Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Toxicological information

Developmental toxicity / teratogenicity

Currently viewing:

Administrative data

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
16-11-2004 to 17-12-2004
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Cross-reference
Reason / purpose for cross-reference:
reference to other study
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Reason / purpose for cross-reference:
reference to other study
Qualifier:
no guideline required
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
other: Crl:CD®(SD)IGS VAF/Plus®
Details on test animals or test system and environmental conditions:
Forty Crl:CD®(SD)IGS BR VAF/Plus® presumed pregnant female rats were randomly assigned to five dosage groups (Groups I through V), eight rats per group.
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
Suspensions of the test article, alpha-iso-Methylionone, and/or the vehicle, corn oil, were administered via oral gavage once daily on days 7 through 17 of presumed gestation (DGs 7 through 17) at dosages of 0, 1.25, 2.5, 5 and 10 mg/kg/day. The dosage volume was 10 mL/kg, adjusted daily on the basis of the individual body weights recorded immediately before administration.
Analytical verification of doses or concentrations:
yes
Details on mating procedure:
not applicable
Duration of treatment / exposure:
Rats were administered the test article and/or vehicle once daily on DGs 7 through 17.
The dosage volume was adjusted daily on the basis of the individual body weights recorded before administration. The rats were administered the test article at approximately the same time each day.
Frequency of treatment:
Rats were administered the test article and/or vehicle once daily.
Duration of test:
21 days
Dose / conc.:
1.25 mg/kg bw/day (actual dose received)
Dose / conc.:
2.5 mg/kg bw/day (actual dose received)
Dose / conc.:
5 mg/kg bw/day (actual dose received)
Dose / conc.:
10 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
8 females
Control animals:
yes, concurrent vehicle
Maternal examinations:
Checks for viability were made twice daily. Clinical observations were recorded daily before dosage. On the first day of dosage administration, post-dosage observations were recorded hourly for the first four hours and at the end of the working day; all subsequent observations were recorded at 60±10 minutes after dosage administration. These observations were also recorded once daily during the post-dosage period. Body weight values were recorded on DG 0 and daily during the dosage and post-dosage periods. Feed consumption values were recorded on DGs 0, 7, 10, 12, 15, 18 and 21.
Ovaries and uterine content:
All rats were sacrificed on DG 21 and were examined for the number and distribution of corpora lutea, implantation sites and uterine contents.
Fetal examinations:
A gross necropsy of the thoracic, abdominal and pelvic viscera was performed. Fetuses were weighed and examined for gross external alterations and sex.
Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
Dead fetuses:
no effects observed
Changes in number of pregnant:
no effects observed
Dose descriptor:
NOAEL
Remarks:
Maternal toxicity
Effect level:
>= 10 mg/kg bw/day
Based on:
test mat.
Remarks on result:
not determinable due to absence of adverse toxic effects
Dose descriptor:
NOAEL
Remarks:
Reproductive toxicity
Effect level:
>= 10 mg/kg bw/day
Based on:
test mat.
Remarks on result:
not determinable due to absence of adverse toxic effects
Abnormalities:
no effects observed
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
no effects observed
Changes in litter size and weights:
no effects observed
External malformations:
no effects observed
Visceral malformations:
no effects observed
Dose descriptor:
NOAEL
Remarks:
Developmental toxicity
Effect level:
>= 10 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects
Abnormalities:
no effects observed
Developmental effects observed:
no
Conclusions:
Under the conditions of the DRF study in rats (GLP), the NOAEL for maternal and developmental toxicity was at least 10 mg/kg bw/day.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2005
Report date:
2005

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
3-methyl-4-(2,6,6-trimethyl-2-cyclohexen-1-yl)-3-buten-2-one
EC Number:
204-846-3
EC Name:
3-methyl-4-(2,6,6-trimethyl-2-cyclohexen-1-yl)-3-buten-2-one
Cas Number:
127-51-5
Molecular formula:
C14H22O
IUPAC Name:
3-methyl-4-(2,6,6-trimethylcyclohex-2-en-1-yl)but-3-en-2-one
Test material form:
liquid

