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EC number: 201-210-7 | CAS number: 79-50-5
- 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
Repeated dose toxicity: oral
Administrative data
- Endpoint:
- short-term repeated dose toxicity: oral
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2002-09-23 to 2003-04-29
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: OECD guideline compliant GLP compliant
Cross-reference
- Reason / purpose for cross-reference:
- reference to same study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 003
- Report date:
- 2003
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
- GLP compliance:
- yes
- Limit test:
- no
Test material
- Reference substance name:
- (±)-dihydro-3-hydroxy-4,4-dimethylfuran-2(3H)-one
- EC Number:
- 201-210-7
- EC Name:
- (±)-dihydro-3-hydroxy-4,4-dimethylfuran-2(3H)-one
- Cas Number:
- 79-50-5
- Molecular formula:
- C6H10O3
- IUPAC Name:
- (±)-dihydro-3-hydroxy-4,4-dimethylfuran-2(3H)-one
- Details on test material:
- - Name of test material (as cited in study report): dl-Lactone
- Physical state: white crystalline mass
- Analytical purity: 99.8 %
- Purity test date: not reported
- Lot/batch No.: BX226
- Expiration date of the lot/batch: 31 May 2003
- Stability under test conditions: Stable under storage conditions
- Storage condition of test material: At room temperature in the dark
Constituent 1
Test animals
- Species:
- rat
- Strain:
- Wistar
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Strain: Wistar, Crl: (WI) BR (outbred, SPF-Quality).
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: 10 weeks
- Weight at study initiation: not reported
- Fasting period before study: no
- Housing: upon arrival 5 animals/sex/cage in suspended stainless steel cages, during the mating females and males caged together 1:1 in
suspended stainless steel cages with wire mesh floors; mated females and males individually housed in labelled polycarbonate cages containing sawdust (SAWI bedding, Jelu Werk, Rosenberg, Germany); from arrival until mating, males and females were housed in separate rooms; offspring kept with the dam until termination; during the final stage of the pregnancy period (from approximately day 16 of gestation onwards) and during lactation,
paper (Enviro-dri, BMI, Helmond, The Netherlands) provided as nesting material;
- Diet (e.g. ad libitum): ad libitum, standard pelleted laboratory animal diet (from Altromin (code VRF 1), Lage, Germany)
- Water (e.g. ad libitum): ad libitum, tap water
- Acclimation period: 5 d
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17.1-24.2
- Humidity (%): 33 -76%
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 / 12
Temporary deviations from the maximum level for relative humidity (with a maximum of 6%) and light/dark cycle (with a maximum of 1 hour) occurred due to cleaning procedures or performance of functional observations in the room. Based on laboratory historical data these deviations are considered not to affect the study integrity.
IN-LIFE DATES:
Acclimatisation: 18 September 2002
Start treatment: 23 September 2002
Start mating: 07 October 2002
Necropsy males: 21 October 2002
Necropsy females and pups: 21 October - 18 November 2002
Administration / exposure
- Route of administration:
- oral: gavage
- Vehicle:
- water
- Remarks:
- Water (Milli-Q) of 37°C adjusted to pH 4. Acetic acid was added to milli-Q water to obtain pH 4.
- Details on oral exposure:
- PREPARATION OF DOSING SOLUTIONS:
23 September -01 October 2002:
Sufficient ammounts of Milli-Q water of 37°C adjusted to pH 4 was added to the weighted test item. Formulations (w/w) were prepared daily within 4 hours prior to dosing and were homogenised to a visually acceptable level. The formulations were kept continuously at 37°C.
02 October -17 November 2002:
The test item was heated at 100°C, melted and divided into seven portions. For every week of study, milli-Q water at pH 4 and of 37°C was added to one of these portions. For every dose group, a stock-solution was prepared. The stock solutions and formulations were stirred and kept continuous at 37°C. From 08 October 2002 onwards, the stock-solution of group 1 was not heated, but was kept at room temperature due to practical reasons. Formulations (w/w) were prepared daily within 4 hours prior to dosing and were homogenised to visually acceptable levels.
Storage conditions:
At 37°C, except for the stock-solution of group 1 which was kept at room temperature from 08 October 2002 onwards.
