Registration Dossier
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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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
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: 834-894-6 | CAS number: 113601-85-7
- 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
Genetic toxicity: in vivo
Administrative data
- Endpoint:
- in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 08 May 2013 to 13 May 2013
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Justification for type of information:
- In-Vivo study carried out as substance is intended for global registration where In-Vivo data is required.
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 013
- Report date:
- 2013
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
- Deviations:
- yes
- Remarks:
- See other information on method section for details
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.12 (Mutagenicity - In Vivo Mammalian Erythrocyte Micronucleus Test)
- Deviations:
- yes
- Remarks:
- See other information on method section for details
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- mammalian erythrocyte micronucleus test
Test material
- Reference substance name:
- Reaction products of 1,4-Benzenedimethanol and 1-naphthol
- EC Number:
- 834-894-6
- Cas Number:
- 113601-85-7
- Molecular formula:
- C10H8O to C82H64O5
- IUPAC Name:
- Reaction products of 1,4-Benzenedimethanol and 1-naphthol
- Test material form:
- solid
- Details on test material:
- Name: CAS 113601-85-7
Chemical name: 1,4-Benzenedimethanol, polymer with 1-naphthalenol
Batch/Lot number: 0950986
Description: Brown, solid
Purity: 100%
Expiry date: 08 February 2020
Storage conditions: Controlled room temperature (15-25°C ≤ 70% relative humidity)
Safety precautions: Routine safety precautions (gloves, goggles, face mask, lab coat) for unknown materials were applied to ensure personnel health and safety. Causes skin irritation and serious eye irritation.
Constituent 1
- Specific details on test material used for the study:
- No further details specified in the study report
Test animals
- Species:
- mouse
- Strain:
- NMRI
- Details on species / strain selection:
- The NMRI mouse is one of the standard animals used internationally in this type of mutagenicity testing
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- Species and strain: RjHan: NMRI mice
Source: Elevage Janvier
Route des Chènes Secs B.P. 4105
53940 LE GENEST-ST-ISLE, France
Justification of species/strain: The NMRI mouse is one of the standard animals used internationally in this type of mutagenicity testing
Hygienic level: SPF at arrival; standard laboratory conditions during the study
Number of animals: Preliminary experiment: 8 males + 8 females
4 groups, 2 animals/sex/group
Main test: 35+2 males
Positive control group: 5 mice
Negative control group: 10 mice
High-dose group: 10+2 mice
Low- and mid-dose groups: 5 mice
Age of animals: approximately 7 weeks (at the treatment)
Body weight: 35.2 – 37.5 g (males, preliminary experiment)
25.0 - 26.5 g (females, preliminary experiment)
32.7 – 35.5 g (males, main test)
Acclimatisation period: at least 5 days
Note: The weight variation did not exceed ± 20 percent of the mean weight/sex at the start of the treatment.
Husbandry
Animal health: Only animals in acceptable health condition were used for the test. Health status was certified by the veterinarian.
Housing/Enrichment: Group caging (5 animals/cage or 2 animals/cage) to allow social interaction, and with deep wood sawdust bedding, to allow digging and other normal rodent activities.
Cage type: II. type polypropylene/polycarbonate
Light: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 19.4 – 24.3°C
Relative humidity: 30 – 70 %
Ventilation: 15 – 20 air exchanges/hour
Animal room: 244/2 (preliminary experiment), 240 (main test)
The environmental parameters were recorded twice daily during the acclimatisation period and experimental phases of the study.
Food and Water Supply
Animals received ssniff® SM R/M-Z+H "Autoclavable complete diet for mice and rats – breeding and maintenance" (Batch number: 445 8440 / 175 8935, Expiry date: May 2013 / August 2013) produced by ssniff Spezialdiäten GmbH (D-59494, Soest, Germany) and tap water from municipal supply, as for human consumption from 500 ml bottle ad libitum.
The food is considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study. The contents of the standard diet are detailed in Appendix 8. The supplier provided an analytical certificate for the batch used.
