Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Remarks:
based on test type (migrated information)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
22 March 2011 - 17 October 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Compliant to GLP and testing guidelines; adequate consistence between data, comments and conclusions.
Qualifier:
according to
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Deviations:
yes
Remarks:
some microscopic slides of testes and epididymides were stained in hematoxylin-eosin by excess
GLP compliance:
yes (incl. certificate)
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Breeder: Charles River Laboratories France, L’Arbresle, France
- Age at study initiation: at the beginning of the treatment period, the males were approximately 10 weeks old
- Mean body weight at study initiation: the males had a mean body weight of 397 g (range: 354 g to 427 g) and the females were approximately
9 weeks old and had a mean body weight of 213 g (range: 183 g to 234 g)
- Fasting period before study: no
- Housing: 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: at least 7 days before the beginning of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 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 (7:00 - 19:00).

IN-LIFE DATES: 05 April 2011 to 17 October 2011.
Route of administration:
oral: gavage
Vehicle:
water
Remarks:
drinking water (treated by reverse osmosis)
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
Dose-levels were calculated in terms of 2-[(1-methylpropyl)amino]ethanol (CAS No. 35265-04-4) which is the form supplied by the Sponsor: there was no correction
factor.

The test item was administered as a solution in the vehicle. The formulations were stirred for 30 minutes in order to ensure the solubilisation of the
test item in the vehicle. No aluminium equipment was used for the preparation of the dosage forms.
The test item dosage forms were prepared on a weekly basis, stored at room temperature prior to use and delivered in brown flasks.

VEHICLE
- Concentration in vehicle: 1, 5 and 10 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg/day.
Details on mating procedure:
- M/F ratio per cage: 1/1
- Length of cohabitation (mating period): until mating occurs or 14 days has elapsed
- 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: High Performance Liquid Chromatography with tandem Mass Spectrometry detection (LC/MS MS) analytical method for the determination of 2-[(1-methylpropyl)amino]ethanol in dosage form samples was used.
Test item concentrations: the test item concentrations in the administered dosage forms analyzed in weeks 1, 5 and 7 remained within an acceptable range of -1.2% to +4.2% when compared to the nominal values (± 10%) on a weekly basis according to a stability (CIT/Study No. 37512 AHS).
Duration of treatment / exposure:
In the males:
− 2 weeks before mating,
− during the mating period (up to 3 weeks),
− until sacrifice (i.e. at least 5 weeks in total),

In the females:
− 2 weeks before mating,
− during the mating period (up to 3 weeks),
− during pregnancy,
− during lactation until day 4 post-partum inclusive,
− until sacrifice for females which had not delivered (day 25 p.c.).
Frequency of treatment:
Daily.
Details on study schedule:
- No F1 parents (only one generation mated)
- Age at mating of the mated animals in the study: 12 weeks for males, 11 weeks for females, approximately.
Remarks:
Doses / Concentrations:
10, 50 and 100 mg/kg/day
Basis:
actual ingested
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 based on the results of the CIT/Study No. 37513 TSR.

- Rationale for animal assignment: computerized stratification procedure.
Positive control:
No (not required).
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS:
- Time schedule: at least twice a day during the treatment period.

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

BODY WEIGHT (GAIN):
- 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 (or until sacrifice), on Days 0, 7, 14 and 20 post-coitum and days 1 and 5 post-partum.

FOOD CONSUMPTION:
- Time schedule: once a week until sacrifice, with the exception of the mating period (not recorded).

REPRODUCTION (apart from indices):
- Pre-coital time and duration of gestation were recorded.
Oestrous cyclicity (parental animals):
The estrous cycle stage was determined from a fresh vaginal lavage (stained with methylene blue), each morning during the pairing period, until the
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)

Testes and epididymides were observed microscopically. Qualitative testis staging did not indicate any abnormalities in the integrity of the various
cell types present within the different stages of the spermatogenic cycle.
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
The males were sacrificed after at least 5 weeks of treatment. The body and selected organs were weighed and a complete macroscopic post-mortem examination was performed. A microscopic examination was performed on the kidneys, stomach with forestomach, epididymis, testes and thymus
from the males in the control- and high dose groups, liver from all groups and and on all macroscopic lesions.

The dams were sacrificed on day 5 p.p., the body and selected organs were weighed and a complete macroscopic examination was performed.
A microscopic examination was performed on the kidneys and the stomach with forestomach, in the control and high-dose groups and, on the liver, ovaries (with oviducts) and thymus in all groups and on all macroscopic lesions.

GROSS NECROPSY
On completion of the treatment period all surviving F0 males and females were deeply anesthetized by an intraperitoneal injection of sodium
pentobarbital and sacrificed by exsanguination.
- males: after the end of the pairing period (at least 5 weeks of treatment in total),
- females: on day 5 p.p.,
- females which had not delivered: on day 25 p.c. (after a body weight recording to check for a possible un-noticed delivery),
- mothers with litter dying entirely: as appropriate.

Pups were sacrificed by an intraperitoneal injection of sodium pentobarbital:
- surviving pups: on day 5 p.p..

HISTOPATHOLOGY
A microscopic examination was performed on:
- all tissues listed in the tissue procedure table in the males and females of the control- and high-dose groups (groups 1 and 4) sacrificed at the end of the treatment period and for female that were sacrificed prematurely,
- all macroscopic lesions of all the animals of the low- and intermediate-dose groups (groups 2 and 3) sacrificed on completion of the treatment
period,
- thymus of males from control and high-dose groups (groups 1 and 4) and thymus of females from control, low-, mid- and high-dose groups
(groups 1 to 4),
- all females sacrificed because of no delivery to investigate possible causes.