VEHICLE
- Justification for use and choice of vehicle (if other than water): Based on trial formulations performed at NOTOX and on information provided by the sponsor. During NOTOX Project 257568 (Determination of the hydrolysis of dl-lactone as a function of pH) it was determined that dllactone was hydrolytically stable at pH 4 and 50°C.
- Concentration in vehicle: 0, 8, 40, 200 mg/mL
- Amount of vehicle (if gavage): 5 ml/kg bw
- Stability: Formulations in Milli-Q water are stable for 4 hours at room temperature and formulations in Milli-Q water adjusted to pH 4 are stable for 4 hours at room temperature and for 8 days at 37°C (determined during this project). - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- The concentration of DL-Lactone was determined by High Performance Liquid Chromatography with Mass Spectrometry detection (LC-MS-MS):
Column: Lichrospher 100 RP-18, (250 * 4 (I.D.) mm; dp=5 pm (Merck, Darmstadt, Germany)
Mobile phase: 20/80/0.1 (v/v/v) Acetonitrile / Milli-Q water/ formic acid
Flow: 1 ml/min
Split: 1:1.5
Detection: SCIEX MSMS system API-300 mass spectrometer (Perkin Elmer, Biosystems, Foster City, Canada),
Interface: Turbo ionspray at 350°C; N2 flow rate of 6000 ml/min.; operated in positive ion mode
Monitored masses: MRM test substance m/z 131.1 --> 113.1, MRM internal standard m/z 127.1 --> 99.0
Injection volume: 100 µL
Column temperature: 20 °C
Autosampler temperature: 4 °C
- Detection limits (LOD, LOQ) (indicate method of determination/calculation):
linear relationship between response and concentration in the concentration range of 0.995 25. 0 mg/1 (r= 0.9989, n=6, weighted 1/concentration²)
LOD = 0.28 mg/L at an injection volume of 100 µl (i.e. 0.28 ng absolute) - Duration of treatment / exposure:
- males:
2 weeks prior to mating, during mating, and upto termination (28 d for all males).
females:
2 weeks prior to mating, during mating, during post-coitum, and at least 4 days of lactation. The mean duration of treatment of females: 43 d, minimum: 28 d, maximum: 56 d - Frequency of treatment:
- 5 d/wk, single daily oral application via gavage
Doses / concentrations
- Remarks:
- Doses / Concentrations:
0, 40, 200, 1000 mg/kg bw/d
Basis:
actual ingested
- No. of animals per sex per dose:
- 10
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale: based on the results of a dose range finding study
- Rationale for animal assignment: computer-generated random algorithm
- Section schedule rationale: not reported - Positive control:
- no
Examinations
- Observations and examinations performed and frequency:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Mortality - twice daily, clinical signs: once daily
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily, in addition once prior to start of treatment and once a week thereafter, this was also performed outside the home cage in a standard arena during the pre-mating period.
BODY WEIGHT: Yes
- Time schedule for examinations: on the first day of exposure and weekly thereafter; mated females on days 0, 7, 14 and 21 of gestation, and during lactation on days 1 and 4.
FOOD CONSUMPTION: Yes
- Time schedule for examinations: Weekly, for males and females; suspended during the mating; food consumption of mated females measured on gestation days 0, 7, 14 and 21, and during lactation on days 1 and 4.
FOOD EFFICIENCY: No
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: Subjective appraisal during the study, but no quantitative investigation introduced as no effect was suspected.
OPHTHALMOSCOPIC EXAMINATION: No
HAEMATOLOGY: Yes
- Time schedule for collection of blood: immediately prior to scheduled post mortem examination,
- Anaesthetic used for blood collection: Yes, isoflurane
- Animals fasted: Yes, over night
- How many animals: 5 males and 5 females, randomly selected
- Parameters checked in table 1 were examined.
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood:
- Animals fasted: Yes / No / No data
- How many animals:
- Parameters checked in table 1 were examined.
URINALYSIS: No
NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations:
assigned males were tested during week 4 of treatment
assigned females were tested during lactation (all before blood sampling).
