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EC number: 214-787-5 | CAS number: 1194-65-6
- 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
Toxicity to reproduction
Administrative data
- Endpoint:
- two-generation reproductive toxicity
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 30 July 1987 to 21 April 1989
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Referenceopen allclose all
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 989
- Report date:
- 1988
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 992
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 993
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- EPA OPP 83-4 (Reproduction and Fertility Effects)
- Deviations:
- yes
- Remarks:
- (actual relative humidity was 20-94%; three incorrect pups were selected for the F1 generation; in addition, the following protocol deviation was also reported; on three occasions test diets were prepared in advance and stored frozen)
- GLP compliance:
- yes
- Limit test:
- no
Test material
- Reference substance name:
- Dichlobenil
- EC Number:
- 214-787-5
- EC Name:
- Dichlobenil
- Cas Number:
- 1194-65-6
- Molecular formula:
- C7H3Cl2N
- IUPAC Name:
- 2,6-dichlorobenzonitrile
- Test material form:
- solid: particulate/powder
- Details on test material:
- - Physical state: solid
- Storage condition of test material: in the dark at ambient room temperature
Constituent 1
Test animals
- Species:
- rat
- Strain:
- other: Crl:CD(SD)BR
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Age at study initiation: (P) approxaimtely 6 wks; (F1) nominally 4 wks
- Weight at study initiation: (P) Males: 139.7-217.4 g; Females: 107.7-149.9 g;
- Housing: stainless steel wireless cages
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 12 days
ENVIRONMENTAL CONDITIONS
- Temperature: 19-25 °C
- Humidity: 40-70 %
- Air changes: minimum 15 per hour
- Photoperiod: 12 hours light/12 hours dark
Administration / exposure
- Route of administration:
- oral: feed
- Vehicle:
- unchanged (no vehicle)
- Details on exposure:
- DIET PREPARATION
- Rate of preparation of diet (frequency): weekly (excpet on three occassions when the diet was prepared in advance and frozen) - Details on mating procedure:
- - M/F ratio per cage: 1/1
- Length of cohabitation: up to 15 days
- Proof of pregnancy: sperm in vaginal smear - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- Samples were taken from the diet formulations prepared in weeks 1, 13, 26 and 36 and analysed by shaking samples with hexane and water (low dose groups) or hexane only (high dose groups). Hexane solutions are further diluted and analysed using GC.
- Duration of treatment / exposure:
- Two generations
- Frequency of treatment:
- Daily
- Details on study schedule:
- - F1 parental animals not mated until 10 weeks after selected from the F1 litters.
Doses / concentrationsopen allclose all
- Dose / conc.:
- 60 ppm
- Dose / conc.:
- 350 ppm
- Dose / conc.:
- 2 000 ppm
- No. of animals per sex per dose:
- Thirty (P); twenty-five (F1 generation)
- Control animals:
- yes, plain diet
Examinations
- Parental animals: Observations and examinations:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily
BODY WEIGHT: Yes
- Time schedule for examinations: weekly except for mated females which were weighed on days 0, 6, 12, 15 and 20 of gestation and days 1, 4, 7, 14 and 21 of lactation.
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined: Yes (weekly)
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes - Litter observations:
- STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: yes
- If yes, maximum of 8 pups/litter (sex distribution as equal as possible); excess pups were killed and discarded.
PARAMETERS EXAMINED
The following parameters were examined in F1 / F2 offspring:
stillbirths, live births, postnatal mortality, litter weights, and clinical condition of pups
GROSS EXAMINATION OF DEAD PUPS:
yes - Postmortem examinations (parental animals):
- SACRIFICE
- Male animals: All surviving animals (at time of weaning of F1 generation).
- Maternal animals: All surviving animals (at time of weaning of F1 generation).
GROSS NECROPSY
- Gross necropsy was performed.
HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues were removed: ovaries, uterus, cervix, vagina, lesions, pituitary, testes, epididymides, seminal vesicles, prostate and coagulating gland. - Postmortem examinations (offspring):
- SACRIFICE
- The F1 offspring not selected as parental animals and all F2 offspring were sacrificed at time of weaning.
