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:
November 2011-March 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was performed according to OECD and US EPA guidelines and according to GLP principles.
Reason / purpose:
reference to same study
Qualifier:
according to
Guideline:
other: US EPA OPPTS 870.3550
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Inc., Portage, MI or Raleigh, NC
- Age at study initiation: (P) 10 wks
- Weight at study initiation: (P) Males: 331-391 g; Females: 211-291 g
- Fasting period before study: no
- Housing: until pairing: individually in stainless steel wire-mesh cage; paired for mating in male cage; after mating males were housed in suspended wire-mesh cages; females were transferred to plastic maternity cages with nesting material, ground corncob bedding.
- Use of restrainers for preventing ingestion (if dermal): yes/no
- Diet: ad libitum PMI Nutrition International, LLC Certified Rodent LabDiet 5002
- Water: ad libitum (reverse-osmosis-purified on-site drinking water)
- Acclimation period: minimum of 10 days for males and 15 days for females

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.2-21.4
- Humidity (%): 42.9-48.8
- Air changes (per hr): at least 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 2 December 2011 To: 22 January 2012
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
Test substance formulations were prepared approximately weekly for each dosage level, divided into daily aliquots and stored refrigerated. Test substance concentrations were 0, 4, 8, 16 and 32 mg/mL.
Dosage volume for all groups was 5 mL/kg.

VEHICLE
- Lot/batch no.: 2AD0465
Details on mating procedure:
- M/F ratio per cage: 1/1
- Proof of pregnancy: vaginal plug or the presence of sperm following a vaginal lavage referred to as day 0 of pregnancy
- After successful mating each pregnant female was caged: individually into plastic maternity cages with nesting material, ground corncob bedding.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Samples were collected from the middle stratum of each dosing formulation (including control group) prepared during the first and the last week of dose administration. Analyses were conducted by means of a validated gas chromatography method using flame ionization detection. Mean concentrations ranged from 95.3% to 108% of target concentration.
Duration of treatment / exposure:
males: 14 daily doses prior to mating; throughout mating period for a total of 31 doses
females: 14 daily doses prior to pairing, dosed through lactation day 3 (females that delivered) or post-mating day 25 (females that failed to deliver) for a total of 39-51 doses
Frequency of treatment:
daily
Remarks:
Doses / Concentrations:
0, 20, 40, 80, 160 mg/kg bw/day
Basis:
actual ingested
No. of animals per sex per dose:
12
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: based on the results of the 28-day OECD 407 study, where a NOAEL of 150 mg/kg bw/day was observed (no significant effects observed). For the present screening study a top dose of 160 mg/kg bw/day was included to increase the probability for detecting some adverse effects, without risking the possibility of excess toxicity due to the longer duration.
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily
- for moribundity and mortality, and for signs of toxicity 2 hours following dose administration. Females expected to deliver were observed for dystocia or other difficulties.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: weekly

BODY WEIGHT: Yes
- Time schedule for examinations: weekly for males and weekly for females until evidence of copulation. Female body weights were recorded on GD 0, 4, 7, 11, 14, 17 and 20 and on lactation days 0, 1 and 4.

FOOD CONSUMPTION: Yes
- Food consumption for each animal determined as g food/kg body weight/day: Yes

WATER CONSUMPTION: No
Sperm parameters (parental animals):
Parameters examined in P male parental generations:
testis weight, epididymides weight, seminal vesicle weight; microscopic examinations of testes, epididymides, seminal vesicles, coagulating glands and prostate gland.
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: no
- all pups

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
litter size, number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities

GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals following completion of the mating period.
- Maternal animals: All surviving animals on lactation day 4 (females that delivered) or on post-mating day 25 (females that failed to deliver)

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including external surface, all orifices and the cranial, thoracic, abdominal, and pelvic cavities, including viscera. Moreover the number and location of corpora lutea and implantation sites were examined , and uteri were investigated for early implantation loss.

HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues according to guidelines were prepared for microscopic examination and weighed, respectively.
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring was sacrificed at 4 days of age and discarded.


