week of treatment, which correlated with the effects on mean body weight gain seen previously and was considered to be adverse. In the second week of the treatment period, mean food consumption became comparable to that of the controls (the slightly lower value was due to one male with low food consumption).
There were no toxicologically important effects on female mean food consumption during pre mating and gestation. During lactation, there was a trend towards a lower mean food consumption when compared with controls, which correlated with the effects on mean body weight gain seen previously, and was considered to be of minor toxicologically relevance.

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:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

There were no macroscopic findings or changes in mean organ weights attributed to the test item
administration. There were no significant microscopic findings noted in the testes, epididymides
from males or in the ovaries or oviducts from pregnant females.


Premature deaths
The cause of moribundity/death of these four animals was not evident at microscopic examination (restricted to macroscopic findings in both sexes, testes and epididymides in males, and ovaries with oviducts in females).
In both males, there were microscopic findings in the stomach which were suggestive of low grade superficial injury. This could have been directly related to the test item or could have been secondary to stress.
In one male sacrificed on day 2, there was a prominent mucus layer overlying the gastric mucosa, which correlated with macroscopic translucent content, and contained a few neutrophils. There were no microscopic correlates to the yellow liquid content in the ileum.
In one male sacrificed on day 11, in the stomach, there were no microscopic correlates to gas distension, however minimal multifocal erosions associated with minimal infiltrate of neutrophils were present in the mucosa of the forestomach. Marked lymphoid atrophy of the thymus correlated with small size at necropsy and was suggestive of stress. Moderate dilation of the lumen correlated with gas distension in the colon. There were no microscopic correlates to the gas distension recorded in the small intestine and cecum.
In one female sacrificed on day 11, there were no microscopic correlates to the gas distension recorded in the ileum.
There were no significant microscopic changes in one female found dead on day 15.

Terminal sacrifice
There were no significant microscopic findings noted in the testes, epididymides from males or in the ovaries or oviducts from pregnant females.
There were no significant microscopic findings in both females sacrificed on day 26 p.c. and not pregnant. In one female given the test item at 100 mg/kg/day, the morphology of the uterine epithelium (large tall columnar epithelium with single cell necrosis but devoid of mitotic figures) was suggestive of early estrus; there were no microscopic correlates to the thickened uterine horns noted at necropsy. In one control female, cells of the corona radiata were present in the lumen of the oviducts, which suggested that ovulation had just happened.

Histopathological findings: neoplastic:

not examined

Other effects:

not examined

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

Description (incidence and severity):

There were no effects on pairing (precoital time) and on mating and fertility indexes.

The mean number of days of pairing before mating was slightly higher in the 100 mg/kg/day group because one pair mated after 14 days instead of within 4 days as for all the other pairs (mean of 2.0 days when excluding this female). This was considered to be incidental.

No evidence of mating was observed for one female (300 mg/kg/day) and its first male and the female was then paired with a second male for several days, but pairing was prematurely stopped because of suspicion of gestation. The female was indeed pregnant and gave birth. Reprotox software automatically back calculated the theoretical pre-coital time taking into account the day of parturition and the usual length of rat gestation (21 days). The theoretical day of mating indeed coincided with the only estrus phase noted for this female.

There were no relevant effects on delivery data.
The only difference from controls was a slightly higher mean post-implantation loss at 1000/600 mg/kg/day, particularly due to one female which had a high post-implantation loss (36%). When compared with Historical Control Data, a test item treatment related effect was considered unlikely.
Gestation duration for the female (300 mg/kg/day) with mating evidence not diagnosed was considered to be similar to the other animals and of 21 days taking into account the parturition date and the estrous day at the beginning of the mating period.

Dose descriptor:

NOAEL

Remarks:

Parental toxicity

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

clinical signs

mortality

body weight and weight gain

food consumption and compound intake

Dose descriptor:

NOAEL

Remarks:

Reproductive performance

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

reproductive performance

Remarks on result:

not determinable due to absence of adverse toxic effects

Critical effects observed:

no

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

There were no test item-related clinical signs in pups.

