Bitumen (Vapour and Aerosol)

Bitumen (vapour and aerosol)
MAK value –
Peak limitation –
Absorption through the skin (2001) H
Sensitization –
Carcinogenicity (2001) Category 2
Prenatal toxicity –
Germ cell mutagenicity –
Synonyms –
Chemical name (CAS) –
CAS number 8052-42-4
Structural formula –
Molecular weight –
Melting point –
Boiling point –
Density –
Vapour pressure –
log Pow* –
This documentation is based on review articles by the ACGIH (2000) and NIOSH
(2000).
* n-octanol/water distribution coefficient

38 Bitumen (vapour and aerosol) Volume 17
1 Composition and Analysis
1.1 Definition and terms
According to DIN 55946 Part 1, bitumen is defined as follows: a product obtained during
the processing of certain types of petroleum, bitumen is a poorly volatile, dark-coloured
mixture of various organic substances whose elasto-viscous properties change with the
temperature (Deutsches Institut für Normung 1983). Bitumen exists as a colloidal system
in which the asphaltenes of the disperse phase are suspended in a coherent phase of high
boiling point maltenes. Natural asphalt formed geologically from petroleum is also a type
of bitumen. Distilled, oxidized and precipitated bitumens are distinguished according to
the production process; the main difference between these products is in their viscosity.
Asphalt, on the other hand, is a naturally occurring or industrially prepared mixture of
bitumen or bituminous binder with mineral substances and perhaps other aggregates or
additives. DIN 55946 differentiates between bitumen and tar. In modern English, the
terms “bitumen” and “asphalt” are not distinguished clearly. In Great Britain and contin-
ental Europe the term bitumen is employed only with reference to the black or brown
petroleum-like substances that are called asphalts in the United States. Bitumen is
however always understood to mean the material used as a binder, bitumen. The term
“cutback” is used for mixtures containing bitumen, which are not identical with the
undiluted material.
In 1977 bitumen was classified as a suspected carcinogen. The TRK (technical expo-
sure limit) for bitumen is currently 10 mg/m3 (AGS 2000).
Most bitumen is processed hot (see Table 1, Appendix p. 72). The determination of
the concentrations of the vapour and aerosols produced in such processes and avoidance
of exposure is the main focus of the primary measures for industrial health and safety. At
present the proportions of dust, fume and mist in such aerosol phases cannot be deter-
mined. Therefore, in the text which follows, all the states of aggregation of bitumen
which occur in the workplace air are subsumed under the term “bitumen fumes”.
1.2 Analysis of PAH in bitumen and its vapour and aerosols
The determination of bitumen fumes and of the levels of polycyclic aromatic hydro-
carbons (PAH) in vapours and aerosols emitted during hot processing of bitumen makes
use of a combined air sampling system with fibre glass filters fitted in front of an adsorp-
tion tube filled with about 2 g of the adsorbent XAD-2 for both personal and static sam-
pling. First the aerosol fraction deposited on the fibre glass filter is weighed. Then the
PAH content is determined by mass spectroscopy. To determine the level of the bitumen
fumes adsorbed as vapour on the XAD-2, the adsorbent is eluted with tetrachloroethylene
and assayed by infra red spectroscopy; calibration curves are prepared with bitumen
condensate. This method is based on that used by the Berufsgenossenschaftlichen Institut
für Arbeitssicherheit (BIA, the trade association’s institute for health and safety at work)
Volume 17 Bitumen (vapour and aerosol) 39
(BIA 1989). The levels of PAH are determined by mass spectroscopy from a sample col-
lected in parallel in a second XAD-2 adsorption tube.
When possible, sampling should take place during the whole work shift. Experience
has shown that it makes sense to collect not just air samples but also samples of the
material being processed and to assay this for PAH as well. Detailed methods for sam-
pling and analysis of such material have been published (HVBG 1997, Josefsson 1983,
Knecht et al. 1987). Studies of the levels of absorption of bitumen aerosol and vapour
through the skin under conditions like those at the workplace have revealed high levels of
skin penetration (Knecht et al. 2001).
1.3 Bitumen vapour and aerosol levels during hot processing
Table 2 (p. 72) lists the currently available analytical data for bitumen fumes in various
work processes. Table 3 (p. 73) lists the concentrations of PAH in various kinds of com-
mercial bitumen in current use (Knecht et al. 1999). Analytical data for the type of
bitumen most used at present (B 65) are available specifically for the production of
asphalt mixtures and for transport of rolled asphalt B 65. The results obtained for the
PAH levels in the aerosol and vapour phase are shown in Tables 4 and 5 (p. 74). More
information and more analytical results for the levels of PAH in bitumen vapour and
aerosols may be found in the literature (Bergmann et al. 1989, Burstyn et al. 2000, Kitto
et al. 1997, Schmidt 1992, Watts et al. 1998, Wichert 1993, Wolff et al. 1989).
2 Toxic Effects and Mode of Action
Because the composition of bitumen vapour and aerosols depends on the nature of the
material in use and the temperature, the biological effects also vary. Irritation can
develop in the respiratory tract after inhalation and also on the skin after skin contact.
The effective concentrations differ widely depending on the experimental design and the
workplace conditions. To date no specific inflammatory reactions have been detected in
the respiratory tract. Epidermal application of condensed bitumen vapour, however, has
been shown to result in keratosis and hyperplasia.
The most important aspect is, however, the carcinogenicity and genotoxicity of the
components of bitumen vapour and aerosols. Epidemiological studies have yielded evi-
dence of an increased lung cancer risk. Individual studies have also revealed increases in
the risks of developing cancer of the stomach, skin and bladder, and leukaemia. In animal
studies, various carcinogenic effects have been detected. Under certain test conditions,
bitumen extracts and condensate of bitumen fumes are genotoxic in the Ames test and in
in vitro tests with mammalian cells. DNA adducts which are ascribed mainly to PAH of
the kind detected in bitumen extracts have been found in exposed workers. Similar
results have been obtained in animal studies with bitumen extracts and condensates.
However, more recent studies have shown that not only PAH-induced DNA adducts are
present but also adducts of substances which have not yet been identified. Further
evidence of PAH exposure is provided by the detection of hydroxypyrene in the urine of
exposed workers. Application of bitumen extracts and bitumen vapour condensates to the
skin of experimental animals results in the development of skin papillomas and car-
cinomas. The few studies available to date suggest that both the PAH present in the bitu-
men and also other substances may be responsible for the carcinogenic effects. That the
substances are available systemically after epidermal application is indicated by the
formation of DNA adducts in the lungs and peripheral lymphocytes.
There are no data available for the mechanism of action of bitumen vapour and aerosols.
3 Toxicokinetics and Metabolism
A Spanish research group (Lafuente and Mallol 1987) studied the excretion of thioethers
in the urine as an indicator of exposure to electrophilic and thus potentially mutagenic
substances. They reported an initial increase in the thioether level in the urine of both
smokers and non-smokers working in road building or in bitumen production. The
increased values, however, decreased during the course of the working week, especially
in heavy smokers, although the level of bitumen exposure was unchanged. Reduced glu-
tathione concentrations or glutathione transferase activity at the beginning of the expo-
sure phase was suggested as an explanation for this phenomenon.
In one Turkish study (Burgaz et al. 1988) and one Italian study (Pasquini et al.
1989a), thioether excretion by road builders was increased significantly only in smokers.
Another study by Burgaz together with researchers from Dutch universities revealed
increased thioether and 1-hydroxypyrene concentrations in the urine of road builders
exposed to “class 1 steam refined bitumens”, bitumens with “75–100 penetration values”
and small amounts of “class 3 cutback bitumens” (Burgaz et al. 1992a).
A German pilot study (Triebig et al. 1986) yielded no evidence of increased thioether
excretion by persons working in road building or in an asphalt mixing plant.
A British-Australian research group did not find any significant differences in the
levels of thioethers and glucaric acid in the urine of road builders and roof construction
workers at the beginning and end of the work shift. The levels determined were also not
significantly different from those obtained for a collective of craftsmen with a lower
average level of exposure to PAH (Hatjian et al. 1995).
Because of the different exposure situations in the different countries and because of a
series of conceivable confounders (e.g. exposure to tar, diesel motor emissions), it is not
possible to evaluate these results conclusively.
The excretion of 1-hydroxypyrene in the urine of persons working with bitumen
products has been studied by several research groups. The level of 1-hydroxypyrene in
the urine of persons exposed to bitumen vapour and aerosols at work tended to be higher
than that in the urine of not exposed control groups or higher than the levels found for the
persons before the beginning of exposure, but the differences were not always significant
(Boogaard and van Sittert 1994, 1995, Burgaz et al. 1992a, 1992b, Hatjian et al. 1995,
1997, Jongeneelen et al. 1988, Levin et al. 1995, Levin and Järvholm 1999, Zhou 1997).
4 Effects in Man
There are no data available for the effects in man of single exposures to bitumen vapour
and aerosols nor for the reproductive or developmental toxicity of the material.
4.1 Repeated exposures
Numerous scientific publications report effects of bitumen vapour and aerosols on the
human airways and kidney. The published results are, however, inconsistent.
Tiredness, reduced appetite, irritation in the larynx and pharynx, coughing and eye
irritation were observed in road construction workers. The differences from a control
group could not be accounted for by effects of smoking habits, number of hours worked
in the previous week or habituation (Norseth et al. 1991).
Overheating of fluorescent lamps in an office complex resulted in the release of
vapour from filling material containing bitumen. Persons working in the immediate area
suffered especially from headaches, irritation of the eyes and throat, and blocked nose.
The symptoms disappeared when the rest of the melted bitumen was removed (Tavris et
al. 1984). Similar symptoms (irritation of the eyes and upper airways, nausea) were
described by persons working in an office block when the bituminous material on the
roof was renewed. Symptoms which persisted in some of the affected persons even after
work on the roof was finished included airway irritation, coughing and sinus infections
(Lynch and Kipen 1998).
22 Italian cable insulaters who worked with hot bitumen complained of irritation in
the nose and throat and of coughing, sputum and hoarseness. Clinical examination
revealed acute rhinitis and bronchitis in most of these persons (Zeglio 1950).
A study of persons who had worked for at least 5 years in any of 25 US American
asphalt refineries revealed an increase in the incidence of respiratory diseases. Anam-
nestic records revealed 31 persons with non-neoplastic lung disorders among 360 work-
ers (control group: 12/277) whereas at the time of the study 40 of 462 asphalt workers
were shown to have such disorders (control group: 24/379). The large majority of the
persons suffered from chronic bronchitis; there were occasional cases of asthma or
emphysema (Baylor and Weaver 1968). The study is poorly documented.
In an equally poorly documented study from the Federal Republic of Germany, it is
reported that neither the incidence of changes in lung function nor the incidence of bron-
chial or pulmonary disorders was increased in 34 municipal road construction workers
relative to the values found for a control group of graveyard workers (Bittighofer et al.
1983).
50 Israeli workers exposed in bitumen production or during use of bitumen for road or
roof construction suffered from irritation of the respiratory passages which was mani-
fested in a feeling of dryness and in coughing. In most cases the persons developed
symptoms of chronic bronchitis. In a control group of 15 factory metal-workers, such
symptoms were not found. Lung function tests, however, revealed no pathological find-
ings either in the persons exposed to bitumen vapour or in the control persons (Schaffer
et al. 1985).
Six roofing workers from the former German Democratic Republic who had special-
ized for more than 20 years in the production of insulated roofs and the insulation of wet
rooms complained of respiratory difficulties. In 4 of the patients, chronic-obstructive
bronchitis was diagnosed and recognized as an occupational disease. The toxic sub-
stances considered to have caused the damage were the pyrolysis products released on
heating “bituminous masses”. All the affected persons had smoked for many years. A
non-smoking colleague was found to have simple chronic bronchitis (Maintz et al. 1987).
The spirometric values determined for 231 Swedish asphalt workers (road and roof
construction) were not significantly different from those for a control group matched for
smoking habits and other factors. The incidence of complaints of bronchitis symptoms
increased, however, with increasing period of exposure (Nyqvist 1978).
170 persons in the USA who were exposed to bitumen vapour and aerosols during
production and loading of bitumen, and road and roof construction work were subjected
to lung function tests before and after the work shift and examined for disorders of the
respiratory system. The exposure situation was described as follows: “total particulate”:
< 1.5–6.2 mg/m3, “respirable particulate”: < 0.6–1.4 mg/m3, “benzene-soluble fraction of
total particulate”: < 0.6–1.3 mg/m3, “volatile hydrocarbons collected in a charcoal tube”:
< 8–19.8 mg/m3. No consistent association could be established between acute impair-
ment of lung function or respiratory symptoms and exposure to bitumen vapour; nor
could associations with exposure to ozone, heat stress, cigarette consumption or shift
length be demonstrated (Gamble et al. 1999).
The causes of death of 4849 persons who carried out maintenance work on the high-
ways in Minnesota were investigated in a cohort study. The collective was subdivided
into town and country cohorts and the results compared with those for the town and
country populations of the state. The standard mortality ratio (SMR) for chronic kidney
failure was significantly increased (p < 0.05) in long-term workers in country regions.
The authors, however, considered an association with workplace exposure to be ques-
tionable, especially as only 3 deaths were involved (Parker et al. 1989).
An Australian cross-sectional study published only as a summary in a letter to the
editor found an increased incidence of kidney disorders in road construction workers
exposed to bitumen. The control groups comprised office and quarry workers. The study
was carried out as the result of the diagnosis of glomerulonephritis in a 36-year-old road
construction worker (Douglas and Carney 1998). Another Australian cross-sectional
study which compared 41 road construction workers (“hydrocarbon-exposed”) with 35
quarry workers (“non-exposed”) found no significant differences between the collectives
with respect to the parameters of renal function which were studied (Carney and
Ferguson 1998).
4.2 Local effects on skin and mucous membranes
Hot bitumen causes burns on the skin (Baruchin et al. 1997). Prolonged skin contact
causes dermatitis, acne-like changes and keratosis but the effects are less severe than
those caused by tar (Schwartz et al. 1957).
Health monitoring of 50 Israeli workers who had been exposed 8 hours daily for at
least 6 months without a break to “bitumen-asphalt-vapour” during production or during
road or roof construction work yielded the following dermatological findings: sequelae
of burns (9.7 %), hyperkeratosis on the hand (14.6 %), xeroderma (12.6 %), contact der-
matitis (7.8 %), nail damage (4.8 %), infections and mycoses (21.5 %), basocellular,
mainly premalignant epitheliomas (solar keratosis) (14.5 %) including 2 cases of epi-
thelioma basalis, and dermatitis resulting from photosensitization (14.5 %). In 62 % of
the exposed workers, a “rather severe form of irritative conjunctivitis” was diagnosed
and described by the authors as being “a direct result of exposure to asphalt vapour”
(Schaffer et al. 1985).
A questionnaire study carried out in Finland found that 44 % of the studied roofing
workers and 31 % of the road construction workers complained of skin irritation caused
by their work. Of the road construction workers, 22 % claimed to suffer often or some-
times from “dermatitis”, of the roof construction workers 15 %. The affected persons did
not ascribe their health problems only to the exposure to hot and cold bitumen but, for
example, also to man-made mineral fibres (roof construction) and products containing
amines (road construction) (Riala et al. 1998).
The case was described of a man who developed facial discoid lupus erythematosus
one month after accidental skin contact with cold liquid bitumen (cutback, solvents: tolu-
ene and benzene). Initially only mild irritation of the skin and conjunctiva was observed
(Lübbe et al. 1996). Dockyard workers (28) developed symptoms of reversible photo-
contact dermatitis on the eyes and on areas of skin which had been contaminated with a
bitumen-containing paint (Davies 1996).
Norwegian road construction workers who were exposed on average to bitumen aero-
sol concentrations of 0.36 mg/m3 complained of irritation of the eyes, larynx and pharynx
and of reduced appetite and tiredness. The symptoms were recorded much more often at
higher concentrations. The concentration of bitumen in the vapour phase was mostly
below 1 ml/m3 (about 5 mg/m3). A dependence of the effects on health on the total con-
centration of volatile bitumen emissions could not be demonstrated (Norseth et al. 1991).
In studies of persons exposed for 8 hours in inhalation chambers to total bitumen vapour
concentrations as high as 20 mg/m3, irritative effects were not observed (Knecht et al.
2001). Levin and Järvholm (1999) have pointed out that low molecular weight poly-
amines, which are present as contaminants in certain adhesion-promoting additives,
could also contribute to the irritative effects of the bitumen used in road construction.
Employees of a factory manufacturing fluorescent ballast boxes and coils for fluores-
cent and high intensity lighting developed nausea, headaches, tiredness, skin rashes and
irritation of the nose, throat and eyes after a new bitumen product (used at 270°C) had
been introduced into the process. Personal sampling and analysis yielded “asphalt” levels
of 0.50 to 1.30 mg/m3 (n = 6, average 0.83, median 0.75). In 11 of the 27 employees who
were examined, the average volume of platelets in the blood was significantly increased
above the values found in the control group (“macrothrombocytosis”), but decreased
after reduction of the level of exposure to “asphalt” fumes (Chase et al. 1994).
4.3 Allergenic effects
The possibility that sensitization may be caused by individual components of bitumen

