Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

Martell AB (2013) performed a combined repeated dose toxicity study with reproduction/developmental toxicity screening test in rats according to OECD guideline 422 (GLP). A NOAEL of 300 mg/kg bw/day was derived.

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2012-05-15 to 2012-07-08
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
Well documented GLP study performed according to OECD Guideline 422. However, dose formulations were not analyzed.
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
yes
Remarks:
no analytical verification of the dose is performed
GLP compliance:
yes (incl. certificate)
Limit test:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: # 1D403

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature protected from humidity and light

OTHER SPECIFICS:
- Name of test material (as cited in study report): Jeffcat DMDEE
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: BIOAGRI Laboratorios
- Age at study initiation: 10 - 11 weeks old
- Weight at study initiation: males: 324.4 - 341.5 grams; females: 204.3 - 217.3 grams; satellite males: 336.0 - 371.0 grams; satellite females: 207.4 - 208.0 grams
- Fasting period before study: no
- Housing: Each animal was housed individually, except during cohabitation. After acclimation, one male was placed into each female cage for pairing. After pairing, females that presented vaginal smears with the presence of sperm were considered mated and housed individually. The rats were housed in polypropylene cages (41x34x19 cm) with wire mesh tops and bedding material (wood shavings). Clean cages were provided twice weekly for all animals. The cages with the test animals were arranged on the racks in such a way that uniform experimental conditions (ventilation and light) were ensured.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.2-22.6°C
- Humidity (%): 40.5-69.9%
- Air changes (per hr): 10-20 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
For each dosage group, the appropriate amount of Jeffcat DMDEE was weighed into a pre-calibrated beaker. The vehicle (deionized water) was added in sufficient quantity to achieve the desired concentration. Each solution was stirred and dispensed into individual containers properly identified. A sufficient quantity of the vehicle was similarly dispended for administration to control animals. The prepared solutions were stored at room temperature.
Details on mating procedure:
Premating:
At the end of the acclimation period, animals were randomly assigned to the experimental groups, housed and the treatment started.

Mating:
After a premating period of 2 weeks, females were cohabited with an assigned male (1 female: 1 male) from the same dose level until evidence of copulation was observed, or 2 weeks had elapsed. Care was taken to avoid sibling mating. Vaginal smears were collected daily during mating period and examined for the presence of sperm. Day 0 of gestation was defined as the day a sperm is found in the vaginal smear. One female that showed no evidence of copulation was assumed pregnant at the end of the mating period, and housed accordingly. Males were euthanized after completing a dosing period of 41 to 42 days.

Gestation:
During gestation period all females were housed individually in the cages. One female without copulation date was housed in an individual cage at the end of the mating period. Female showing no evidence of copulation was euthanized 24-25 days after the last day of mating period.

Lactation:
The day when delivery is completed was designated day 0 of lactation (postnatal day 0). On day 0 of lactation the number of alive and dead pups/sex were recorded. Dams with offspring were euthanized on day 4 postnatal. All pups were euthanized at day 4 postnatal.
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
Parental animals (males and females) were treated with a volume of 4 mL/kg bw, starting at 10-11 weeks old and ending when the animals were euthanized. Satellite animals (5 animals per sex of the control and 5 animals per sex of the high dose group) were kept for 14 days after the scheduled necropsy of parental animals without treatment, for observation of reversibility, persistence or delayed occurrence of toxic effects. Satellite animals were not mated and, consequently, were not used for assessment of reproduction/developmental toxicity.
Frequency of treatment:
7-day per-week-basis
Dose / conc.:
0 mg/kg bw/day (nominal)
Dose / conc.:
50 mg/kg bw/day (nominal)
Dose / conc.:
150 mg/kg bw/day (nominal)
Dose / conc.:
300 mg/kg bw/day (nominal)
No. of animals per sex per dose:
12 in the main study
5 animals per sex of the control group and 5 animals per sex of the high dose group for the satellite group
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale:
The dosage levels (in mg/kg body weight/day) were selected following consultation with the Sponsor.
Parental animals: Observations and examinations:
DETAILED CLINICAL OBSERVATIONS: Yes
All animals underwent a daily clinical observation for overt signs of ill health. These include, but are not limited to, changes in skin and fur, eye and mucous membranes, respiratory, circulatory, autonomic and central nervous system, motor activity and behavioral patterns.