Water quality control analysis is performed once every three months and microbiological assessment is performed monthly by Veszprém County Institute of State Public Health and Medical Officer Service (ÁNTSZ, H-8201 Veszprém, József A.u.36., Hungary). The quality control results are retained in the archives at CiToxLAB Hungary Ltd.
Bedding
Bedding of certified wood chips especially designed to keep animals in the best natural environment was provided for animals during the study. Lignocel 3/4-FASERN Hygienic Animal Bedding produced by J. Rettenmaier & Söhne GmbH & Co.KG (Holzmühle 1, 73494 Rosenberg, Germany) was available to animals during the study.
Identification
Animals were identified by a number unique within the study, written with indelible ink on the tail and cross-referenced to the Animal Master File at CiToxLAB Hungary Ltd. The cages were marked with identification cards, with information about study code, sex, dose group and individual animal numbers.
Randomisation
The animals were assigned to their respective treatment groups by randomization based on body weights. Animals were randomly allocated to the negative and positive control groups based on the most recent actual body weight; SPSS/PC+ software was used in order to verify homogeneity/variation among/within groups. Females and males were randomized separately.
Administration / exposure
- Route of administration:
- oral: gavage
- Vehicle:
- PEG-400 was used for vehicle of the study.
- Details on exposure:
- Preliminary toxicity test:
A preliminary toxicity test was performed to identify the appropriate maximum dose level for the main test. The preliminary toxicity test also determined whether there are large differences in toxicity between the sexes or not. Groups of two male and female mice were treated at one occasion by oral gavage at the dose levels of 2000, 1000, 500 and 250 mg/kg body weight.
The treatment volume was 10 mL/kg body weight. Animals were examined regularly for toxic signs and mortalities. The surviving mice were euthanized 48 hours after treatment. No bone marrow smears were prepared in the preliminary experiment.
Main test:
Based on the results of the preliminary toxicity test, 2000 mg/kg body weight (the maximum recommended dose) was selected for the highest dose level of the test item in the main micronucleus test. Two lower dose levels separated by a factor of two (1000 and 500 mg/kg body weight) were also included in the main test. The dose levels were expressed in terms of the test item as received.
The main test was performed using male animals only because the toxic effect of the test item was similar in both sexes in the preliminary toxicity test. - Duration of treatment / exposure:
- 48 hours
- Frequency of treatment:
- Dose of 2000, 1000 and 500 mg/kg body weight by oral gavage administered at the start of the study. No further treatment was administered for the duration of the study.
- Post exposure period:
- In the low and mid dose groups, furthermore in the positive control group the sampling was made once at 24 hours after treatment. In the high dose group and vehicle control group, sampling was made 24 and 48 hours after treatment. Five male animals per dose group were used for sampling on each one occasion.
Doses / concentrationsopen allclose all
- Dose / conc.:
- 2 000 mg/kg bw (total dose)
- Dose / conc.:
- 1 000 mg/kg bw (total dose)
- Dose / conc.:
- 500 mg/kg bw (total dose)
- No. of animals per sex per dose:
- 5 animals per dose
- Control animals:
- yes, concurrent vehicle
- Positive control(s):
- Cyclophosphamide monohydrate was used as positive control material for the study. It was dissolved in sterile physiological saline solution for treatment. Routine safety precautions (lab coat, gloves, safety glasses and face mask) for the positive control material were applied to assure personnel health and safety. The positive control formulations were prepared immediately before treatment in the Central Dispensary Unit of CiToxLAB Hungary Ltd.
Data of the chemical used as positive control substance are shown below:
Name: Cyclophosphamide monohydrate
Abbreviation: CP
Supplier: Sigma-Aldrich Co.
Lot No.: 120M1253V
Appearance: White powder
Expiry date: 31 December 2013
Storage condition: Refrigerated (2-8 °C)
Examinations
- Tissues and cell types examined:
- Erythrocytes obtained from the bone marrow of the femur.
- Details of tissue and slide preparation:
- The bone marrow was flushed out of each pair of femurs with foetal bovine serum (5 mL) using a syringe and needle into a sterile centrifuge tube. After mixing, the cell suspension was concentrated by a gentle centrifugation and the supernatant was discarded. Smears of the cell pellet were made on standard microscope slides (at least 2 slides / animal). Slides were air-dried at room temperature for approximately 24 hours.