- Dose groups that were examined: all
- Battery of functions tested: sensory activity / grip strength / motor activity
The following tests were performed in 5 males and 5 females, randomly selected from each group:
- hearing ability (score 0 = normal/present, 1 = abnormal/absent).
- pupillary reflex (score 0 = normal/present, 1 = abnormal/absent).
- static righting reflex (score 0 = normal/present, 1 = abnormal/absent).
- grip strength (score 0 = normal/present, 1 = abnormal/absent).
In order to avoid hypothermia of pups, dams were removed from the pups for not more than 30-40 minutes.
- motor activity test (recording period: 12 hours during overnight for individual animals, using a computerised monitoring system, Pearson Technical Services, Debenham, Stowmarket, England). During the motor activity test, males were caged individually and females were caged with their offspring.
OTHER: - Sacrifice and pathology:
- GROSS PATHOLOGY: Yes, see table 2
HISTOPATHOLOGY: Yes, see table 2
- Of the selected 5 males/group of the control and high dose group: additional slides of the testes prepared to examine staging of spermatogenesis; testis processed, sectioned at 3-4 microns, and stained with PAS/hematoxylin.
- The following slides were examined by a pathologist:
The preserved organs and tissues of the selected animals of groups 1 and 4.
The additional slides of the testes of the selected 5 males/group of groups 1 and 4 to examine staging of spermatogenesis
The preserved organs and tissues of the animals of all dose groups which died spontaneously or were killed in extremis all gross lesions of all animals (all dose groups)
The preserved organs and tissues of all non-pregnant females and animals suspected of infertility - Other examinations:
- Observations offspring:
- numbers of live and dead pups at the First Litter Check (= check at day 1 of lactation) and daily thereafter (if possible, defects or cause of death evaluated)
- individual weight of all live pups on days 1 and 4 of lactation
- sex of all pups (by assessment of the ano-genital distance)
- number of pups with physical or behavioural abnormalities daily
Macroscopic examination of offspring:
- all offspring sexed and externally examined if practically possible
- examination of stomach for the presence of milk
- descriptions of all macroscopic abnormalities recorded; defects or cause of death were evaluated, if possible
- no pups preserved - Statistics:
- If the variables could be assumed to follow a normal distribution, the Dunnett-test (many-to-one t-test) based on a pooled variance estimate was applied for the comparison of the treated groups and the control groups.
The Steel-test (many-to-one rank test) was applied instead of the Dunnett-test if the data could not be assumed to follow a normal distribution.
The exact Fisher-test was applied for 2x2 tables if variables could be dichotomized without loss of information.
All tests were two-sided and in all cases p < 0.05 was accepted as the lowest level of significance.
Results and discussion
Results of examinations
- Clinical signs:
- effects observed, treatment-related
- Description (incidence and severity):
- Aggressive and restless behaviour in females during days 5 to 15 of treatment;
- Mortality:
- mortality observed, treatment-related
- Description (incidence):
- Aggressive and restless behaviour in females during days 5 to 15 of treatment;
- Body weight and weight changes:
- no effects observed
- Food consumption and compound intake (if feeding study):
- no effects observed
- Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- not examined
- Haematological findings:
- no effects observed
- Clinical biochemistry findings:
- effects observed, treatment-related
- Description (incidence and severity):
- serum potassium level of males of the highest dose group statistical significant increased
- Urinalysis findings:
- not examined
- Behaviour (functional findings):
- no effects observed
- Organ weight findings including organ / body weight ratios:
- effects observed, treatment-related
- Description (incidence and severity):
- , but no dose response, therefore regarded as not treatment related
- Gross pathological findings:
- no effects observed
- Histopathological findings: non-neoplastic:
- no effects observed
- Histopathological findings: neoplastic:
- not examined
- Details on results:
- CLINICAL SIGNS AND MORTALITY
- Females of the highest dose group showed aggressive and restless behaviour during days 5 to 15 of treatment.
BODY WEIGHT AND WEIGHT GAIN
- body weights and body weight gain unaffected by treatment up to and including 1000 mg/kg body weight/day.