Seven pups with dehydration and thin appearance from birth to scheduled sacrifice were from one litter (1000/600 mg/kg/day) and this was considered incidental.
Other incidental clinical signs of test item pups included limb or tail necrosed, hematoma (head), scab and/or wound (abdomen, head), dehydration. The nature and incidence of these signs can be commonly observed in pups of this age or was observed with low or similar incidence as controls, and were considered not toxicologically relevant.

Dermal irritation (if dermal study):

not examined

Mortality / viability:

no mortality observed

Description (incidence and severity):

There were no test item-related deaths or clinical signs in pups.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

There were no relevant effects on mean pup body weights and mean pup body weight changes.

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):

not examined

Nipple retention in male pups:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

There were no test item-related macroscopic findings at pup necropsy.
Incidental macroscopic findings of pups included autolysis (2, 0, 1, 1 dead pups in groups of 0, 100, 300 and 1000/600 mg/kg/day respectively), absence of milk in stomach (0, 4, 11, 3 dead and scheduled sacrificed pups) and an irregular color of the liver was observed in one pup (1000/600 mg/kg/day).

Histopathological findings:

not examined

Other effects:

not examined

Behaviour (functional findings):

not examined

Developmental immunotoxicity:

not examined

Dose descriptor:

NOAEL

Remarks:

Toxic effects on progeny

Generation:

F1

Effect level:

600 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Remarks on result:

not determinable due to absence of adverse toxic effects

Critical effects observed:

no

Reproductive effects observed:

no

Conclusions:

The test item was administered daily by oral gavage to male and female Sprague-Dawley rats, for 2 weeks before pairing, during pairing, and for females, during gestation and until day 4 p.p., at dose-levels of 100, 300 or 1000/600 mg/kg/day in the reproduction screening test (OECD 421).
 There were adverse findings in animals treated with 1000/600 mg/kg/day: three test item-related deaths, marked clinical signs (piloerection, hypoactivity, emaciated appearance, breathing difficulties), lower body weight gain, body weight losses and lower food consumption. There were no toxicologically important effects on reproductive, developmental and pathology parameters.

Based on the experimental conditions of this study:
. the No Observed Adverse Effect Level (NOAEL) for parental toxicity was considered to be 300 mg/kg/day,
. the NOAEL for reproductive performance (mating and fertility) was considered to be 600 mg/kg/day,
. the NOAEL for toxic effects on progeny was considered to be 600 mg/kg/day.

Executive summary:

The objective of this study was to evaluate the potential toxic effects of the test item following daily oral administration (gavage) to male and female rats from before mating, through mating and, for females, through gestation until day 4 post-partum (p.p.), based on the OECD guideline No. 421, 27 July 1995.

This study provided information on male and female reproductive performance, such as gonadal function, mating behavior, conception, development of the conceptus and parturition.

The study was conducted in compliance with the principles of Good Laboratory Practice Regulations.

 

Methods

Three groups of ten male and ten female Sprague-Dawley rats received the test item daily by oral (gavage) from before pairing, through pairing and, for the females, through gestation until day 4 p.p. The test item was administered as a solution in the vehicle,corn oil, at dose-levels of 100, 300 or 1000/600 mg/kg/day. The high-dose animals was first treated at 1000 mg/kg/day until day 15 of treatment and then at 600 mg/kg/day from day 16 following strong clinical signs and premature deaths in some animals.

Another group of ten males and ten females received the vehicle alone under the same experimental conditions and acted as a control group. A constant dosage-volume of 5 mL/kg/day was used.

The concentration of the dose formulation was checked in study weeks 2, 3 and 6.

The animals were checked daily for mortality, morbidity and clinical signs. Body weight and food consumption were recorded approximatelyonce a week. The animals were paired for mating after 2 weeks of treatment and the dams were allowed to litter and rear their progeny until day 5 p.p. The total litter sizes and numbers of pups of each sex were recorded after birth. The pups were observed daily for clinical signs, abnormal behaviour and external abnormalities and weighed on days 1 and 5 p.p.