Statistics:
Analyses were conducted using two-tailed tests for minimum significance levels of 1% and 5%, comparing each test substance-treated group to the control group by sex.
Parental mating, fertility, conception and copulation indices were analyzed using the Chi-square test with Yates' correction factor. A parametric one-way ANOVA to determine intergroup differences was applied to mean parental BW, BW changes and food consumption, offspring BW and BW changes, gestation length, numbers of implantation sites, number of pups born, live litter size on PND 0, corpora lutea, organ weights (absolute and relative) and pre-coital intervals. If the ANOVA revealed significant (p<0.05) intergroup variance, Dunnett's test was used to compare the test substance-treated groups to the control group. Kruskal-Wallis nonparametric ANOVA was used to determine intergroup-differences for mean litter proportions of males at birth and postnatal survival. If the ANOVA revealed significant (p<0.05) intergroup variance, Dunn's test was used to compare the test substance-treated groups to the control group. Histopathological findings of treated groups were compared to the control group by the Fisher's Exact test.
Reproductive indices:
Mating, fertility, and copulation/conception indices were calculated as follows:

Male (Female)
Mating Index (%) = (No. of Males (Females) with Evidence of Mating (or Confirmed Pregnant)/Total No. of Males (Females) Used for Mating) x 100

Male Fertility
Index (%) = (No. of Males Siring a Litter/Total No. of Males Used for Mating) x 100

Male Copulation Index (%) = (No. of Males Siring a Litter/No. of Males with Evidence of Mating (or Females with Confirmed Pregnancy)) x 100

Female Fertility
Index (%) = (No. of Females with Confirmed Pregnancy/Total No. of Females Used for Mating) x 100

Female Conception Index (%) = (No. of Females with Confirmed Pregnancy/ No. of Females with Evidence of Mating (or Confirmed Pregnancy)) x 100
Offspring viability indices:
Litter parameters were defined as follows:

Mean Live Litter Size = Total No. of Viable Pups on PND 0/ No. of Litters with Viable Pups PND 0

Postnatal Survival Between Birth and PND 0 or PND 4 (Pre-selection) (% Per Litter) =
(Sum of (Viable Pups Per Litter on PND 0 or PND 4 [Pre-selection]/No. of Pups Born Per Litter)/No. of Litters Per Group) x 100

Postnatal Survival for All
Other Intervals (% Per Litter) = (Sum of (Viable Pups Per Litter at End of Interval N/Viable Pups Per Litter at Start of Interval N)/No. of Litters Per Group) x 100

Where N= PND 0-1 and 1-4
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
non-dose related salivation
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
slightly decreased food intake and BW gain in males only
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
slightly decreased food intake and BW gain in males only
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
increased liver (m/f) and kidney (m) weights
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
male liver and kidneys
Other effects:
not examined
Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS):
Salivation prior to dosing and clear material around nose and mouth are considered test substance related but not to be adverse.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS):
High dose males showed slightly lower, statistically significant during days 13-20 and 0-31, mean BW gain on day 31 (-6.3%) compared to controls.

ORGAN WEIGHTS (PARENTAL ANIMALS):
Mean (absolute and relative) liver weights were statistically significant increased in high dose females by 16 and 19.8%, respectively. In high dose males relative liver weight was statistically significant increased (17%) and relative kidney weight was increased by 12.7%. These observations are considered non-adverse.