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows:
GROSS NECROPSY
- Gross necropsy was performed.
- Number of implantation sites were counted and recorded for F1 females. - Statistics:
- Using nested analysis of variance for normally distributed errors or by non-parametric techniques for non-normally distributed errors.
The standard deviation obtained from analysis of variance was used for 't' tests between control and treatment groups. Where necessary, data were transformed before analysis.
Non-parametric testing was performed using the Kruskal-Wallis test for a treatment-related response.
Significant differences between control and treatment groups were determined using the Wilcoxon rank sum test. - Reproductive indices:
- Median pre-coital time = time by which half the females had mated
Fertility index = (no. of pregnant females / no. of paired females) x 100
Fecundity index = (no. of pregnant females / no. of mated females) x 100
Gestation index = (no. of females with live pups / no. of pregnant females) x 100 - Offspring viability indices:
- Post-implantation survival index (F1 only) = (no. of pups born / no. of implantation sites) x 100
Live birth index = (no. of pups alive on day 1 / no. of pups born) x 100
Viability index 1 = (no. of pups alive on day 4 before culling / no. of pups alive on day 1) x 100
Viability index 2 = (no. of pups alive on day 7 / no. of pups alive on day 4 after culling) x 100
Viability index 3 = (no. of pups alive on day 14 / no. of pups alive on day 7) x 100
Viability index 4 = (no. of pups alive on day 21 / no. of pups alive on day 14 x 100)
Results and discussion
Results: P0 (first parental generation)
General toxicity (P0)
- Clinical signs:
- effects observed, treatment-related
- Description (incidence and severity):
- Clinical observations of P generation adults were restricted to those commonly seen in this strain of rat in these laboratories e.g. occasional instances of rough hair coat, slight hair loss, fur staining and chronnodacryorrhoea. Incidence of fur staining and slight hair loss was slightly increased at 2000 ppm in P generation females during late gestation.
- Mortality:
- mortality observed, non-treatment-related
- Description (incidence):
- Treatment did not adversely affect mortality. One P generation male was found dead during week 14; necropsy revealed a liver mass. This death was not attributable to treatment. There were no other adult deaths.
- Body weight and weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- Body weight gains were lower in males and females at 2000 ppm; body weight gain in females during gestation was also lower at 2000 ppm.
- Food consumption and compound intake (if feeding study):
- effects observed, treatment-related
- Description (incidence and severity):
- Food consumption was lower than controls in males and females at 2000 ppm. Initial lowering of food consumption in P generation animals was so marked as to suggest adverse palatability of the test material in the diet.
- Food efficiency:
- no effects observed
- Description (incidence and severity):
- Food conversion efficiency was similar in all groups, with the exception of week 1 for the highest dose level.
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- not examined
- Haematological findings:
- not examined
- Clinical biochemistry findings:
- not examined
- Urinalysis findings:
- not examined
- Behaviour (functional findings):
- not examined
- Immunological findings:
- not examined
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Histopathological findings: non-neoplastic:
- no effects observed
- Description (incidence and severity):
- There were no significant histopathological effects on either the pituitary or reproductive tract attributable to test material administration.
- Histopathological findings: neoplastic:
- not examined
- Other effects:
- not examined
Reproductive function / performance (P0)
- Reproductive function: oestrous cycle:
- no effects observed
- Reproductive function: sperm measures:
- no effects observed
- Reproductive performance:
- no effects observed
- Description (incidence and severity):
- Mating performance, fertility, fecundity, pregnancy rate and duration of gestation were not adversely affected by treatment.
Effect levels (P0)
- Key result
- Dose descriptor:
- NOEL
- Effect level:
- 60 ppm
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- body weight and weight gain
- other: equivalent to 3 mg/kg bw/day Based on lower bodyweight gain in offspring in both adults and pups, no effects on fertility were noted
Results: F1 generation
General toxicity (F1)
- Clinical signs:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Clinical observations of F1 generation adults were restricted to those commonly seen in this strain of rat in these laboratories e.g. occasional instances of rough hair coat, slight hair loss, fur staining and chronnodacryorrhoea.