BODY WEIGHT: Yes
Males were weighed on the first day of dosing and weekly thereafter (including mating and post-mating periods). Females were weighed on first day of dosing and once a week during premating and mating periods, on days 0, 7, 14 and 20 of gestation, and during lactation on the same days as the weighing of litters (on days 0 and 4 postnatal).

FOOD CONSUMPTION
Food consumption was determined on the same day of body weight determination during premating and lactation periods, except on day 0. During the gestation period food consumption was determined on days 3, 6, 9, 12, 15, 18 and 20. After the mating period, food consumption of males was determined weekly. Food consumption was not determined for exposed animals during the mating period.

OTHER:
Organ weights: At scheduled necropsy, testes and epididymides of all males were weighed. Organ weights were obtained for the following organs from 5 animals/sex/group:
liver, kidneys, adrenals, thymus, spleen, brain, heart
Sperm parameters (parental animals):
testis weight, epididymides weight
Litter observations:
Live pups were counted, sexed and weighed on days 0 and 4 postnatal.

Postmortem examinations (parental animals):
GROSS PATHOLOGY: Yes
At termination, all parental animals were examined macroscopically for any abnormalities or pathological changes. The animals were euthanized in a carbon dioxide chamber. The number of implantation sites and corpora lutea were recorded. The animals were disposed in biological garbage and then incinerated. All pups were grossly examined for abnormalities of the oral, thoracic and abdominal cavities.

HISTOPATHOLOGY: Yes
At the scheduled necropsy, the following organs of all animals were preserved:
testes, epididymides, ovaries, prostate, semincal vesicle and coagulating gland, bulbourethral gland, organs showing alterations

The following organs and tissues of 5 animals/sex/group were preserved:
adrenals (right and left), bone marrow (femur), brain (cerebrum, cerebellum and pons), esophagus, heart, intestine (duodenum, jejunum, ileum - including Peyer's patches, cecum, colon, rectum/anus), kidneys (right and left), liver (3 lobes), lungs, lymph nodes (mesenteric and submaxillary), pancreas, peripheral nerve (sciatic), spinal cord (cervical, midthoracic and lumbar sections), spleen, stomach (glandular and non-glandular), trachea, thymus, thyroid/parathyroid, urinary bladder, uterus, all gross lesions

Full histopathology of the preserved organs and tissues listed above were performed in highest dose and control animals. The examination of the thymus was extended to females of other dosage groups and satellite females, because changes were observed in the highest dose group.
Postmortem examinations (offspring):
SACRIFICE
All pups were euthanized at day 4 postnatal