HISTOPATHOLOGY (PARENTAL ANIMALS)
In high dose males increased hepatocellular vacuolation (minimal and mild) were observed; kidneys of high dose males showed an increase in basophilic tubules and an increase in minimal tubular dilatation. These observations are considered non-adverse.
Dose descriptor:
NOAEL
Effect level:
80 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: overall effects slightly lower bw gain in high dose males.
Dose descriptor:
NOAEL
Remarks:
reproduction
Effect level:
>= 160 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: see 'Remark'
Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings:
not examined
OTHER FINDINGS (OFFSPRING):
Lower mean number of pups born (11.9 and 11.3 pups per dam) were observed in the 80 and 160 mg/kg bw/day groups, respectively (control group 14.5 pups per dam). However, the results from the 80 mg/kg bw/day group are comparable to the historical minimum values observed in WIL historical control data, 11.7 pups per dam. Furthermore, the lower mean number of pups born in the high dose group were primarily attributed to one female (single pup born). Excluding this female , -mean number of pups born was 12.2, which is well within WIL historical data ranges. The observed lower mean values for pups born in the 80 and 160 mg/kg bw/day groups are considered therefore not toxicologiccally relevant or test-substance related.
Dose descriptor:
NOAEL
Remarks:
developmental
Generation:
F1
Effect level:
>= 160 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No developmental toxicity was observed up to the highest dose level tested (160 mg/kg bw/day).
Reproductive effects observed:
not specified
Conclusions:
Treatment with test item by oral gavage in male and female Sprague-Dawley rats at dose levels of 20, 40, 80 and 160 mg/kg bw/day according to EU/EPA test guidelines resulted in slightly lower body weight gain in high dose male rats. Based on these observations in high dose animals, the NOAEL for parental systemic toxicity is 80 mg/kg bw/day. No reproduction and developmental toxicity was observed for treatment up to 160 mg/kg bw/day. A reproduction and developmental NOAEL of at least 160 mg/kg bw/day was derived.
Executive summary:

Test Guidance

According to OECD 421 and US EPA OPPTS 870.3550 guidelines

Method and materials

Test item was administrated by daily oral gavage to male and female Sprague-Dawley rats at dose levels of 20, 40, 80 and 160 mg/kg bw/day. Males were exposed for 2 weeks prior to mating, during mating and up to termination (total of 31 doses). The females were exposed for 2 weeks prior to mating, during mating, during post-coitum, and during 3 days of lactation (39 -51 doses).

Results

Analysis demonstrated mean concentration ranges from 95.3% to 108% of target concentrations.

Parental toxicity was observed at the high dose only, and consisted of slightly lower body weight gain in high dose male rats at 160 mg/kg bw/day. The observed statistically significant increased relative liver weights in males and females, and increased absolute liver weights in females, together with increased hepatocellular vacuolation and increased minimal tubular dilation in kidneys observed in high dose males are considered non-adverse. Based on these observations in high dose animals, the NOAEL for parental systemic toxicity is 80 mg/kg bw/day. Maternal toxicity was at least 160 mg/kg bw/day, as no adverse effects were observed in dams.

Conclusions

No reproduction and developmental toxicity was observed for treatment up to 160 mg/kg bw/day. A reproduction and developmental NOAEL of at least 160 mg/kg bw/day was derived.
Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
160 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
Satisfactory
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 key screening study for reproduction and developmental effects was performed according to OECD 421 and US EPA OPPTS 870.3550 guidelines

The test item was administrated by daily oral gavage to male and female Sprague-Dawley rats at dose levels of 20, 40, 80 and 160 mg/kg bw/day. Males were exposed for 2 weeks prior to mating, during mating and up to termination (total of 31 doses). The females were exposed for 2 weeks prior to mating, during mating, during post-coitum, and during 3 days of lactation (39 -51 doses).

Analysis demonstrated mean concentration ranges from 95.3% to 108% of target concentrations.

Parental toxicity was observed at the high dose only, and consisted of slightly lower body weight gain in high dose male rats at 160 mg/kg bw/day. The observed statistically significant increased relative liver weights in males and females, and increased absolute liver weights in females, together with increased hepatocellular vacuolation and increased minimal tubular dilation in kidneys observedin high dose males are considered non-adverse. Based on these observations in high dose animals, the NOAEL for parental systemic toxicity is 80 mg/kg bw/day. Maternal toxicity was at least 160 mg/kg bw/day, as no adverse effects were observed in dams.

No reproduction and developmental toxicity was observed for treatment up to 160 mg/kg bw/day. A reproduction and developmental NOAEL of at least 160 mg/kg bw/day was derived.