Dried slides were fixed in methanol for a minimum of 5 minutes and allowed to air-dry. Two slides per animal were stained with 10 % Giemsa solution for 20 minutes then thoroughly rinsed with distilled water, and then air-dried at room temperature for at least 12 hours. After staining, coverslips were mounted on them. - Evaluation criteria:
- Criteria for a positive response:
The test item is considered to have shown genotoxic activity if statistically significant increases in the frequency of micronucleated polychromatic erythrocytes are observed in treated animals compared to the corresponding negative controls, and the increases are dose-related. Historical control data are taken into consideration when evaluating the biological significance of small increases.
Criteria for a negative response:
The test item is concluded to have given a negative response if no reproducible, statistically significant increases are observed above the concurrent and historical negative control values.
Equivocal response:
Results which do not meet the criteria for a positive or negative response are considered to be equivocal. Further investigations or scoring of additional cells may be necessary in case of an equivocal result. - Statistics:
- Not specified.
Results and discussion
Test results
- Key result
- Sex:
- male
- Genotoxicity:
- negative
- Toxicity:
- no effects
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
Any other information on results incl. tables
PRELIMINARY TOXICITY TEST
According to the observation in a short preliminary solubility test, no proper formulation could be made at 200 mg/mL concentration using physiological saline as vehicle. However, the formulation at the same concentration using PEG 400 (Poly(ethylene glycol) 400) as vehicle was suitable for treatment of the animals. Therefore, PEG 400 was selected for vehicle of the study and the following dose groups were examined in the preliminary toxicity test: 2000, 1000, 500 and 250 mg/kg body weight (2 animals/sex/group).
There was no treatment related effect on the body weight in the preliminary experiment. The individual body weights of the animals in the preliminary experiment are shown in Table 3 of Appendix 3. The observed clinical signs are listed in Table 4 of Appendix 3. All animals were free of clinical signs in the preliminary experiment except of one male in the 1000 mg/kg body weight dose group showing piloerection at one time point, but this observation was not considered to represent a dose-related toxicity.
Based on the results of the preliminary toxicity test, dose levels of 2000, 1000 and 500 mg/kg body weight were selected for the micronucleus test. As there were no differences between male and female animals in the preliminary experiment, the main experiment was performed using male mice only.
MOUSE MICRONUCLEUS TEST
Groups of five male mice were treated with the test item at 2000, 1000 and 500 mg/kg body weight or with the vehicle (PEG 400) in the main experiment (two replacement animals were also treated in the high dose group). All mice in the negative (vehicle) control and test item groups were dosed by oral gavage using a dose volume of 10 mL/kg body weight. Animals of the positive control group were treated by intraperitoneal injection with Cyclophosphamide at 60 mg/kg body weight using a dose volume of 10 mL/kg body weight.
No marked effect of test item treatment on the body weight of the mice was observed in the main test. Marked body weight loss (>10%) was detected for one negative control animal but since it had normal bone marrow results, this was not considered to be a significant finding
No mortality or signs of systemic toxicity were observed during the study. Piloerection and wheal at the left thorax ventral area was observed for one animal in the negative control group, since it had normal bone marrow results, this was not considered to be a significant finding (Appendix 5). The animals in the test item treated and positive control groups were symptom-free during the whole observation period (Appendix 5).
Two thousand polychromatic erythrocytes (PCEs) were scored per animal* to assess the micronucleated cells. The frequency of micronucleated cells was expressed as percent of micronucleated cells based on the first 2000 PCEs counted in the optic field. The proportion of immature among total erythrocytes was determined for each animal by also counting mature erythrocytes (NCEs) until a total of 1000 PCEs+NCEs had been observed.
*Note: During the initial analysis, two animals in the group treated with 2000 mg/kg bw at the 48-hour sampling point had less than 2000 PCEs on the slides. Although sufficient cells were subsequently found on further slides which were sent later, the slides of additional 2 replacement animals were then also analysed. Thus, there are 7 animals in this treatment group.