Test animals

Species:
rat
Strain:
other: Crl:CD®(SD)IGS VAF/Plus®
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Inc., Raleigh, North Carolina
- Age at study initiation: 67 days
- Weight at study initiation: 213 - 241
- Housing: Rats were individually housed in stainless steel, wire-bottomed cages, except during the cohabitation period. During cohabitation, each pair of male and female rats was housed in the male rat’s cage. All cage sizes and housing conditions were in compliance with the Guide for the Care and Use of Laboratory Animals.
- Diet: Rats were given ad libitum access to Certified Rodent Diet® #5002 (PMI® Nutrition International, Inc., St. Louis, Missouri) in individual feeders.
- Water: Local water that had been processed by passage through a reverse osmosis membrane (R.O. water) was available to the rats ad libitum from an automatic watering access system and/or individual water bottles attached to the cages. Chlorine was added to the processed water as a bacteriostat.
- Acclimation period: five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): targeted at 64°F to 79°F (18°C to 26°C);
- Humidity (%): targeted at 30% to 70%
- Air changes (per hr): ten changes per hour of 100% fresh air that had been passed through 99.97% HEPA filters
- Photoperiod (hrs dark / hrs light): An automatically controlled 12-hours light:12-hours dark fluorescent light cycle was maintained. Each dark period began at 1900. The lights were turned on 9 minutes early on 15 December 2004, to facilitate the laboratory schedule.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
Suspensions of the test article were prepared daily at the Testing Facility.

VEHICLE
- Lot/batch no.: 103K0107 and 074K0025
- Purity: Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to have been present in the vehicle that would have interfered with the results of this study. The expiration dates are October 2007 (lot 103K0107) and November 2008 (lot 074K0025).
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The results of all concentration and homogeneity analyses were within ±10% of target concentration and ≤5% RSD, respectively. The 3 mg/mL concentration level was determined to be stable for ten days when stored at ambient conditions, protected from light. The results of these analyses are available. Stability at the same conditions covering a range of 0.125 to 1 mg/mL was verified in CR-DDS, Argus Division Protocol TIF00001.
Details on mating procedure:
After acclimation, 140 virgin female rats were placed into cohabitation with 140 breeder male rats, one male rat per female rat. The cohabitation period consisted of a maximum of five days. Female rats with spermatozoa observed in a smear of the vaginal contents and/or a copulatory plug observed in situ were considered to be DG 0 and assigned to individual housing. Rats were observed for viability at least twice each day of the study and for general appearance weekly during the acclimation period and on DG 0.
Duration of treatment / exposure:
Gestation Days 7-17
Frequency of treatment:
once daily
Duration of test:
21 days
Doses / concentrationsopen allclose all
Dose / conc.:
3 mg/kg bw/day (actual dose received)
Dose / conc.:
10 mg/kg bw/day (actual dose received)
Dose / conc.:
30 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
25 females
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Dosages were selected on the basis of a dosage-range study (Argus Protocol TIF00001), in which dosages of 0, 1.25, 2.5, 5 and 10 mg/kg/day were administered daily on DGs 7 to 17. In that range-finding study, no adverse clinical observations were considered testarticle related. Although not dosage dependent, body weight gains were increased during the entire dosage period (defined as DGs 7 through 18), the gestation period following the initiation of dosage administration (DGs 7 to 21), and the complete gestation period (DGs 0 to 21), in comparison with the vehicle control group values. During the postdosage period (DGs 18 to 21), body weight gains remained increased in the 1.25, 2.5, 5 and 10 mg/kg/day dosage groups, as compared to the vehicle control group value. Absolute (g/day) and relative (g/kg/day) feed consumption values during the dosage period (DGs 7 to 18) were comparable between the five dosage groups. All rats were pregnant. There were no litters consisting of all resorbed conceptuses, and there were no dead fetuses. All Caesarean-sectioning and litter observations were comparable between the five dosage groups, and no fetal gross alterations were noted. Based on these data, dosages of 0 (Vehicle), 3, 10 and 30 mg/kg/day of alpha-iso-Methylionone were selected for the developmental toxicity study in rats. The 10 mg/kg/day dosage was expected to be a no-observable-adverse-effect-level (NOAEL) for both maternal and embryo-fetal toxicity.