cannot be excluded (Baylor and Weaver 1968).
4.4 Genotoxicity
In collectives of roof construction workers, an increased incidence of DNA adducts in

the peripheral lymphocytes was detected by means of 32P-postlabelling methods (Herbert
et al. 1990; see also Table 6, p. 75). DNA adducts with PAH were found in the DNA of
the white blood cells in 10 of 12 roofing workers (83 %) but in only 2 of 17 control per-
sons (17 %). In the chromatographic analysis, the Rf values of the adducts were similar to
or identical with those of the benzo[a]pyrene-diol epoxide control adduct. The occu-
pational exposure of the roofing workers and control persons was monitored with per-
sonal sampling tubes and also skin swabs. The results for the roofing workers showed
that the levels of polycyclic compounds in the air and in the skin swabs were correlated
with the levels of DNA adducts.
One research group demonstrated a significantly higher level of alkali-induced DNA
strand breaks in peripheral mononuclear blood cells of roofing workers than in road con-
struction workers (Fuchs et al. 1996).
Hatjian et al. (1997) carried out comparative examinations of 16 road construction
workers, 13 roofing workers and 21 control persons who were not exposed to bitumen.
The authors established that the concentrations of 9 selected PAH in the air inhaled by
the road workers correlated with the amount of 1-hydroxypyrene excreted in the urine. In
addition, the incidence of sister chromatid exchange in peripheral lymphocytes of both
roofing and road workers increased in parallel with the exposure concentrations of the 9
PAH.
It can, however, not be decided whether or not the effects described above result only
from the occupational exposure to bitumen fumes. Although the effects of smoking were
excluded—especially for the roofing workers—exposure to tar during removal of old tar-
containing surface materials is conceivable. Likewise other confounders such as diesel
motor or chimney emissions could have played a role.
4.5 Carcinogenicity
The development of malignant melanomas after skin burns caused by bitumen has been
reported (Peila et al. 1999).
All the currently available epidemiological studies of other end points are summarized
in Table 7 (p. 79) and Table 8 (p. 84).
In 1994, the IARC published a meta-analysis of the results of carcinogenicity studies