The group treated with 2000 mg/kg bw, which gave the highest number of micronuclei at the 24-hour sampling point, were compared with the relevant vehicle control group using the Kruskal Wallis test. This gave a value of H = 2.635, which is non-significant. The average number of micronuclei observed at 2000 mg/kg bw at the 48-hour sampling point was lower than the corresponding negative control group. Therefore there was a negative response at both times.
The positive and negative control results were also compared, and gave a value of H = 6.990 (p<0.01). The positive control treatment therefore caused a very substantial increase, demonstrating the sensitivity of the test system.
The positive and negative control* data are considered to give adequate data to confirm the validity of the study.
*Note: For one animal in the vehicle control group at the 48-hour sampling point, the number of micronuclei was slightly higher than the upper limit of the historical control range. However, the difference was minor and the mean value of this group was within the historical control range, so this fact was considered to be acceptable.
The frequency of micronucleated polychromatic erythrocytes of the negative (solvent) control group was within the range of historical laboratory control data; the positive control item produced a biologically and statistically relevant increase in the number of micronucleated polychromatic erythrocytes; each treated and control group included at least 5 analysable animals; therefore, the test was considered to be valid.
Results of the Preliminary Experiment
Individual Body Weights for all Animals with Group Means
Animal Number |
Gender |
Dose (mg/kg/bw |
Body weight (%) |
Change (%) |
||
Day 1 |
Day 2 |
Day 3 |
||||
1980 |
M |
2000 |
36.3 |
34.6 |
36.3 |
0.0 |
1984 |
M |
2000 |
36.7 |
36.1 |
37.2 |
1.4 |
|
|
Mean |
36.5 |
35.35 |
36.75 |
0.7 |
1978 |
M |
1000 |
36.8 |
36.1 |
36.4 |
-1.1 |
1985 |
M |
1000 |
36.2 |
35.1 |
36.0 |
-0.6 |
|
|
Mean |
36.5 |
35.60 |
36.20 |
-0.8 |
1979 |
M |
500 |
36.8 |
36.4 |
36.7 |
-0.3 |
1986 |
M |
500 |
35.8 |
34.2 |
34.6 |
-3.4 |
|
|
Mean |
36.30 |
35.30 |
35.65 |
-1.8 |
1981 |
M |
250 |
37.5 |
36.7 |
36.5 |
-2.7 |
1989 |
M |
250 |
35.2 |
34.7 |
35.0 |
-0.6 |
|
|
Mean |
36.35 |
35.7 |
35.75 |
-1.7 |
1995 |
F |
2000 |
26.0 |
25.5 |
26.3 |
1.2 |
1997 |
F |
2000 |
26.2 |
25.7 |
27.4 |
4.6 |
|
|
Mean |
26.10 |
25.6 |
26.85 |
2.9 |
1991 |
F |
1000 |
26.3 |
27.1 |
27.5 |
4.6 |
1996 |
F |
1000 |
25.9 |
25.6 |
26.1 |
0.8 |
|
|
Mean |
26.1 |
26.35 |
26.8 |
2.7 |
1993 |
F |
500 |
26.5 |
25.7 |
25.9 |
-2.3 |
2001 |
F |
500 |
25.0 |
25.0 |
25.0 |
0.0 |
|
|
Mean |
25.75 |
25.35 |
25.45 |
-1.2 |
1999 |
F |
250 |
25.0 |
24.4 |
24.8 |
-0.8 |
2000 |
F |
250 |
26.5 |
26.3 |
27.0 |
1.9 |
|
|
Mean |
25.75 |
25.35 |
25.9 |
0.6 |
Notes:
1. M: male, F: female
2. Change: [Body Weight (Day 3) – Body Weight (Day 1)] / Body Weight (Day 1) x 100
Clinical observations
DOSE LEVEL: 2000 mg/kg body weight/day SEX: MALE
ID Number |
Animal Number |
Observations |
Time points |
Frequency |
|||||||
After treatment |
|||||||||||
30’ |
1h |
2h |
3h |
4h |
5h |
24h |
48h |
||||
1980 |
3 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
1984 |
7 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
DOSE LEVEL: 1000 mg/kg body weight/day SEX: MALE
ID Number |
Animal Number |
Observations |
