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EC number: 219-291-2 | CAS number: 2403-88-5
- Life Cycle description
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- Ecotoxicological Summary
- Aquatic toxicity
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- Short-term toxicity to fish
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- Long-term toxicity to aquatic invertebrates
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- Toxicity to microorganisms
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- Toxicological Summary
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Endpoint summary
Administrative data
Description of key information
oral: NOEL < 60 mg/kg/day (GLP-OECD Guideline study, MHW, 1998)
Key value for chemical safety assessment
Repeated dose toxicity: via oral route - systemic effects
Link to relevant study records
- Endpoint:
- short-term repeated dose toxicity: oral
- Remarks:
- combined repeated dose and reproduction / developmental screening
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Guideline study (OECD)
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
- GLP compliance:
- yes
- Remarks:
- Biosafety Research Center, Food, Drugs and Pesticides (An-Pyo Center), Japan
- Limit test:
- no
- Species:
- rat
- Strain:
- Crj: CD(SD)
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Age at study initiation: 10 weeks old for female and male animals
- Weight at study initiation: 359 - 400g for males; 227 - 282g for females - Route of administration:
- oral: gavage
- Vehicle:
- water
- Analytical verification of doses or concentrations:
- not specified
- Duration of treatment / exposure:
- Males: for 48 days (from 2 weeks prior to mating)
Females: for 41 - 52 days (from 14 days before mating to day 3 of lactation) - Frequency of treatment:
- once daily
- Remarks:
- Doses / Concentrations:
0, 60, 200, 600 mg/kg/day
Basis:
actual ingested - No. of animals per sex per dose:
- 12
- Control animals:
- yes, concurrent vehicle
- Observations and examinations performed and frequency:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: once a day
DETAILED CLINICAL OBSERVATIONS: Yes (see table)
BODY WEIGHT: Yes (see table)
- Time schedule for examinations: On the first day, the last day of the administration, the day of sacrifice and once a week during the administration period. For pregnant females: on the day 0, 14, and 20 of gestation and on day 0 and 4 of lactation.
FOOD CONSUMPTION: was determined on the same dy when body weight was determined for 24 hours.
OPHTHALMOSCOPIC EXAMINATION: No
HAEMATOLOGY: Yes
- Time schedule for collection of blood: at time of necropsy
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: only males
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at time of necropsy
- Animals fasted: No data
- How many animals: only males
URINALYSIS: No
- Time schedule for collection of urine:
- Metabolism cages used for collection of urine: Yes / No / No data
- Animals fasted: Yes / No / No data
- Parameters checked in table [No.?] were examined.
NEUROBEHAVIOURAL EXAMINATION: No
- Time schedule for examinations:
- Dose groups that were examined:
- Battery of functions tested: sensory activity / grip strength / motor activity / other: - Sacrifice and pathology:
- GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes - Statistics:
- Dunnett's or Scheffe's test for continuous data and Chi square test for quantal data
- Dose descriptor:
- NOAEL
- Effect level:
- < 60 mg/kg bw/day (nominal)
- Sex:
- male/female
- Basis for effect level:
- other: clinical signs: 8/12 of the male and in all female rats blepharoptosis was found
- Critical effects observed:
- not specified
- Conclusions:
- Body weight gain was decreased at more than 200 mg/kg/day in both sexes. In clinical signs, blepharoptosis and mydriasis were observed in all groups of both sexes, and their changes were dose-related. In 60 mg/kg/day group, the incidence for mydriasis was low and was observed concurrently with blepharoptosis at a slight degree and only sporadically. Mortality occurred in one female at 60 mg/kg/day, and in three males and one female at 600 mg/kg/day. At 600 mg/kg/day, the discontinued rats revealed reddish spots in digestive tracts, abnormal foci with gastric ulcer and the vacuolar degeneration in renal tubular epithelium. On the basis of mortality as well as clinical signs, a NOAEL of less than 60 mg/kg/day was set under the conditions of the study.