GROSS NECROPSY
All pups were grossly examined for abnormalities of the oral, thoracic and abdominal cavities.
Statistics:
Quantitative variables such as body weights, food consumption and organ weights were analyzed by One Way Analysis of Variance (ANOVA), followed by Dunnett's test if significance is detected, or by non-parametric test of Kruskal-Wallis, according to the results of tests for normality and homogeneity of variance. For qualitative or non-parametric data such as clinical findings, macroscopic and microscopic findings and fetal findings, comparison between means were carried out using the Fisher's Exact Test or Chi-Square Test. The level of significance was set at 5%.
Reproductive indices:
The precentage of pre-implantation loss, post-implantation loss, mating index, fertility index, gestation index on day 4 post-partum were calculated (for each pregnant animal) according to the following:
% Pre-implantation loss = (Number of corpora lutea - Number of implantation sites x 100)/Number of corpora lutea
% Post-implantation loss = (Number of implantations - Number of live fetuses x 100)/Number of implantations
% Mating index = (Number of females mated x 100)/Number of females paired
% Fertility index = (Number of females pregnant x 100)/Number of mated pairs
% Gestation index = (Number of females with live pups at birth x 100)/Number of females pregnant
Offspring viability indices:
The percentage of live birth index and viability index on day 4 post-partum were calculated (for each pregnant animal) according to the following:
% Live birth index = (Number of live born pups x 100)/Number of delivered pups
% Viability index on day 4 post-partum = (Number of surviving pups on day 4 post-partum x 100)/Number of live born pups
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
Two females and one male at 300 mg/kg/day presented clinical signs between treatment days 25 to 41. These clinical signs were variable in nature, transitory, occurred in only a few animals, and were not observed in the high dose group satellite animals. The significance of these findings is not clear, but they are unlikely to be specific to the test article.
Mortality:
no mortality observed
Description (incidence):
The test substance did not cause mortality during the study
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Body weight, body weight gain and food consumption were unaffected at all dose levels.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Body weight, body weight gain and food consumption were unaffected at all dose levels.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
Decreased diffuse lymphocyte cellularity in thymus was observed in five females exposed to 150 mg/kg/day (animals 71, 74, 76, 77 and 82), while in females at 300 mg/kg/day was noted a thymus involution in rats 95, 98, 102, 104 and 106. These findings were not observed in treated and satellite males or satellite females, terminal thymus weights were substantially decreased in comparison to the female satellite control group. This observation is most likelly due to the nanimals' physiological response to, and following, pregnancy, and is not considered test article related.
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):
None of the mating or gestation parameters were considered to have been affected by treatment with the test item.
Dose descriptor:
NOAEL
Effect level:
300 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: overall effects
Critical effects observed:
not specified
Clinical signs:
no effects observed
Description (incidence and severity):
No clinical signs were observed in pups until day 4 postnatal.
Mortality / viability:
mortality observed, non-treatment-related
Description (incidence and severity):
On post-natal day 0, two pups at 150 mg/kg/day and three pups at 300 mg/kg/day were found dead. These findings were considered incidental and not related to the test item since they were observed only in a few pups and were not statistically significant. Pup mortality on post-natal day 4 was higher in control than treated groups.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
The mean body weight of pups on days 0 and 4 postnatal was similar in all groups
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 specified
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Description (incidence and severity):
The necropsy evaluation did not reveal any treatment-related findings in pups.
Histopathological findings:
not examined
Other effects:
not specified
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
300 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: embryo-fetal toxicity
Critical effects observed:
not specified
Reproductive effects observed:
not specified
Conclusions:
Under the experimental conditions of the study, the No Observed Adverse Effect Level (NOAEL) of the test item in Wistar rats was 300 mg/kg/day for males and females and 300 mg/kg/day for maternal-embryo-fetal toxicity.
Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
300 mg/kg bw/day
Study duration:
subacute
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 combined repeated dose toxicity study with the reproduction/developmental toxicity screening test is performed in rats, in which male and female rats are exposed to 0 (vehicle), 50, 150, 300 mg/kg bw/d via gavage (OECD 422).

The NOAEL in Wistar rats is considered to be 300 mg/kg bw/day for males and females and 300 mg/kg/day for maternal-embryo-fetal toxicity.

In this study, several findings were observed in males and females at mid and high dose levels. Two females and one male at high dose presented clinical signs between treatment days 25 and 41. These clinical signs were variable in nature, transitory and occurred in only a few animals. These clinical signs were not observed in the high dose group satellite animals. During the functional observational battery, hyperreflexia was observed in three females at the 300 mg/kg/day dose level. The incidence of these findings was not statistically significant when compared to controls. The significance of these findings is not clear, but they are unlikely to be specific to the test article. In males exposed to mid and high dose, total protein, globulin and albumin were statistically significant lower with a dose related trend, while potassium in females at high dose level was higher than control. However, these findings were not observed in any other high dose group, either the satellite group.

Females at mid-dose showed decreased diffuse lymphocyte cellularity in thymus and at high dose displayed a thymus involution. In both cases, these lesions were correlated with a lower thymus weight, although with statistical reach only at high dose level. These findings were not observed in treated and satellite males or satellite females. It is important to note that for all treated females, including the control group females, terminal thymus weights were substantially decreased in comparison to the female satellite control group. This observation is most likely due to the animals' physiological' response to, and following, pregnancy, and is not considered test article related.

Despite the above findings in parental animals, no effects considered to be test item related were observed in the pups.