Time points |
Frequency |
|||||||
After treatment |
|||||||||||
30’ |
1h |
2h |
3h |
4h |
5h |
24h |
48h |
||||
1978 |
1 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
- |
7/8 |
Piloerection |
- |
- |
- |
- |
- |
- |
- |
+ |
1/8 |
||
1985 |
8 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
DOSE LEVEL: 500 mg/kg body weight/day SEX: MALE
ID Number |
Animal Number |
Observations |
Time points |
Frequency |
|||||||
After treatment |
|||||||||||
30’ |
1h |
2h |
3h |
4h |
5h |
24h |
48h |
||||
1979 |
2 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
1986 |
9 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
DOSE LEVEL: 250 mg/kg body weight/day SEX: MALE
ID Number |
Animal Number |
Observations |
Time points |
Frequency |
|||||||
After treatment |
|||||||||||
30’ |
1h |
2h |
3h |
4h |
5h |
24h |
48h |
||||
1981 |
4 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
1989 |
12 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
DOSE LEVEL: 2000 mg/kg body weight/day SEX: FEMALE
ID Number |
Animal Number |
Observations |
Time points |
Frequency |
|||||||
After treatment |
|||||||||||
30’ |
1h |
2h |
3h |
4h |
5h |
24h |
48h |
||||
1995 |
18 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
1997 |
20 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
DOSE LEVEL: 1000 mg/kg body weight/day SEX: FEMALE
ID Number |
Animal Number |
Observations |
Time points |
Frequency |
|||||||
After treatment |
|||||||||||
30’ |
1h |
2h |
3h |
4h |
5h |
24h |
48h |
||||
1991 |
14 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
1996 |
19 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
DOSE LEVEL: 500 mg/kg body weight/day SEX: FEMALE
ID Number |
Animal Number |
Observations |
Time points |
Frequency |
|||||||
After treatment |
|||||||||||
30’ |
1h |
2h |
3h |
4h |
5h |
24h |
48h |
||||
1993 |
16 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
2001 |
24 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
DOSE LEVEL: 250 mg/kg body weight/day SEX: FEMALE
ID Number |
Animal Number |
Observations |
Time points |
Frequency |
|||||||
After treatment |
|||||||||||
30’ |
1h |
2h |
3h |
4h |
5h |
24h |
48h |
||||
1999 |
22 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
2000 |
23 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
Individual Body Weight Data
Mice sacrificed 24 hours after dosing
Treatment |
ID Number |
Animal number |
Body weight (g) |
||
Day 1 |
Day 2 |
Change |
|||
Group 1 Negative (vehicle) control (PEG 400) |
2327 |
2 |
34.2 |
33.3 |
-2.6 |
2343 |
18 |
32.7 |
28.3 |
-13.5 |
|
2354 |
29 |
34.6 |
33.3 |
-3.8 |
|
2356 |
31 |
34.1 |
33.1 |
-2.9 |
|
2370 |
45 |
34.7 |
34.7 |
0.0 |
|
Mean |
34.1 |
32.5 |
-4.5 |
||
Group 2 SN-475N (500 mg/kg bw) |
2334 |
9 |
34.9 |
33.5 |
-4.0 |
2337 |
12 |
34.1 |
34.4 |
0.9 |
|
2338 |
13 |
34.2 |
33.7 |
-1.5 |
|
2340 |
15 |
34.6 |
33.5 |
-3.2 |
|
2360 |
35 |
32.7 |
31.7 |
-3.1 |
|
Mean |
34.1 |
33.4 |
-2.2 |
||
Group 3 SN-475N (1000 mg/kg bw) |
2333 |
8 |
33.4 |
33.1 |
-0.9 |
2341 |
16 |
34.9 |
34.8 |
-0.3 |
|
2358 |
33 |
34.0 |
34.1 |
0.3 |
|
23361 |
36 |
34.2 |
33.8 |
-1.2 |
|
2366 |
41 |
34.5 |
34.5 |
0.0 |
|
Mean |
34.2 |
34.1 |
-0.4 |
||
Group 4 SN-475N (2000 mg/kg bw) |
2331 |
6 |
34.5 |
32.9 |
-4.6 |
2332 |
7 |
34.3 |
32.8 |
-4.4 |
|
2346 |
21 |
34.0 |
32.7 |
-3.8 |
|
2349 |
24 |
35.0 |
35.6 |
1.7 |
|
2365 |
