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EC number: 200-001-8
CAS number: 50-00-0
Aqueous solutions of formaldehyde (ca 40%) induced skin corrosion in rabbits. Skin irritant effects are expected at concentrations > 3%.No studies are available on eye irritation, however, formaldehyde has skin corrosive properties (no testing required).Eye irritation in humans due to exposure to gaseous formaldehyde is the most sensitive endpoint. From studies in volunteers it is concluded that the NOAEL for objective eye irritation was 0.7 ppm in case of a constant exposure level and 0.4 ppm with peaks of 0.8 ppm. The odor threshold in humans is 0.145 ppm (50-percentile; WHO 1989) or 0.110 ppm (Berglund et al. 2012). When comparing thresholds for sensory irritation and odor it has to be taken into account that a differentiation between both on a subjective basis is hardly possible. Olfaction confounds sensory irritation. It is well known that there are differences in susceptibilities for odor and sensory irritation to eyes and nose. For example, Cometto-Muniz & Cain (1990, IUCLID Section 7.12) could demonstrate for 11 chemicals that the threshold for odor was clearly lower than the threshold for irritation. The odor threshold was determined in normosmic and the irritation threshold in anosmic subjects. Similarly, eye irritation thresholds were well above odor thresholds (Cometto-Muniz & Cain, 1995, IUCLID Section 7.12). A similar conclusion can be derived for formaldehyde (Cain et al., 1986): 33 persons were exposed for two 30 min sessions, interrupted by an exposure free interval of 15 min, to 0, 0.27 0.46, 1.1, and 2.05 ppm. Sensory irritation to eyes, nose, throat and odor perception were investigated by questionnaire. Subjective scaling of odor perception was stronger than perceived irritation at all concentrations. However, exposure to room air led to a higher odor perception as compared to perceived irritation. In a review by Greenberget al. (2013) it was concluded that perception of odor cannot be used as a surrogate marker for chemical exposure.
Skin irritation in 2 rabbits
Exposure duration 20 h
24 h after initiation
Superficial Ne & B
48 h after initiation
72 h after initiation
Ne (leather like)
Ne (black, hardened)
Ne: necrosis; B: bleeding
Comparable to guideline study (BASF Test) with acceptable restrictions
(exposure period 20 h; occlusive dressing; application of a patch soaked
with ca. 1 mL formalin; partly limited documentation, no details about
the test substance).
Two White Vienna rabbits were exposed for 20 h under occlusive dressing
to a patch (2.5 x 2.5 cm²) soaked with 40% formalin. Readings were
performed 24, 48, 72 h and 6 and 8 days after application. Beside edema
and erythema superficial necrosis was obvious 24 h after initiation of
exposure. In both rabbits full thickness necrosis were found the
Conclusion: A solution of 40% formalin induced full thickness necrosis
in 2 rabbits after 20 h occlusive exposure.
In a study comparable to OECD Guideline 404 (BASF AG, 1973 & 1974)
with acceptable restrictions (exposure period 20 h; occlusive dressing;
application of a patch soaked with ca. 1 mL formalin; partly limited
documentation, no details about the test substance) two White Vienna
rabbits were exposed for 20 h under occlusive dressing to a patch (2.5 x
2.5 cm²) soaked with 40% formalin.40% formalin, no further details
Readings were performed 24, 48, 72 h and 6 and 8 days after application.
Besides edema and erythema, superficial necrosis was obvious 24 h after
initiation of exposure. In both rabbits full thickness necrosis were
found the following days.
In supporting studies on rats (Sekizawa et al., 1994) authors
reported “erosion” of the skin at 7-9% aqueous formaldehyde solution; no
effects were seen at this concentration in mice and guinea pigs using a
similar experimental design. In rats, however, no skin effects were
reported at a concentration of 3% (Sekizawa et al., 1994).