Reference
-Mortality and time to death: In the 600 mg/kg group, one and two males died after 6 and 9 days, respectively. Furthermore one high-dose female each died after 3 and 16 days. In the high-dose rats which died prematurely, the mortality was considered as compound-related. However, the cause of death could not be elucidated
for the dead female rat of the 60 mg/kg/day-group, since no histopathological correlate was detected.
-Body weights: For males at 200 mg/kg, a tendency for low body weight gain during administration period was observed and statistically significant difference from controls was noticed for the treatment period 29 - 43 days (Dunnets test p ≤ 0.05). Low body weight gain during the gestation period in females at 200 mg/kg was also observed (Dunnets test p ≤ 0.01).
Body weight change of male rats:
Dose level (mg/kg) | 0 | 60 | 200 | 600 |
No. of animals | 12 | 12 | 12 | 12 (N) |
Days of experiment | ||||
1 | 385±12 | 385±11 | 385±11 | 385±12 (12) |
8 | 403±17 | 399±23 | 395±13 | 368±41 (11) |
15 | 422±21 | 419±23 | 409±20 | 402±21 (9)* |
22 | 445±20 | 436±28 | 428±19 | 416±17 (9)** |
29 | 473±20 | 463±29 | 448±19* | 441±20 (9)** |
36 | 493±21 | 484±32 | 463±23* | 455±22 (9)** |
43 | 503±18 | 494±29 | 474±24* | 465±24 (9)** |
49 | 482±19 | 475±32 | 450±26** | 438±19 (9)** |
Gain 1 - 43 | 118±14 | 109±22 | 89±20** | 81±19 (9)** |
Significant difference from control group; *: P ≤ 0.05 **: P ≤ 0.01; Mean ± S.D. Unit: gram
Body weight change of female rats:
Dose level (mg/kg) | 0 | 60 | 200 | 600 |
Before mating period | ||||
No. of animals | 12 | 12 | 12 | 12 (N) |
Days of before mating | ||||
1 | 251±14 | 252±13 | 253±15 | 252±14 (12) |
8 | 259±16 | 257±13 | 253±18 | 255±12 (11) |
15 | 268±14 | 266±13 | 261±19 | 270±17 (11) |
Gain 1- 15 | 17±8 | 14±9 | 8±8 | 17±14 (11)* |
Dose level (mg/kg) | 0 | 60 | 200 | 600 |
Gestation period | ||||
No. of animals | 12 | 11 | 12 | 10 |
Day of gestation | ||||
0 | 281±15 | 276±22 | 274±18 | 280±19 |
7 | 310±16 | 310±18 | 301±22 | 301±22 |
14 | 350±17 | 349±15 | 339±23 | 336±19 |
21 | 458±20 | 457±22 | 432±30* | 418±27** |
Gain 0-21 | 176±12 | 181±11 | 158±18** | 138±15** |
Dose level (mg/kg) | 0 | 60 | 200 | 600 |
Lactation period | ||||
No. of animals | 12 | 11 | 12 | 10 (N) |
Days of lactation | ||||
0 | 315±21 | 323±27 | 293±32 | 288±25 |
4 | 332±23 | 327±21 | 308±26 | 312±26 (9) |
Gain 0-4 | 18±24 | 4±21 | 15±24 | 20±24 (9) |
Significant difference from control group; *: P ≤ 0.05 **: P ≤ 0.01; Mean ± S.D. Unit: gram
Food/water consumption: For males at 600 mg/kg, a tendency for increase in food consumption during administration period was observed and statistically significant differences from controls were noticed on day 8 -48 (Dunnets test p ≤ 0.01). For females at 600 mg/kg, a tendency for increase in food consumption during the premating period (Dunnets test p ≤ 0.01) and gestation period (Dunnets test p ≤ 0.05) was observed. However, the statistically significant difference from controls was not observed during the lactation period in all groups.