40 |
33.4 |
32.4 |
-3.0 |
|
Mean |
34.2 |
33.3 |
-2.8 |
||
Group 5 Positive control (Cyclophosphamide 60 mg/kg bw)
|
2330 |
5 |
34.5 |
33.7 |
-2.3 |
2342 |
17 |
34.0 |
32.5 |
-4.4 |
|
2350 |
25 |
33.5 |
31.8 |
-5.1 |
|
2351 |
26 |
35.1 |
33.0 |
-6.0 |
|
2363 |
38 |
34.3 |
32.5 |
-5.2 |
|
Mean |
34.3 |
32.7 |
-2.6 |
Note:
Change = [Terminal Body Weight (Day 2) – Initial Body Weight (Day 1)] / Initial Body Weight (Day 1) x 100
Individual Body Weight Data
Mice sacrificed 48 hours after dosing
Treatment |
ID Number |
Animal number |
Body weight (g) |
||
Day 1 |
Day 2 |
Change |
|||
Group 1 Negative (vehicle) control (PEG 400) |
2326 |
1 |
34.5 |
34.2 |
-0.9 |
2329 |
4 |
32.9 |
32.5 |
-1.2 |
|
2348 |
23 |
35.3 |
35.3 |
0.0 |
|
2359 |
34 |
33.9 |
32.9 |
-2.9 |
|
2364 |
39 |
34.3 |
34.2 |
-0.3 |
|
Mean |
34.2 |
33.8 |
-1.1 |
||
Group 4 SN-475N (2000 mg/kg bw) |
2344 |
19 |
35.5 |
34.1 |
-3.9 |
2347 |
22 |
33.6 |
33.8 |
0.6 |
|
2353 |
28 |
34.4 |
32.5 |
-5.5 |
|
2355 |
30 |
33.6 |
33.2 |
-1.2 |
|
2369 |
44 |
34.3 |
33.3 |
-2.9 |
|
2328* |
3* |
35.2 |
34.7 |
-1.4 |
|
2367* |
42* |
33.5 |
33.5 |
0.0 |
|
Mean |
34.3 |
33.6 |
-2.1 |
Notes:
Change = [Terminal Body Weight (Day 3) – Initial Body Weight (Day 1)] / Initial Body Weight (Day 1) x 100
*: replacement animal
Clinical Observations
Mice sacrificed 24 hours after dosing
DOSE LEVEL: Negative(vehicle) control SEX: MAL
ID Number |
Animal Number |
Observations |
Time points |
Frequency |
||||||
After treatment |
||||||||||
30’ |
1h |
2h |
3h |
4h |
5h |
24h |
||||
2327 |
2 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
7/7 |
2343 |
18 |
Symptom free |
+ |
+ |
+ |
- |
- |
- |
- |
3/7 |
Piloerection |
- |
- |
- |
- |
- |
- |
+ |
1/7 |
||
Wheal (left thorax ventral) |
- |
- |
|
+ |
+ |
+ |
+ |
4/7 |
||
2354 |
29 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
7/7 |
2356 |
31 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
7/7 |
2370 |
45 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
7/7 |
DOSE LEVEL: 500 mg/kg body weight SEX: MALE
ID Number |
Animal Number |
Observations |
Time points |
Frequency |
||||||
After treatment |
||||||||||
30’ |
1h |
2h |
3h |
4h |
5h |
24h |
||||
2334 |
9 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
7/7 |
2337 |
12 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
7/7 |
2338 |
13 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
7/7 |
2340 |
15 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
7/7 |
2360 |
35 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
7/7 |
DOSE LEVEL: 1000 mg/kg body weight SEX: MALE
ID Number |
Animal Number |
Observations |
Time points |
Frequency |
||||||
After treatment |
||||||||||
30’ |
1h |
2h |
3h |
4h |
5h |
24h |
||||
2333 |
8 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
7/7 |
2341 |
16 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
7/7 |
2358 |
33 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
7/7 |
2361 |
36 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
7/7 |
2366 |
41 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
7/7 |
DOSE LEVEL: 2000 mg/kg body weight SEX: MALE
ID Number |
Animal Number |
Observations |
Time points |
Frequency |
||||||
After treatment |
||||||||||
30’ |
1h |
2h |
3h |
4h |
5h |
24h |
||||
2331 |
6 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
7/7 |
2332 |