The results from studies on sensitization give information on max.
non-irritant concentrations of aqueous solutions in the GPMT. A 2%
solution was non-irritant in 4 independent studies (challenge
concentration in non-induced controls) and concentration used for
topical induction (minimal irritant concentration) was 5 or 10% solution
(see Section Skin sensitization; Bayer AG, 1987; Kimber et al., 1991;
Hoechst AG, 1983; Hilton et al., 1996). In the study of Hoechst AG
(1983) even a concentration of 4% resulted in no irritation in 10
controls (24 h, occlusive; 1st reading). Sensitization studies in humans
(see WHO, 1989 in Section Sensitization data (humans)) suggested a
non-irritant concentration at 1%; in diagnostic patch testing for
sensitization concentrations of formaldehyde not leading to irritation
have to be used (i.e. many authors proposed 1%).
A literature search after the last IUCLID update was carried
out up to April 20, 2015 and provided the following new information:
Saito et al. (2011, supporting) investigated the reaction of the
skin of mice after application of 2, 5, and 10% formaldehyde solutions
in acetone, once per week over 5 weeks. 25 µl were painted to the dorsal
and ventral surface of the ear lobes. Skin thickness was significantly
increased by 10% and 5% and numerically by 2%. Histopathological
alterations were observed, like infiltration of inflammatory cells or
skin hypertrophy, by the 5 and 10% solutions. The effect was
significantly suppressed by capsazeine, a TRPV-1 antagonist, showing
that the TRPV-1 nociceptive receptor is involved in the skin response.
The involvement of the TRPV-1 receptor was further substantiated by
investigating TRPV-1 gene-knockout mice which showed an attenuated
response in the ear swelling test as compared to wild type animals
(Usuda et al., 2012, supporting). And finally, also Tian et al. (2009,
supporting) showed that this nociceptive receptor is involved in vivo in
formaldehyde induced pain after injection of a 4% formaldehyde solution
into the plantar surface of rat hind paws, as well as in vitro by
electrophysiological measurement of the response in dorsal root ganglion
cells. The in vivo and in vitro effects, both were attenuated by
In summary, irritant effects are expected at concentrations > 3%.
These effects are mediated by the TRPV-1 nocireceptor in the skin.
In support of this recommendation, a recent study on microvascular
leakage of rat skin after dermal application for 40 minutes showed that
skin damage was demonstrated at concentrations >= 2.5% formaldehyde
(increases in the amount of Evans blue dye extravagated into the skin),
the NOAEC was 1% (Futamura et al., 2009).
No data are available according to current guidelines. Studies on
skin irritation/corrosion brought evidence for corrosive properties (no
testing on eye irritation required). Data presented by Carpenter & Smith
(1946, limited validity) suggested corrosive properties of formaldehyde
solutions (5µL of 15% solution) after exposure to the rabbit eye.
Additionally, in the study of Sekizawa et al. (1994), only 0.01 mL of
7-9% formaldehyde solution in water resulted in opaque cornea (other
parameters of eye irritation were not scored).
Therefore, corrosive properties of formaldehyde solution in the
eye at concentration in the range of 7% are to be expected.
Lai et al. (2013, supporting) studies the effect of aqueous
formaldehyde solutions on rabbit corneal cells in vivo and in vitro in a
non-guideline study. In vivo, the eyes were exposed to small filter
discs soaked with 0, 20, 100, 200, or 300 ppm of formaldehyde in water
for 5 min. 1, 3, 7 and 10 days after exposure filter strips were
introduced into the rabbit’s eye to measure moisture according to the
Schirmer’s test. In vitro rabbit corneal cell preparations were exposed
to aqueous solutions containing 5-600 ppm formaldehyde over 3-5 min and
cell morphology, death and proliferation were determined as well as
changes of mitochondria. The authors concluded that in vitro already 5
ppm induced damage to the corneal cells that became apparent only after
prolonged observation period. Higher concentrations led to effects
already after shorter periods. In vivo, tear production was increased at
all exposure concentrations and observation times.
Irritation in humans exposed to gaseous formaldehyde
The exposure to gaseous formaldehyde resulted in irritation of
eyes, nose and throat. There is evidence that eye irritation in humans
is the most sensitive endpoint. Slight discomfort due to irritation
(mainly eyes) was noted in some individuals even at a concentration of
0.3 mg/m³ (0.25 ppm; variation at all dose levels within +-20% target
concentration). Although subjects were asked for rating of discomfort
due to the degree of airway irritation in the results section only
ratings for discomfort are presented without differentiation for
irritation to the eyes or respiratory tract. Even at the highest
concentration tested (2 mg/m³) the average discomfort never exceeded the
middle of the “slight discomfort” range (Andersen & Molhave, 1983). A
main drawback of this study is the lack of a sham exposure group.