for bitumen (Partanen and Boffetta 1994). Occupational exposure to bitumen occurs in a
variety of jobs. The workers were subdivided into groups called “roofers”, “road con-
struction workers” and “other workers”. In all, 20 studies were analysed. These 20 stud-
ies which were included in the meta-analysis are marked in Tables 7 and 8. The reasons
why certain studies were not included in the analysis were numerous and are also men-
tioned in the tables.
The relative risks (called simply “risk” in the text that follows and in Table 7) found
in the meta-analysis are listed below. For the “roofers”, increased incidence of cancer
was found for the localizations lungs (RR = 1.8; 95 % CI: 1.5–2.1), stomach (RR = 1.7;
95 % CI: 1.1–2.5), skin (RR = 4.0; 95 % CI: 0.8–12), and leukaemia (RR = 1.7; 95 % CI:
0.9–2.9). For the road workers the relative risks were all lower than for roofing workers:
lungs (RR = 0.9; 95 % CI: 0.8–1.0), stomach (RR = 1.1; 95 % CI: 0.8–1.5), bladder
(RR = 1.2; 95 % CI: 0.7–1.8), skin (RR = 2.2; 95 % CI: 1.2–3.7) and leukaemia (RR =
1.3; 95 % CI: 0.9-1.8). The increased risk of skin cancer was caused by the results of one
study. Workers in other jobs had an increased risk of developing lung cancer (RR = 1.5;
95 % CI: 1.2–1.8). The evaluation of these results is made more difficult by the fact that
in many cases the persons were also conceivably exposed to tar products. Because the
IARC was of the opinion that the data are inadequate for evaluation of the carcinogen-
icity of bitumen, a multi-national study was initiated. To date, results from Germany and
Norway are available and a provisional summary from the IARC (2000).
Some of the studies deserve particular attention. On the one hand, in some studies
very high risks were observed; on the other, the publications of Hansen (1989a, 1989b,
1991) have been repeatedly criticised in the literature. Some of these “conspicuous”
studies and the results available to date from the IARC study are presented below.
A) Bender et al. (1989)
In this cohort study, the cancer mortality of 4849 workers who were employed by the
Department of Transportation in Minnesota (USA) for at least one year between 1945
and 1984 was investigated. In the group of workers employed for a period between 30
and 39 years, the leukaemia risk was significantly increased (n = 7 deaths; SMR = 4.25,
95 % CI: 1.71–8.76). The study was carried out because of a cluster of 6 cases of leu-
kaemia in a small community in western Minnesota where one case would have been
expected. Five of these six cases were employed as “highway maintenance workers”. It is
noteworthy that the leukaemia risk is only increased for the workers employed for a
period between 30 and 39 years. This finding could be a chance result because of the
large number of comparisons which were carried out. The leukaemia risk for workers
employed for less than 30 years was 0.79. The increased leukaemia risk could also con-
ceivably have been caused by exposure to herbicides containing 2,4-D (2,4-dichloro-
phenoxyacetic acid) and 2,4,5-T (2,4,5-trichlorophenoxyacetic acid).
B) Hansen (1989a, 1989b, 1991)
In this Danish study, both cancer incidence and mortality were reported. Some of the
observed risks were markedly increased. The cancer incidence, for example, is twice that
found in the general population of Denmark. The lung cancer risk is 3.44 times that in the
general population. The analysis of mortality revealed similar risks. The study has been
criticised for potential selection bias and for confounding by alcohol consumption and
smoking. In a subsequent analysis, the risks were adjusted for smoking habits and for
town/country differences. The SMR for lung carcinoma for workers more than 40 years
old was reduced from 2.64 (95 % CI: 1.71–3.90) to 2.46 (95 % CI: 1.59–3.63) when
smoking was taken into account and further to 2.24 (95 % CI: 1.45–3.30) when adjusted
for smoking and town/country differences. Thus the risk remains increased even when
these two factors are taken into account. The other criticism of this study, that the
persons had perhaps been exposed to tar products, remains unanswered and impedes
interpretation of the results.
C) IARC study
C1) Germany
In this study the cancer risk of bitumen workers in Germany is investigated (Frentzel-
Beyme et al. 2000). Included in the study were a total of 10679 workers who had been
employed by 138 road construction firms and asphalt mixing plants for at least 2 months.
The sub-collective of male workers employed for at least 12 months (n = 7886) was
analysed. 470 workers have died since the beginning of the study. For 135 of the persons,
a tumour was given as the cause of death. Increased mortality from lung cancer, cancer of
the upper respiratory tract and the oesophagus is reported. The risk is, however, also
increased in the group of workers not exposed to bitumen (see Table 7, p. 83). None of
the differences documented for the tumour localizations are statistically significant. Thus,
for example, the difference between the SMR values for lung carcinoma in all persons
exposed to bitumen (SMR = 1.97) and all non-exposed persons (SMR = 1.52) is not
significant.
Simultaneous exposure to other substances such as diesel soot, quartz, asbestos or
cigarette smoke has been suggested as a possible explanation. These factors have not
been studied to date. As a whole, the study does not reveal significantly increased tumour
risks for persons exposed to bitumen in comparison to those for non-exposed persons. It
is, however, conspicuous that the risks of tumours at certain localizations are increased in
the whole collective.
C2) Norway
The Norwegian results were presented at an occupational medical congress (Randem et
al. 2000). The cohort includes about 9000 persons working with asphalt in 11 different
firms. During the study period, tumours were diagnosed in 383 of the workers (SIR =
0.90; 0.81–1.0). For 72 workers lung carcinoma was diagnosed (SIR = 1.34; 1.05–1.69)
The lung (carcinoma) was the only localization with a significantly increased risk. It is
unclear whether the increased risk is a result only of exposure to bitumen.
C3) Preliminary results
At a congress in September 2000, first results of the IARC study from 8 countries were
presented (IARC 2000). The SMR for lung cancer was higher for the bitumen workers
than for the non-exposed building workers (SMR = 1.17, 95 % CI: 1.04–1.32 compared
with SMR = 0.94, 95 % CI: 0.82–1.07). Internal comparison of the two groups yielded a
relative risk of 1.01 (95 % CI: 0.82–1.24). The conclusion drawn by the IARC was that
workers exposed to bitumen in Europe do not seem to have an increased risk of devel-
oping lung cancer.
Evaluation
When all the studies are examined, the lung (carcinoma) is the localization with the most
significant results. When interpreting the results for this tumour localization, some bitu-
men-specific problems should be considered. One problem is the potential co-exposure
to tar products. Another is associated with the composition of the asphalt surfacing ma-
terial and the temperature at which the asphalt is used. The higher the temperature, the
easier it is to use the asphalt but the higher are the concentrations of the fumes.
Another problem is that the workers do not work in jobs involving exposure to bitu-
men for the whole of the year. In the winter months the exposed persons are unemployed
or do other jobs. Details are not given in the publications.
The exposed workers have an increased risk of developing cancer. But because of the
limitations mentioned above, it is not possible to decide from the available results
whether bitumen should be classified as a human carcinogen. The results of the European
study by the IARC suggest, however, than the lung cancer risk is either not increased by
bitumen or increased only very slightly.
5 Animal Experiments and in vitro Studies
There are no data available for the reproductive or developmental toxicity of bitumen
vapour and aerosols in experimental animals.
5.1 Acute toxicity
Rats (Sprague-Dawley) were treated by intratracheal instillation either with sterile physi-
ological saline (0.25 ml) or with one of three doses (0.1 mg, 0.5 mg, 2 mg in 0.25 ml
sterile physiological saline) of a condensate of road asphalt fumes (taken from a tank). A
third group of rats was treated by intratracheal instillation of 0.25 ml 1 % DMSO. Rats
from all the groups were killed either one day or three days after treatment. To determine
whether the effects of exposure can be cumulative, animals were treated by intratracheal
instillation on three subsequent days and then killed. To detect cell damage, extracellular
lactate dehydrogenase (LDH) and protein were determined in the first bronchial lavage
fluid. Chemiluminescence and determination of tumour necrosis factor-α (TNF-α) and
interleukin-1 (IL-1) production were used to assay alveolar macrophage function. The
results demonstrated that exposure of rats to condensate of road asphalt fumes neither
caused acute pulmonary inflammation or damage nor did it significantly alter alveolar
macrophage function (Ma et al. 2000).
Residues of vacuum distillation (vacuum residuum samples API 81-13, API 81-14;
CAS 64741-56-6) were dissolved in corn oil and administered to groups of 5 male and
female Sprague-Dawley rats in doses of 5000 mg/kg body weight; mortality during the
14-day observation period was not increased. Hypoactivity and diarrhoea were observed
(API 1982a, 1982b).
Distillation residues (vacuum residuum samples API 81-13, API 81-14; CAS 64741-
56-6) were warmed in a water bath and applied occlusively for 24 hours to the shaved or
abraded skin of 2 male and 2 female New Zealand White rabbits in doses of 2000 mg/kg
body weight. Diarrhoea was seen in one female animal on day 1 and in one other female
on days 6 and 7 after treatment with sample API 81-13. During the 14-day observation
period mortality was not increased (API 1982a, 1982b).
5.2 Subacute, subchronic and chronic toxicity
5.2.1 Inhalation
In mice which had inhaled a bitumen-water aerosol for a period of 16.5 months, emphy-
sema, bronchiolar dilation and, in some animals, pneumonitis and severe localized bron-
chitis were detected. Mice exposed in whole animal exposure chambers to asphalt fumes
(asphalt heated to 121°C) for 21 months developed bronchitis with peribronchial infiltra-
tion of large round cells, epithelial hyperplasia, loss of cilia, flattening and necrosis of the
bronchial epithelium, loss of respiratory epithelium, and atrophy and necrosis of the
alveolar epithelium (Simmers 1964; study described in Table 14, p. 114). In a 2-year
inhalation study (vapour from asphalt heated to 121–135°C) severe chronic fibrotic
pneumonitis with peribronchial adenomatosis, epithelial metaplasia in the bronchial
mucosa and bronchiectasis were observed in the female rats and in guinea pigs (Hueper
and Payne 1960; study described in Table 14, p. 114).
5.2.2 Ingestion
Groups of four pigs (40 kg body weight) were given daily doses of 10 g bitumen (Hun-
garian products, one distillation bitumen, one extraction bitumen) for 63 or 71 days.
Appetite, body weight gains and state of health of the animals were unaffected. At the
end of the study, histological examination of the livers and kidneys of the pigs revealed
no pathological changes. In another group of animals treated for the same period with
daily doses of 10 g tar pitch from brown coal, appetites were impaired by the treatment.
In the livers, the amount of interlobular connective tissue was slightly increased (Köves
and Zakar 1959).
5.2.3 Dermal absorption
Mice treated dermally with the substance developed pneumonitis and chronic dermatitis
(Simmers 1965a; study described in Table 13, p. 100) or hyperkeratosis and inflamm-
ation of the skin (Kireeva 1968; study described in Table 13, p. 102). Wallcave et al.
(1971) reported not only epidermal hyperplasia but also the formation of ulcers and small
abscesses. In addition, amyloidosis was detected in the spleen and kidneys of mice
treated epicutaneously with asphalt (study described in Table 13 p. 103). Sivak et al.
(1997) observed no effects of epicutaneous application of various asphalt samples and
their fractions on the body weights of mice (study described in Table 13 p. 109).
Two distillation residues (vacuum residuum samples API 81-13, API 81-14; CAS
64741-56-6) were applied undiluted to the skin of New Zealand White rabbits in patch
tests (groups of 5 male and 5 female animals, 200, 1000, 2000 mg/kg body weight, 6
hours daily, 3 times weekly for 4 weeks). Of the animals treated with API 81-13, one
male from the 2000 mg/kg dose group died on day 9 and one control group female on
day 3; of the animals treated with API 81-14, one male from the 200 mg/kg dose group
died on day 13 and one female from the 2000 mg/kg dose group on day 13. An associa-
tion with the treatment was not assumed. Two other animals from the API 81-13 series
(one male control animal on day 6, one female from the 1000 mg/kg dose group on day
10) were killed with paralysis of the hind limbs. This effect was considered to result from
the handling of the animals (fixation during treatment). During the treatment, reduced
food consumption, desquamation of the skin, and wheezing were observed. In the
animals of the highest API 81-13 dose group on day 21, body weight gains were
significantly lower than in the control group. It was generally not possible to assess skin
reddening because the sample material could not be wiped off. Oedema formation was
slight to moderate. Microscopic examination of the skin revealed minimal to moderate
dermatitis and hyperkeratosis. The haematological and clinicochemical parameters were
unaffected (API 1983a, 1983b).
5.3 Local effects on skin and mucous membranes

The irritative effects of two distillate residues (vacuum residuum samples API 81-13,
API 81-14; CAS 64741-56-6) were studied by application to the skin of 6 male New
Zealand White rabbits. The backs and flanks of the animals were shaved and immedi-
ately before the start of the study some skin areas were abraded. The samples (0.5 ml,
temperature not specified) were applied occlusively for 24 hours on both shaved and
abraded skin areas on each animal and were then wiped off. The Draize scores for skin
reactions were assessed after 24, 72 and 96 hours and after 7 and 14 days. At the end of
the observation period, erythema was still present (score 0.5–0.8). The primary irritation
index determined after 24 and 72 hours on intact and abraded skin was 0.2 and 0.4,
respectively, for both samples. Autopsy revealed no effects (API 1982a, 1982b).
Two distillate residues (vacuum residuum samples API 81-13, API 81-14; CAS
64741-56-6) were tested by introduction of 0.1 ml samples (temperature not specified)
into the conjunctival sac of the right eye of each of 6 New Zealand White rabbits. The
eyes of three other similarly treated animals were rinsed with warm water 20 to 30 sec-
onds after treatment. The Draize scores were determined 1, 24, 48 and 72 hours and 7
days after treatment, after 24 hours being 4.0 for API 81-13 and 4.7 for API 81-14. After
7 days and 72 hours, respectively, no visible effects remained (API 1982a, 1982b).
5.4 Allergenic effects
The allergenic effects of two distillate residues (vacuum residuum samples API 81-13,
API 81-14; CAS 64741-56-6) were studied with male Hartley guinea pigs by means of
the occlusive patch test. Induction was carried out by application of 0.4 ml of the undi-
luted test material once weekly for 6 hours. This treatment was carried out for 3 weeks.
Provocation, 14 days after the last induction treatment, was carried out with 0.4 ml un-
diluted test material. A positive control was included. The vacuum distillates proved to
be inactive (API 1984a, 1984b).
5.5 Genotoxicity
Genotoxicity studies have been carried out with bitumen solutions, extracts and conden-
sates from various sources and processes (see Tables 9 to 11, pp. 88–95). Because of
differences in the methods used, the results cannot readily be compared and an overall
assessment is difficult.
5.5.1 In vitro
The results obtained in the Salmonella mutagenicity test with bitumen solutions and
extracts were mainly negative or not unambiguously positive even though occasionally a
special variant of the test was used, a method developed for testing mineral oils and
stated by the authors to be particularly sensitive, with hamster liver microsomes and a
higher concentration of NADPH (Blackburn et al. 1986). In contrast, condensates of
bitumen fumes mostly produced an increase in the numbers of mutations in Salmonella
typhimurium especially after metabolic activation (Table 10, p. 90).
In a forward mutation assay with the mouse lymphoma cell line L5178Y, the mutation
frequency was significantly increased in the presence of rat liver S9 mix by a petroleum
vacuum distillation residue dissolved in acetone, but not without metabolic activation
(API 1984c, 1984d).
In the presence of a metabolic activation system, bitumen vapour condensate pro-
duced adducts with dissolved DNA (Table 9, p. 88).
In cultured eukaryotic cells, bitumen solutions produced DNA adducts and DNA-
protein cross-links. Bitumen vapour condensates increased the incidence of micronuclei
in V79 cells but did not induce chromosomal aberrations in cultured CHO cells (a cell
line from Chinese hamster ovary).
5.5.2 In vivo
In experimental animals given doses of bitumen solutions, extracts or condensates, local