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

At the request of by the Sponsor and based upon the microscopic results of the high-dose group, liver and ovaries (with oviducts) of the low- and
intermediate-dose groups were examined.

ORGAN WEIGHTS:
The body weight of each animal sacrificed as scheduled was recorded before sacrifice, and the organs specified in the Tissue Procedure Table were
weighed (wet) as soon as possible after dissection.
The ratio of organ weight to body weight (recorded immediately before sacrifice) was calculated.
Postmortem examinations (offspring):
SACRIFICE: on day 5 post-partum

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

HISTOPATHOLOGY: No

ORGAN WEIGTHS: No
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 concepti) / Number of implantations
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:
no effects observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
at 100 mg/kg bw/d in females during pregnancy period
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
at 100 mg/kg bw/d in females during pregnancy period
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
at 100 mg/kg bw/d in females
Other effects:
not examined
Reproductive function: oestrous cycle:
no effects observed
Reproductive function: sperm measures:
not examined
Reproductive performance:
effects observed, treatment-related
Description (incidence and severity):
at 100 mg/kg/day
MORTALITY:
There were no unscheduled deaths.

CLINICAL SIGNS:
Males
There were no treatment-related clinical signs.
The few isolated observations (soft feces, chromodacryorrhea, cutaneous lesion or area of air loss) recorded in controls or treated groups are commonly observed in this species and strain.

Females (premating period)
There were no findings during the premating period.

Females (pregnancy period)
There were no treatment-related clinical signs.
One out of 10 and 6/10 females in the 50 and 100 mg/kg/day groups, respectively, were sacrificed for absence of delivery on day 25 p.c..
All females that did not deliver were pregnant with the exception of one high-dose animal.
An isolated observation (soft feces) recorded in the 10 mg/kg/day group is commonly observed in this species and strain.

Females (lactation period)
There were no treatment-related clinical signs at 10 and 50 mg/kg/day.
One out of four females in the 100 mg/kg/day group with a dead litter was prematurely sacrificed on day 3 p.p..

BODY WEIGHT (GAIN):
Males
There were no treatment-related effects on mean body weights or mean body weight changes.
Isolated reductions in mean body weight were recorded on study day 15 (-7.0% vs. controls, p<0.05) and study day 22 (-6.7% vs. controls, p<0.05) in animals treated at 10 mg/kg/day. In absence of a dose-relationship, an association with the treatment by the test item was considered unlikely.
Isolated reductions or increases in mean body weight change were recorded on periods of days 1 to 8 (-31.0% vs. controls, p<0.05) and 8 to 15
(-41.4% vs. controls, p<0.05) at 10 mg/kg/day or on period of days 22 to 29 at 50 or 100 mg/kg/day (+64.7% and +52.9% vs. controls, p<0.05).
In absence of a dose-relationship, an association with the treatment by the test item was considered unlikely.

Females (premating period)
There were no effects on mean body weight or mean body weight change during the premating period.

Females (pregnancy period)
On day 20 p.c. and at 100 mg/kg/day, there was a statistically significant reduction in mean body weight (-21.3% vs. controls, p<0.001).
Mean body weight change was also markedly reduced at 100 mg/kg/day on periods of days 7 to 14 (-33.3% vs. controls, p<0.05) and on periods of days 14 to 20 p.c. ( 92.4% vs. controls, p<0.001), resulting in an overall reduction of -55.6% vs. controls (p<0.001) on period of days 0 to 20 p.c..
Taking into account both the amplitude of the effects and their presence in the high-dose group only, these finding were considered to be related to the treatment by the test item.

Females (lactation period)
There were no significant effects on mean body weight during the lactation period. However, mean body weight change was reduced in the three remaining dams treated at 100 mg/kg/day on periods of days 1 to 5 p.p. ( 68.2% vs. controls, p<0.01). It was correlated with lower food consumption at this dose-level.

FOOD CONSUMPTION:
Males
There were no treatment-related effects on mean food consumption.

Females (premating period)
There were no effects on mean food consumption during the premating period.

Females (pregnancy period)
There were no effects on mean food consumption during the pregnancy period up to 50 mg/kg/day.
At 100 mg/kg/day, mean food consumption was reduced on days 14 to 20 (-16.1 vs. controls, p<0.05). This effect was considered to be related to the treatment by the test item.

Females (lactation period)
There was a lower mean food consumption in the three remaining dams treated at 100 mg/kg/day on period of days 1 to 5 p.p. (37.2% vs. controls,
p<0.001). This effect was probably secondary to the low number of suckling pups.

REPRODUCTIVE PERFORMANCE:
Mating and fertility data
There were no effects of treatment with the test item on mating and fertility parameters. Only one female (W22203; 100 mg/kg/day) was not pregnant.

Estrous cycle
There were no indications of abnormal estrous cycles when compared with those of the control females in any test item-treated females.