The males were sacrificed after 5 weeks of treatment and the dams on day 5 p.p. Final body weights and selected organs weights (epididymides, testes, ovaries) were recorded and a macroscopic post-mortem examination of the principal thoracic and abdominal organswas performed, with particular attention paid to the reproductive organs. A microscopic examination was performed on selected organs (epididymides, testes, ovaries) from the control and high-dose groups and on all macroscopic lesions.

The pups were sacrificed on day 5 p.p. and submitted for a macroscopic post-mortem examination of the principal thoracic and abdominal organs.

Results 

The test item concentrations in the administered dose formulations analyzed in weeks 2, 3 and 6 were within the acceptance criteria (± 10% of the nominal concentrations).

Mortality

At 1000 mg/kg/day, there were three premature deaths which were ascribed to the test item treatment. The affected animals had similar clinical signs as the surviving animals and lost weight before death. However, the cause of moribundity was not determined at macroscopic and microscopic examinations for these prematurely sacrificed animals for ethical reasons. At necropsy, changes in the stomach (gas distension/dilation with translucent content) and intestines (gas distension/yellow liquid) seen in these animals were considered to be most likely related to the test item. In both males, there were microscopic findings in the stomach (prominent mucus/forestomach erosion and neutrophilic infiltrate) which were suggestive of low grade superficial injury. This could have been directly related to the test item or could have been secondary to stress. In one male, lymphoid atrophy of the thymus was also suggestive of stress. There was also at 1000 mg/kg/day one found dead female on dayrelationship with the test item treatment was unclear.

 

Clinical signs

Piloerection, hypoactivity, emaciated appearance and breathing difficulties noted at 1000 mg/kg/day in 14/16 surviving animals were considered to be test item-related and adverse. Ptyalism was observed with dose-related incidence and duration from 100 mg/kg/day.

 

Body weight, body weight gain and food consumption

At 1000 mg/kg/day and when compared with controls, males had a lower mean body weight gain during the first week of treatment (+10 g, vs.+39 g, p<0.001) associated with a lower mean food consumption (-19%, p<0.01). These effects were considered to be adverse. In the second half of the treatment period, there were dose-related higher mean body weights in males from test item groups (up to +48%, p<0.05). There were no toxicologically important effects on female mean body weight, mean body weight gainand mean food consumption during premating and gestation periods. In lactation, females given 1000/600 mg/kg/day had lower mean body weight gain (+3 g,vs.+17 g; p<0.05) and mean food consumption (-11%) than controls.

 

Reproductive data

There were no effects on pairing, mating and fertility data.There were no toxicological relevant effects on mean delivery data.

 

Histopathology

There were no macroscopic findings or changes in mean organ weights attributed to the test item administration. There were no significant microscopic findings noted in the testes, epididymides from males or in the ovaries or oviducts from pregnant females.

 

Pups

There were no test item-related deaths or clinical signs in pups andno toxicological relevant effects on mean pup body weights, mean pup body weight gains oron the percentage of male pups at birth. There were no test item-related macroscopic findings at pup necropsy.

Conclusion

The test item was administered daily by oral gavage to male and female Sprague-Dawley rats, for 2 weeks before pairing, during pairing, and for females, during gestation and until day 4 p.p., at dose-levels of 100, 300 or 1000/600 mg/kg/day.

There were adverse findings in animals treated with 1000/600 mg/kg/day: three test item-related deaths, marked clinical signs (piloerection, hypoactivity, emaciated appearance, breathing difficulties), lower body weight gain, body weight losses and lower food consumption. There were no toxicologically important effects on reproductive, developmental and pathology parameters.

 

Based on the experimental conditions of this study:

. the No Observed Adverse Effect Level (NOAEL) for parental toxicity was considered to be 300 mg/kg/day,

. the NOAEL for reproductive performance (mating and fertility) was considered to be 600 mg/kg/day,

. the NOAEL for toxic effects on progeny was considered to be 600 mg/kg/day.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

600 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

This screening test is reliable, performing according to the OECD guideline n°421 (klimisch score of 1).