In another study with 10 volunteers no eye irritation occurred at
0.5 ppm (analytical and target concentration very close together) but
odor was detected; dose dependent eye irritation was reported at ≥ 1 ppm
(Kulle, 1993, exposure period 3 h,).
There are numerous other studies available with similar findings.
A detailed documentation and evaluation of ocular and respiratory
irritation in humans are presented in reviews by Paustenbach et al.
(1997) and Arts et al. (2006). A summary of selected studies on this
endpoint is tabulated in Section Exposure related observations in humans
- Direct observations, (Summary chemosensory irritation).
Both reviews (Paustenbach et al., 1997; Arts et al., 2006)
primarily relied on human volunteer studies, since according to the
authors investigations on subjective irritant effects at the workplace
may be compromised by two major problems:
- Fluctuations of the exposure concentrations with higher peak
exposures that may lead to an important recall bias for subjective
reporting of irritant effects;
- Presence of contaminants at the workplace other than
The analysis by Paustenbach et al.(1997) was based on the
construction of a concentration effect curve for eye irritation. In the
review of Arts et al. (2006) a benchmark analysis was carried out in
addition for the studies of Andersen and Mölhave (1983) and Kulle
(Kulle, 1993; Kulle et al., 1987) taking into account the possible
non-zero response at 0 ppm exposures.
Even when concentrating only on controlled volunteer studies both
review groups nevertheless noted for all studies one or more of the
– reliance only on subjective scorings without objective
measurements for irritation (apart from Weber-Tschopp, 1977)
– such subjective reporting do not reliably differentiate between
irritation and odor perception.
– often zero exposure groups are missing to allow for an estimate
of the background of subjective reporting for irritancy (up to 15-20
– missing criteria for severity of effects to differentiate
between perception / annoyance / adversity
Both of these reviews basically came to the same conclusions:
– irritation to eyes is more sensitive than that to nose and throat
– concurrent control groups show that 15-20% of the
exposed volunteers will report slight subjective eye irritation even at
zero exposure (sham exposure data measured in 7 controlled studies,
Paustenbach et al. (1997)
– subjective eye irritation at low exposure levels rapidly
subsides after termination of exposure
– appreciable / moderate / annoying eye irritation will start at
or above 1 ppm
– between 0 and 0.3 ppm there is no increase in eye irritation
above a background level of 10- 20% of volunteers reporting effects at
– reports of irritation below 0.3-0.5 ppm are too unreliable to
attribute the irritation solely to formaldehyde
– finally, Arts et al. (2006) concluded that sensory irritation in
humans can be strongly influenced by subjective feelings and odor may
confound reporting of irritation.(Dalton et al. cited in Arts et al.,
An important shortcoming of all studies (but one) reviewed by
Paustenbach et al. (1997) and Arts et al. (2006) was that only
subjective reporting for irritation were given. The only “old” study
investigating objective eye irritation parameters found an increase in
eye-blinking frequency at a high concentration of 1.7 ppm (Weber-Tschopp
et al., 1977 as referenced by Paustenbach et al., 1997).
If only subjective ratings on irritation are available a careful
interpretation is necessary to avoid misinterpretation due to bias or
confounding factors. There are several important factors that need to be
considered before subjective findings can be interpreted as true adverse
health effects (Dalton, 2002; cited in Arts et al., 2006):
- exposure history: workers in comparison to naïve volunteers;
- expectations and beliefs: prior information obtained for the
- bias from odor perception: problems in differentiation between
irritation and odor;
- social factors: influence by behaviour of bystanders or study
- personality variables: influence on expectation by positive or
In the case of only subjective symptoms being reported clear to
moderate subjective eye irritation should be selected as POD rather than
very slight to slight irritation as this level of response is often
already is reported at near zero exposure (Paustenbachet al, 1997;
Artset al, 2006b).