Delivery data
All treatment-related effects were observed at 100 mg/kg/day, only. Statistically significant differences from controls included:
- an increase in mean duration of gestation (22.3 days vs. 21.1 days in controls, p<0.001) in females which delivered,
- a lower mean number of corporea lutea (10.2 vs. 15.6 in controls, p<0.001) in pregnant females,
- a lower mean number of implantations sites (9.3 vs. 15.5 in controls, p<0.001) in pregnant females,
- a lower mean number of pups delivered (3.0 vs. 13.7 in controls, p<0.001).

In addition, at 100 mg/kg/day in pregnant females, there were increased mean pre implantation loss (8.3 vs. 0.7% in controls, p<0.05) and increased mean post-implantation loss (75.1 vs. 12.0% in controls).

ORGAN WEIGHTS:
Treatment-related lower mean absolute and relative liver weights were noted at 100 mg/kg/day, females being more affected than males. It correlated microscopically with lower glycogen content in hepatocytes of treated animals.
High absolute and relative mean thymus weights were noted in females given the test item at 100 mg/kg/day. In the absence of microscopic correlates, a relationship to treatment was ruled out.

GROSS PATHOLOGY:
No treatment-related macroscopic changes were noted in males and females given the test item.
The few macroscopic findings noted at the end of the treatment period were of those commonly recorded in the Sprague Dawley rat and none were
considered to be related to the test item administration.

HISTOPATHOLOGY:
In the liver, a lower mean glycogen content was observed in males and females given the test item at 100 mg/kg/day. There was a trend to dose-related decreased glycogen storage in females from 10 mg/kg/day. The effect was more severe in males than in females. Taking into account the amplitude of the change, this effect was not considered to be adverse.
Focal to multifocal subcapsular necrosis were observed in the liver of some treated females without dose relationship. As the change was often focal, at minimal severity, and could be observed spontaneously, a relationship to treatment was ruled out.

Reproductive organs
Males
Testes and epididymides were observed microscopically. Qualitative testis staging did not indicate any abnormalities in the integrity of the various cell types present within the different stages of the spermatogenic cycle.

Females
Treatment-related microscopic change was observed in the ovaries of animals given 100 mg/kg/day. It consisted of a higher incidence and severity
of increased size of follicular structures. Follicles were larger, with constant internal cavities (late stage follicles). It was observed in 1/10 control females, 1/10 females given 10 mg/kg/day, 2/10 animals treated at 50 mg/kg/day and 7/10 females given 100 mg/kg/day. Although marginal, the increased incidence and severity of the change in females given 100 mg/kg/day was considered to be likely related to the test item.
Dose descriptor:
NOAEL
Remarks:
Parental toxicity
Effect level:
50 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: Reduced body weight gain in females during gestation.
Dose descriptor:
NOAEL
Remarks:
Parental toxicity
Effect level:
>= 100 mg/kg bw/day (actual dose received)
Sex:
male
Basis for effect level:
other: No treatment-related effect was observed in male rats.
Dose descriptor:
NOAEL
Remarks:
Reproductive performance
Effect level:
50 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: Pre- and post-implantation losses and high incidence of follicular changes at 100 mg/kg/day.
Clinical signs:
no effects observed
Mortality / viability:
mortality observed, treatment-related
Description (incidence and severity):
at 100 mg/kg bw/d
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
at 100 mg/kg bw/d, not directly linked to treatment
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings:
not examined
VIABILITY (OFFSPRING)
The number of mortalities was similar between the control and treated groups at 10 and 50 mg/kg/day. It was considered that there were no effects of treatment with the test item at these dose levels. At 100 mg/kg/day, the low number of litters and pups delivered preclude drawing any conclusion.

CLINICAL SIGNS:
There were no treatment-related findings.

BODY WEIGHT (GAIN):
Both male and female pups from the group treated at 100 mg/kg/day had a higher mean body weight than pups from the control group on days 1 and 5 p.p. and gained more body weight during the lactation period. The difference was statistically significant in males. These values were attributed to the low mean number of pups per litter in the high-dose group at birth (3.0 vs. 13.7 in the control group, p<0.001).
There were no effects at 10 or 50 mg/kg/day.

GROSS PATHOLOGY:
There were no treatment-related findings.
Dose descriptor:
NOAEL
Remarks:
Developmental toxicity
Generation:
F1
Effect level:
>= 50 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No direct effect on the progeny during the lactation period. However, the low number of litters and pups delivered at 100 mg/kg/day, preclude drawing any conclusion at this dose-level
Reproductive effects observed:
not specified
Conclusions:
The test item, 2-[(1-methylpropyl)amino]ethanol (99.7% purity), was administered daily by oral gavage to male and female Sprague Dawley rats, for 2 weeks before mating, during mating, gestation and until day 5 post partum, at dose-levels of 10, 50 or 100 mg/kg/day.

At the dose-levels of 10 and 50 mg/kg/day, there was no treatment-related effect.
At the dose-level of 100 mg/kg/day, no treatment-related effect was observed in male rats. Six out of ten female rats were sacrificed for absence of delivery on day 25 p.c. and one out of four females with a dead litter was prematurely sacrificed on day 3 p.p.. Mean body weight gain was markedly reduced during the whole gestation and mean food consumption was reduced on days 14 to 20 of gestation. An increase in mean duration of gestation, a lower mean number of corporea lutea, a lower mean number of implantations sites and a lower mean number of pups delivered were observed at this dose-level. In addition, there were increased mean pre implantation loss and increased mean post-implantation loss. High absolute and relative mean thymus weights and a higher incidence and severity of increased size of follicular structures in ovaries were noted in females.