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

Screening study on the reproduction toxicity (OECD 421):

The objective of this study was to evaluate the potential toxic effects of the test item following daily oral administration (gavage) to male and female rats from before mating, through mating and, for females, through gestation until day 4 post-partum (p.p.). This study provided information on male and female reproductive performance, such as gonadal function, mating behavior, conception, development of the conceptus and parturition.

Three groups of ten male and ten female Sprague-Dawley rats received the test item daily by oral (gavage) from before pairing, through pairing and, for the females, through gestation until day 4 p.p.The test item was administered as a solution in the vehicle,corn oil, at dose-levels of 100, 300 or 1000/600 mg/kg/day. The high-dose animals was first treated at 1000 mg/kg/day until day 15 of treatment and then at 600 mg/kg/day from day 16 following strong clinical signs and premature deaths in some animals. Another group of ten males and ten females received the vehicle alone under the same experimental conditions and acted as a control group. A constant dosage-volume of 5 mL/kg/day was used.

 

At 1000 mg/kg/day, there were three premature deaths which were ascribed to the test item treatment. The affected animals had similar clinical signs as the surviving animals and lost weight before death. However, the cause of moribundity was not determined at macroscopic and microscopic examinations for these prematurely sacrificed animals for ethical reasons. At necropsy, changes in the stomach (gas distension/dilation with translucent content) and intestines (gas distension/yellow liquid) seen in these animals were considered to be most likely related to the test item. In both males, there were microscopic findings in the stomach (prominent mucus/forestomach erosion and neutrophilic infiltrate) which were suggestive of low grade superficial injury. This could have been directly related to the test item or could have been secondary to stress. In one male, lymphoid atrophy of the thymus was also suggestive of stress. There was also at 1000 mg/kg/day one found dead female on dayrelationship with the test item treatment was unclear.

Piloerection, hypoactivity, emaciated appearance and breathing difficulties noted at 1000 mg/kg/day in 14/16 surviving animals were considered to be test item-related and adverse. Ptyalism was observed with dose-related incidence and duration from 100 mg/kg/day.

At 1000 mg/kg/day and when compared with controls, males had a lower mean body weight gain during the first week of treatment (+10 g,vs.+39 g, p<0.001) associated with a lower mean food consumption (-19%, p<0.01). These effects were considered to be adverse. In the second half of the treatment period, there were dose-related higher mean body weights in males from test item groups (up to +48%, p<0.05). There were no toxicologically important effects on female mean body weight, mean body weight gainand mean food consumptionduring premating and gestation periods. In lactation, females given 1000/600 mg/kg/day had lower mean body weight gain (+3 g,vs.+17 g; p<0.05) and mean food consumption (-11%) than controls.

There were no macroscopic findings or changes in mean organ weights attributed to the test item administration. There were no significant microscopic findings noted in the testes, epididymides from males or in the ovaries or oviducts from pregnant females.

 

There were no effects on pairing, mating and fertility data.There were no toxicological relevant effects on mean delivery data.

There were no test item-related deaths or clinical signs in pups andno toxicological relevant effects on mean pup body weights, mean pup body weight gains or on the percentage of male pups at birth. There were no test item-related macroscopic findings at pup necropsy.

 

Based on the experimental conditions of this study:

. the No Observed Adverse Effect Level (NOAEL) for parental toxicity was considered to be 300 mg/kg/day,

. the NOAEL for reproductive performance (mating and fertility) was considered to be 600 mg/kg/day,

. the NOAEL for toxic effects on progeny was considered to be 600 mg/kg/day.

Effects on developmental toxicity

Description of key information

A developmental toxicity study in rat is proposed in this dossier.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no study available (further information necessary)

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Justification for classification or non-classification

Based on the results of the screening test on reproduction, no classification for Diethylamine modified ethoxylated trimethylolpropane triacrylate is required for reprotoxicity according to the Regulation EC n°1272/2008.