The deficiencies of the former studies on local sensory irritation
were overcome by a recent investigation of Lang et al. (2007). In
contrast to many of the former investigations this study included
objective measures for sensory irritancy besides subjective ratings,
peak exposures, masking of olfactory effects by a strong odorant (ethyl
acetate) and physical activity. Further, the evaluation of subjective
symptoms took into account positive / negative affectivity traits of the
participants as determined by the PANAS (Positive And Negative Affect
Schedule) questionnaire. For example a negative affectivity trait is
indicated if a subject claims to be “distressed” or “irritable”, while
the positive affectivity counterpart would be “active” or “proud”.
In short, the following results were obtained:
- Blinking frequency and conjunctival redness as objective
symptoms for eye irritation, ranging from slight to moderate, were
significantly increased by short-term peak exposures of 1.0 ppm on a
baseline exposure of 0.5 ppm. The NOEL for these objective parameters
was 0.3 ppm with 15 min peaks of 0.6 ppm formaldehyde or a constant
exposure to 0.5 ppm.
- Subjective ratings of eye and respiratory irritation and
olfactory symptoms occurred already at concentrations of 0.3 ppm, but
not at 0.15 ppm; however, exposure to ethyl acetate only was also
(wrongly) perceived as irritating.
-Volunteers who rated their personality as “anxious” by PANAS
tended to report more pronounced complaints. When “negative affectivity”
was used as covariate, the level of 0.3 ppm was no longer an effect
level, but 0.5 ppm with peaks of 1 ppm was. This corresponds to the
effect level obtained by objective parameters.
- Whenever increased symptom reporting occurred, these scores were
reversed 16 h after the end of exposure.
These results clearly show that subjective reporting on irritation
is strongly influenced by
- the interaction with odor perception as demonstrated by
co-exposure to ethyl acetate and
- personal affectivity traits as demonstrated by using PANAS as
Therefore, taking into account the variety of possible confounders
on subjective symptoms preferential it is suggested that objective
findings like eye blinking frequency or conjunctival redness should be
used as reliable indicators for irritation to the eyes.
In conclusion, this study provides two well-founded NOELs for eye
irritation being the most sensitive effect parameter for formaldehyde,
namely 0.3 ppm plus 15 min. peaks of 0.6 ppm or 0.5 ppm continuously.
Recently a new study on chemosensory effects in human volunteers
became available from the same institute (Müller et al., 2013). In
comparison to Lang et al. (2007) the new investigation included the
following new features:
- increase of the number of participants from 21 (Lang et al.,
2007) to 41.
- Refinement of the NOAEC/LOAEC by using the following exposure
concentrations: 0 ppm; 0.3 ppm + peaks of 0.6 ppm; 0.4 ppm + peaks of
0.8 ppm; 0.5 ppm; 0.7 ppm each over 4 h.
- An important additional aspect was allocation of the
participants according to their sensitivity for trigeminal nasal
irritation (CO2threshold) into subgroups of hyper- and hyposensitives
(20 persons each) and extremely hyper- and hyposensitives (10 persons
Eye blinking frequency and subjective ratings of symptoms and
complaints (SPES) were determined before and within the last 15 min of
exposure, and the following parameters were examined before and shortly
- conjunctival redness
- tearfilm break-up time
- nasal resisstance and flow
- olfactory functions (n-butanol threshold, sniffing sticks for
odor discrimination and identification).
Similar to the study of Lang et al. (2007) personal affectivity
traits were taken into account by PANAS.
None of the objective parameters showed an effect indicative for
sensory irritation up to the highest exposure concentrations of 0.7 ppm
or 0.4 ppm + peaks of 0.8 ppm:
- conjunctival redness. In comparison, Lang et al. (2007) found an
increased conjunctival redness during peak exposure to 1.0 ppm. The
findings of both studies together indicate that the NOAEL is in the
range between 0.7 – 1.0 ppm and any effect caused by peak exposures
around 1 ppm will rapidly subside.
- Eye blinking frequency even tended to decrease in the total
group as well as in the different sensitivity subgroups. This leads to
the same conclusion as for conjunctival redness when comparing the
present data to those of Lang et al., (2007).
- For nasal flow and resistance no exposure related effect was
- Tearfilm break-up time rather showed a trend for an increase but
not for a decrease as would have to be expected for substances leading
to eye irritation.