7 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
7/7 |
2346 |
21 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
7/7 |
2349 |
24 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
7/7 |
23655 |
40 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
7/7 |
DOSE LEVEL: Positive control (CP, 60 mg/kg body weight) SEX: MALE
ID Number |
Animal Number |
Observations |
Time points |
Frequency |
||||||
After treatment |
||||||||||
30’ |
1h |
2h |
3h |
4h |
5h |
24h |
||||
2330 |
5 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
7/7 |
2342 |
17 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
7/7 |
23250 |
25 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
7/7 |
2351 |
26 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
7/7 |
2363 |
38 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
7/7 |
Clinical Observations
Mice sacrificed 48 hours after dosing
DOSE LEVEL: Negative (vehicle) control SEX: MALE
ID Number |
Animal Number |
Observations |
Time points |
Frequency |
|||||||
After treatment |
|||||||||||
30’ |
1h |
2h |
3h |
4h |
5h |
24h |
48h |
||||
2326 |
1 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
2329 |
4 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
2348 |
23 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
2359 |
34 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
2364 |
39 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
DOSE LEVEL: 2000 mg/kg body weight/day SEX: MALE
ID Number |
Animal Number |
Observations |
Time points |
Frequency |
|||||||
After treatment |
|||||||||||
30’ |
1h |
2h |
3h |
4h |
5h |
24h |
48h |
||||
2344 |
19 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
2347 |
22 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
2353 |
28 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
2355 |
3 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
2369 |
44 |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
2328* |
3* |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
2367* |
42* |
Symptom free |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
+ |
8/8 |
*: replacement animal
Micronucleus Data
Mice sacrificed 24 hours after dosing
Treatment |
ID Number |
Animal number |
MNPCE/ 2000 PCE |
PCE/1000 PCE + NCE |
Group 1 Negative (vehicle) control (PEG 400) |
2327 |
2 |
4 |
346 |
2343 |
18 |
1 |
238 |
|
2354 |
29 |
4 |
289 |
|
2356 |
31 |
3 |
591 |
|
2370 |
45 |
4 |
321 |
|
Mean |
3.2 |
357.0 |
||
SD |
1.30 |
136.89 |
||
Group 2 SN-475N (500 mg/kg bw) |
2334 |
9 |
9 |
344 |
2337 |
12 |
2 |
221 |
|
2338 |
13 |
1 |
431 |
|
2340 |
15 |
5 |
450 |
|
2360 |
35 |
3 |
283 |
|
Mean |
4.0 |
345.8 |
||
SD |
3.16 |
97.00 |
||
Group 3 SN-475N (1000 mg/kg bw) |
2333 |
8 |
5 |
225 |
2341 |
16 |
8 |
245 |
|
2358 |
33 |
2 |
513 |
|
23361 |
36 |
3 |
300 |
|
2366 |
41 |
5 |
459 |
|
Mean |
4.6 |
348.4 |
||
SD |
2.30 |
129.99 |
||
Group 4 SN-475N (2000 mg/kg bw) |
2331 |
6 |
5 |
278 |
2332 |
7 |
3 |
371 |
|
2346 |
21 |
8 |
209 |
|
2349 |
24 |
9 |
268 |
|
2365 |
40 |
4 |
313 |
|
Mean |
5.8 |
287.8 |
||
SD |
2.59 |
59.70 |
||
Group 5 Positive control (Cyclophosphamide 60 mg/kg bw)
|
2330 |
5 |
54 |
182 |
2342 |
17 |
66 |
266 |
|
2350 |
25 |
68 |
450 |
|
2351 |
26 |
120 |
330 |
|
2363 |
38 |
62 |
353 |
|
Mean |
74.0 |
316.2 |
||
SD |
26.27 |
99.97 |
MNPCE: Number of Micronucleated Polychromatic Erythrocytes referring to counts of 2000 PCE.
