Registration Dossier
Registration Dossier
Diss Factsheets
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 403-830-5 | CAS number: 89331-94-2 B 290; BK 400; CK 34; DIBUTYL-N-102; DX-20; FAT NR. 40391/A; FLUORAN BLACK BD 869; FLUORAN SCHWARZ BD 869; NOIR FLUORANE BD 869; ODB-2; PSD-290; SENOR-2; TG-31; TH-108; WINCON-2
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data

Developmental toxicity / teratogenicity
Administrative data
- Endpoint:
- developmental toxicity
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2020-02-04 to 2020-03-03
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 020
- Report date:
- 2020
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 414 (Prenatal Developmental Toxicity Study)
- Version / remarks:
- 25 June 2018
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Limit test:
- no
Test material
- Reference substance name:
- 6'-(dibutylamino)-3'-methyl-2'-(phenylamino)spiro[isobenzofuran-1(3H),9-(9H)-xanthen]-3-one
- EC Number:
- 403-830-5
- EC Name:
- 6'-(dibutylamino)-3'-methyl-2'-(phenylamino)spiro[isobenzofuran-1(3H),9-(9H)-xanthen]-3-one
- Cas Number:
- 89331-94-2
- Molecular formula:
- C35H36N2O3
- IUPAC Name:
- 6'-(dibutylamino)-3'-methyl-2'-(phenylamino)-3H-spiro[2-benzofuran-1,9'-xanthen]-3-one
Constituent 1
Test animals
- Species:
- rat
- Strain:
- Wistar
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Toxi-Coop Zrt. 1103 Budapest, Cserkesz u. 90., Hungary
- Age at study initiation: 9-10 weeks
- Weight at study initiation: 189-236 g
- Housing:
Before mating: 1-2 females per cage
1-2 males per cage
During mating: 1 male and 1-2 females / cage
During gestation: 2 sperm positive females per cage, if not possible 1 sperm positive female per cage
- Diet: ad libitum, ssniff® SM R/M " breeding and maintenance autoclavable complete feed (ssniff Spezialdiäten GmbH, 59494 Soest, Germany)
- Water: ad libitum, tap water from municipal supply
- Acclimation period: 6 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.2 – 24.1
- Humidity (%): 30 - 52
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12
Administration / exposure
- Route of administration:
- oral: gavage
- Vehicle:
- other: Methylcellulose
- Details on exposure:
- PREPARATION OF DOSING SOLUTIONS: Formulations were prepared in the formulation laboratory of the Test Facility not longer than 3 days before the administration. Formulations were stored in the refrigerator.
VEHICLE
- Justification for use and choice of vehicle: The test item was not soluble in water therefore 1% Methylcellulose was used.
- Concentration in vehicle: 6.25, 25, 100 mg/mL
- Amount of vehicle: 10 mL/kg bw
- Lot/batch no.: 1908-4485 - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- Five samples from different places were taken from each concentration. Similarly, five samples were taken from the vehicle (Control
group) and analyzed. The suitability of the chosen vehicle for the test item at the intended concentrations was verified up front. WinCon-2 recovery was 104 and 101 % of nominal concentrations at 5 and 100 mg/mL in 1% Methylcellulose, respectively. WinCon-2 proved to be stable at room temperature for 24 hours (mean recovery was 102 % of starting concentration at 5 mg/mL and 103 % at 100 mg/mL) and at 5 ± 3°C for 3 days (recovery was 102 % of starting concentration at 5 mg/mL and 99 % at 100 mg/mL). A separate analytical report provided these results. - Details on mating procedure:
- - Impregnation procedure: cohoused
- M/F ratio per cage: 1/2
- Length of cohabitation: 2 - 4 hours
- Verification of same strain and source of both sexes: yes
- Proof of pregnancy: vaginal plug and/or sperm in vaginal smear referred to as day 0 of pregnancy - Duration of treatment / exposure:
- from gestational day 5 to 19
- Frequency of treatment:
- daily
Doses / concentrationsopen allclose all
- Dose / conc.:
- 62.5 mg/kg bw/day
- Dose / conc.:
- 250 mg/kg bw/day
- Dose / conc.:
- 1 000 mg/kg bw/day
- No. of animals per sex per dose:
- 25 to 26
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale: The dose setting was based on the results obtained in a study with the same test item 6' -(Dibutylamino)-3' -methyl-2' -phenylaminospiro (isobenzofuran-l(3H),9'-(9H)-xanthene)-3-one) “ODB-2 a study in the rat of reproductive function on one generation by oral administration” performed at Huntingdon Research Centre Ltd. 1993 (Guideline for Testing of Chemicals, No. 415, adopted May 1983.) where the dosages of 62.5, 250 or 1000 mg/kg/day did not lead to any effect on the growth and reproductive capacity of male and female rats or development of their offspring. The dosage of the test item at which no signs of toxicity was recorded was therefore considered to be 1000 mg/kg/day which is the limit dose according to the OECD 414 test guideline.