Examinations

Maternal examinations:
Female rats with spermatozoa observed in a smear of the vaginal contents and/or a copulatory plug observed in situ were considered to be DG 0 and assigned to individual housing. Rats were observed for viability at least twice each day of the study and for general appearance weekly during the acclimation period and on DG 0. The rats were also examined for clinical observations, abortions, premature deliveries and deaths before and approximately 60 ± 10 minutes after dosage administration and once daily during the postdosage period. Body weights were recorded weekly during the acclimation period, on DG 0, and daily during the dosage and postdosage periods. Feed consumption values were recorded on DGs 0, 7, 10, 12, 15, 18 and 21.
Ovaries and uterine content:
All female rats were sacrificed by carbon dioxide asphyxiation on DG 21, Caesarean-sectioned and a gross necropsy of the thoracic, abdominal and pelvic viscera was performed. Uteri of apparently non-pregnant rats were examined while being pressed between glass plates to confirm the absence of implantation sites. Uteri and ovaries of apparently non-pregnant rats were retained in neutral buffered 10% formalin will be discarded when authorized by the Study Director. The number and distribution of corpora lutea were recorded. The uterus of each rat was excised and examined for pregnancy, number and distribution of implantation sites, live and dead fetuses and early and late resorptions. An early resorption was defined as a conceptus in which organogenesis was not grossly evident. A late resorption was defined as one in which the occurrence of organogenesis was grossly evident. A live fetus was defined as a term fetus that responded to stimuli. Non-responding term fetuses were considered to be dead (there were no dead fetuses). Dead fetuses and late resorptions were differentiated by the degree of autolysis present; marked to extreme autolysis indicated that the fetus was a late resorption. Placentae were examined for size, colour and shape.
Fetal examinations:
Each fetus was removed from the uterus, placed in an individual container and individually identified with a tag noting the study number, litter number, uterine distribution and fixative. Each fetus was subsequently weighed and examined for sex and gross lesions. Live fetuses were sacrificed by an intraperitoneal injection of sodium pentobarbital. Approximately one-half of the fetuses in each litter were examined for soft tissue alterations, using an adaptation of Wilson’s sectioning technique. These fetuses were fixed in Bouin’s solution; sections were retained in alcohol. The remaining fetuses (approximately one-half of the fetuses in each litter) were eviscerated, cleared, stained with alizarin red S and examined for skeletal alterations. The fetuses were initially fixed in alcohol. Skeletal preparations were retained in glycerin with thymol added as a preservative.
Statistics:
- Data generated during the course of this study were recorded either by hand or using the Argus Automated Data Collection and Management System and the Vivarium Temperature and Relative Humidity Monitoring System. All data were tabulated, summarized and/or statistically analyzed using the Argus Automated Data Collection and Management System, the Vivarium Temperature and Relative Humidity Monitoring System, Microsoft® Excel (part of Microsoft® Office 97/2000/XP), Quattro Pro 8 and/or The SAS System (version 6.12).
- Averages and percentages were calculated. Litter values were used where appropriate.
- Clinical observations and other proportional data were analyzed using the Variance Test for Homogeneity of the Binomial Distribution.
- Continuous data (e.g., body weights, body weight changes, feed consumption values and litter averages for percent male fetuses, percent resorbed conceptuses, fetal body weights and fetal anomaly data) were analyzed using Bartlett’s Test of Homogeneity of Variances and the Analysis of Variance, when appropriate [i.e., Bartlett’s Test was not significant (p>0.001)]. If the Analysis of Variance was significant (p≤0.05), Dunnett’s Test was used to identify the statistical significance of the individual groups. If the Analysis of Variance was not appropriate [i.e., Bartlett’s Test was significant (p≤0.001)], the Kruskal-Wallis Test was used, when less than or equal to 75% ties were present. In cases where the Kruskal-Wallis Test was statistically significant (p≤0.05), Dunn’s Method of Multiple Comparisons was used to identify the statistical significance of the individual groups. If there were greater than 75% ties, Fisher’s Exact Test was used to analyze the data.
- Count data obtained at Caesarean-sectioning of the dams were evaluated using the procedures described above for the Kruskal-Wallis Test.