- Dermal irritation (if dermal study):
- not examined
- Mortality / viability:
- mortality observed, non-treatment-related
- Description (incidence and severity):
- Offspring survival was not adversely affected by treatment. One F1 generation male was killed during week 12 following accidental injury to the snout. This death was not attributable to treatment. There were no other adult deaths.
- Body weight and weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- Body weight gains were lower in males and females at 2000 ppm; body weight gain in females during gestation was also lower at 2000 ppm. Body weight gain was lower than controls in F1 males at 350 ppm.
- Food consumption and compound intake (if feeding study):
- effects observed, treatment-related
- Description (incidence and severity):
- Food consumption was lower than controls in males and females at 2000 ppm.
- Food efficiency:
- no effects observed
- Description (incidence and severity):
- Food conversion efficiency was similar in all groups.
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- not examined
- Haematological findings:
- not examined
- Clinical biochemistry findings:
- not examined
- Urinalysis findings:
- not examined
- Sexual maturation:
- no effects observed
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Gross pathological findings:
- no effects observed
- Description (incidence and severity):
- Necropsy findings of weaned offspring were not adversely affected by treatment.
- Histopathological findings:
- effects observed, treatment-related
- Description (incidence and severity):
- There were no significant histopathological findings for either the pituitary or reproductive tract attributable to test material administration. Hyaline droplet nephropathy was recorded in the kidneys of three high dose male. This finding was considered related to test material administration.
- Other effects:
- not examined
Developmental neurotoxicity (F1)
- Behaviour (functional findings):
- not examined
Developmental immunotoxicity (F1)
- Developmental immunotoxicity:
- not examined
Effect levels (F1)
open allclose all
- Key result
- Dose descriptor:
- NOEL
- Generation:
- F1
- Effect level:
- 60 ppm
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- body weight and weight gain
- other: equivalent to 3 mg/kg bw/day Based on lower bodyweight gain in offspring in both adults and pups, no effects on fertility were noted
- Key result
- Dose descriptor:
- NOAEL
- Generation:
- F1
- Effect level:
- 2 000 ppm
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: No observed adverse effects
Target system / organ toxicity (F1)
- Critical effects observed:
- not specified
Overall reproductive toxicity
- Reproductive effects observed:
- not specified
Any other information on results incl. tables
Table 1: Summary of adult performance
Dose group (ppm) | ||||||||
0 | 60 | 350 | 2000 | |||||
P | F1 | P | F1 | P | F1 | P | F1 | |
Males | ||||||||
In group | 30 | 25 | 30 | 25 | 30 | 25 | 30 | 25 |
Dead: post pairing | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 1 |
Failing to induce pregnancy | 1 | - | 2 | - | 2 | - | 1 | - |
Females | ||||||||
In group | 30 | 25 | 30 | 25 | 30 | 25 | 30 | 25 |