and systemic DNA adducts were induced. Studies by Genevois et al. (1996) with BD4
rats revealed that the adduct pattern found could not be explained entirely by the unsub-
stituted PAH present in the epicutaneously applied condensates of bitumen fumes.
Repeated oral administration to rats of large doses of residues from the vacuum dis-
tillation of petroleum did not increase the incidence of structural chromosomal aberra-
tions in the bone marrow of the animals (API 1984c, 1984d).
5.6 Carcinogenicity
Introductory remarks: The most important long-term animal studies of the carcino-
genicity of “asphalt” (or bitumen) have been carried out in the USA (see Tables 13–15,
pp. 99–118). In our presentation and evaluation of these studies, the question arises as to
the relationship of biological effect and chemical identity of the “asphalt” or “bitumen”
sample being tested.
Because of the inconsistency in the use of the terms “asphalt” and “bitumen” in the
literature, in Tables 13 to 15 and in the text below the “asphalt” and “bitumen” samples
used in the animal studies are described with the terms used in the original publications.
It must be pointed out in this context that neither the American “asphalt” nor the Euro-
pean “bitumen” are classified in terms of their chemical composition but merely on the
basis of their physical properties such as viscosity, softening temperature, flow behav-
iour, and resistance to penetration. Different kinds of “asphalt” and bitumen are available
commercially, defined only by their physical properties and intended for a variety of
purposes; the materials are often also modified to make them suitable for special appli-
cations (IARC 1985).
Because the (physical) product specification is not directly correlated with the chemi-
cal composition, it is not possible to conceive a generally applicable standard test sub-
stance for which the carcinogenic effects should be determined.
The chemical composition of “asphalt” and bitumen varies both with the source of the
petroleum and the production process (injection of steam or air, later admixture of oils
and other components, etc.). Thus, for example, two samples of bitumen with the same
physical specification and production history can differ in their carcinogenic activity if
the starting material originated from different oil fields. The carcinogenicity of two
samples can be different even when the oil field and physical properties are the same if
different production processes or additives were used to achieve the specification. Not
even the products of a single producer can be standardized biologically because all pro-
ducers buy their crude petroleum starting material from different countries, depending on
the current prices.
The carcinogen content of the bitumens to which people are exposed at work has not
been defined. The producer can see no reason to provide his product with a certificate
giving the results of a chemical analysis for known carcinogens. The estimation of the
carcinogenic activity of a bitumen sample is made more difficult still by the fact that
these complex mixtures can also contain still unidentified carcinogens.
The situation described above makes clear why it was not possible in the carcino-
genicity studies described below to use “asphalt” samples which had been subjected to
sufficient chemical analysis to yield substance-effect relationships.
In summary: There are numerous kinds of “asphalt” or bitumen which vary in their
chemical composition. The content of carcinogens varies too. Nonetheless, scientifically
documented analyses (see Tables 3 and 5; Niemeier et al. 1988, Table 13a; NIOSH
1981) demonstrate that a series of carcinogenic substances are common to a number of
kinds of bitumen or “asphalt”. The data available at present indicate that these are mainly
PAH and heterocyclic substances. Likewise, these carcinogens are common to the
aerosols produced in hot processing of the bitumens. On this basis it seems to be
reasonable to apply the results obtained in animal studies with, for example, “roofing
asphalt, type I” also to the bitumens used in road construction.
5.6.1 Dermal application
Characterization of the animal studies

In the early long-term animal studies (up till about 1980) the experiments were carried
out with mixtures of heated “road paving asphalt”, with solutions of this and similar
materials in benzene or toluene and with crude fractions (e.g. “saturates”, “aromatics”).
The results of these studies are inconsistent. Taken as a whole, however, the early animal
studies document a weak tumour-inducing effect of the various “asphalt” samples on the
skin of mice of several strains. But the methods used in most of these early studies were
inadequate. For example, the doses were inadequately defined, or too many of the treated
animals died prematurely, or the tumour yield was not given as the number of animals
with tumours but as the total number of tumours in the group of animals.
In the later long-term animal studies, on the other hand, the “asphalt” samples used
were defined—as well as possible—and the tumours recorded in minute detail. Exem-
plary and accordingly informative are three large series of studies carried out for and
designed together with the US National Institute for Occupational Safety and Health
(NIOSH) (Niemeier et al. 1988, NIOSH 1981, Sivak et al. 1997). The study material
comprised condensates of the fumes from two commercial USA roofing bitumens
(“roofing asphalt” from Exxon) which met the specifications of the American Society for
Testing and Materials (ASTM). One of the samples was for use on flat roofs (“dead
level”, “type I”), the other for pitch roofs (“steep”; “type III”). For these commercial
products, the producer issued official recommendations as to the temperature at which
they should be applied on the roof. The temperatures used to produce the fumes in the
laboratory were taken from these recommendations. The experimental variables during
testing of the fume condensates included: two different temperatures for fume produc-
tion, three different strains of mice (C3H/HeJ, CD-1 and Sencar), the presence and ab-
sence of sunlight (UV-B), fractionation of the condensate by high performance liquid
chromatography (HPLC) and testing of the fractions alone or in combination, testing
certain fractions for tumour-promoting activity in two-stage experiments, and positive
and negative controls.
In addition, the levels of 16 aromatics identified by the NIOSH and the US Environ-
mental Protection Agency as “suspect carcinogens” were determined in the condensates
by gas chromatography/mass spectrometry (GC/MS). The clearly positive results for a
tumourigenic effect of the condensates of the fumes from the two “roofing asphalts” were
based on data obtained with more than 100 groups of animals (30 to 50 animals per
group).
The publications summarized above are now discussed in more detail below. More
details may be seen in Table 13 (p. 99).
Simmers et al. (1959) determined the skin carcinogenic potential of a mixture of three
steam-treated and three air-treated “asphalt” samples from Californian refineries (see
Table 13, p. 99). The semi-solid mixture was liquefied with benzene so that it could be
applied topically. This “asphalt”-benzene suspension (concentration not specified) was
applied twice weekly with a glass rod to the skin between the shoulder blades of each of
32 male and 36 female C57 Black mice. Groups of 31 male and 32 female animals
treated with benzene alone served as the controls. In the treated group, 12 squamous cell
carcinomas developed at the application site; five (other?) treated animals developed
papillomas, also at the application site. The squamous cell tumours were classified
histologically as malignant. Before the carcinomas appeared, hair loss, desquamation and
papilloma formation were seen on the treated skin area. Metastases did not develop. No
skin tumours were detected in the control group; at most slight hair loss and desqua-
mation were observed.
The authors did not specify how many animals in the treated group had a tumour. In a
later study (Simmers 1965a) reference is made to the first publication (Simmers et al.
1959) and it is stated that 22 % of the treated mice developed squamous cell carcinomas.
That would mean that of the 68 animals, 15 developed tumours. Simmers et al. (1959)
concluded that the squamous cell carcinomas were caused by components of the
“asphalt”.
To exclude the possibility that the benzene had contributed to the tumour formation
and to clarify the toxicological differences between steam-treated and air-treated
“asphalt”, Simmers (1965a) carried out further studies. He used two mixtures, in each
case of three similarly pretreated samples (“steam-refined” and “air-refined”), and
applied these (single doses: about 75 to 100 mg) after warming in a water bath directly to
the (once) shaved interscapular skin of the C57 Black mice (Table 13, p. 100). Groups of
50 mice (25 male and 25 female) were used but no control animals. After 7 weeks the
groups had been reduced by pneumonitis to 32 (the mixture of “air-refined asphalts”) and
27 animals (the mixture of “steam-refined asphalts”). Of these 27 animals, only 6
remained at the end of the year. As these animals had unspecific skin reactions, the group
was stocked up with another 13 animals.
Results with “air-refined asphalt”: at the end of the study only one of the 32 treated ani-
mals had developed a papilloma, another a tumour originating in a skin appendage and a
third a lung adenoma. The author reported that the applied material hardened to a crust
which the animals tore off. Chronic dermatitis in the application area is also reported.
Results with “steam-refined asphalt”: of the 19 animals which survived until the end of
the study, 3 had developed a squamous cell carcinoma at the application site. In addition,
two papillomas were identified.
Simmers reported that also the steam-refined material did not maintain good skin
contact. Therefore for a further series of experiments, toluene was chosen as diluent. The
mixture of “air-refined asphalts” was diluted with toluene in a ratio of 10:1 and the sus-
pension applied to 10 male and 10 female mice three times weekly for two years (Table
13, p. 101). In this way on average 284 applications of 20 to 30 mg of the material per
application were carried out. The exact “asphalt” concentration is not given because the
toluene evaporated and had to be replaced from time to time. The control group com-
prised 5 male and 10 female animals treated three times weekly with pure toluene but
only for 19 months so that a maximum of 230 applications was achieved. Autopsy of the
animals treated with the asphalt-toluene mixture revealed 9 squamous cell carcinomas of
the skin. One of these carcinomas originated in a skin appendage and was macroscopi-
cally visible after as few as 147 applications. One squamous cell carcinoma had infil-
trated a regional lymph node and had replaced all the lymphoid cells. Another squamous
cell carcinoma had infiltrated the shoulder blade. In two mice, chronic dermatitis was
diagnosed. In the 15 control mice (toluene alone), one small papilloma was found in only
one animal. In all other animals, hair loss, desquamation and slight thickening of the
dermis and epidermis were observed. The author stated that he had found no evidence in
the literature of co-carcinogenic effects of topically applied toluene (or benzene).
The large difference in tumour yields produced by pure “air-refined asphalt” (2
tumours) and the same material diluted with toluene (9 squamous cell carcinomas) was
explained by the better skin contact achieved with the toluene suspension. In comparison,
“steam-refined asphalt”, which stuck to the skin a little better than did “air-refined
asphalt”, induced three squamous cell carcinomas and two papillomas.
In an attempt to associate the carcinogenic effects of the complex “steam-refined
asphalt” mixture with single classes of substances, Simmers (1965b) divided the mixture
into four fractions by chromatography on silica gel and adsorption to clay. The fractions
were designated according to their main components: asphaltenes, aromatics, saturated
compounds and resins. Two of the fractions were fluorescent under UV light. These two
were pooled and tested biologically in the expectation that the fluorescent substances
were the carcinogens. The viscous yellowish-green fluorescent material was rubbed into
the interscapular fur of 25 male and 25 female C57 Black mice three times weekly with a
glass rod (Table 13, p. 101). The average single dose was 33.4 mg. The minimum num-
ber of applications was 72, the maximum 242. Controls were not included because spon-
taneous tumours had never been observed in the interscapular area. Autopsy was carried
out on 40 of the 50 animals. Since a tumour was not detectable macroscopically on 10 of
these 40 animals (but only dry, scaly and hairless skin in the application area), these 10
animals were not subjected to histological examination. In the remaining 30 animals not
only hair loss but also irregularly thickened skin and deeply ingrown hair follicles were
seen. Skin cancer was diagnosed in 13 animals. The histological examination revealed
squamous cell carcinomas in 7 animals (including a squamous cell carcinoma of the
anus; in this animal a leiomyosarcoma of the small intestine was also found), basal cell
carcinomas in 5 animals and in one other animal a carcinoma which had developed from
a sebaceous gland. One squamous cell carcinoma had produced pulmonary metastases. In
addition, 13 papillomas were recorded, one of which had been penetrated by a
leiomyosarcoma. The yield of animals with tumours was 32.5 % of the 40 animals
subjected to microscopic examination and 26 % of the 50 animals present at the start of
the experiment.
Simmers concluded from his results that the “asphalt” samples which he had studied
certainly contained substances which were carcinogenic for mouse skin. He concluded
that the carcinogenic activity of the combined saturated and aromatic fractions of the
“steam-refined asphalts” was higher than that of the same material diluted with benzene
(Simmers et al. 1959) and than that of the undiluted materials (Simmers 1965a).
Hueper and Payne (1960) studied the skin carcinogenic potential of four “road asphalts
that were steam distillation products” from petroleum fields in Mississippi, California,
Venezuela and Oklahoma (Table 13, p. 100). Each of the samples was diluted with acet-
one until it could be applied dropwise to the neck skin of the experimental animals. For
the tests with each of the three US American samples, 50 C57 Black mice (25 males and
25 females) were used, for the Venezuelan sample 100 animals. The “asphalt” solutions
were applied twice weekly for 2 years. Of the 250 treated mice, 1 developed a skin carci-
noma, 2 others a papilloma. In addition, 7 animals developed leukaemia. The authors
consider that the “asphalt” was responsible not only for the skin tumours but also for the
leukaemia which developed in animals treated with the material from Venezuela (4
cases), Oklahoma (2) and Mississippi (1). The authors are convinced that the test sam-
ples contained weak to moderately powerful (skin) carcinogens. In addition they suggest
that components of the topically applied “asphalt” had systemic effects and stimulated
the development of leukaemia.
Kireeva (1968) tested three “sludge asphalts (straight distillation)” produced from Rus-
sian petroleum (residues BN-5, BN-3, BN-2) and suitable for road construction. The
material was diluted with benzene (40 %) and applied once weekly to the skin of groups
of 40 to 50 C57 White mice (Table 13, p .102). The control group (23 animals) was
treated with benzene alone. Details of the doses are not given in the publication. Of the
50 animals treated with residue BN-5, one developed a cornified squamous cell carci-
noma another a sebaceous gland adenoma. In 5 animals, lung adenomas and adeno-
carcinomas were found. Of the 50 animals treated with residue BN-3, one developed a
subcutaneous fibrosarcoma, another a papilloma. In addition, tumours of the internal
organs were found in 4 animals: one lung adenoma, 2 lymphoreticular sarcomas and one
liver haemangioma. None of the 40 animals treated with residue BN-2 developed skin
tumours and only one a lung adenoma. In the 23 animals of the control group there were
no skin tumours; one animal had lung adenomas.
The following non-tumourigenic effects were seen frequently in the animals treated
with “sludge asphalt”: thin fur, partial or total atrophy of the skin papilla, focal hyper-
plasia of the epidermis and the follicle epithelia, hyperkeratosis with acute and chronic
inflammation. Atrophy of the sebaceous glands, the hair follicles and the epidermis were
also seen in some of the control animals.
Wallcave et al. (1971) carried out studies with 8 “road paving grade asphalts” in an
attempt to find a correlation between skin tumour-inducing effect and content of PAH
(Table 13, p. 103). The samples used were derived from vacuum distilled crude oils from
the USA, South America and the Middle East and their ASTM penetration specification
was 85/100. For the biological tests, 10 % solutions in benzene were prepared and these
applied in 25 µl aliquots (corresponding to 2.5 mg “asphalt”) twice weekly to a 2.5 cm2
area of the shaved dorsal skin of Swiss albino mice (about 30 animals per group, 15
males and 15 females). The control group animals were treated topically with 25 µl ben-
zene alone.
Autopsy and histological examination of the animals treated with “asphalt” revealed
hyperplasia of the epidermis often accompanied by infiltration of the dermis with inflam-
matory cells and ulceration, amyloidosis especially of the spleen and kidneys. In the 218
treated animals which survived until the end of the study, 6 skin tumours were found
(2.7 %); one tumour was found in the control group (3.8 %). In addition, both subcuta-
neous tumours and tumours of the internal organs were found but in no case was the
incidence higher than in the control group. The authors concluded from their results that,
although the observed tumour yield corresponded to that found by Hueper and Payne
(1960), the total number of tumours was too small for an association between PAH con-
tent and skin tumourigenic effect to be established.
In an extensive series of studies, the NIOSH (1981) studied the tumour-inducing effects
on mouse skin of the volatile components of “roofing asphalts” (Table 13, p. 104). In all,
13 groups of 50 male C3H/HeJ mice and 13 groups of 50 male CD-1 mice were used.
The test materials were fume condensates from two commercial samples of “asphalt”
which differed in their physical properties, especially in their softening temperatures, in
accordance with their intended use (“type I, dead level” for flat roofs; “type III, steep”
for pitch roofs). These “roofing asphalts”—produced by Exxon—met the physical speci-
fications of the ASTM. To obtain the condensates, 10 kg portions of the samples were
heated to 232°C and 316°C, respectively, while stirring. Because the NIOSH study (and
those of other authors) are frequently criticized for the high temperatures used to produce
the fumes, the reasons for this choice of temperatures are discussed here briefly.
The temperatures were based on the recommendations of the producer. Thus, the
temperature recommended for “type I asphalt” during application to the roof is 177–
204°C; the kettle temperature is typically about 30°C higher. The temperature recom-
mended for the application of “type III asphalt” is 199–246°C (and the corresponding
typical kettle temperature would be 230–276°C). In the study, however, “type III as-
phalt” was heated to only 232°C, rather a low temperature for this kind of material. The
second temperature used to produce condensates, 316°C, was chosen intentionally high
to imitate the mostly uncontrolled hot kettle temperatures found under real working con-
ditions.
The fumes were frozen out in cold traps at –80°C. Negative controls (treatment with
solvent alone, no treatment) and positive controls (treatment with benzo[a]pyrene)
yielded the background values for determining tumour yield. To simulate the situation
during exposure of workers, who are often exposed simultaneously to bitumen fumes and
sunlight, all the samples were tested on mouse skin both with and without exposure to
artificial sunlight (UV-B produced with a xenon arc lamp). The results are described
below (Table 13, p. 104).
1) 25 mg condensate applied twice weekly for 18 months to the dorsal skin of the mice
induced skin papillomas and squamous cell carcinomas in all test groups. Male
C3H/HeJ mice developed more tumours than did male CD-1 mice and also a higher
proportion of malignant tumours.
2) The combination of condensate with UV-B irradiation (for 30 to 45 minutes after