Examinations
- Maternal examinations:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily (signs of morbidity and mortality were checked twice daily)
- Cage side observations included check of behavior and general condition, duration and severity of the clinical signs and observations for signs of morbidity and mortality.
BODY WEIGHT: Yes
- Time schedule for examinations: gestation days 0, 3, 5, 8, 11, 14, 17 and 20
POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day #20
- Organs examined: Uterus with cervix and ovaries, organ weight of thyroid glands together with the parathyroid glands
OTHER:
Examination of placental signs: All sperm positive animals were examined for vaginal bleeding (placental sign of gestation) on the 13th gestational day. If negative on day 13, the examination was repeated on day 14 of gestation.
Blood Collection and Determination of Serum Levels of TSH, FT3 and FT4: Blood samples collected for TSH and Thyroid Hormones (FT3 and FT4) measurements were drawn from all sperm positive females from the
sublingual vein in the morning on the day of necropsy within a short timeframe (not exceeding two hours). - Ovaries and uterine content:
- The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes - Fetal examinations:
- - External examinations: Yes: all per litter
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
- Head examinations: Yes: all per litter - Statistics:
- Data were individually recorded on data sheets, transferred, and compiled by computer or compiled manually. The statistical evaluation of data was performed with the program package SPSS PC+4.0. The homogeneity of variance between groups was checked by Bartlett’s homogeneity of variance test. Where no significant heterogeneity is detected a one-way analysis of variance (ANOVA) was carried out. If the obtained result is significant Duncan’s Multiple Range test was used to assess the significance of intergroup differences. If the result of the Bartlett’s test was significant, the Kruskal-Wallis analysis of variance was used and the inter-group comparisons were performed using Mann-Whitney U-test.
Dams or litters were excluded from the data evaluation in cases of:
- Non pregnant females (no implantation, no corpora lutea), total exclusion
- Disease or death of the dam unrelated to the treatment (total exclusion)
Although these animals were excluded from the data evaluation the Study Report contains all data of these animals, too. A male/female fetus was considered as retarded in body weight, when its weight was below the average minus twofold standard deviation of the control male/female fetuses. - Historical control data:
- The results were compared to the laboratory's historical control data.
Results and discussion
Results: maternal animals
General toxicity (maternal animals)
- Clinical signs:
- no effects observed
- Mortality:
- no mortality observed
- Body weight and weight changes:
- no effects observed
- Clinical biochemistry findings:
- no effects observed
- Description (incidence and severity):
- The free thyroid hormone (FT3 and FT4) and TSH levels were similar in all dose groups.
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Description (incidence and severity):
- Thyroid weight was not effected by treatment.
- Gross pathological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- A dilated renal pelvis was found in two females each in the 62.5, 250 and 1000 mg/kg bw/day dose groups and one female in the control group. This finding was not attributed to the treatment considering the lack of dose response. Diaphragmatic hernia as a developmental disorder (hence not related to the treatment) was observed in one female in the 1000 mg/kg bw/day group. The uterine horn was filled with fluid to distention in one female in the 1000 mg/kg bw/day group. Considering the low incidence, this was not attributed to an effect of the test item. Blood was found in the uterine horn of one dam in the 62.5 and one in the 250 mg/kg bw/day dose group.
- Histopathological findings: non-neoplastic:
- no effects observed
- Description (incidence and severity):
- According to the expert’s evaluation, there were no test item related lesions observed upon histological examinations of the thyroid tissue. Cyst forming was recorded for the thyroid gland of one control female.
Maternal developmental toxicity
- Number of abortions:
- no effects observed
- Pre- and post-implantation loss:
- no effects observed
- Total litter losses by resorption:
- no effects observed
- Early or late resorptions:
- no effects observed
- Dead fetuses:
- no effects observed
- Changes in pregnancy duration:
- no effects observed
- Changes in number of pregnant:
- no effects observed
Effect levels (maternal animals)
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- > 1 000 mg/kg bw/day
- Based on:
- test mat.