Pregnant/% | 29/96.7 | 25/100 | 28/93.3 | 20/80 | 28/93.3 | 24/96 | 29/96.7 | 25/100 |
With total resorption | 1 | - | 0 | - | 0 | - | 0 | - |
Total litter loss | 4 | 4 | 3 | 4 | 3 | 4 | 5 | 2 |
With live pups at day 21 post-partum | 24 | 21 | 25 | 16 | 25 | 20 | 24 | 23 |
Table 2: Group mean body weight gain (g)
Dose group (ppm) | |||||||||
0 | 60 | 350 | 2000 | ||||||
Time interval | Sex | P | F1 | P | F1 | P | F1 | P | F1 |
0 -5 | M | 199.9 | 260.6 | 192.0 | 256.7 | 192.7 | 242.6 | 148.1 | 201.9 |
F | 90.5 | 120.5 | 91.9 | 122.1 | 88.0 | 111.5 | 67.1 | 98.7 | |
6 -10 | M | 88.9 | 104.6 | 98.8 | 113.6 | 83.5 | 93.8 | 68.8 | 71.0 |
F | 32.1 | 40.3 | 31.0 | 37.7 | 34.5 | 39.4 | 23.8 | 33.3 | |
11 -13 | M | 33.2 | 44.7 | 23.3 | 43.8 | 29.5 | 37.6 | -6.1 | 41.7 |
F | 100 | 112 | 109 | 115 | 109 | 108 | 88 | 85 | |
14 -16 | M | 29.8 | 31.8 | 29.3 | 41.8 | 30.7 | 48.3 | 53.7 | 42.6 |
F | 29 | 46 | 40 | 36 | 53 | 45 | 65 | 60 |
*14-17 weeks for males of the F1 generation
Table 3: Group mating data
Dose group (ppm) | ||||||||
0 | 60 | 350 | 2000 | |||||
Mating indices | P | F1 | P | F1 | P | F1 | P | F1 |
Number of paired females | 30 | 25 | 30 | 25 | 30 | 25 | 30 | 25 |
Number of mated males | 30 | 25 | 30 | 22 | 29 | 24 | 29 | 25 |
Number of pregnant females | 29 | 25 | 28 | 20 | 28 | 24 | 29 | 25 |
Fertility index (%) | 96.7 | 100 | 93.3 | 80 | 93.3 | 96 | 96.7 | 100 |
Fecundity index (%) | 96.7 | 100 | 93.3 | 90.9 | 96.6 | 100 | 100 | 100 |
Table 4: Group mean pup weights (g)
Dose group (ppm) | ||||||||
0 | 60 | 350 | 2000 | |||||
Day | P | F1 | P | F1 | P | F1 | P | F1 |
1 (combined) | 6.1 | 5.7 | 5.7 | 5.9 | 5.5 | 5.9 | 5.6 | 5.8 |
4 (combined) | 8.6 | 7.9 | 7.9 | 8.4 | 7.2 | 7.7 | 6.9 | 7.8 |
7 (combined) | 13.8 | 13.2 | 13.1 | 13.7 | 12.1 | 12.8 | 10.4 | 12.0 |
14 (combined) | 29.8 | 30.0 | 29.1 | 29.8 | 27.4 | 27.4 | 21.9 | 24.4 |
21 (combined) | 49.8 | 50.9 | 49.5 | 49.7 | 46.7 | 45.98 | 38.2 | 39.9 |
21 (males) | 51.2 | 51.8 | 50.3 | 51.0 | 49.1 | 47.2 | 39.5 | 40.6 |
21 (females) | 49.6 | 49.6 | 48.7 | 48.5 | 44.7 | 45.9 | 37.9 | 38.9 |
% weight changes | 716 | 793 | 4768 | 742 | 749 | 678 | 582 | 588 |
Table 5: Group incidence histopathology findings
P generation | F1 generation | ||||||||
Dose group* and sex | Dose group* and sex | ||||||||
0 ppm M | 2000 ppm M | 0 ppm F | 2000 ppm F | 0 ppm M | 2000 ppm M | 0 ppm F | 2000 ppm F | ||
Animals examined | 30 | 30 | 30 | 30 | 25** | 24** | 25** | 25** | |
Animals unremarkable | 10 | 10 | 19 | 23 | 8 | 9 | 15 | 17 | |
Skin | Acanthosis | 0 | 1 | 3 | 3 | 1 91) | 1 (2) | 9 (9) | 7 (7) |
Dermatitis | 2 | 0 | 3 | 0 | 0 (1) | 1 (2) | 2 (9) | 0 (7) | |
Adnexal atrophy | - | - | - | - | 0 (1) | 0 (2) | 0 (9) | 1 (7) | |
Liver | Necrosis (lobar) | 0 | 1 | 0 | 0 | 1 (1) | - | - | - |
Kidneys | Focal nephropathy | 0 | 10 | 0 | 0 | - | - | - | - |
Hydroneophrosis | 2 | 0 | 0 | 0 | 1 (1) | 1 (3) | - | - | |
Nephroblastoma | 0 | 0 | 0 | 1 | - | - | - | - | |
Hyaline droplets | - | - | - | - | 0 (1) | 3 (3) | - | - | |
Basophilic tubules | - | - | - | - | 0 (1) | 3 (3) | - | - | |
Casts | - | - | - | - | 0 (1) | 3 (3) | - | - | |
Cyst | - | - | - | - | 0 (1) | 1 (3) | - | - | |
Pituitary | Altered cell focus | 0 | 2 | 0 | 0 | - | - | - | - |