application of the condensate) delayed the appearance of the tumours. The tumour
yield was also (slightly) reduced. In no case did the UV-B irradiation increase the
tumour yield above that induced by the condensate alone.
3) The carcinogenicity of the condensate increased with the temperature used to produce
the fumes. Thus the condensate produced at 316°C induced more tumours in both
strains of mice than did that produced at 232°C.
The conclusions which Niemeier et al. (1988) drew from their results are detailed below.
1) The risk associated with occupational exposure to “asphalt” fumes is greater than had
been documented at that time in the literature.
2) When the condensates of the two kinds of “asphalt” were produced at the same tem-
perature and tested in the same mouse strain, the tumourigenic effects were the same.
3) The carcinogenicity can increase with the temperature used for fume production. (For
this reason, the authors suggest that the recommended kettle temperature should not
be exceeded.)
4) The carcinogenicity of the “asphalt” fume condensates is (unlike that of coal tar pitch)
not even approximately accounted for by their content of benzo[a]pyrene and other
PAH because effect and PAH content were not correlated.
5) The carcinogenicity of the condensates is probably also caused by co-carcinogens and
tumour promoters, for example, by the aliphatic hydrocarbons which are also present
in large quantities.
McKee et al. (1986) studied the carcinogenic potential of “bitumen” from Athabasca tar
sands (Table 13, p. 107). The material which is separated from solids (mostly sand) and
impurities with hot water, steam and sodium hydroxide solution (“syncrude process”) is
suitable for use in road construction without any other treatment.
The bitumen was applied to the interscapular skin of 50 male C3H/HeJ mice in 25 µl
aliquots of a 75 % suspension, three times weekly for the whole life of the animals (aver-
age survival period about 76 weeks). Another group of 50 C3H/HeJ mice was treated
with the solvent toluene alone as control. All animals were examined daily for the devel-
opment of tumours. One animal in the group treated with bitumen developed a squamous
cell carcinoma in the application area, another animal a benign tumour. The authors
pointed out that although the treatment with bitumen had induced only two tumours—
that is, a non-significant number—one of these tumours was a squamous cell carcinoma,
a tumour which the authors had never seen in the interscapular region of control animals.
McKee and Lewis (1987) also tested “raw bitumen” from the deposits at Cold Lake Oil
Sands in Alberta, Canada, another material produced with the “syncrude process” (Table
13, p. 108). In a similar study (McKee et al. 1986), this crude bitumen proved to have
much more carcinogenic activity than the material from Athabasca sands. It induced skin
tumours in 26 % of the treated animals, carcinomas in 8 animals and papillomas in 5. In
the control group treated with toluene alone, no tumours were detected. The authors
concluded that “Cold Lake” bitumen was a moderately powerful epidermal carcinogen
which was not less dangerous than other bitumens produced by the usual fractionation of
petroleum.
Sivak et al. (1997) heated “type III roofing asphalt” (“standard commercial type, steep”,
of the same specification as used by Niemeier et al. (1988)) to 316°C, produced conden-
sates of the fumes and fractionated these by HPLC to yield five fractions (A to E). The
chemical composition of the individual fractions was determined by GC/MS. The fol-
lowing materials were tested for carcinogenicity on mouse skin: “raw asphalt”, “heated
asphalt less fume” (material heated to 316°C while the fumes were permitted to escape),
“heated asphalt plus fume”, “neat asphalt fume” (native fumes), the HPLC fractions
(single fractions and various combinations), “asphalt” fumes reconstituted from the sin-
gle fractions as well as a series of negative and positive controls (Table 13, p. 109).
Benzo[a]pyrene in three different concentrations served as the positive control. In addi-
tion, the HPLC fractions A, D and E were tested both for co-carcinogenic effects (with
benzo[a]pyrene) and also for tumour-promoting effects (initiator: benzo[a]pyrene in
three different initial doses). These fractions were selected for testing for co-carcinogenic
and tumour-promoting effects because they contained potential promoters such as long-
chain alkanes and phenols.
In all, 40 groups of 30 male C3H/HeJ mice were used. The materials were dissolved
in 50 µl cyclohexane/acetone (1:1) and applied to the shaved dorsal skin twice weekly.
The concentrations of the solutions prepared from the HPLC fractions A to E were cho-
sen to correspond to their proportions by weight in the fumes (A: 64.1%, B: 8.3%, C:
10.5%, D: 11.5%, E: 5.6%). In addition, unfractionated fumes (“neat fume”) was tested
on Sencar mice to detect potential differences in sensitivity from the C3H/HeJ mice.
In Table 13 (p. 109) are shown the most important groups, the numbers of animals
with tumours, the total numbers of papillomas and carcinomas per group, and the average
survival of the animals.
a) Condensed fumes of “type III asphalt” increased significantly the yield of papillomas
and carcinomas in both mouse strains. Thus unfractionated fumes induced skin carci-
nomas in 20 of 30 C3H/HeJ mice whereas “raw asphalt” induced these tumours in
only 3 of 30 C3H/HeJ mice. If “type III asphalt” was heated to 316°C without trap-
ping the fumes, the residue had no effects. That means: the tumourigenic activity of
the tested “roofing asphalt” is mostly transferred to the fumes.
b) Of the HPLC fractions, B and C proved to be most active; they induced carcinomas in
10 and 17 animals (of 30 in each case). The constituents of fraction B included: ben-
zothiophenes, other aromatic thiophenes and their oxidation products (e.g. sulfones),
phenanthrenes, benzofurans and dibenzofurans. Some of these substances have been
shown to be mutagens or carcinogens. Thus the effects obtained with fraction B are
understandable. This is not the case for fraction C which may have contained mainly
cycloalkenones, cycloalkadienones, dihydrofuranones, dihydroindenones, pyrenes and
fluoranthenes. The authors consider that the tumourigenic effects of fraction C are dif-
ficult to understand because the main components of this fraction are either not or
only very weakly carcinogenic. It is, however, pointed out that fraction C contained
small amounts of methylated four ring or five ring aromatics which are powerful car-
cinogens. Also oxidation products of such aromatic compounds (e.g. hydroxymethyl
derivatives) are conceivable active components for they too have high mutagenic
potential.
Fractions A, D and E had no detectable carcinogenic effects. Combinations of frac-