PCE: Polychromatic Erythrocyte
NCE: Normochromatic Erythrocyte
Micronucleus Data
Mice sacrificed 48 hours after dosing
Treatment |
ID Number |
Animal number |
MNPCE/ 2000 PCE |
PCE/1000 PCE + NCE |
Group 1 Negative (vehicle) control (PEG 400) |
2326 |
1 |
3 |
126 |
2329 |
4 |
8 |
291 |
|
2348 |
23 |
5 |
393 |
|
2359 |
34 |
7 |
195 |
|
2364 |
39 |
5 |
324 |
|
Mean |
5.6 |
265.8 |
||
SD |
1.95 |
105.78 |
||
Group 4 SN-475N (2000 mg/kg bw) |
2344 |
19 |
4 |
303 |
2347 |
22 |
2 |
351 |
|
2353 |
28 |
8 |
178 |
|
2355 |
30 |
5 |
161 |
|
2369 |
44 |
4 |
208 |
|
2328* |
3* |
1 |
232 |
|
2367* |
42* |
3 |
217 |
|
Mean |
3.9 |
235.7 |
||
SD |
2.27 |
68.15 |
MNPCE: Number of Micronucleated Polychromatic Erythrocytes referring to counts of 2000 PCE.
PCE: Polychromatic Erythrocyte
NCE: Normochromatic Erythrocyte
*: replacement animal
Applicant's summary and conclusion
- Conclusions:
- No induction of micronuclei in bone marrow erythrocytes was observed following administration of SN-475N to mice at up to and including 2000 mg/kg bw/day; thus, there was no evidence of any genotoxic activity of the test item under the conditions of this study.
- Executive summary:
The objective of the study was to determine whether SN-475N test item caused genotoxic effects resulting in the formation of micronuclei in erythrocytes of treated mice in accordance with the OECD guideline “Mammalian Erythrocyte Micronucleus Test”, No. 474 (1997).
In the preliminary toxicity test, groups of two male and two female mice were treated with the test item formulated in PEG 400 at 2000, 1000, 500 and 250 mg/kg body weight by oral gavage. No treatment related effect was observed in the preliminary experiment. Therefore, the highest dose level selected for the main test was 2000 mg/kg body weight which is the maximum recommended dose level for materials of low toxicity. Only male mice were used in the main test as observations in the preliminary test showed that there was no substantial difference in the toxicity of the test item between the sexes.
In the main test, groups of male mice were treated with the vehicle (PEG 400) or the test item at 2000, 1000 and 500 mg/kg body weight by oral gavage or the positive control item (Cyclophosphamide dissolved in physiological saline) at 60 mg/kg body weight administered by intraperitoneal injection (two replacement animals were also treated in the high dose group). Five mice from each group were examined 24 hours after dosing, and a further five mice dosed with the vehicle or test item at 2000 mg/kg body weight were examined 48 hours after dosing. Bone marrow smears were prepared on glass slides for each of the mice, stained, and scored. Two thousand polychromatic erythrocytes (PCEs) were scored per animal to assess the micronucleated cells.
Test item treatment showed no marked effect on body weight in the main test. No mortality or signs of systemic toxicity were observed during the study. The animals in the test item treated and positive control groups were symptom-free during the whole observation period.
No biologically or statistically significant increases in the frequency of micronucleated polychromatic erythrocytes were seen in mice treated with the test item, compared to the negative (vehicle) control values at any of the sampling time points. The positive control treatment caused a large, clearly positive response demonstrating the sensitivity of the test system.
In conclusion, no induction of micronuclei in bone marrow erythrocytes was observed following administration of SN-475N to mice at up to and including 2000 mg/kg bw/day; thus, there was no evidence of any genotoxic activity of the test item under the conditions of this study.
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.