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
All clinical observations were considered to be unrelated to treatment because: 1) the number of affected rats was not dosage dependent; and/or 2) the number of rats with an observation did not significantly differ from the control group values. Clinical observations included chromorhinorrhea, excess salivation, sparse hair coat, localized alopecia on the limbs, urine-stained abdominal fur, red or yellow perioral substance, rales, decreased motor activity, cold to touch, ungroomed coat, soft or liquid faeces, the presence of a scab on the head, and the presence of a red perivaginal substance.
Mortality:
no mortality observed
Description (incidence):
All female rats survived to scheduled sacrifice.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Body weights and body weight gains were unaffected at dosages as high as 30 mg/kg/day. Body weight gains in the 3, 10 and 30 mg/kg/day dosage group were 99.0%, 99.9% and 101.2%, respectively, of the control group value for the entire dosage period (calculated as DGs 7 to 18). Maternal body weights on DG 21 in the 3, 10 and 30 mg/kg/day dosage group were 99.2%, 99.8% and 99.1%, respectively, of the control group value.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Absolute (g/day) and relative (g/kg/day) feed consumption values were unaffected by dosages as high as 30 mg/kg/day. Absolute feed consumption values in the 3, 10 and 30 mg/kg/day dosage groups were 101.0%, 102.1% and 102.6%, respectively, of the control group value for the entire dosage period (calculated as DGs 7 to 18).
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
All necropsy observations were considered to be unrelated to treatment because: 1) the number of affected rats was not dosage dependent; and/or 2) the number of rats with an observation did not significantly differ from the control group values. One 3 mg/kg/day dosage group rat had urinary tract lesions [slight dilation of the pelvis of both kidneys, thickened urinary bladder walls, presence of renal calculi (ureter and bladder)]. Incidences of these lesions are common in this strain of rat. The only other gross lesion observed was a tan mass in the thoracic cavity that adhered to all the thoracic viscera in one 10 mg/kg/day dosage group rat. No other gross lesions were observed.

Maternal developmental toxicity

Pre- and post-implantation loss:
no effects observed
Description (incidence and severity):
The litter averages for implantations were comparable among the four dosage groups and did not significantly differ.
Total litter losses by resorption:
no effects observed
Description (incidence and severity):
No dam had a litter consisting of only resorbed conceptuses.
Early or late resorptions:
no effects observed
Description (incidence and severity):
The litter averages for early and late resorptions, percentage of dead or resorbed conceptuses were comparable among the four dosage groups and did not significantly differ.
Dead fetuses:
no effects observed
Description (incidence and severity):
There were no dead fetuses.
Changes in number of pregnant:
no effects observed
Description (incidence and severity):
Pregnancy occurred in 21 to 25 female rats per dosage group.
Other effects:
no effects observed
Description (incidence and severity):
No Caesarean-sectioning or litter parameters were affected by dosages as high as 30 mg/kg/day. The litter averages for corpora lutea were comparable among the four dosage groups and did not significantly differ. All placentae appeared normal.

Effect levels (maternal animals)

open allclose all
Dose descriptor:
NOAEL
Remarks:
Maternal toxicity
Effect level:
>= 30 mg/kg bw/day
Based on:
test mat.
Remarks on result:
not determinable due to absence of adverse toxic effects
Dose descriptor:
NOAEL
Remarks:
Reproductive toxicity
Effect level:
>= 30 mg/kg bw/day
Based on:
test mat.
Remarks on result:
not determinable due to absence of adverse toxic effects

Maternal abnormalities

Abnormalities:
no effects observed

Results (fetuses)