tions caused tumours only when they contained either fraction B or C; inactive frac-
tions had no modifying effects and neither decreased nor increased the tumour yields
of the active fractions B or C. The combination of all the fractions yielded the same
number of tumours as did the unfractionated fumes of “type III asphalt”.
c) The fractions A, B and E were inactive in the tests for both co-carcinogenicity and
tumour promoting activity.
d) The tumour yield obtained in Sencar mice was the same as that in C3H/HeJ mice. CD-
1 mice were shown to be less sensitive.
5.6.2 Inhalation
Hueper and Payne (1960) exposed 30 guinea pigs (“strain 13”) and 65 “Bethesda Black
rats” in an exposure chamber to fumes of “roofing asphalt, blown from residual oil of
asphaltic type” (Table 14, p. 114). The fumes were produced by heating 1 kg portions of
the material to 121°C to 135°C within the exposure chamber. During each 5-hour daily
exposure, 2 to 10 g of the material was evaporated. The animals were exposed on 4 days
per week for 2 years. Each week a new 1 kg portion of “roofing asphalt” was used. None
of the animals exposed to the fumes developed carcinomas of the lungs or the skin.
However, the following pathological changes were seen in both guinea pigs and rats:
marked chronic fibrotic pneumonitis, peribronchial adenomatosis with proliferation of
the bronchiolar epithelium. In addition in the rats, epithelial metaplasia of the bronchial
mucosa was detected. Bronchiectasis was also observed. Hueper and Payne account for
the lack of lung tumours by suggesting that the carcinogenic substances were destroyed
by oxidation when the fumes were mixed with air.
Simmers (1964) exposed 20 C57 Black mice (10 male and 10 female animals) to an
aerosol of “asphalt droplets” in damp air (the material was described as a “pooled sample
from six California refineries ... contained both steam and air-blown samples”) (Table
14, p. 114). The aerosol was produced in two steps: a) by vigorous shaking of hot “as-
phalt” in hot water, b) nebulization of the initially formed droplets. To avoid whole body
exposure, the experimental animals were placed in plastic tubes and these were arranged
radially around an exposure chamber so that the nose of the animal extended past the
front end of the plastic tube through a hole into the exposure chamber. The animals were
exposed in this way for 30 minutes daily, 5 days weekly for 16.5 months. Of the 20 ani-
mals used, only 3 survived until the end of the study, that is, 3 animals survived 410 30-
minute exposures. Ten mice survived 280 and more exposures. Autopsy was carried out
on 17 animals. In one animal a papillary adenoma was found in the lungs. Histological
examination of the lungs revealed further changes: occasional severe but localized bron-
chitis, dilation of the bronchioles, emphysema-like modification of lung areas.
In another experiment, Simmers (1964) exposed 30 C57 Black mice to fumes from
the same mixture of 3 “air-blown” and 3 “steam-blown samples” (material description as
above) produced by heating about 300 g portions to 121°C. The fumes were conducted
through a chamber containing initially 6 cages each with 5 mice and their food. The ani-
mals were exposed for 6 to 7.5 hours daily, 5 times weekly for 21 months. During the
study a total of 2.25 kg fumes were blown through the chamber. Nine mice survived 401
exposures, 15 survived 300. As the experiment progressed, the inside of the chamber
became covered with a layer of yellowish-brown fluorescent oily material. The food was
also contaminated, but this did not stop the animals eating and their body weight gains
were normal. No gastrointestinal tumours were observed. The control group comprised 6
male mice. Autopsy was carried out on 21 of the 30 exposed animals; a bronchial aden-
oma was found in one animal. The other findings included bronchitis, pneumonitis,
abscesses, loss of cilia from the bronchial epithelium, epithelial atrophy with flattening of
the cells, and thickened alveolar septa. Occasionally local emphysema and peribronchial
infiltration of round cells was seen. Epithelial hyperplasia was also observed. The author
pointed out that mice exposed to cigarette smoke or other air contaminants show similar
symptoms. He suggested that the observed changes represent general effects of air
contaminants containing PAH.
NIOSH scientists (1977b) have attempted to estimate the concentrations in the expo-
sure chamber in Simmers’ (1964) experiment. On the basis of an assumed air flow of 1 to
10 m3/h they calculated exposure concentrations in the range between 74 and 929 mg/m3.
That means that the animals were exposed to much higher amounts of the substances than
a worker in the asphalt industry could be during the whole of his working life. For the
assessment of these studies, knowledge of the experimental conditions described below is
relevant.
Bonnet et al. (2000) and Brandt et al. (2000) have developed an inhalation system
consisting of a bitumen vapour generator and an exposure chamber. In this system the
authors produced bitumen vapour in (total) concentrations of 5 mg/m3 and 50 mg/m3 and
determined the concentrations of 20 selected PAH in this vapour. Arithmetic suggests
that all the components in the 50 mg/m3 atmosphere would be present at 10 times their
concentration in the 5 mg/m3 atmosphere. However, this was true for only few sub-
stances. In particular the highly volatile polycyclic compounds with high saturation
vapour pressures, e.g. acenaphthene, fluorene, phenanthrene, anthracene, fluoranthene,
and naphthalene, were present in the 50 mg/m3 atmosphere at concentrations 25 to 2000
times those in the 5 mg/m3 atmosphere. Under real exposure conditions at the workplace,
however, the authors did not find such differences. They determined the concentration
profiles of polycyclic compounds at 6 mg/m3 (workers exposed at the hot kettle) and at
23 mg/m3 (application of bitumen with a trowel) and found that the profiles were very
similar when the expected (total) concentration differences were taken into account. The
reason for the differences found in the exposure system appears to be the fact that the
substances could not evaporate freely and so the more volatile components reached their
saturation pressure more rapidly. Therefore, in the exposure system an “artificial”
atmosphere is produced which is not like that produced when working with bitumen
outdoors. Such conditions were very likely also present in the exposure chamber used by
Simmers (1964).
5.6.3 Subcutaneous or intramuscular injection
In studies in which bitumen was painted on the skin, the carcinogenic potential of the
substance after subcutaneous or intramuscular injection was also investigated. Simmers
et al. (1959) tested a mixture of 6 different samples of “steam-blown asphalt” and “air-
blown asphalt” from Californian refineries (Table 15, p. 116). From this material, a 1 %
suspension in olive oil was produced and 0.2 ml of the suspension was injected twice
weekly subcutaneously into the interscapular region of 33 male and 27 female C57 Black
mice. The control group comprised 32 male and 28 female mice treated in the same way
with olive oil alone. After 41 weeks the injection frequency had to be reduced to one per
week because too much subcutaneous material accumulated. Autopsy revealed 8
sarcomas, one rhabdomyosarcoma and 7 fibrosarcomas at the injection site. Metastases
were not observed. The authors concluded from their results that the sample of “pooled
petroleum asphalts” contained substances which induced sarcomas after subcutaneous
injection.
In another study, Simmers (1965a) injected mixtures of 3 undiluted “air-refined
asphalts” and “steam-refined asphalts” from “West coast crude petroleums” (Table 15,
p. 117). The material was injected from a syringe whose cylinder was heated via a water-
jacket. The plunger was moved with a calibrated screw. With this apparatus, a single
initial dose of 200 mg of the two asphalt mixtures was injected subcutaneously in the
interscapular region. Groups of 50 C57 Black mice (25 males and 25 females) were used
for each “asphalt” mixture and treated as described above. Animals in which no depot of
the material could be felt after 3 to 4 months were given a second injection. Autopsy was
carried out on 38 of the animals treated with “steam-refined asphalt”; only 15 of these
animals were subjected to histological examination. A lung adenoma was found in one
animal. Malignant tumours were not detected. However, “asphalt” was found in five mice
at locations distant from the injection site, for example, in the abdominal cavity (1
animal), in the thorax (2), near a kidney (1) and in the liver (1). The material at these
locations was enclosed in a thin coating of connective tissue made up of relatively few
cells. Autopsy was also carried out on 38 of the animals treated with “air-refined asphalt”
and 24 of these animals were subjected to histological examination. Five malignant
tumours and five lung adenomas were found. A large rhabdomyosarcoma had developed
at the injection site on one animal. This tumour was centred around some of the injected
“asphalt” material and had formed adhesions with the muscle of the right thigh. At the
same time, metastases were found in the lungs and liver. A second animal (killed after 22
months) had developed a rhabdomyosarcoma of the abdominal wall, also with central
“asphalt” inclusions. A third animal (killed after 23 months) had developed a tumour at
the injection site which originated in skin appendages (hair follicles, sebaceous glands).
This tumour had practically completely replaced the left lung. Simmers ascribed the
tumour-inducing effect of “air-refined asphalt” to the various substances produced by the
air injection although this treatment reduced the level of aromatic compounds in the
asphalt.
Hueper and Payne (1960) carried out studies like those of Simmers (1965a) and used
the same strain of mouse, C57 Black. These studies also demonstrated that the injection
of dissolved bitumen (solvent: tricaprylin) into the subcutis can cause the development of
sarcomas both locally and in distant lymph nodes (Table 15, p. 116).
5.7 Other effects
The proportion of CD4+ T cells, the ratio of the T cell subsets CD4+/CD8+, the monocyte
count and the serum IgG level were significantly increased in 16 Turkish road construc-
tion workers above the values found in a control group of 12 persons who were not
exposed to PAH. The authors considered that these results demonstrate effects of chronic
PAH exposure on the immune system (Karakaya et al. 1999).
The activity of the 5 “asphalt” fractions studied by Sivak et al. (1997) was investi-
gated in an in vitro test for inhibition of intercellular communication. Toraason et al.
(1991) reported that the asphalt fumes and the fractions B, C, D and E inhibited meta-
bolic cooperation in the V79 assay in a concentration-dependent manner. DMSO extracts
of the asphalt fume condensate and all five fractions caused concentration-dependent
inhibition of the communication of human epidermal keratinocytes (Wey et al. 1992).
In an in vitro study, primary alveolar rat macrophages were incubated for 24 hours at
37°C with various concentrations of a condensate of road asphalt fumes (withdrawn from
a tank). Lactate dehydrogenase activity was determined in the culture medium as a
measure of cytotoxicity. Tumour necrosis factor-α (TNF-α) and interleukin-1 (IL-1)
were determined as markers of macrophage function, as was the chemiluminesence
activity (production of reactive oxygen radicals). After exposure of macrophages to con-
centrations below 200 µg/ml, a slight but significant increase in the release of TNF-α
was detected. The in vitro exposure of macrophages to condensate of road asphalt fumes
did not impair the vitality of the cells nor induce extensive release of cytokines or oxi-
dants (Ma et al. 2000).
6 Manifesto (MAK value/classification)
In 1977 bitumen was classified in Carcinogen category III B because of the “numerous
carcinogenic hydrocarbons which it contains”. At that time, animal studies showing that
application of bitumen extracts to the skin caused tumour development were already
available. It was, however, not known whether or not the carcinogenic substances also
occurred in the bitumen fumes formed during work with bitumen and whether the sub-
stances in this form had carcinogenic potential. An association between the exposure of
workers to bitumen fumes or aerosols and the development of tumours could not be
detected.
Since then numerous new publications have presented better analytical data for the
composition of the bitumen itself and of the fumes (vapour and aerosols) formed during
its processing. Studies of the systemic PAH doses taken up by workers during exposure
to bitumen have also been published. The carcinogenicity of such exposures to bitumen
fumes has been investigated in cohort studies and case-control studies. In addition,
numerous animal studies have been carried out, both with bitumen itself and with the
fumes produced during work with bitumen. Analyses have revealed not only PAH but
also naphthalene and other substances with carcinogenic potential. The epidemiological
studies have yielded evidence of carcinogenic effects, especially of an association with
lung carcinoma. However, as was clear at the time of writing the first documentation for
bitumen, co-exposure to PAH in tar, which is known to have carcinogenic effects, means
it is impossible to make a definitive statement about the carcinogenic effects of bitumen
on its own. On the other hand, the DNA damage which was found in exposed workers,
especially in peripheral lymphocytes, was not completely identical with that seen after
exposure to PAH.
The animal studies have yielded no evidence for a tumourigenic effect of inhaled
bitumen fumes, but the fume condensates were tumourigenic when applied to the skin.
On the skin both papillomas and carcinomas developed. The condensates used were not
obtained at the workplace but may be considered qualitatively comparable in composi-
tion. After epidermal application of condensates of bitumen fumes, DNA damage was
induced also in animal studies and also in organs other than the skin. These bitumen
condensates prepared in the laboratory were also genotoxic in various test systems but
only weakly so or only in certain kinds of tests. This could be accounted for by the
presence in the condensates of numerous non-genotoxic substances which inhibited the
genotoxicity of others.
The available studies do not make it clear whether or not bitumen vapour and aerosols
have carcinogenic effects in man. Numerous studies of this question are currently in
progress. Animal studies have yielded unequivocally positive results which suggest that
bitumen vapour and aerosols should be classified in Carcinogen category 2. Although the
positive carcinogenicity data were all obtained with one species, three strains of this
species were involved. The data are also supported by the fact that the fumes to which
workers are exposed contain various components known to be carcinogenic in both
animals and man and that these fumes cause DNA damage in vivo in both workers and
exposed animals. In addition, they are supported by the known genotoxicity of bitumen
vapour and aerosol. A conceivable objection to these arguments is that the exposure
situation in the animal studies which yielded positive results was not identical with the
exposure found at the workplace (ACGIH 2000, Reinke et al. 2000). Nonetheless,
because bitumen itself and the bitumen vapour and aerosols which are inhaled contain
carcinogenic substances and have genotoxic and carcinogenic effects in experimental
animals, bitumen vapour and aerosols are classified in Carcinogen category 2.
Bitumen, vapour and aerosols, is designated with an “H” because skin penetration by
the carcinogenic substances has been demonstrated in animal studies.
To minimize the risk when working with bitumens, the exposure not only to carcino-
genic PAH but also to other carcinogenic substances such as thioarenes, acridines and
carbazoles should be reduced.
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Appendix: Tables 1–15