Based on the results of this study:
- the No Observed Adverse Effect Level (NOAEL) for parental toxicity was considered to be 50 mg/kg/day (based on the reduced body weight gain in females during gestation),
- the NOAEL for reproductive performance (mating and fertility) was considered to be 50 mg/kg/day (based on pre- and post-implantation losses and high incidence of follicular changes at 100 mg/kg/day),
- the NOAEL for effects on the progeny was considered to be equal or higher than 50 mg/kg/day (no direct effect on the progeny during the lactation period). However, the low number of litters and pups delivered at 100 mg/kg/day, preclude drawing any conclusion at this dose-level.
Executive summary:

The objective of this study was to evaluate the potential toxic effects of the test item, 2-[(1-methylpropyl)amino]ethanol following daily oral administration (gavage) to male and female rats from before mating, through mating and gestation and until day 4 post-partum (p.p.).

This study provides initial information on possible toxicological effects likely to arise from repeated exposure on male and female reproductive performance, such as gonadal function, mating behavior, conception, development of the conceptus and parturition.

This study was conducted as a reproduction/developmental screening test according to OECD 421 guideline and GLP.

Three groups of 10 male and 10 female Sprague-Dawley rats received the test item, 2-[(1-methylpropyl)amino]ethanol(purity 99.7%), by oral (gavage) administration once a day, 2 weeks before mating, through mating and gestation and until day 4 p.p..The dose-levels were 10, 50 or 100 mg/kg/day. Another group of 10 males and 10 females received the vehicle, drinking water treated by reverse osmosis, alone, under the same experimental conditions and acted as a control group. The dosing volume was 10 mL/kg/day.

 

Clinical signs and mortality were checked once and twice a day during the treatment period, respectively. Body weight and food consumption were recorded weekly and at determined intervals for females during gestation and lactation.

The animals were paired for mating after 2 weeks of treatment. The estrous cycles were monitored during the mating period. 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. Pups were observed daily for clinical signs and pup body weights were recorded on days 1 and 5p.p..

 

The males were sacrificed after at least 5 weeks of treatment. The body and selected organs were weighed and a complete macroscopic post-mortem examination was performed. A microscopic examination was performed on the kidneys, stomach with forestomach, epididymis, testes and thymus from the males in the control- and high‑dose groups, liver from all groups and and on all macroscopic lesions.

 

The dams were sacrificed on day 5 p.p., the body and selected organs were weighed and a complete macroscopic examination was performed. A microscopic examination was performed on the kidneys and the stomach with forestomach, in the control and high-dose groups and, on the liver, ovaries (with oviducts) and thymus in all groups and on all macroscopic lesions.

 

Pups were sacrificed on day 5 p.p.and, including those found dead, were carefully examined for gross external abnormalities.

The test item concentrations in the administered dosage forms analyzed in weeks 1, 5 and 7 remained within an acceptable range of variation when compared to the nominal values.

F0 generation

There were no unscheduled deaths in males. However,1/10 and 6/10 females in the 50 and 100 mg/kg/day groups, respectively, were sacrificed for absence of delivery on day 25post-coitum(p.c.). All females which did not deliver were pregnant with the exception of one high-dosed animal

There were no treatment-related clinical signs in males and in females during the premating, pregnancy or lactation periods.

 

There were no treatment-related effects on mean body weights or mean body weight changes in males and in females during the premating period.

In females at 100 mg/kg/day only and onday 20p.c.,there was a markedly and statistically significant reduction in mean body weight (-21.3% vs.controls, p < 0.001). Mean body weight change was also markedly reduced at 100 mg/kg/day on periods of days 7 to 14 (-33.3% vs. controls, p > 0.05) and days 14 to 20 p.c. (‑92.4% vs. controls, p < 0.001), resulting in an overall reduction of -55.6% vs. controls (p < 0.001) on period of days 0 to 20 p.c. Taking into account both the amplitude of the effects and their presence in the high-dose group only, these finding were considered to be related to the treatment by the test item.

There were no treatment-related effects on mean food consumption in male. At 100 mg/kg/day, in females and during the pregnancy period only,mean food consumption was moderately reduced on days 14 to 20 (-16.1 vs. controls, p < 0.05). This effect was considered to be related to the treatment by the test item.

There were no effects of treatment with the test item on mating.

 

There were no indications of abnormal estrous cycles when compared with those of the control females in any test item-treated female.

 

All treatment-related effects on mean reproductive parameters were observed at 100 mg/kg/day, only. Statistically significant differences from controls included:

.         an increase in mean duration of gestation (22.3 days vs. 21.1 days in controls),

.         a lower mean number ofcorporea lutea(12.3 vs. controls),

.         a lower mean number of implantations sites (12.3 vs. controls).

 

In addition, there was a marked increase in mean implantation loss at 100 mg/kg/day (9.7 vs. controls).

 

F1 generation

The number of pup mortalities was similar between the control and treated groups.

 

There were no treatment-related clinical signs in pups.

 

Both male and female pups from the group treated at 100 mg/kg/day gained more body weight during the lactation period. The difference was statistically significant in males. These values were attributed to the low mean number of pups per litter in the high-dose group (3.0 vs. the control group).

There were no effects at 10 or 50 mg/kg/day.

 

There were no treatment-related effects on sex-ratio and on gross external examination.