Further studies to investigate reproduction toxicity are considered to be scientifically unjustified.

EC 276-743-1 has a relatively low molecular weight of 192.3 gm/mol, the divalent form having a molecular weight of 290 gm/mol. The substance is slightly soluble in water( 42.1 mg/L), with a relatively high octanol/water partition coefficient (log Kow = 4.5) and a low vapour pressure (0.26Pa @ 25oC).

Although the physical chemical properties suggest that EC 276-743-1 is of adequate molecular size to participate in endogenous absorption mechanisms within the mammalian gastrointestinal tract should that material be ingested, acute oral, a 28-day repeated-dose oral gavage toxicity and reproduction/developmental toxicity screening studies identified that systemic toxicity was secondary to the local irritation effects on the forestomach of rats. The forestomach in rodents is an organ that is not relevant to humans and consequently such local irritant effects, and any secondary effects related to local irritation in the forestomach, are not relevant to human exposure.

EC 276-743-1 was also tested for acute toxicity following dermal application. The single-dose dermal application of the test material resulted in no manifestations of systemic toxicity that would suggest systemic absorption through cutaneous barriers.Dermal absorption studies on alkyl phenols and structural analogues have shown thatexposure of human skin to concentrations of up to 10% NPE result in minimal exposure. The percentage dose absorbed was concentration dependant (ca. 1% for a 0.1% solution, 0.1% for a 1% solution and 0.01% for a 10% solution). There were no large species differences for absorption seen in the study. Therefore systemic absorption through the cutaneous barrier is unlikely for the proposed uses.

The potential for inhalation toxicity was not measured. However, the EC 276-743-1 vapour pressure indicates a very low propensity to enter atmospheric air in a respirable form (predicted to be ca. 0.1 ppm under ambient conditions). Thus, respiratory absorption under normal use and handling of this material is expected to be inconsequential.

The corrosive/irritant nature of the test material at low concentrations also indicates that any effects observed in long-term repeat dose studies such as those for reproduction toxicity would also be attributable to the local irritant effect prior to observation of any systemic effect as noted in the existing studies via the oral route. It may be concluded that adverse systemic toxicity only occurs at or above dose levels that cause severe local irritation

A screening study for reproductive and developmental toxicity has been performed with the substance at dose levels derived from the 28-day repeat oral dose study in rats. At the highest dose level tested (160 mg/kg bw/day) indications of irritation effects were observed (weight loss in males) but these were not considered to be adverse. At the higher dose levels used in the 28-day repeat oral dose study, effects on reproductive organs (seminal vesicles) were seen. Additional tissues to investigate the effects on male reproduction organ parameters were included in the reproduction/developmental toxicity screening but the effects seen in the 28-day repeat oral dose study were not reproduced in the screening study at the lower dose levels where irritation effects were classed as non-adverse.

Therefore, further studies to investigate reproduction toxicity are considered to be scientifically unjustified.


Short description of key information:
Based on observations in high dose animals, the NOAEL for parental systemic toxicity is 80 mg/kg bw/day. Maternal toxicity was at least 160 mg/kg bw/day, as no adverse effects were observed in dams.
No reproduction and developmental toxicity was observed for treatment up to 160 mg/kg bw/day. A reproduction and developmental NOAEL of at least 160 mg/kg bw/day was derived.