completed 01.03.2001
Appendix
Table 1. Use of bitumen in Germany in 1998 (Bau-Berufsgenossenschaft 2001)
Use Tonnage per year (1000 t) %
rolled asphalt 2500 74.5

bitumen sheeting 700 20.9
cold bitumen 100 3.0
mastic asphalt, manual application 32 1.0
mastic asphalt, mechanical application 17 0.5
hot bitumen 4 0.1
Total 3353 100.0
Table 2. Concentrations of bitumen fumes (vapour and aerosol) produced during hot processing of
bitumen in various jobs: the concentrations given must be seen as maximum values because the
analytical procedure also detects other hydrocarbons (e.g. from road traffic) (Bau-Berufsgenossen-
schaft 2001)
Job Concentration (mg/m3)
production of bitumen 2.6

production and transport of asphalt
control room 0.8
outdoor working area 0.7
transport of asphalt 4.3
road construction
drivers of finishing machines 12.2
asphalting foremen 10.1
drivers of road rollers 2.9
application of hot bitumen
foam glass installation 19.3
production of bitumen sheeting 4.3
roof construction work
heat welding of bitumen sheeting 8.8
hot casting of bitumen 27.7
work with mastic asphalt
loading outdoors 15.0
loading indoors 38.0
transport with wheelbarrows 53.2
transport with buckets 12.8
smoothing indoors 35.9
smoothing outdoors 8.2
mechanical processing 41.8
Table 3. Levels of polycyclic aromatic hydrocarbons (PAH) in commercially available bitumen products in mg/kg (mean ± standard deviation)
Volume 17
(Knecht et al. 1999)
Level of PAH (mg/kg)
Bitumen product HB 90/100 B 45 B 65 B 80 B 200 B 85/25 B 95/35

Number of batches(n) (4) (8) (5) (9) (6) (4) (4)
naphthalene 0.47 ± 0.19 0.32 ± 0.22 0.29 ± 0.11 0.23 ± 0.20 0.31 ± 0.26 0.74 ± 0.45 1.47 ± 1.31
benzo[b]thiophene 0.14 ± 0.08 0.20 ± 0.26 0.28 ± 0.11 0.19 ± 0.22 0.19 ± 0.14 0.17 ± 0.13 0.23 ± 0.12
acenaphthylene 0.02 ± 0.00 0.03 ± 0.02 0.17 ± 0.25 0.03 ± 0.03 0.10 ± 0.11 0.08 ± 0.08 0.15 ± 0.16
acenaphthene 0.09 ± 0.04 0.15 ± 0.13 0.18 ± 0.07 0.08 ± 0.06 0.14 ± 0.14 0.19 ± 0.11 0.29 ± 0.27
fluorene 1.22 ± 0.52 0.45 ± 0.32 0.69 ± 0.33 0.36 ± 0.38 0.21 ± 0.29 1.76 ± 0.82 2.10 ± 0.93
dibenzothiophene 0.80 ± 0.31 0.98 ± 0.94 0.90 ± 0.83 0.55 ± 0.70 0.75 ± 1.09 2.06 ± 0.81 13.29 ± 18.73
phenanthrene 1.30 ± 0.28 1.59 ± 1.34 1.42 ± 0.78 0.57 ± 0.36 0.93 ± 1.02 5.60 ± 3.40 10.75 ± 7.77
anthracene 0.14 ± 0.05 0.11 ± 0.09 0.14 ± 0.08 0.05 ± 0.04 0.20 ± 0.15 0.68 ± 0.52 1.99 ± 1.88
fluoranthene 0.42 ± 0.21 0.32 ± 0.28 0.29 ± 0.12 0.21 ± 0.16 0.22 ± 0.19 1.37 ± 0.82 2.96 ± 1.40

pyrene 0.69 ± 0.21 0.98 ± 0.80 0.76 ± 0.33 0.63 ± 0.69 1.14 ± 0.68 3.18 ± 1.99 5.08 ± 3.42
benzo[b]naphtho[2,1-d]thiophene 2.34 ± 1.54 3.63 ± 2.65 3.51 ± 1.73 3.48 ± 2.76 4.18 ± 2.60 6.07 ± 4.70 15.06 ± 13.13
benz[a]anthracene 6.15 ± 3.86 1.42 ± 1.15 1.11 ± 0.73 1.97 ± 2.04 1.97 ± 0.80 3.68 ± 1.98 6.22 ± 2.22
chrysene 0.53 ± 0.18 2.59 ± 2.18 4.18 ± 2.17 2.50 ± 2.69 2.53 ± 1.02 4.63 ± 2.23 6.24 ± 2.75
benzofluoranthene a 2.04 ± 1.04 2.48 ± 1.62 1.82 ± 1.22 2.94 ± 2.95 3.12 ± 2.26 3.82 ± 1.61 5.85 ± 2.75
benzo[e]pyrene 5.39 ± 2.51 7.06 ± 3.43 5.18 ± 3.76 5.94 ± 5.02 7.30 ± 3.07 8.92 ± 3.73 10.63 ± 4.52
benzo[a]pyrene 1.20 ± 0.51 2.08 ± 0.95 1.71 ± 1.35 1.41 ± 1.12 1.78 ± 0.67 1.68 ± 0.43 2.74 ± 0.92
indeno[1,2,3-cd]pyrene 0.67 ± 0.22 0.72 ± 0.57 0.43 ± 0.15 0.56 ± 0.39 0.74 ± 0.60 0.98 ± 0.81 1.28 ± 0.65
dibenz[a,h]anthracene 1.47 ± 0.36 1.38 ± 0.83 0.83 ± 0.45 1.53 ± 1.21 2.48 ± 0.95 2.17 ± 0.43 2.04 ± 0.79
benzo[ghi]perylene 4.88 ± 0.36 3.28 ± 1.92 2.85 ± 1.05 2.31 ± 1.33 3.84 ± 1.95 4.38 ± 3.01 5.18 ± 1.61
a
determined as the sum of benzo[b]fluoranthene, benzo[j]fluoranthene and benzo[k]fluoranthene
73
Table 4. Concentrations of bitumen aerosols and PAH in bitumen fumes during production and
a
transport of rolled asphalt B 65 in an asphalt mixing plant (Knecht 2000)
Asphalt mixing plant process Aerosol PAH in aerosol PAH in vapour Total PAH
(processing temperature) (mg/m3) (µg/m3) (µg/m3) (µg/m3)
mixer platform (165°C) 6.25 4.50 79.10 83.60

transport (165°C) 0.35 1.50 4.10 5.60
mixer platform outlet (170°C) 5.94 1.01 80.45 81.46
transport (170°C) 0.41 – 11.30 11.30
a
analyses carried out by the Steinbruch-Berufsgenossenschaft in 1998
Table 5. Concentrations of PAH (µg/m3) in bitumen fumes (aerosols and vapour) during produc-
a
tion and transport of rolled asphalt B 65 (Knecht 2000)
PAH concentration [µg/m3]

Mixer platform Transport Mixer platform Transport
outlet
PAH components Aerosol Vapour Aerosol Vapour Aerosol Vapour Vapour
naphthalene 0.06 44.71 0.11 2.21 48.54 8.58

1-benzothiophene 3.56 0.02 1.31 0.14
acenaphthylene 1.16 0.13 0.57 0.18
acenaphthene 0.10 6.52 0.09 0.25 12.25 0.73
fluorene 0.29 10.36 0.34 0.44 8.18 0.74
dibenzothiophene 0.07 3.01 0.13 0.97 0.18
phenanthrene 0.32 6.91 0.31 0.28 5.20 0.54
anthracene 0.31 0.73 0.07 0.07 1.54
fluoranthene 0.32 0.62 0.31 0.31 0.13 1.22 0.11
pyrene 0.32 0.51 0.27 0.28 0.11 0.67 0.10
benzo[b]naphtho 0.61 0.38 0.17
[2,1-d]thiophene
benz[a]anthracene 0.45 0.36
chrysene 0.57 0.29 0.17
b
benzofluoranthene 0.46 0.23
benzo[e]pyrene 0.08 0.13
benzo[a]pyrene 0.13 0.08
indeno[1,2,3-cd]pyrene
dibenz[a,h]anthracene 0.31
benzo[ghi]perylene 0.19
Total PAH 4.50 79.10 1.50 4.12 1.01 80.45 11.30
a
analyses carried out by the Steinbruch-Berufsgenossenschaft in 1998
b
determined as the sum of benzo[b]fluoranthene, benzo[j]fluoranthene and benzo[k]fluoranthene
Volume 17
Table 6. Studies of markers of bitumen exposure in groups of exposed persons
Collective Method Exposure situation Results Comments References
12 roofing workers detection of B[a]P concentration in roofing workers: 5.2 ± 4.0 fmol/µg; weak correlation with Lee et al.
(collective as in Herbert albumin-PAH air: 0.6–1.3 µg/m3 controls: 3.3 ± 2.1 fmol/µg DNA adducts 1991
et al. 1990), adducts in serum (0.05 < p < 0.1); increased about (0.05 < p < 0.1)
12 control persons; with ELISA and 1.6-fold; “… of borderline signifi-
matched for age, sex, the monoclonal cance”; in contrast, level of DNA-
smoking; antibody 8E11 adducts in the peripheral white blood
USA cells of the same collective about 10
times higher than in the controls
(Herbert et al. 1990)
12 male roofing workers detection of DNA B[a]P concentration in DNA adducts with aromatics in 10 for exposed persons sig- Herbert et
during removal of an old adducts in white air: 0.6–2.0 µg/m3; roofing workers (up to 10 adducts/ nificant correlation with al. 1990
pitch roof and installation blood cells by 32P PAH concentration in 108 nucleotides) and two control PAH level (p = 0.01) and
of a new “asphalt” roof; postlabelling air: 6.1–32.4 µg/m3 persons (0.2–0.3 adducts/108 nucleo- B[a]P level (p = 0.04) in
12 control persons tides) skin swabs after the shift
(hospital staff or patients) but not with the corres-

matched for age, sex, ponding concentrations in
smoking; the workplace air;
USA no association with
smoking habits
25 “roofers” exposed to assay of DNA air monitoring and results as % of “tail” DNA: some of the “roofers” Reid et al.
hot asphalt, strand breaks in assay of average values at the start of the week (n = 18) were also exposed 2000
12 “construction peripheral blood 1-hydroxypyrene in in “roofers” 15.1 ± 0.6 and in control to tar; the % “tail” DNA
workers” not exposed to leukocytes at the urine (no data) persons 14.3 ± 1.1; values were lower in the
asphalt during the beginning and end at the end of the week 16.7 ± 0.6 in “roofers” who were not
previous 5 years of the working the “roofers” and 14.5 ± 1.0 in the also exposed to tar than in
week by means of control persons those who were.
the comet assay
75
76
Table 6. continued