Cyst | 0 | 3 | 0 | 0 | 1 | 0 | 0 | 0 | |
No sample | 0 | 0 | 1 | 0 | - | - | - | - | |
Oral cavity | Necropsy finding/ no equivalent finding | 2 | 1 | 2 | 2 | - | 1 (1) | - | - |
Stomatitis | - | - | - | - | - | 1 (1) | - | - | |
Nasal cavity | Rhinitis | - | - | - | - | - | 1 (1) | - | - |
Abdominal cavity | Necropsy finding/ no equivalent finding | 0 | 1 | 0 | 0 | - | - | - | - |
Lungs | Inflammatory cell foci | - | - | - | - | 1 (1) | - | - | - |
Congestion/heamorrhage | 2 | 1 | 0 | 1 | 1 (1) | - | - | - | |
Foamy histiocytes | 2 | 1 | 0 | 0 | - | - | - | - | |
Lymphoid hyperlasia | 1 | 1 | 0 | 0 | - | - | - | - | |
Pneunomitis | 2 | 1 | 0 | 0 | - | - | - | - | |
Skull | Exostoses | 1 | 1 | 0 | 0 | - | - | - | - |
Foot | Arthritis | 1 | 0 | 0 | 0 | - | - | - | - |
Dermatitis | 0 | 1 | 0 | 0 | - | - | - | - | |
Tail | Dermatitis/folliculitis | 0 | 3 | 0 | 0 | 1 (1) | - | - | - |
Mandiubular lymph node | Hyperplasia | - | - | - | - | ||||
Ear | Chondritis | - | - | - | - | ||||
Testis | Atrophy | 1 | 1 | - | - | 2 | 2 | - | - |
Sperm granuloma | - | - | - | - | 0 | 1 | - | - | |
Epididymis | Inflammatory cell foci | 0 | 1 | - | - | 1 | 0 | - | - |
Oligospermia | 0 | 1 | - | - | 0 | 2 | - | - | |
Seminal vesicle | Not remarkable | 30 | 30 | - | - | 24 | 25 | - | - |
Coagualting gland | One sample | 4 | 3 | - | - | 0 | 1 | - | - |
Protstate | Inflammatory cell foci | - | - | - | - | 6 | 6 | - | - |
Prostatitis | 10 | 10 | - | - | 3 | 4 | - | - | |
Ovary | Not remarkable | - | - | 30 | 30 | - | - | 25 | 25 |
One sample | - | - | - | - | - | - | 1 | 0 | |
Uterus | Arteritis | - | - | 1 | 0 | - | - | 0 | 0 |
Oestrus stage | - | - | 1 | 1 | - | - | 0 | 0 | |
Cervix | Not remarkable | - | - | - | - | - | - | 0 | 0 |
Vagina | Not remakrable | - | - | 30 | 30 | - | - | 0 | 0 |
Cause of death | Liver lesion | 1 | |||||||
Upper respiratory infection/ stomatitis | 1 |
If less animals were examined, the real number examined is presented between brackets.
Applicant's summary and conclusion
- Conclusions:
- Under the conditions of the test, administration of the test material to rats over two generations at dietary concentrations up to 2000 ppm did not adversely affect adult reproduction or offspring survival. Body weight gains were lower in adults and offspring at 2000 ppm and to a lesser extent at 350 ppm. Food consumption was also reduced in adults at 2000 ppm. No adverse effects of treatment were seen at the low level of 60 ppm which is considered to be the NOEL (equivalent to 3 mg/kg bw/day).
- Executive summary:
In a GLP compliant study conducted in line with standardised guideline EPA OPP 83-4, the effect of the test material on reproductive toxicity was determined in the rat.
Rats were administered the test material by oral (diet) at 0, 60, 350 and 2000 ppm.
Adult reproduction or offspring survival was not adversely affected. Body weight gains were lower in adults and offspring at 2000 ppm and to a lesser extent at 350 ppm. Food consumption was also reduced in adults at 2000 ppm. No adverse effects of treatment were seen at the low level of 60 ppm which is considered to be the NOEL (equivalent to 3 mg/kg bw/day).
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