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

Effects on developmental toxicity

Description of key information
Based on observations in high dose animals, the NOAEL for parental systemic toxicity is 80 mg/kg bw/day. Maternal toxicity was at least 160 mg/kg bw/day, as no adverse effects were observed in dams.
No reproduction and developmental toxicity was observed for treatment up to 160 mg/kg bw/day. A reproduction and developmental NOAEL of at least 160 mg/kg bw/day was derived.
Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
November 2011-March 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was performed according to OECD and US EPA guidelines and according to GLP principles.
Reason / purpose:
reference to same study
Qualifier:
according to
Guideline:
other: US EPA OPPTS 870.3550
Deviations:
no
Qualifier:
according to
Guideline:
other: OECD guideline 421 (Reproduction/Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Inc., Portage, MI or Raleigh, NC
- Age at study initiation: (P) 10 wks
- Weight at study initiation: (P) Males: 331-391 g; Females: 211-291 g
- Fasting period before study: no
- Housing: until pairing: individually in stainless steel wire-mesh cage; paired for mating in male cage; after mating males were housed in suspended wire-mesh cages; females were transferred to plastic maternity cages with nesting material, ground corncob beding.
- Use of restrainers for preventing ingestion (if dermal): yes/no
- Diet: ad libitum PMI Nutrition International, LLC Certified Rodent LabDiet 5002
- Water: ad libitum (reverse-osmosis-purified on-site drinking water)
- Acclimation period: minimum of 10 days for males and 15 days for females

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.2-21.4
- Humidity (%): 42.9-48.8
- Air changes (per hr): at least 10
- Photoperiod (hrs dark / hrs light): 12/12
IN-LIFE DATES: From: 2 December 2011 To: 22 January 2012
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
Test substance formulations were prepared approximately weekly for each dosage level, divided into daily aliquots and stored refrigerated. Test substance concentrations were 0, 4, 8, 16 and 32 mg/mL.
Dosage volume for all groups was 5 mL/kg.


VEHICLE
- Lot/batch no.: 2AD0465
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Samples were collected from the middle stratum of each dosing formulation (including control group) prepared during the first and the last week of dose administration. Analyses were conducted by means of a validated gas chromatography method using flame ionization detection. Mean concentrations ranged from 95.3% to 108% of target concentration.
Details on mating procedure:
- M/F ratio per cage: 1/1
- Proof of pregnancy: vaginal plug or the presence of sperm following a vaginal lavage referred to as day 0 of pregnancy
- After successful mating each pregnant female was caged: individually into plastic maternity cages with nesting material, ground corncob bedding.
Duration of treatment / exposure:
males: 14 daily doses prior to mating; throughout mating period for a total of 31 doses
females: 14 daily doses prior to pairing, dosed through lactation day 3 (females that delivered) or post-mating day 25 (females that failed to deliver) for a total of 39-51 doses
Frequency of treatment:
daily
Duration of test:
until lactation day 4
Remarks:
Doses / Concentrations:
0, 20, 40, 80, 160 mg/kg bw/day
Basis:
actual ingested
No. of animals per sex per dose:
12
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: based on the results of the 28-day OECD 407 study, where a NOAEL of 150 mg/kg bw/day was observed (no significant effects observed). For the present screening study a top dose of 160 mg/kg bw/day was included to increase the probabilty for detecting some adverse effects, without risking the possibility of excess toxicity due to the longer duration.
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily
- for moribundity and mortality, and for signs of toxicity 2 hours following dose administration. Females expected to deliver were observed for dystocia or other difficulties.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: weekly

BODY WEIGHT: Yes
- Time schedule for examinations: weekly for males and weekly for females until evidence of copulation. Female body weights were recorded on GD 0, 4, 7, 11, 14, 17 and 20 and on lactation days 0, 1 and 4.
FOOD CONSUMPTION
- Food consumption for each animal determined as g food/kg body weight/day: Yes

WATER CONSUMPTION: No

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on lactation day #4
- Organs examined: according to guidelines
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes / No / No data
Examinations included:
- Gravid uterus weight: No
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: No (dams delivered)
- Number of late resorptions: No (dams delivered)
Fetal examinations:
- External examinations: Yes, Fo animals delivered and the dams and her litter were euthanized on LD/PND 4. All pups were examined.
Statistics:
Analyses were conducted using two-tailed tests for minimum significance levels of 1% and 5%, comparing each test substance-treated group to the control group by sex.
A parametric one-way ANOVA to determine intergroup differences was applied to mean parental BW, BW changes and food consumption, offspring BW and BW changes, gestation length, numbers of implantation sites, number if pups born, live litter size on PND 0, corpora lutea, organ weights (absolute and relative) and pre-coital intervals. If the ANOVA revealed significant (p<0.05) intergroup variance, Dunnett's test was used to compare the test substance-treated groups to the control group. Kruskal-Wallis nonparametric ANOVA was used to determine intergroup differences for mean litter proportions of males at birth and postnatal survival. If the ANOVA revealed significant (p<0.05) intergroup variance, Dunn's test was used to compare the test substance-treated groups to the control group. Histopathological findings of treated groups were compared to the control group by the Fisher's Exact test.
Indices:
Litter parameters were defined as follows:

Mean Live Litter Size = Total No. of Viable Pups on PND 0/ No. of Litters with Viable Pups PND 0

Postnatal Survival Between Birth and PND 0 or PND 4 (Pre-selection) (% Per Litter) =
(Sum of (Viable Pups Per Litter on PND 0 or PND 4 [Pre-selection]/No. of Pups Born Per Litter)/No. of Litters Per Group) x 100

Postnatal Survival for All
Other Intervals (% Per Litter) = (Sum of (Viable Pups Per Litter at End of Interval N/Viable Pups Per Litter at Start of Interval N)/No. of Litters Per Group) x 100

Where N= PND 0-1 and 1-4
Details on maternal toxic effects:
Maternal toxic effects:yes

Details on maternal toxic effects:
In high dose females a statistically significant increased relative liver weight (m/f) and increased absolute liver weight (f) were observed. These effects are considered non-adverse.
Dose descriptor:
NOAEL
Effect level:
>= 160 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: developmental toxicity
Dose descriptor:
NOAEL
Effect level:
>= 160 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: maternal toxicity
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects
Abnormalities:
not specified
Developmental effects observed:
not specified
Conclusions:
Treatment with test item by oral gavage in male and female Sprague-Dawley rats at dose levels of 20, 40, 80 and 160 mg/kg bw/day according to EU/EPA test guidelines did not result in adverse effects in the dams. Based on the absence of adverse effects in the dams, the NOAEL for maternal systemic toxicity is at least 160 mg/kg bw/day. No reproduction and developmental toxicity was observed for treatment up to 160 mg/kg bw/day. A reproduction and developmental NOAEL of at least 160 mg/kg bw/day was derived.
Executive summary:

Test Guidance

According to OECD 421 and US EPA OPPTS 870.3550 guidelines

Method and materials

Test item was administrated by daily oral gavage to male and female Sprague-Dawley rats at dose levels of 20, 40, 80 and 160 mg/kg bw/day . Males were exposed for 2 weeks prior to mating, during mating and up to termination (total of 31 doses). The females were exposed for 2 weeks prior to mating, during mating, during post-coitum, and during 3 days of lactation (39 -51 doses).

Results

Analysis demonstrated mean concentration ranges from 95.3% to 108% of target concentrations.

Parental toxicity was observed at the high dose only, and consisted of slightly lower body weight gain in high dose male rats at 160 mg/kg bw/day. The observed statistically significant increased relative liver weights in males and females, and increased absolute liver weights in females, together with increased hepatocellular vacuolation and increased minimal tubular dilation in kidneys observed in high dose males are considered non-adverse. Based on these observations in high dose animals, the NOAEL for parental systemic toxicity is 80 mg/kg bw/day. Maternal toxicity was at least 160 mg/kg bw/day, as no adverse effects were observed in dams.

Conclusions

No reproduction and developmental toxicity was observed for treatment up to 160 mg/kg bw/day. A reproduction and developmental NOAEL of at least 160 mg/kg bw/day was derived.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
160 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
Satisfactory
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

A key screening study for reproduction and developmental effects was performed according to OECD 421 and US EPA OPPTS 870.3550 guidelines

The test item was administrated by daily oral gavage to male and female Sprague-Dawley rats at dose levels of 20, 40, 80 and 160 mg/kg bw/day. Males were exposed for 2 weeks prior to mating, during mating and up to termination (total of 31 doses). The females were exposed for 2 weeks prior to mating, during mating, during post-coitum, and during 3 days of lactation (39 -51 doses).