Two-generation reproductive toxicity study/extended one generation reproductive toxicity study: The study does not need to be conducted since in the available reproduction/developmental toxicity screening test, the substance did not cause any adverse effect on reproductive organs and tissues. In addition, in a 2-year inhalation repeated dose toxicity study in rats performed with morpholine (read-across substance), no adverse effect on reproductive organs and tissues were observed. An extended one-generation reproductive toxicity study with this analogue substance morpholine will be performed (requested by ECHA in final desicion, period 2017-2018). Therefore it is concluded that a two-generation study/EOGRTS with the test substance DMDEE is scientifically unjustified.  

Effects on developmental toxicity

Description of key information
In a developmental toxicity study (BASF AG, 2009), Morpholine hydrochloride (97%) was administered as an aqueous solution to 25 time-mated female Wistar rats (Crl:WI[Han]) by gavage at doses of 75; 250 and 750 mg/kg bw/day on gestation days (GD) 6 through 19. The developmental NOAEL is 750 mg/kg bw/day. No adverse fetal findings of toxicological relevance were evident at any dose. The developmental toxicity study is classified as acceptable and satisfies the guideline requirement for a developmental toxicity study (OECD 414) in rat.
No further testing (OECD 414) is suggested for DMDEE as read across to the above mentioned study can be performed.
Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2007-10-30 to 2009-02-13
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Guideline study according to OECD GL 414 (“Prenatal Developmental Toxicity Study”)
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no
Species:
rat
Strain:
Wistar
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH
- Age at study initiation: about 10-12 weeks
- Weight at study initiation: 143.5-197.9 g
- Fasting period before study: no data
- Housing: individually in type M III Makrolon cages supplied by BECKER & CO., Castrop-Rauxel, Germany
- Diet: ad libitum (ground Kliba maintenance diet mouse/rat “GLP”)
- Water: ad libitum (drinking water)
- Acclimation period:


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24 °C
- Humidity (%): 30 - 70 %
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12


IN-LIFE DATES: From: 05.11.2007 To: 20.11.2007
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
The test substance solutions were prepared at the beginning of the administration period and thereafter at intervals of 7 days, which took into account the analytical results of the stability verification. For the preparation of the solutions, appropriate amounts of the test substance were weighed
in a beaker, topped up with drinking water and subsequently thoroughly mixed using a magnetic stirrer.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Analyses of the test substance preparations were carried out as a separate study compliance with the Principles of Good Laboratory
Practice at GKA Competence Center Analytics, BASF SE, Ludwigshafen, Germany. As analytical method .....

Samples of the test substance solutions were sent to the analytical laboratory twice during the study period for verification of the concentrations. Analytical verifications of the stability of the test substance in drinking water for a period of at least 7 days at room temperature were carried out before the study was initiated.
Details on mating procedure:
The animals were paired by the breeder (“time-mated”) and supplied on GD 0 (= detection of vaginal plug/sperm). The animals arrived on the same day (GD 0) at the experimental laboratory. The following day was designated as “GD 1”.
Duration of treatment / exposure:
Test substance was administered on gestation days (GD) 6 through 19.
Frequency of treatment:
The test substance solutions were administered to the animals orally by gavage, once a day, always at approx. the same time in the morning.
A standard dose volume of 10 mL/kg bw was used for each test group. At terminal sacrifice on GD 20, all females had implantation sites.
No. of animals per sex per dose:
25 female rats
Control animals:
yes, concurrent vehicle
Maternal examinations:
MORTALITY: Yes
- Twice a day on working days or once a day on Saturdays, Sundays or on public holidays (GD 0-20)

CAGE SIDE OBSERVATIONS/DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: at least once daily for any signs of morbidity, pertinent behavioral changes and signs of overt toxicity.
- Time schedule: If such signs occurred, the animals were examined several times daily (GD 0-20)

BODY WEIGHT: Yes
- Time schedule for examinations: GD 0, 1, 3, 6, 8, 10, 13, 15, 17, 19 and 20
- Furthermore, the corrected bw gain was calculated after terminal sacrifice (terminal bw on GD 20 minus weight of the unopened uterus minus bw on GD 6).