- No exposure related effect was observed for olfactory function
(n-butanol threshold, odor discrimination or identification.
For subjective symptoms (subdivided into the subgroups of“eye
irritation”, “nasal irritation”, “olfactory symptoms” and “perception of
impure air”) the following results were obtained:
- there were no statistically significant changes of subjective
symptoms or dose related trends for the categories “eye irritation” and
“nasal irritation” in all subgroups of participants.
- for “olfactory symptoms” there was a statistically significant
increase for the total group at >= 0.3 ppm with 0.6 ppm peaks, when
compared with the pre-exposure examination, and at >=0.4 ppm with 0.8
ppm peaks, when compared to 0 ppm exposure. However, there was no clear
dose-response relationship. Generally, hypersensitive volunteers
reported more often, partly significantly, “olfactory symptoms” than
hyposensitive subjects, especially in the extreme subgroups.
- there was a statistically significant increase for the SPES item
“perception of impure air” in the total group and in “hypersensitive”
and “extremely hypersensitive” volunteers compared to “hyposensitive”
participants. However, there was no significant change of “perception of
impure air” in comparison with the control (zero concentration), since
volunteers exposed to zero concentration did also report an increase of
their “perception of impure air”.
The findings for the symptom subgroups “olfactory symptoms” and
“perception of impure air” cannot be taken as adverse effects in the
sense of subjective sensory irritation, but they are rather indicative
for annoyance. Furthermore, they point to the difficulty to clearly
differentiate between olfaction and sensory irritation.
The findings for eye and nasal irritation are in contrast to
results of Lang et al. (2007). By statistical analysis taking into
account negative personality traits obtained by PANAS questionnaire,
Lang et al. (2007) gave strong evidence that these effects were related
to personality factors but were not due to clear subjective sensory
irritation. This is strongly supported by Müller et al. (2010): they
compared the PANAS scores of both studies and found that the
participants of the Lang study had a statistically significantly higher
negative affectivity than those of the Müller study. Thus the subjective
scoring for sensory irritation was driven by personality traits in the
In conclusion, the study of Meuller et al. (2013) showed that
1. formaldehyde concentrations of 0.7 ppm for 4 hours and peak
concentrations of 0.8 ppm formaldehyde for 15 minutes superimposed to
0.4 ppm did not cause adverse chemosensoric effects, neither for
objective parameters of irritation nor for subjective complaints for
2. there were no significant differences between hypo- and
3. the NOAEL was 0.7 ppm without peak exposure and 0.4 ppm with
0.8 ppm peaks
4. there was only an increase of subjective olfactory symptoms
starting at 0.3 ppm with peaks of 0.6 ppm, which is not interpreted as
IARC (2006) presented a summary on toxic effects in animals (see
Section Additional toxicological information) as well as a summary on
toxic effects in humans including data on local irritation.
Mueller et al. (2013, key) further refined the results obtained by
Lang et al. (2008). 41 male volunteers (non-smokers, age ±9.9 years)
were exposed in a randomised schedule to 0, 0.5, 0.7 ppm and to 0.3 ppm
with 4 15 min peaks of 0.6 ppm and to 0.4 ppm with peaks of 0.8 ppm.
During exposure 4 cycle ergometer units at 80 W were performed for 15
min. Subjective pain perception induced by nasal application of carbon
dioxide served as indicator for sensitivity to sensory nasal irritation
to define subjects hyper- and hyposensitive for irritation. The
following parameters were examined before and after exposure: subjective
rating of symptoms and complaints (Swedish Performance Evaluation
System), conjunctival redness, eye-blinking frequency, self-reported
tear film break-up time and nasal flow rates. In addition, the influence
of personality factors on the volunteer’s subjective scoring was
examined (Positive And Negative Affect Schedule; PANAS). No indications
for subjective or objective indications of sensory irritation were
obtained under these exposure conditions. There was a statistically
significant differences for olfactory symptoms, especially for the
‘perception of impure air’ when comparing the subjective symptoms under
formaldehyde exposure with zero exposure. These subjective complaints
were more pronounced in hypersensitive subjects. When comparing the
studies of Lang and Mueller, Lang et al. (2008) observed subjective
symptoms of eye irritation already at 0.3 ppm while these effects were
not found by Mueller et al. (2013) even at higher exposures. This is
explained by Mueller et al. (2013) by the fact, that negative
affectivity was significantly higher in the subjects exposed in the Lang
study as compared to those of Mueller. The increased ‘perception of
impure air’ was attributed by the authors impairment of well-being
caused by situational and climatical conditions in the exposure chamber,
because a statistically significant difference in symptom scores between
FA exposures and control condition was missing, and hypersensitive
subjects reported statistically significantly higher complaints even
after exposure to 0 ppm. The NOAEC for sensory irritation was 0.7 ppm
over 4 h and 0.4 ppm with peaks of 0.8 ppm.