Fetal body weight changes:
no effects observed
Description (incidence and severity):
The litter averages for fetal body weights were comparable among the four dosage groups and did not significantly differ.
Reduction in number of live offspring:
no effects observed
Description (incidence and severity):
The litter averages for live fetuses were comparable among the four dosage groups and did not significantly differ.
Changes in sex ratio:
no effects observed
Description (incidence and severity):
No Caesarean-sectioning or litter parameters were affected by dosages as high as 30 mg/kg/day. The litter averages for percentage of live male fetuses were comparable among the four dosage groups and did not significantly differ.
Changes in litter size and weights:
no effects observed
Description (incidence and severity):
The litter averages for litter sizes, fetal body weights were comparable among the four dosage groups and did not significantly differ.
External malformations:
no effects observed
Description (incidence and severity):
All fetal gross external (malformations or variations) were considered unrelated to treatment because: 1) the fetal or litter incidence was not dosage-dependent; 2) the incidences did not significantly differ from the control group values; and/or 3) the incidences were within the ranges observed historically at the Testing Facility. In Groups 0, 3, 10, 30 mg/kg bw, litters with fetuses with alterations numbered 5 (20.8%), 7 (28.0%), 7 (29.2%) and 2 (9.5%), respectively. The numbers of fetuses with any alteration observed were 6 (1.7%), 7 (1.9%), 14 (4.2%) and 5 (1.7%) and the percentages of fetuses with any alteration per litter were 1.6%, 1.9%, 3.8% and 1.8% in these same respective dosage groups.
Skeletal malformations:
no effects observed
Description (incidence and severity):
All fetal skeletal alterations (malformations or variations) were considered unrelated to treatment because: 1) the fetal or litter incidence was not dosage-dependent; 2) the incidences did not significantly differ from the control group values; and/or 3) the incidences were within the ranges observed historically at the Testing Facility. Skeletal ossification averages did not differ among the groups. In Groups 0, 3, 10, 30 mg/kg bw, litters with fetuses with alterations numbered 5 (20.8%), 7 (28.0%), 7 (29.2%) and 2 (9.5%), respectively. The numbers of fetuses with any alteration observed were 6 (1.7%), 7 (1.9%), 14 (4.2%) and 5 (1.7%) and the percentages of fetuses with any alteration per litter were 1.6%, 1.9%, 3.8% and 1.8% in these same respective dosage groups.
Visceral malformations:
no effects observed
Description (incidence and severity):
All fetal soft tissue alterations (malformations or variations) were considered unrelated to treatment because: 1) the fetal or litter incidence was not dosage-dependent; 2) the incidences did not significantly differ from the control group values; and/or 3) the incidences were within the ranges observed historically at the Testing Facility. Skeletal ossification averages did not differ among the groups. In Groups 0, 3, 10, 30 mg/kg bw, litters with fetuses with alterations numbered 5 (20.8%), 7 (28.0%), 7 (29.2%) and 2 (9.5%), respectively. The numbers of fetuses with any alteration observed were 6 (1.7%), 7 (1.9%), 14 (4.2%) and 5 (1.7%) and the percentages of fetuses with any alteration per litter were 1.6%, 1.9%, 3.8% and 1.8% in these same respective dosage groups.

Effect levels (fetuses)

Dose descriptor:
NOAEL
Remarks:
Developmental toxicity
Effect level:
>= 30 mg/kg bw (total dose)
Based on:
test mat.
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects

Fetal abnormalities

Abnormalities:
no effects observed

Overall developmental toxicity

Developmental effects observed:
no

Applicant's summary and conclusion

Conclusions:
Under the conditions of the oral developmental toxicity study in rats (OECD 414, GLP), the NOAEL for maternal and developmental toxicity was at least 30 mg/kg bw/day.
Executive summary:

In a developmental toxicity study performed according to OECD 414 and GLP, the substance, dissolved in corn oil, was administered to 25 rats per group by gavage at gestation days (GD) 7 through 17 at dosages of 0, 3, 10 and 30 mg/kg bw/day. Viabilities, clinical observations, body weights and feed consumption values were recorded. All rats were sacrificed on GD 21, Caesarean-sectioned and examined for the number and distribution of corpora lutea, implantation sites and uterine contents. A gross necropsy of the thoracic, abdominal and pelvic viscera was performed. Fetuses were weighed and examined for gross external, visceral and skeletal alterations and sex.

All female rats survived to scheduled sacrifice. All clinical and necropsy observations were considered to be unrelated to treatment. Maternal body weights, body weight gains and absolute and relative feed consumption values were unaffected at dosages as high as 30 mg/kg bw/day. Pregnancy occurred in 21 to 25 rats in each dosage group. Caesarean-sectioning and litter parameters were not affected by dosages of the test article as high as 30 mg/kg bw/day. No fetal alterations occurred that were considered associated with treatment. On the basis of these data, the maternal and developmental NOAEL is greater than 30 mg/kg bw/day.