FOOD CONSUMPTION: Yes
- With the exception of day 0, the consumption of food was determined on the same days as bw

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Organs examined: Liver, Kidneys, Thyroid glands (with parathyroid glands)
- Cesarean Section: uteri and the ovaries were removed and following data were recorded:
Weight of the unopened uterus, Number of corpora lutea, Number and distribution of implantation sites classified as live fetuses/dead implantations
(cf. "Ovaries and uterine content")

OTHER: CLINICAL PATHOLOGY prior to sacrifice on gestation day 20,
- Blood was taken from the retro-orbital venous plexus from not fasted animals
- Examinations:
- Hematology - WBC, RBC, HGB, HCT, MCV, MCH, MCHC, PLT, Differential blood count, Reticulocytes
- Clinical chemistry - enzymes: ALT, AST, ALP, GGT
- Clinical chemistry - blood chemistry parameter: INP, CA, UREA, CREA, GLUC, TBIL, TPROT, ALB, GLOB TRIG, CHOL
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number and distribution of implantations sites: Yes
- Number of live fetuses: Yes
- Number of dead implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Number of dead fetuses: Yes


Fetal examinations:
- External examinations: Yes
- Each fetus was weighed, sexed, and external tissues and all orifices were examined macroscopically
- Furthermore, the viability of the fetuses and the condition of placentae, umbilical cords, fetal membranes, and fluids were examined.
- Individual placental weights were recorded
- Fetuses were sacrificed: Approximately one half of the fetuses per dam were eviscerated, skinned and placed in ethanol,
the other half was placed in Harrison’s fluid for fixation
- Soft tissue examinations: Yes, fetuses fixed in Harrison’s fluid
- Skeletal examinations: Yes, fetuses fixed in ethanol
- Head examinations: No data
Statistics:
CLINICAL and FETAL EXAMINATIONS:
Food consumption, bw, bw change, corrected bw gain (net maternal bw change), carcass weight, weight of unopened uterus, number of corpora lutea, number of implantations, number of resorptions, number of live fetuses, proportions of preimplantation loss,
proportions of postimplantation, loss, proportions of resorptions, proportion of live fetuses in each litter, litter mean fetal body weight, litter mean placental weight; Female mortality, females pregnant at terminal sacrifice, number of litters with fetal findings; Proportions of fetuses with malformations, variations and/or unclassified observations in each litter.
PATHOLOGY:
Means and standard deviations of each test group were calculated. Weight parameters were evaluated.
Indices:
- Conception rate (in %) [number of pregnant animals/number of fertilized animals x 100]
- Preimplantation loss (in %) [(number of corpora lutea – number of implantations)/number of corpora lutea x 100]
Historical control data:
Gestational parameters of animals of this strain and age.
Mortality:
no mortality observed
Description (incidence):
There were no substance-related or spontaneous mortalities in any of the groups.
Description (incidence and severity):
Additionally, transiently reduced mean food consumption and bw gain as well as affection of liver cells and liver cell metabolism was noted at the 750 mg/kg bw/d dose level.
Description (incidence and severity):
Additionally, transiently reduced mean food consumption and bw gain as well as affection of liver cells and liver cell metabolism was noted at the 750 mg/kg bw/d dose level.
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
The oral administration of 250 and 750 mg/kg bw/d Morpholine hydrochloride caused a mild, regenerative anemia in the dams, along with increased liver weights.
Description (incidence and severity):
The oral administration of 250 and 750 mg/kg bw/d Morpholine hydrochloride caused a mild, regenerative anemia in the dams, along with increased liver weights.
Details on maternal toxic effects:
The oral administration of Morpholine hydrochloride to the dams at 75, 250 and 750 mg/kg bw/day had no influence on gestational parameters.
Dose descriptor:
NOAEL
Effect level:
75 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
haematology
other: maternal toxicity
Dose descriptor:
NOAEL
Effect level:
750 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: developmental toxicity
Details on embryotoxic / teratogenic effects:
Fetal examinations revealed no influence of the test compound on sex distribution of the fetuses and fetal body weights. Morpholine hydrochloride shows no direct and specific effect on the respective morphological structures.
Remarks on result:
not determinable due to absence of adverse toxic effects
Abnormalities:
not specified
Developmental effects observed:
not specified

Fetal findings in this study were primarily limited to a slight increase in delayed ossification in the mid- and high-dose groups.These specific skeletal variations mirror common minor effects on fetal morphology which are considered to be transient in nature, being obviously secondary to maternal toxicity. Thus, these findings were regarded to be of no toxicological relevance and are not classified as adverse effects.