Pathology (F0 generation)

Treatment-related lower liver weights were noted in males and females at 100 mg/kg/day and correlated microscopically with lower glycogen content in hepatocytes of treated animals. There was a trend to dose-related decreased glycogen storage in females from 10 mg/kg/day. This change was not considered to be adverse.

No treatment-related macroscopic changes were noted in males and females (including the prematurely sacrificed one) given the test item.

 

Qualitative testis staging did not indicate any abnormalities in the integrity of the various cell types present within the different stages of the spermatogenic cycle.

Treatment-related microscopic increased size of follicular structures was observed in the ovaries of animals given 100 mg/kg/day. Although marginal, the increased incidence and severity of the ovarian change in females given 50 mg/kg/day was considered to be likely related to the test item.

The test item, 2-[(1-methylpropyl)amino]ethanol (99.7% purity), was administered daily by oral gavage to male and female Sprague‑Dawley rats, for 2 weeks before mating, during mating, gestation and until day 5 post‑partum, at dose-levels of 10, 50 or 100 mg/kg/day.

 

At the dose-levels of 10 and 50 mg/kg/day, there was no treatment-related effect.

At the dose-level of 100 mg/kg/day, no treatment-related effect was observed in male rats. Six out of ten female rats weresacrificed for absence of delivery on day 25 p.c.and one out of four females with a dead litter was prematurely sacrificed on day 3 p.p.. Mean body weight gain was markedly reduced during the whole gestation and mean food consumption was reduced on days 14 to 20 of gestation. An increase in mean duration of gestation, a lower mean number of corporea lutea, a lower mean number of implantations sites and a lower mean number of pups delivered were observed at this dose-level.In addition, there were increased mean pre‑implantation loss and increased mean post-implantation loss. High absolute and relative mean thymus weights and a higher incidence and severity of increased size of follicular structures in ovaries were noted in females.

 

Based on the results of this study:

. the No Observed Adverse Effect Level (NOAEL) for parental toxicity was considered to be 50 mg/kg/day (based on the reduced body weight gain in females during gestation),

. the NOAEL for reproductive performance (mating and fertility) was considered to be 50 mg/kg/day (based on pre- and post-implantation losses and high incidence of follicular changes at 100 mg/kg/day),

. the NOAEL for effects on the progeny was considered to be equal or higher than 50 mg/kg/day (no direct effect on the progeny during the lactation period). However, the low number of litters and pups delivered at 100 mg/kg/day, preclude drawing any conclusion at this dose-level.

Effect on fertility: via oral route
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
50 mg/kg bw/day
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

The potential toxic effects of the test item, sec-butylaminoethanol , following daily oral administration (gavage) to male and female rats from before mating, was evaluated through mating and gestation and until day 4 post-partum (p.p.) (Spezia, 2011a).

This study provides initial information on possible toxicological effects likely to arise from repeated exposure on male and female reproductive performance, such as gonadal function, mating behavior, conception, development of the conceptus and parturition.

This study was conducted as a reproduction/developmental screening test according to OECD 421 guideline and GLP.

Three groups of 10 male and 10 female Sprague-Dawley rats received the test item, 2-[(1-methylpropyl)amino]ethanol(purity 99.7%), by oral (gavage) administration once a day, 2 weeks before mating, through mating and gestation and until day 4p.p..The dose-levels were 10, 50 or 100 mg/kg/day. Another group of 10 males and 10 females received the vehicle, drinking water treated by reverse osmosis, alone, under the same experimental conditions and acted as a control group. The dosing volume was 10 mL/kg/day.

Clinical signs and mortality were checked once and twice a day during the treatment period, respectively. Body weight and food consumption were recorded weekly and at determined intervals for females during gestation and lactation.

The animals were paired for mating after 2 weeks of treatment. The estrous cycles were monitored during the mating period. The dams were allowed to litter and rear their progeny until day 5p.p.. The total litter sizes and numbers of pups of each sex were recorded after birth. Pups were observed daily for clinical signs and pup body weights were recorded on days 1 and 5p.p..

The males were sacrificed after at least 5 weeks of treatment. The body and selected organs were weighed and a complete macroscopicpost-mortemexamination was performed. A microscopic examination was performed on the kidneys, stomach with forestomach, epididymis, testes and thymus from the males in the control- and high‑dose groups, liver from all groups and and on all macroscopic lesions.

The dams were sacrificed on day 5p.p.,the body and selected organs were weighed and a complete macroscopic examination was performed. A microscopic examination was performed on the kidneys and the stomach with forestomach, in the control and high-dose groups and, on the liver, ovaries (with oviducts) and thymus in all groups and on all macroscopic lesions.

Pups were sacrificed on day 5p.p.and, including those found dead, were carefully examined for gross external abnormalities.The test item concentrations in the administered dosage forms analyzed in weeks 1, 5 and 7 remained within an acceptable range of variation when compared to the nominal values.

F0 generation

There were no unscheduled deaths in males. However,1/10 and 6/10 females in the 50 and 100 mg/kg/day groups, respectively, were sacrificed for absence of delivery on day 25post-coitum(p.c.). All females which did not deliver were pregnant with the exception of one high-dosed animal

There were no treatment-related clinical signs in males and in females during the premating, pregnancy or lactation periods.

There were no treatment-related effects on mean body weights or mean body weight changes in males and in females during the premating period.