Food consumption change of female rats:
Dose level (mg/kg) | 0 | 60 | 200 | 600 | |
Before mating period | |||||
No. of animals | 12 | 12 | 12 | 11 | |
Days of before mating | 1-8 | 21±1 | 22±1 | 21±2 | 21±1 |
8-10 | 21±2 | 23±1 | 24±2 | 27±2** | |
Cumulative consumption | 1-15 | 305±21 | 310±19 | 314±24 | 341±25** |
Gestation period | |||||
No of dams | 12 | 11 | 12 | 10 | |
Day of gestation | 0-7 | 27±2 | 28±1 | 28±2 | 29±4 |
7-14 | 28±2 | 31±1 | 31±3 | 31±3* | |
14-24 | 29±39 | 31±2 | 29±2 | 29±4 | |
Cumulative consumption | 0-21 | 539±12 | 626±23 | 614±41 | 627±57 |
Lactation period | |||||
No of dams | 12 | 11 | 12 | 9 | |
Day of lactation | 0-4 | 35±10 | 32±8 | 33±8 | 35±10 |
Significant difference from control group; *: P ≤ 0.05 **: P ≤ 0.01; Mean ± S.D. Unit: gram
Food consumption change of male rats:
Dose level (mg/kg) | 0 | 60 | 200 | 600 | |
No of animals | 12 | 12 (N) | 12 | 12 (N) | |
Days of experiment | 1-8 | 28±2 | 29±4 | 28±2 | 24±7 |
8-15 | 30±2 | 31±2 | 31±2 | 35±3 (9)** | |
22-29 | 31±2 | 31±2 (11) | 31±2 | 34±3 (9)** | |
29-36 | 32±2 | 32±3 | 32±2 | 35±2 (9)** | |
36-43 | 30±2 | 31±2 | 31±2 | 34±2 (9)** | |
43-48 | 30±2 | 31±2 | 30±3 | 34±3 (9)** | |
Cumulative consumption | 1-15 | 410±28 | 416±36 | 413±30 | 428±36 (9) |
22-48 | 798±38 | 810±56 (11) | 811±49 | 889±51 (9)** |
Significant difference from control group; *: P ≤ 0.05 **: P ≤ 0.01; Mean ± S.D. Unit: gram
- Clinical signs: Blepharoptosis and mydriasis were observed in all groups of both sexes in a dose-related fashion. In the 60 mg/kg/day group, mydriasis was observed only in one male and one female, on the other hand blepharoptosis was observed in eight of 12 males (8/l2) and all females (12/12). Either of the clinical signs was of a slight degree and occurred only sporadically.
Clinical observation in male rats:
Dose level (mg/kg) | Days experiment | ||||||||
mydriasis | 1-7 | 8-14 | 15-21 | 22-28 | 29-35 | 36-42 | 43-49 | Total | |
0 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | |
60 | 0/12 | 1/12 | 0/12 | 0/12 | 0/12 | 1/12 | 0/12 | 1/12 | |
200 | 4/12 | 4/12 | 0/12 | 0/12 | 0/12 | 8/12 | 2/12 | 9/12 | |
600 | 12/12 | 11/11 | 7/9 | 9/9 | 9/9 | 9/9 | 8/9 | 12/12 | |
blepharoptosis | 0 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 |
60 | 3/12 | 2/12 | 1/12 | 1/12 | 0/12 | 6/12 | 0/12 | 8/12 | |
200 | 12/12 | 12/12 | 12/12 | 12/12 | 12/12 | 12/12 | 12/12 | 12/12 | |
600 | 12/12 | 10/11 | 9/9 | 9/9 | 9/9 | 9/9 | 9/9 | 12/12 | |
dead | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
60 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | |
200 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | |
600 | 1 | 2 | 0 | 0 | 0 | 0 | 0 | 3 |
Clinical observation in female rats (before and during mating period):
Dose level (mg/kg) | Days experiment | ||||||||