- males of the 200 mg/kg bw group:
statistical significant decreased body weights on day 1 of the mating period, but not toxicological relevant due to lack of dose response relationship
- males of the 1000 mg/kg bw group:
on day 8 of the pre-mating period, a very slight statistical significant increased body weight gain, incidental and not toxicological relevant
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
unaffected by treatment up to and including 1000 mg/kg bw/day.
HAEMATOLOGY
- not affected by treatment
CLINICAL CHEMISTRY
- males of the 1000 mg/kg bw group:
statistical significant increse of serum potassium levels, but only slight and therefore of no toxicological significance
effects on glucose and inorganic phosphate without dose-response and therefore of no toxicological significance
NEUROBEHAVIOUR
- no changes were observed in hearing ability, pupillary reflex, static righting reflex and grip strength in the treated animals
- variation in motor activity did not indicate a relation with treatment
- females of the highest dose group showed a decreased motor activity at the low sensors; but absence of similar changes of the high sensors; therefore considered to be of no toxicological relevance.
ORGAN WEIGHTS
- no treatment-related changes
- males of the 200 mg/kg bw group: statistically significantly decreased absolute and relative adrenals weight
- females of the 40 mg/kg bw group: statistically significantly increased relative adrenals weight
both considered to be not related to treatment due to lack of dose response
GROSS PATHOLOGY
- No treatment related alterations
HISTOPATHOLOGY: NON-NEOPLASTIC
- No treatment related alterations
- The assessment of the integrity of the spermatogenetic cycle did not provide any evidence of impaired spermatogenesis.
Effect levels
- Dose descriptor:
- NOAEL
- Effect level:
- 200 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: absence of any adverse effect at this dose level
Target system / organ toxicity
- Critical effects observed:
- not specified
Any other information on results incl. tables
- Table 1: Animal body weight development
Males |
||||||
PRE-MATING |
|
GROUP 1 CONTROL |
GROUP 2 40 MG/KG |
GROUP 3 200 MG/KG |
GROUP 4 1000 MG/KG |
|
DAY 1 |
MEAN |
332 |
333 |
--- |
326 |
|
WEEK 1 |
ST.DEV. |
9.7 |
9.5 |
--- |
11.4 |
|
|
N |
10 |
10 |
0 x |
10 |
|
DAY 8 |
MEAN |
354 |
359 |
344 |
356 |
|
WEEK 2 |
ST.DEV. |
13.0 |
11.0 |
17.5 |
15.6 |
|
|
N |
10 |
10 |
10 |
10 |
|
MATING |
|
|
|
|
|
|
DAY 1 |
MEAN |
380 |
383 |
361 * |
381 |
|
|
ST.DEV. |
16.4 |
15.9 |
15.3 |
16.1 |
|
|
N |
10 |
10 |
10 |
10 |
|
DAY 8 |
MEAN |
--- |
407 |
--- |
380 |
|
|
ST.DEV. |
--- |
16.0 |
--- |
--- |
|
|
N |
0 |
3 |
0 |
1 |
|
POST MATING |
|
|
|
|
|
|
|
MEAN |
398 |
401 |
383 |
386 |
|
|
ST.DEV. |
16.5 |
12.2 |
18.9 |
27.3 |
|
|
N |
10 |
7 |
10 |
9 |
|
|
MEAN |
417 |
425 |
405 |
405 |
|
|
ST.DEV. |
19.3 |
9.4 |
22.4 |
25.1 |
|
|
N |
10 |
7 |
10 |
9 |
|
Females |
||||||
PRE-MATING |
|
GROUP 1 CONTROL |
GROUP 2 40 MG/KG |
GROUP 3 200 MG/KG |
GROUP 4 1000 MG/KG |
|
DAY 1 |
MEAN |
235 |
231 |
237 |
235 |
|
WEEK 1 |
ST.DEV. |
9.3 |
14.5 |
10.2 |
11.4 |
|
|
N |
10 |
10 |
10 |
10 |
|
DAY 8 |
MEAN |
242 |
245 |
239 |
239 |
|
WEEK 2 |
ST.DEV. |
9.0 |
10.5 |
9.8 |
11.4 |
|
|
N |
10 |
10 |
10 |
10 |
|
MATING |
|
|
|
|
|
|
DAY 1 |
MEAN |
253 |
255 |