In females at 100 mg/kg/day only and onday 20p.c.,there was a markedly and statistically significant reduction in mean body weight (-21.3%vs.controls, p < 0.001). Mean body weight change was also markedly reduced at 100 mg/kg/day on periods of days 7 to 14 (-33.3%vs.controls, p > 0.05) and days 14 to 20p.c.(‑92.4%vs.controls, p < 0.001), resulting in an overall reduction of -55.6%vs.controls (p < 0.001) on period of days 0 to 20p.c.Taking into account both the amplitude of the effects and their presence in the high-dose group only, these finding were considered to be related to the treatment by the test item.

There were no treatment-related effects on mean food consumption in male. At 100 mg/kg/day, in females and during the pregnancy period only,mean food consumption was moderately reduced on days 14 to 20 (-16.1vs.controls, p < 0.05). This effect was considered to be related to the treatment by the test item.

There were no effects of treatment with the test item on mating.

There were no indications of abnormal estrous cycles when compared with those of the control females in any test item-treated female.

All treatment-related effects on mean reproductive parameters were observed at 100 mg/kg/day, only. Statistically significant differences from controls included:

.         an increase in mean duration of gestation (22.3 daysvs.21.1 days in controls),

.         a lower mean number ofcorporea lutea(12.3vs.controls),

.         a lower mean number of implantations sites (12.3vs.controls).

In addition, there was a marked increase in mean implantation loss at 100 mg/kg/day (9.7vs.controls).

F1 generation

The number of pup mortalities was similar between the control and treated groups.

There were no treatment-related clinical signs in pups.

Both male and female pups from the group treated at 100 mg/kg/day gained more body weight during the lactation period. The difference was statistically significant in males. These values were attributed to the low mean number of pups per litter in the high-dose group (3.0vs.the control group).

There were no effects at 10 or 50 mg/kg/day.

There were no treatment-related effects on sex-ratio and on gross external examination.

Pathology (F0 generation)

Treatment-related lower liver weights were noted in males and females at 100 mg/kg/day and correlated microscopically with lower glycogen content in hepatocytes of treated animals. There was a trend to dose-related decreased glycogen storage in females from 10 mg/kg/day. This change was not considered to be adverse.

No treatment-related macroscopic changes were noted in males and females (including the prematurely sacrificed one) given the test item.

Qualitative testis staging did not indicate any abnormalities in the integrity of the various cell types present within the different stages of the spermatogenic cycle.

Treatment-related microscopic increased size of follicular structures was observed in the ovaries of animals given 100 mg/kg/day. Although marginal, the increased incidence and severity of the ovarian change in females given 50 mg/kg/day was considered to be likely related to the test item.

The test item, 2-[(1-methylpropyl)amino]ethanol (99.7% purity), was administered daily by oral gavage to male and female Sprague‑Dawley rats, for 2 weeks before mating, during mating, gestation and until day 5post‑partum, at dose-levels of 10, 50 or 100 mg/kg/day.

At the dose-levels of 10 and 50 mg/kg/day, there was no treatment-related effect.

At the dose-level of 100 mg/kg/day, no treatment-related effect was observed in male rats. Six out of ten female rats weresacrificed for absence of delivery on day 25p.c.and one out of four females with a dead litter was prematurely sacrificed on day 3p.p.. Mean body weight gain was markedly reduced during the whole gestation and mean food consumption was reduced on days 14 to 20 of gestation. An increase in mean duration of gestation, a lower mean number ofcorporea lutea, a lower mean number of implantations sites and a lower mean number of pups delivered were observed at this dose-level.In addition, there were increased mean pre‑implantation loss and increased mean post-implantation loss. High absolute and relative mean thymus weights and a higher incidence and severity of increased size of follicular structures in ovaries were noted in females.

Based on the results of this study:

. the No Observed Adverse Effect Level (NOAEL) for parental toxicity was considered to be 50 mg/kg/day (based on the reduced body weight gain in females during gestation),

. the NOAEL for reproductive performance (mating and fertility) was considered to be 50 mg/kg/day (based on pre- and post-implantation losses and high incidence of follicular changes at 100 mg/kg/day),

. the NOAEL for effects on the progeny was considered to be equal or higher than 50 mg/kg/day (no direct effect on the progeny during the lactation period). However, the low number of litters and pups delivered at 100 mg/kg/day, preclude drawing any conclusion at this dose-level.


Short description of key information:
Based on an OECD 421 study:
. the No Observed Adverse Effect Level (NOAEL) for parental toxicity was considered to be 50 mg/kg/day (based on the reduced body weight gain in females during gestation),
. the NOAEL for reproductive performance (mating and fertility) was considered to be 50 mg/kg/day (based on pre- and post-implantation losses and high incidence of follicular changes at 100 mg/kg/day),
. the NOAEL for effects on the progeny was considered to be equal or higher than 50 mg/kg/day (no direct effect on the progeny during the lactation period). However, the low number of litters and pups delivered at 100 mg/kg/day, preclude drawing any conclusion at this dose-level.