mydriasis | 1-7 | 8-14 | 15-21 | 22-28 | 29-35 | 36-42 | 43-49 | Total | |
0 | 0/12 | 0/12 | 0/12 | 0/0 | 0/0 | 0/0 | 0/0 | 0 | |
60 | 0/12 | 1/12 | 0/12 | 0/0 | 0/0 | 0/0 | 0/0 | 1 | |
200 | 3/12 | 1/12 | 0/12 | 0/0 | 0/0 | 0/0 | 0/0 | 4 | |
600 | 12/12 | 11/11 | 10/11 | 2/1 | 1/1 | 1/1 | 1/1 | 12 | |
blepharoptosis | 0 | 0/12 | 0/12 | 0/12 | 0/0 | 0/0 | 0/0 | 0/0 | 0 |
60 | 11/12 | 12/12 | 8/12 | 0/0 | 0/0 | 0/0 | 0/0 | 12 | |
200 | 12/12 | 12/12 | 12/12 | 0/0 | 0/0 | 0/0 | 0/0 | 12 | |
600 | 12/12 | 11/11 | 11/11 | 2/1 | 1/1 | 1/1 | 1/1 | 12 | |
dead | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
60 | 0 | 0 | 1 | 0 | 0 | 0 | 0 | 1 | |
200 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | |
600 | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 1 |
Clinical observation in female rats (Gestation period):
Dose level (mg/kg) | Days experiment | Total | ||||||||||||||||||||||||
0 | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | 15 | 16 | 17 | 18 | 19 | 20 | 21 | 22 | 23 | 0-23 | ||
mydriasis | 0 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0 |
60 | 0/11 | 0/11 | 0/11 | 0/11 | 0/11 | 0/11 | 0/11 | 0/11 | 0/11 | 0/11 | 0/11 | 1/11 | 0/11 | 0/11 | 0/11 | 0/11 | 0/11 | 0/11 | 0/11 | 0/11 | 0/11 | 0/11 | 0/11 | 0/11 | 1 | |
200 | 1/12 | 3/12 | 2/12 | 1/12 | 1/12 | 3/12 | 3/12 | 3/12 | 0/12 | 1/12 | 1/12 | 1/12 | 1/12 | 0/12 | 0/12 | 0/12 | 0/12 | 1/12 | 1/12 | 1/12 | 2/12 | 2/12 | 0/12 | 0/12 | 11 | |
600 | 8/10 | 7/10 | 6/10 | 7/10 | 8/10 | 9/10 | 9/10 | 8/10 | 9/10 | 7/10 | 7/10 | 8/10 | 6/10 | 5/10 | 5/10 | 5/10 | 4/10 | 7/10 | 9/10 | 9/10 | 9/10 | 9/10 | 4/10 | 0/10 | 10 | |
blepharoptosis | 0 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0 |
60 | 5/11 | 5/11 | 5/11 | 6/11 | 6/11 | 4/11 | 7/11 | 3/11 | 4/11 | 4/11 | 3/11 | 2/11 | 3/11 | 2/11 | 1/11 | 4/11 | 3/11 | 2/11 | 6/11 | 5/11 | 0/11 | 5/11 | 1/11 | 0/11 | 10 | |
200 | 12/12 | 12/12 | 12/12 | 12/12 | 12/12 | 12/12 | 12/12 | 11/12 | 11/12 | 12/12 | 12/12 | 12/12 | 12/12 | 12/12 | 12/12 | 11/12 | 12/12 | 12/12 | 12/12 | 12/12 | 10/12 | 11/12 | 6/12 | 0/12 | 12 | |
600 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 10/10 | 4/10 | 0/10 | 10 |
Clinical observation in female rats (Lactation period):
Dose level (mg/kg) | Days experiment | Total | |||||
mydriasis | 0 | 1 | 2 | 3 | 4 | 0-4 | |
0 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0 | |
60 | 0/11 | 0/11 | 0/11 | 0/11 | 0/11 | 0 | |
200 | 10/12 | 10/12 | 8/12 | 2/12 | 0/12 | 12 | |
600 | 9/10 | 8/10 | 9/10 | 8/9 | 0/9 | 10 | |
blepharoptosis | 0 | 0/12 | 0/12 | 0/12 | 0/12 | 0/12 | 0 |
60 | 3/11 | 1/11 | 1/11 | 1/11 | 0/11 | 3 | |
200 | 12/12 | 12/12 | 12/12 | 12/12 | 0/12 | 12 | |
600 | 10/10 | 10/10 | 9/10 | 9/9 | 0/9 | 10 |
- Haematology: Males: No dose-related changes in haematology.