252 |
254 |
|
|
ST.DEV. |
10.0 |
10.7 |
9.2 |
12.4 |
|
|
N |
10 |
10 |
10 |
10 |
|
DAY 8 |
MEAN |
--- |
292 |
--- |
290 |
|
|
ST.DEV. |
--- |
10.1 |
--- |
--- |
|
|
N |
0 |
3 |
0 |
1 |
|
POST COITUM |
|
|
|
|
|
|
DAY 0 |
MEAN |
252 |
264 |
250 |
251 |
|
|
ST.DEV. |
15.0 |
24.4 |
5.5 |
9.0 |
|
|
N |
8 |
9 |
8 |
9 |
|
DAY 7 |
MEAN |
284 |
295 |
280 |
284 |
|
|
ST.DEV. |
15.5 |
23.4 |
8.0 |
15.7 |
|
|
N |
8 |
9 |
8 |
9 |
|
DAY 14 |
MEAN |
318 |
334 |
324 |
317 |
|
|
ST.DEV. |
15.3 |
26.5 |
10.3 |
21.6 |
|
|
N |
8 |
9 |
8 |
9 |
|
DAY 21 |
MEAN |
409 |
440 |
429 |
404 |
|
|
ST.DEV. |
33.3 |
40.1 |
22.4 |
36.3 |
|
|
N |
8 |
9 |
8 |
9 |
|
LACTATION |
|
|
|
|
|
|
DAY 1 |
MEAN |
312 |
317 |
313 |
313 |
|
|
ST.DEV. |
22.4 |
29.0 |
11.6 |
23.0 |
|
|
N |
8 |
9 |
8 |
9 |
|
DAY 4 |
MEAN |
312 |
325 |
319 |
318 |
|
|
ST.DEV. |
10.8 |
30.1 |
8.8 |
22.5 |
|
|
N |
7 |
9 |
8 |
8 |
|
x: The body weight determination on day 1 of the pre-mating period of the males of the 200 mg/kg dose group was performed incorrectly.
* / ** : Dunnett-Test based on pooled variance significant at 5% (*) or 1% (**) levelApplicant's summary and conclusion
- Conclusions:
- RS-Pantolactone (named DL-LACTONE in the study report) was tested for it's repated dose toxicity in rats according to OECD TG 422.
In conclusion, gavage treatment of male and female Wistar rats with RS-Pantolactone at dose levels of 40, 200 or 1000 mg/kg body weight/day for at least 28 days (during premating ,mating, post-coitum, and lactation), revealed slight parental toxicity in animals receiving 1000 mg/kg b.w./day. - Executive summary:
RS-Pantolactone (named DL-LACTONE in the study report) was tested for it's repated dose toxicity in rats according to OECD TG 422. The test item was administered by daily oral gavage to male and female Wistar rats at dose levels of 40, 200 or 1000 mg/kg body weight/day. The males were exposed for 2 weeks prior to mating, during mating, and up to termination (28 days for all males). The females were exposed for 2 weeks prior to mating, during mating, during post-coitum, and at least 4 days of lactation. The mean duration of treatment of females was 43 days, with a minimum of 28 days and a maximum of 56 days.
Parental toxicity was assessed by observing mortality, clinical signs, body weights, food consumption, functional observations, clinical laboratory investigations, macroscopic examination, organ weights, and microscopic examination.
At 40 mg/kg b.w./day, no parental toxicity was observed.
At 200 mg/kg b.w./day, no parental toxicity was observed.
At 1000 mg/kg b.w./day, parental toxicity consisted of clinical symptoms (aggressive and restless behaviour) in females during days 5 to 15 of treatment, and increased serum potassium level in males.
In conclusion, gavage treatment of male and female Wistar rats with RS-Pantolactone at dose levels of 40, 200 or 1000 mg/kg body weight/day for at least 28 days (during premating ,mating, post-coitum, and lactation), revealed slight parental toxicity in animals receiving 1000 mg/kg b.w./day.
Based on the results in this combined repeated dose toxicity study with reproduction/developmental screening test, the definitive parental No Observed Adverse Effect Level (NOAEL) was established as being 200 mg/kg body weight/day.
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