Justification for selection of Effect on fertility via oral route:
GLP guideline study

Effects on developmental toxicity

Description of key information
No developmental toxicity was observed in pregnant rats exposed to 150 ppm (460 mg/m3) of a structural analogue substance, N-methylethanolamine.
Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study well documented, meets generally accepted scientific principles, acceptable for assessment
Principles of method if other than guideline:
N-Methylethanolamine was vaporized and administered to approximately 15 pregnant rats in one to three concentrations for 7 hr/day on gestation days 7 to 15, and dams were sacrificed on day 20. Fetuses were individually weighed, and two-thirds of them were fixed in Bouin's solution and examined for soft-tissue anomalies. The other one-third were fixed in alcohol, stained with Alizarin Red and examined for skeletal defects.
GLP compliance:
no
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Breeding Laboratories, Wilmington, MA
- Age at study initiation: no data
- Weight at study initiation: no data
- Fasting period before study:
- Housing: alone in 38 x 33 x 17-cm polycarbonate cages with filter tops
- Diet: Purina Lab Chow ad libitum
- Water: tap water ad libitum
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 24 +/- 2
- Humidity (%): 40 +/- 20
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
inhalation: vapour
Type of inhalation exposure (if applicable):
whole body
Vehicle:
clean air
Details on exposure:
Pregnant females were transported from the animal quarters to the inhalation chambers in their home cages with filter tops (Hazleton Systems, Aberdeen, MD). They were placed individually in 13 x 25 x 189-cm stainless steel wire mesh cages within exposure chambers.
Air flow through the chambers provided approximately four air changes per minute.
Exposures were conducted sequentially in one or two chambers, with a third chamber for sham exposure of control subjects.
Control animals were placed in similar chambers for the same hours as the exposed animals; a pooled group of controls (N = 34) served as the comparison group for the first three chemicals examined. Another group of 15 controls served as the comparison group for the last two chemicals examined, as these groups were exposed at a later time (approximately 6 months later) than the first three.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The concentrations within the exposure chambers as measured by the infrared analyzer were relatively close to those obtained from gas chromatography.
Details on mating procedure:
Males weighing over 300 g were placed individually into a cage with three females weighing 200 to 300 g. Vaginal smears were taken each morning, and the presence of sperm marked day zero of gestation.
Duration of treatment / exposure:
Exposures conducted 7 hr/day, and the animals were left in the chamber for at least one additional hour blow-off time after vapor generation terminated. They were then returned in their individual housing cages to the animal quarters, where water bottles were replaced.
Frequency of treatment:
Exposures were conducted on gestation days 7-15.
Duration of test:
On day 20 of gestation, dams were sacrificed.
Remarks:
Doses / Concentrations:
150.0 +/- 15.2 ppm (460 +/- 46 mg/m3)
Basis:
analytical conc.
No. of animals per sex per dose:
approximately 15 pregnant rats
Control animals:
other: three solvents were compared with a pooled group (N = 34) of sham-exposed controls, and the remaining two were compared with a group of 15 controls.
Maternal examinations:
Feed and water intake and maternal weight were recorded weekly (i.e., on days 7, 14, and 21); any other signs of maternal toxicity were noted daily.
On day 20 of gestation, the females were individually weighed and euthanized by chloroform asphyxiation.
Ovaries and uterine content:
The entire uterus was removed and numbers of resorption sites (classified as early, middle or late) and live fetuses wvere determined.
Fetal examinations:
Fetuses were serially removed, blotted of excess fluids, weighed, examined for external malformations and external sex determined.
One third of the fetuses were randomly selected and placed in 95% ethanol, and the remainingfetuses were placed in Bouin's solution. After being in
the Bouin's solution for at least 1 week, these fetuseswere examined for visceral abnormalities using Wilson's razor blade sectioning technique. The viscera wereexamined with the aid of a dissecting microscope. A representative sample of sections with malformations was identified by dam number and saved in 70% alcohol.

Fetuses were examined for skeletal defects by using a modified Staples technique. They were fixed in 95% alcohol, eviscerated and macerated in 2% KOH/Alizarin Red S solution. The fetuses were further macerated and cleared in the appropriate solutions of 2% KOH/glycerin (60:40, 40:60, 20:80) and stored in 100% glycerin. A crystal of thymol was added to each storage vial to retard fungal growth. Storage vials were individually identified by dam number.
Statistics:
Numbers of implants and proportions of resorptions were independently analyzed by using a Kruskal-Wallis test corrected for ties, with subsequent multiple comparisons to determine where the differences occurred. Analysis of pup weights involved a mixed model analysis of covariance (with the number of live pups in the litter as the covariate) using maximum likelihood estimation. The model was mixed, since there was both within-litter and between-litter variation. Subsequently, pairwise comparisons between the pooled control group and each treatment group were performed. Incidence oftotal defects and of total variants were compared using a Kruskal-Wallis test with multiple comparisons with the litter as the experimental unit and the level of significance at p< 0.05.
Details on maternal toxic effects:
Maternal toxic effects:no effects

Details on maternal toxic effects:
Effects upon pregnancy outcome:
Number pregnant/mated - 15/16 (Control); 18/24 (Treated)
Implants/dam - 13.1 (Control); 11.9 (Treated)
Litters with resorptions - 10 (67%; Control); 10 (56%; Treated)
Live fetuses/litter - 11.6 (Control); 11.8 (Treated)

Dose descriptor:
NOAEC
Effect level:
>= 150 other: ppm (analytical) (460 mg/m3)
Basis for effect level:
other: maternal toxicity
Dose descriptor:
NOAEC
Effect level:
>= 150 other: ppm (analytical) (460 mg/m3)
Basis for effect level:
other: developmental toxicity
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
Live fetal body weights - Female, 3.36 (Control); 3.59 (Treated); Male, 3.33 (Control); 3.69 (Treated)
Number of litters with abnormal litter - 5 (Control); 7 (Treated)