The importance to clearly differentiate between sensory irritation
and olfaction was demonstrated by Berglund et al. (2012, key). 31
subjects (18–35 years old) were exposed to formaldehyde at
concentrations varying between 6.36 and 1000 ppm corresponding to the
Swedish TLV. Exposure was carried out in a hood exposure system and the
volunteers took one sniff of the atmosphere over 3 sec (3
sniffs/min).P50absolute thresholds (50% correct detection of odor) were
for formaldehyde odor 110 ppb (range 23–505). For sensory irritation the
P50could not be calculated because too few subjects were studied and the
exposure was limited to 1000 ppb. But all thresholds for irritation were
higher than for odor.
In a comprehensive review about factors that may influence
olfaction Greenberg et al. (2013, key) concluded that perception of odor
cannot be used as a surrogate marker for chemical exposure. Odor
perception is affected by the psychological state and bias because odor
is often negatives biased by association with health-related symptom.
Some further studies in humans on self-reported symptoms during
prolonged work with formaldehyde are mentioned here, although not
related to single exposures. As far as subjective symptoms were recorded
these studies are less reliable than those in volunteers under
controlled exposure conditions. The major problems stem from factors
like exposure to mixtures of unknown composition, peak exposures that
were not controlled for, or recall bias to former exposures. The main
subjective symptoms were reported by students of a gross anatomy
dissection course using a more detailed questionnaire (Mori et al.,
2013, supportive). The symptoms were reversible 6 months after the
course. Apart from subjective reportings, again the problem is exposure
assessment. Exposure was reported to be about 0.2 ppm, but this was
measured by area and not by personal sampling for 20 min after start of
the course. Thereby personal peak exposures could not be accounted for.
Some studies reported objective parameters associated with
formaldehyde exposure. Neghab et al. (2011, supporting) carried out a
cross sectional study with 70 workers of a melamine-formaldehyde
producing factory including 24 non-exposed referents. In total 7 air
samples were taken over 40 min. The frequency of some of the
self-reported respiratory symptoms (e.g. cough, chest tightness,
episodes of chest illness associated with cold) was significantly higher
in the exposed population and pre- and post-shift parameters of
pulmonary function showed significant decrements. A recovery of lung
function capacity was observed following temporary cessation of
exposure. Airborne formaldehyde exposures clearly exceeded current
exposure limit values with 0.78 ppm (SD=0.4). Hisamitsu et al. (2011,
supportive) investigated serum IgE levels, olfactory tests and nasal
sensitivity to histamine in 41 medical students before, during and after
an anatomy dissection course. Olfactory anomalities and increased
histamine sensitivity were observe red during and immediately after the
course but the effects were reversible and disappeared after completion
of the course. Formaldehyde concentrations ranged from 0.51-0.97 ppm
(mean 0.67) in the centre of the laboratory.
Wieslander and Norbaeck (2010, supporting) studied symptoms of
upper respiratory tract irritation in 31 indoor painters. Exposure
concentrations over 8 h were measured for formaldehyde, other volatile
organic chemicals and microbial organic chemicals. In comparison to
controls, painters had increased ocular symptoms, lysozyme and
myeloperoxidase in nasal lavage, while tear film break-up time and nasal
patency (by acoustic rhinometry) were reduced. As the daily formaldehyde
levels were below detection limits (30 µg/m³) and other volatile organic
chemicals prevailed, the reported symptoms are not caused by
According to EU Classification,
Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC)
No. 1272/2008, the substance has to be classified as Skin Corr 1B, H314
and Serious eye damage 1, H318.
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.
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