Analysis demonstrated mean concentration ranges from 95.3% to 108% of target concentrations.

Parental toxicity was observed at the high dose only, and consisted ofslightly lower body weight gain in high dose male rats at 160 mg/kg bw/day. The observed statistically significant increased relative liver weights in males and females, and increased absolute liver weights in females, together with increased hepatocellular vacuolation and increased minimal tubular dilation in kidneys observedin high dose males are considered non-adverse. Based on these observations in high dose animals, the NOAEL for parental systemic toxicity is 80 mg/kg bw/day. Maternal toxicity was at least 160 mg/kg bw/day, as no adverse effects were observed in dams.

No reproduction and developmental toxicity was observed for treatment up to 160 mg/kg bw/day. A reproduction and developmental NOAEL of at least 160 mg/kg bw/day was derived.

Further studies to investigate reproduction toxicity are considered to be scientifically unjustified.

EC 276-743-1 has a relatively low molecular weight of 192.3 gm/mol, the divalent form having a molecular weight of 290 gm/mol. The substance is slightly soluble in water( 42.1 mg/L), with a relatively high octanol/water partition coefficient (log Kow = 4.5) and a low vapour pressure (0.26Pa @ 25oC).

Although the physical chemical properties suggest that EC 276-743-1 is of adequate molecular size to participate in endogenous absorption mechanisms within the mammalian gastrointestinal tract should that material be ingested, acute oral, a 28-day repeated-dose oral gavage toxicity and reproduction/developmental toxicity screening studies identified that systemic toxicity was secondary to the local irritation effects on the forestomach of rats. The forestomach in rodents is an organ that is not relevant to humans and consequently such local irritant effects, and any secondary effects related to local irritation in the forestomach, are not relevant to human exposure.

EC 276-743-1 was also tested for acute toxicity following dermal application. The single-dose dermal application of the test material resulted in no manifestations of systemic toxicity that would suggest systemic absorption through cutaneous barriers.Dermal absorption studies on alkyl phenols and structural analogues have shown thatexposure of human skin to concentrations of up to 10% NPE result in minimal exposure. The percentage dose absorbed was concentration dependant (ca. 1% for a 0.1% solution, 0.1% for a 1% solution and 0.01% for a 10% solution). There were no large species differences for absorption seen in the study. Therefore systemic absorption through the cutaneous barrier is unlikely for the proposed uses.

The potential for inhalation toxicity was not measured. However, the EC 276-743-1 vapour pressure indicates a very low propensity to enter atmospheric air in a respirable form (predicted to be ca. 0.1 ppm under ambient conditions). Thus, respiratory absorption under normal use and handling of this material is expected to be inconsequential.

The corrosive/irritant nature of the test material at low concentrations also indicates that any effects observed in long-term repeat dose studies such as those for reproduction toxicity would also be attributable to the local irritant effect prior to observation of any systemic effect as noted in the existing studies via the oral route. It may be concluded that adverse systemic toxicity only occurs at or above dose levels that cause severe local irritation

A screening study for reproductive and developmental toxicity has been performed with the substance at dose levels derived from the 28-day repeat oral dose study in rats. At the highest dose level tested (160 mg/kg bw/day) indications of irritation effects were observed (weight loss in males) but these were not considered to be adverse. At the higher dose levels used in the 28-day repeat oral dose study, effects on reproductive organs (seminal vesicles) were seen. Additional tissues to investigate the effects on male reproduction organ parameters were included in the reproduction/developmental toxicity screening but the effects seen in the 28-day repeat oral dose study were not reproduced in the screening study at the lower dose levels where irritation effects were classed as non-adverse.

Therefore, further studies to investigate reproduction toxicity are considered to be scientifically unjustified.


Justification for selection of Effect on developmental toxicity: via oral route:
GLP Guideline study

Justification for classification or non-classification

No effects on reproduction and development are observed below the limit of parental toxicity.