Visceral observations (litters/fetuses):
Number litters/fetuses examined - 15/116 (Control); 18/143 (Treated)
Malformations - Brain (hydrocephalus, mild), 3/6 (Control); 3/5 (Treated).
Variations (62 controls; 74 treated) - Testicular (undescended testes) - 0 (Control); 1/1 (Treated); Testicular (position variation) - 10/22 (Control); 9/13 (Treated) - Urinary tract (dilated renal pelvis) - 3/ 4 (Control); 5/5 (Treated); Urinary tract (dilated ureter) - 3/6 (Control); 6/18 (Treated); Urinary tract (hydronephrosis) - 2/2 (Control); 0 (Treated); Urinary tract (hydroureter) - 3/3 (Control); 5/6 (Treated); Urinary tract (ectopic kidney) - 1/1 (Control); 0 (Treated).
% Normal (mean ±standard deviation) - 89 ±26% (Control); 92 ±15% (treated)
% Abnormal (mean ±standard deviation) - 11 ±26% (Control); 8 ±15% (treated)

Skeletal observations (litters/fetuses):
Number litters/fetuses examined - 14/58 (Control); 18/69 (Treated)
Malformations - 0 (Control); 0 (Treated)
Variations - Ribs (rudimentary 14th) - 1/1 (Control); 1/2 (Treated); Ribs (rudimentary lumbar) - 0 (Control); 1/1 (Treated); Skull (incomplete ossification of supraoccipital) - 0 (Control); 1/1 (Treated); Skull (incomplete ossification of interparietal) - 1/1 (Control); 1/2 (Treated); Skull (incomplete ossification of parietal) - 1/1 (Control); 0 (Treated); Sternebrae (decreased number) - 1/1 (Control); 1/1 (Treated); Sternebrae (split) - 0 (Control); 1/1 (Treated); Sternebrae (misaligned) - 1/1 (Control); 5/5 (Treated); Centra (misshapen thoracic) - 1/1 (Control); 5/5 (Treated); Pelvic Girdle (pubis missing) - 0 (Control); 2/2 (Treated); Pelvic Girdle (ischium missing or reduced in size) - 0 (Control); 1/1 (Treated).
% Normal (mean ±standard deviation) - 100 ±0% (Control); 100 ±0% (treated)
% Abnormal (mean ±standard deviation) - 0% (Control); 0% (treated)
Abnormalities:
not specified
Developmental effects observed:
not specified
Conclusions:
N-Methylethanolamine showed neither maternal nor fetal toxicity effects.
Executive summary:

N-Methylethanolamine was vaporized and administered at 150 ppm to approximately 15 pregnant rats for 7 hr/day on gestation days 7 to 15, and dams were sacrificed on day 20 of gestation. Fetuses were individually weighed, and two-thirds of them were fixed in Bouin's solution and examined for soft-tissue anomalies. The other one-third were fixed in alcohol, stained with Alizarin Red and examined for skeletal defects. As overall result for the substance N-Methylethanolamine it can be stated that neither maternal nor fetal toxicity effects can be concluded in this study.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
60 mg/kg bw/day
Species:
rat
Quality of whole database:
Screening study insufficient to assess the developmental toxicity.
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEC
Species:
rat
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information

The developmental toxicity of sec-Butyl aminoethanol was investigated in groups of 6 pregnant rats after daily oral administration of 0, 3, 10, 30 and 60 mg/kg bw/d from gestation Day 0 to gestation Day 11 (Cicalese, 2010). The test item was suspended in 0.5% CMC in water and administered at a dose volume of 10 mL/kg from Day 0 to Day 11 post coitum. The following investigations were performed on dams: mortality check, clinical signs, body weight and food consumption. All females were caesarean-sectioned on Day 12 post coitum and subjected to post mortem examination. The number of corpora lutea, implantations, early and late intrauterine deaths, live embryos and uterus weight was reported. No mortality occurred during the study. The number of females with live embryos on gestation Day 12 was 6 in all groups. No signs were recorded in treated animals. No differences of toxicological relevance were seen on the body weight gain. The food consumption was comparable between control and treated groups. No treatment-related effects were seen on the terminal body weight, uterus weight and absolute weight gain. Litter data were unaffected by the treatment. No findings were described in treated animal. sec-Butyl aminoethanol did not induce maternal toxicity, embryo-toxicity or adverse effects on the pre-implantation and post implantation stages of the embryo, when administered to pregnant rats from gestation Day 0 to gestation Day 11.

An analogue substanve, N-Methylethanolamine, was vaporized and administered at 150 ppm to approximately 15 pregnant rats for 7 hr/day on gestation days 7 to 15, and dams were sacrificed on day 20 of gestation (Nelson et al., 1984). Fetuses were individually weighed, and two-thirds of them were fixed in Bouin's solution and examined for soft-tissue anomalies. The other one-third were fixed in alcohol, stained with Alizarin Red and examined for skeletal defects. As overall result for the substance N-Methylethanolamine it can be stated that neither maternal nor fetal toxicity effects can be concluded in this study.


Justification for selection of Effect on developmental toxicity: via oral route:
Test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Justification for selection of Effect on developmental toxicity: via inhalation route:
Read across from methylaminoethanol

Justification for classification or non-classification

No classification is warranted according to CLP and DSD criteria.