- Basis for effect level:
- other: No effects observed
Maternal abnormalities
- Key result
- Abnormalities:
- no effects observed
Results (fetuses)
- Fetal body weight changes:
- 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
- Changes in postnatal survival:
- no effects observed
- External malformations:
- no effects observed
- Skeletal malformations:
- no effects observed
- Visceral malformations:
- no effects observed
Effect levels (fetuses)
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- >= 1 000 mg/kg bw/day
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: no effects observed
Fetal abnormalities
- Key result
- Abnormalities:
- no effects observed
Overall developmental toxicity
- Key result
- Developmental effects observed:
- no
Applicant's summary and conclusion
- Conclusions:
- Oral treatment of pregnant Han: Wistar rats from gestation day 5 up to day 19 (the day before Caesarean section) with WinCon-2 at the dose levels of 62.5, 250 and 1000 mg/kg bw/day did not cause any maternal effects. Fetal examinations did not reveal any adverse effects on the fetuses.
- Executive summary:
WinCon-2 was examined for its possible prenatal developmental toxicity. Groups of 25, 26 and 26 sperm-positive female Han: of Wistar origin rats were treated with WinCon-2 by oral administration daily at three dose levels of 62.5, 250 and 1000 mg/kg bw/day respectively from day 5 up to and including day 19 post coitum. A control group of 26 sperm positive females was included and the animals were given the vehicle 1% methylcellulose. The treatment volume was 10 mL/kg bw. A sufficient stability and homogeneity in the chosen vehicle were verified over the range of relevant concentrations at the appropriate frequency of preparation. WinCon-2 in 1% methylcellulose was stable at room temperature for at least one day and for three days in the refrigerator (5 ± 3 oC) at the concentrations of 5 and 100 mg/mL. Analytical control of dosing solutions was performed during the first and last week of treatment. Concentrations of the test item in the dosing formulations varied in the acceptable range between 102 and 108 % of nominal concentrations at both analytical occasions confirming proper dosing. During the study, mortality was checked and clinical observations were performed. Body weight and food consumption of the dams were also recorded. The day, when sperm was detected in the vaginal smear, was regarded as day 0 of gestation. Blood sampling for determination of thyroid hormones FT3 and FT4 as well as TSH, Caesarean section and gross pathology were performed on gestational day 20. Thyroids were weighed and evaluated histologically. The number of implantations, early and late resorptions, live and dead fetuses in each uterine horn and the number of corpora lutea were recorded. Each fetus was weighed and examined for sex and gross external abnormalities. The placentas were weighed and examined externally. External fetal sex was determined by gross pathological examination and compared with internal (gonadal) sex in all fetuses (examined for both skeletal and soft tissue alterations). The anogenital distance was measured. In addition, indication of incomplete testicular descent / cryptorchidism was noted in male fetuses. About half of each litter was preserved for visceral examination and the other half of the litters were preserved for skeletal evaluation. At visceral examination the bodies were micro dissected by means of a dissecting microscope. The heads were examined by Wilson's free-hand razor blade method. After cartilage-bone staining the skeletons were examined by means of a dissecting microscope. All abnormalities found during the fetal examinations were recorded.
Results
Evaluated litters
In total, on gestation day 20 there were 25, 22, 23 and 22 evaluated litters in the control, 62.5, 250 and 1000 mg/kg bw/day group, respectively.
Mortality, clinical signs, necropsy findings
None of the females died before scheduled necropsy due to the test item and there were no test item related clinical signs recorded in the dose groups. No treatment related necropsy findings were observed.
Food consumption, body weight
Food consumption and body weight development were not affected by the test item.
FT3, FT4 and TSH level
The test item did not influence the thyroid hormone levels in any of the treated animals.
Thyroid weight
There were no test item related differences in thyroid weight among the dosing groups.
Histopathology of thyroid glands
The treatment did not result in histological changes of the thyroid gland in any of the dose groups.
Intrauterine mortality
Number of implantations, intrauterine mortality and sex distribution of the fetuses were not influenced by the treatment.
Fetal examinations
There were no test item related adverse effects on the fetal- and placental weight, ano-genital distance, external and visceral development of fetuses. The number of litters with malformations was one in all groups. There was no increase of variations during fetal examinations indicated.
Conclusion
Oral treatment of pregnant Han: Wistar rats from gestation day 5 up to day 19 (the day before Caesarean section) with WinCon-2 at the dose levels of 62.5, 250 and 1000 mg/kg bw/day did not cause any maternal effects. Fetal examinations did not reveal any adverse effects on the fetuses.
Based on the observations the No Observed Adverse Effect Level (NOAEL) was determined as follows:
NOAEL (maternal toxicity): 1000 mg/kg bw/day
NOAEL (developmental toxicity including teratogenicity): 1000 mg/kg bw/day
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.

EU Privacy Disclaimer
This website uses cookies to ensure you get the best experience on our websites.