Executive summary:

In a developmental toxicity study (BASF AG, 2009), Morpholine hydrochloride (97%) was administered as an aqueous solution to 25 time-mated female Wistar rats (Crl:WI[Han]) by gavage at doses of 75; 250 and 750 mg/kg bw/day on gestation days (GD) 6 through 19. The control group, consisting of 25 females, was dosed with the vehicle (drinking water) in parallel. A standard dose volume of 10 mL/kg body weight was used for each test group. At terminal sacrifice on GD 20, all females (25 per group) had implantation sites. The oral administration of 250 and 750 mg/kg bw/d caused a mild, regenerative anemia in the dams, along with increased liver weights. Additionally, transiently reduced mean food consumption and bw gain as well as affection of liver cells and liver cell metabolism was noted at the high-dose level. The oral administration of Morpholine hydrochloride to the dams up to 750 mg/kg bw/d had no influence on gestational parameters. Conception rate, mean number of corpora lutea, total implantations, resorptions and live fetuses, fetal sex ratio, and the values calculated for pre- and the postimplantation losses were all unaffected.The maternal NOAEL is 75 mg/kg body weight/day based on statistically significant hematological changes in the dams at 250 and 750 mg/kg bw/day. Fetal examinations revealed no influence of Morpholine hydrochloride on sex distribution of the fetuses and fetal body weights.Morpholine hydrochloride shows no direct and specific effect on the respective morphological structures. Fetal findings in this study were primarily limited to a slight increase in delayed ossification in the mid- and high-dose groups. These specific skeletal variations mirror common minor effects on fetal morphology which are considered to be transient in nature, being obviously secondary to maternal toxicity. Thus, these findings were regarded to be of no toxicological relevance and are not classified as adverse events. The developmental NOAEL is 750 mg/kg bw/day. No adverse fetal findings of toxicological relevance were evident at any dose.The developmental toxicity study is classified as acceptable and satisfies the guideline requirement for a developmental toxicity study (OECD 414) in rat.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
750 mg/kg bw/day
Study duration:
subchronic
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

In a developmental toxicity study (BASF AG, 2009), Morpholine hydrochloride (97%) (read across substance) was administered as an aqueous solution to 25 time-mated female Wistar rats (Crl:WI[Han]) by gavage at doses of 75; 250 and 750 mg/kg bw/day on gestation days (GD) 6 through 19. The control group, consisting of 25 females, was dosed with the vehicle (drinking water) in parallel. A standard dose volume of 10 mL/kg body weight was used for each test group. At terminal sacrifice on GD 20, all females (25 per group) had implantation sites. The oral administration of 250 and 750 mg/kg bw/d caused a mild, regenerative anemia in the dams, along with increased liver weights. Additionally, transiently reduced mean food consumption and bw gain as well as affection of liver cells and liver cell metabolism was noted at the high-dose level. The oral administration of Morpholine hydrochloride to the dams up to 750 mg/kg bw/d had no influence on gestational parameters. Conception rate, mean number of corpora lutea, total implantations, resorptions and live fetuses, fetal sex ratio, and the values calculated for pre- and the postimplantation losses were all unaffected. Fetal examinations revealed no influence of Morpholine hydrochloride on sex distribution of the fetuses and fetal body weights. Morpholine hydrochloride shows no direct and specific effect on the respective morphological structures. Fetal findings in this study were primarily limited to a slight increase in delayed ossification in the mid- and high-dose groups. These specific skeletal variations mirror common minor effects on fetal morphology which are considered to be transient in nature, being obviously secondary to maternal toxicity. Thus, these findings were regarded to be of no toxicological relevance and are not classified as adverse events. The developmental NOAEL is 750 mg/kg bw/day. No adverse fetal findings of toxicological relevance were evident at any dose. The developmental toxicity study is classified as acceptable and satisfies the guideline requirement for a developmental toxicity study (OECD 414) in rat.

Justification for classification or non-classification

Based on the available data and according to the CLP criteria, DMDEE should not be classified as toxic to reproduction.