- Gross pathology incidence and severity: No changes in gross pathology in the survival rats. However, the reddish spots of digestive tracts, the abnormal foci with gastric ulcer, and the vacuolar degeneration in renal tubular epithelium were observed in the discontinued rats of the 600 mg/kg group.
- Organ weight changes: Males: Increase of adrenals weight was observed in 600 mg/kg group.
Females: In 600 mg/kg, increase in adrenals and liver weights were observed.
male | female | |||||
Dose level (mg/kg/day) | 0 | 200 | 600 | 0 | 200 | 600 |
Absolute weight | ||||||
Liver (g, mean ± SD) | 13.56±0.08 | 12.94±1.92 | 11.76±1.09** | 14.45±1.82 | 14.56±1.07 | 15.43±2.43 |
Adrenals (mg, mean ± SD) | 66±2 | 67±12 | 71±8 | 80±10 | 862±8 | 87±10 |
Relative weight | ||||||
Liver (mg, mean ± SD) | 2.815±0.086 | 2.867±0.315 | 2.682±0.206 | 4.341±0.396 | 4.741±0.270 | 4.927±0.535** |
Adrenal (mg, mean ± SD) | 13.688±1.934 | 14.865±2.922 | 16.250±1.836* | 24.142±2.978 | 26.647±2.910 | 27.825±2.976** |
- Histopathology: No abnormalities were observed in the survival rats. However, red spots of digestive tracts, abnormal foci with gastric ulcer and vacuolar degeneration of renal tubular epithelium were observed in rats of the 600 mg/kg group which died prematurely.
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed
- Dose descriptor:
- NOAEL
- Study duration:
- subchronic
- Species:
- rat
Repeated dose toxicity: inhalation - local effects
Endpoint conclusion
- Species:
- rat
Additional information
There is only one report of a repeated oral toxicity study using rats. In an oral (via gavage) rat study according to the OECD combined repeated dose and reproductive/development toxicity screening test [OECD TG 422], organ weight and histopathological changes were observed (MHW, Japan, 1998). This MHW study was identified as the key study because it was well conducted and used a current protocol. Details of the study are as follows. The study was conducted at doses of 0, 60, 200, and 600 mg/kg/day. In males of the 600 mg/kg group, 1 and 2 rats died after 6 and 9 days, respectively. One female of the 60 mg/kg as well as one female of the 600 mg/kg group died after 16 and 3 days, respectively. In male rats and female rats (during the gestation period) at 200 mg/kg a tendency for low body weight gain during the administration period was observed and a statistically significant difference from controls was noticed. In both sexes at 600 mg/kg, a remarkable body weight decrease was observed. Blepharoptosis and mydriasis were observed in all dose groups of both sexes in a dose-related manner. In 60 mg/kg/day group, the incidence for mydriasis was low and was observed concurrently with blepharoptosis at a slight degree and only sporadically. There were no evidence of alterations in haematology and biochemistry. Relative adrenal weights in the 600 mg/kg group of both sexes were increased, and relative liver weights in the 600 mg/kg group of females were increased. Only animals that died prematurely revealed histopathological findings. They consisted of reddish spots in the digestive tracts, abnormal foci with gastric ulcer and vacuolar degeneration in the renal tubular epithelium. On the basis of mortality as well as clinical signs, a NOAEL of less than 60 mg/kg/day was set under the conditions of the study.
Conclusion: On the basis of mortaliy and clinical signs observed also at the low-dose, a NOAEL of less than 60 mg/kg/day was set under the conditions of the study.
Justification for classification or non-classification
There is currently no need for classification based upon repeated oral exposure to 2,2,6,6 -tetramethylpiperidin-4 -ol, since none of the observed effects would lead to a hazard relevant for classification.
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