7 “roofers”, assay of alkali- for roof construction roofing workers: a) level of alkali- smokers are under- Fuchs et al.
18 road paving workers, induced DNA work oxidized bitu- induced DNA strand breaks signi- represented in the control 1996
9 bitumen painters; strand breaks mens were used and ficantly higher than in controls group; controls on average
34 control persons (office (alkaline elution) heated to 350°C; b) significantly higher levels of younger than exposed
workers, students); and DNA adducts exposure to products strand breaks on Fridays than on persons; DNA adducts
West Germany; (32P postlabelling) derived from coal is Mondays; assayed in only 14 of the
median age of exposed in peripheral conceivable during road construction workers: the 34 exposed persons;
persons 43 years, mononuclear blood repair of old roofs; for
level of alkali-induced DNA strand
controls 27 years cells painting, the bitumen breaks not significantly increased cigarette smoking has little
was diluted with relative to that in controls but effect on the level of
kerosine or gas oil. increased during the working week alkali-induced DNA strand
bitumen painters: relatively high breaks
level of alkali-induced DNA strand
breaks on Mondays; levels decreased
during the working week (formation
of cross-links?);
number of DNA adducts (max.
0.3/108 nucleotides) increased in road
workers and bitumen painters with
increasing time in the job
18 road construction assay of SCE in work with asphalt SCE incidence in road workers differences in SCE inci- Bittighofer
workers, peripheral accounted for only one 6.6 ± 0.6, in controls 5.0 ± 1.0 dence were also found et al. 1984
12 control persons lymphocytes third of the working (p = 0.001) between the smokers and
(gardeners); hours non-smokers of the two
Germany collectives;
no increase in mutagen-
icity of extracts of urine
Volume 17
from non-smoking road
workers (Salmonella
mutagenicity test with
TA97, TA98, TA100)
Volume 17
Table 6. continued
28 men, 21 employed as assay of SCE and 1-hydroxypyrene incidence of SCE hydroypyrene excretion Burgaz et
rakermen in road paving micronuclei in excretion: bitumen in bitumen workers 5.1 ± 0.6, not correlated with al. 1998
operations, 7 prepared a peripheral workers 0.78 ± 0.46 in controls 4.7 ± 0.7 (p < 0.05); incidence of SCE or
hot mix of stone chips lymphocytes µmol/mol creatinine, incidence of micronuclei micronuclei;
and bitumen in asphalt controls: 0.52 ± 0.44 in bitumen workers 2.3 ± 0.4 ‰,
plant; µmol/mol creatinine in controls 1.8 ± 0.3 ‰, (p < 0.0001) authors’ claim that the
28 controls (university determined values reveal
and hospital employees); significant differences
Turkey seems questionable
28 non-smoking male exposure total PAH in inhaled 1-hydroxypyrene in urine of bitumen motor car exhaust gases Järvholm et
“road pavers”; determined with air 0.2–24 µg/m3; workers 0.96 µmol/l, control persons did not affect the results al. 1999
30 non-smoking control personal samplers; asphalt free of coal tar 0.6 µmol/l; the difference is signifi- because neither coronene
persons; assay of heated to 150–200°C cant; no significant differences nor benzo[ghi]perylene
Sweden 1-hydroxypyrene, between values before and after the were found in the samples
SCE, micronuclei shift; no significant differences

in peripheral between exposed and control persons
lymphocytes in incidence of SCE or micronuclei
16 men “employed in urine concentrates concentrations in the mutagenicity only detectable with Pasquini et
road bituminization”, tested for mutagen- inhaled air: TA98 plus S9; not increased by al. 1989a
27 controls (office icity in Salmonella bitumen aerosols added β-glucuronidase; numbers of
workers); typhimurium 0.6–0.8 mg/m3, mutants expressed per unit of
Italy (TA98, TA100) B[a]P 0.03–0.6 µg/m3 creatinine significantly higher in non-
with and without smoking exposed persons than in
metabolic non-smoking controls
activation
17 “asphalt exposed” assay of DNA levels of DNA adducts higher in 8 of Zhou 1997
16 “controls” adducts in 9 exposed persons than in controls;
desquamated difference not statistically significant
urothelial cells (p > 0.05)
(32P postlabelling)
77
78
Table 6. continued

healthy male workers assay of SCE in see under “Collective” incidence of SCE: no clear correlation of SCE Hatjian et
P1: 6 “pavers” (road peripheral incidence with summed workplace al. 1997
repair work with “hot lymphocytes group P1: 0.059 ± 0.001 concentrations of selected 2-ring
asphalt”) to 5-ring aromatics
P2: 10 “pavers” (repair of group P2: 0.063 ± 0.002 difference between P1 and C
road surfaces with “50 statistically significant (p < 0.05);
penetration grade P2, R1 and R2 all significantly
bitumen”, 150–180oC) increased above both C and M
R1: 9 “roofers” group R1: 0.064 ± 0.003
(waterproofing roofs
“using felt ... and hot
oxidised grade bitumen”,
tank temperature about
300oC)
R2: 4 “roofers” (work group R2: 0.073 ± 0.007
with mastic asphalt, “25
penetration oxidised
blown bitumen”, 190oC)
2 control groups:
M: 15 workers, some group M: 0.054 ± 0.003
with comparable jobs but
“not occupationally
exposed to PAHs”
C: 6 “administrative group C: 0.048 ± 0.001
office workers ... not
Volume 17
occupationally exposed
to PAHs”
B[a]P, benzo[a]pyrene; SCE, sister chromatid exchange; PAH, polycyclic aromatic hydrocarbons
Volume 17
Table 7. Cohort studies of the carcinogenicity of bitumen
Study objective Exposure of sub-groups Results for cancer risks (95 % confidence Included in IARC meta- References
interval) analysis / comments
462 asphalt workers from 25 oil asphalt workers, ≥ no Baylor and

refineries, 379 controls; 5 years in the job no statistical analysis Weaver
USA (average 15.1 years) 1968
cancer mortalitiy (1960–1971); roofers, waterproofers all: SMR = 1.5 (1.3–1.6) yes Hammond
5939 male employees identified lung: SMR = 1.6 (1.3–1.9) smoking habits not recorded, et al. 1976
from union dossiers compared oral cavity, throat, larynx, simultaneous exposure to
with total male population; oesophagus: SMR = 2.00 (1.7–2.8) benzo[a]pyrene
USA stomach: SMR = 1.7 (1.1–2.5)
bladder: SMR = 1.7 (0.9–2.9)
lung cancer on the death roofers (estimated lungs: SMR = 5.0 (2.5–8.9) yes Menck and
certificate and lung cancer population at risk, 2000) Henderson
incidence from the Los Angeles 1976
County Surveillance Program
(white males)

cancer mortality in males roofers and slaters PMR = 1.2 (1.1–1.3) no Milham
(proportional cancer mortality study included non-exposed 1982
1950–1979), persons, increased incidence
Washington State of deaths from silicosis
road graders and pavers, increased PMR for lung cancer
machine operators and
excavators
proportional cancer mortality; 327 highway cancer risks yes Maizlish et
1570 employees of the maintenance workers all: PMR = 1.2 (0.9–1.5) increased skin cancer risk al. 1988
Department of Transportation, digestive organs: PMR = 1.5 (1.0–2.2) possibly caused by exposure
California lungs: PMR = 1.0 (0.6–1.5) to sunlight (California),
persons exposed simul-
taneously to herbicides
79
80
Table 7. continued

cancer morbidity, 1486 men in 11 production of hot-lay all: SMR = 1.5 (0.9–2.4) yes Povarov et
Estonian factories (1974–1984) asphalt, ≥ 3 years in the lungs: SMR = 2.1 al. 1988
job, 10369 person years;
benzo[a]pyrene-
concentration 0.002–
0.007 g/m3 (?)
cancer mortality (1945–1984); highway maintenance significantly increased numbers of deaths yes Bender et
4849 men employed by the workers (total collective) from: leukaemia (30–39 years in the job): no contact with tar since the al. 1989
Minnesota Department of SMR = 4.25 (1.71–8.76) 1930s; significantly reduced
Transportation for at least one cancer of the urinary tract and kidneys incidence of death from lung
year (40–49 years latency period): cancer in the total collective
SMR = 2.92 (1.17–6.02) (SMR = 0.69; 95 %
CI = 0.52–0.90)
cancer mortality (1970–1980); asphalt plants, roofing for the years 1975–1980 significant yes Hansen
1320 unskilled asphalt industry felt plants, one tar plant; increase in the incidence of 1989a
workers compared with 43024 employed at the time of a malignant neoplasms:
“unskilled men in occupations census on 09.11.1970 SMR = 1.59 (1.06–2.28)
without exposure to carcinogens”; and brain tumours:
Denmark SMR = 5.0 (1.03–14.6)
in persons over 45 years old
cancer morbidity (1959–1984); heavily exposed, mean significant increase in tumour incidence in yes Hansen
679 men working with mastic asphalt vapour persons 40 to 89 years old after correction for smoking 1989b
asphalt, (6692 person-years at concentrations: all tumours: SIR = 1.95 (1.53–2.44) habits and town-country
risk), 4 mg/m3 (road workers) oral cancer: SIR = 11.1 (1.35–40.1) differences: SMR=2.14
controls: total male population 20 mg/m3 (floor layers) oesophagus cancer: SIR = 6.98 (1.44–20.4) (respiratory tract cancer) or
Volume 17
(1958–1982), rectal cancer: SIR = 3.18 (1.28–6.56) SMR = 2.06 (digestive tract
Denmark lung cancer: SMR = 2.29 (1.75–2.93 cancer)
Volume 17
Table 7. continued
cancer mortality; 679 men see Hansen (1989b) all tumours (whole collective): yes Hansen
working with mastic asphalt SMR = 2.25 (1.72–1.88) lung cancer (adjusted for 1991
(follow up of the cohort from all data for persons aged 40 to 89 years smoking and town/country
Hansen 1989b until the year lung cancer: differences): SMR = 2.24
1986), 7434 person-years at risk; SMR = 2.90 (1.88–4.29) (95 % CI = 1.45–3.30)
control total male population of cancer except lung cancer:
Denmark 1960–1985 SMR = 2.00 (1.41–2.76)
cancer mortality and incidence; asphalt workers stomach cancer: SIR = 2.1 (0.9–4.1) yes Engholm et
2572 asphalt workers, 704 lung cancer: SIR = 1.2 (0.5–2.4) al. 1991
roofing workers; Sweden lymph vessels and blood: SMR = 0.28,
SIR = 0.69
leukaemia: SIR = 2.12
all kinds of cancer: SIR = 0.86

roofing workers stomach cancer: SMR = 2.30, SIR = 1.98 yes
lung cancer: SMR = 2.79, SIR = 3.62
lymph vessels and blood: SMR = 2.68,
SIR = 1.23
leukaemia: SIR = 2.26
all kinds of cancer: SIR = 1.34
cancer mortality; roofers and slaters lung: SMR = 3.0 (1.2–7.2) yes NIOSH
USA 1992
cancer mortality in men (death roofers significantly increased incidence of cancer no Minder and
register 1979–1982 and census of the oral cavity and larynx: PMR = 3.30 results given only for the oral Beer-
data 1980); (1.21–7.20), cavity Porizek
Switzerland 6 deaths 1992
81
82
Table 7. continued

mortality among 9585 men highway maintenance lung: SMR = 1.3 (1.0–1.5) no Hwang et
(1958–1980) employed for at workers (total collective) rectum: SMR = 1.7 (1.0–2.7) study not published until after al. 1995
least one year by the New York 1994
State Department of co-exposure to solvents and
Transportation; control: total male pesticides
population of New York
109000 cases of cancer in persons roadbuilding hands all tumours: SIR = 1.1 (1.0–1.1) no Pukkala
born in the years 1906–1945, lips: SIR = 1.2 (0.8–1.8) published after 1994 1995
based on 1970 census data with lung: SIR = 1.1 (1.0–1.3)
follow-up 1970–1971;
Finland
lung cancer (men):
SIR = 3.50 (2.07–5.53)
SIR* = 3.25 (1.92–5.13)
all types of cancer (men):
SIR = 2.12 (1.49–2.94)
SIR* = 2.07 (1.45–2.86)
cancer mortality and incidence bitumen construction lung cancer: SMR = 1.4 no Partanen et
among 9643 persons employed workers not published until after al. 1997
for at least 3 months by 6 road non-bitumen construction lung cancer: SMR = 1.4 1994; part of the IARC study
construction firms (1968–1984); workers
Finland exposed to bitumen lung cancer: SIR = 1.4 (0.9–1.9)
fumes
Volume 17
Volume 17
Table 7. continued
analysis of pooled data from two road construction work lung cancer risk significantly increased for no Brüske-
case-control studies of lung with bitumen or asphalt the following groups: published after 1994 Hohlfeld et
cancer in men in East and West ever exposed: OR adjusted for smoking and al. 1998
Germany; 3498 cases, 3541 OR = 1.86 (1.25–2.76) exposure to asbestos; authors
population controls exposed for > 20 calender years: claimed that differentiation
OR = 2.47 (1.05–5.83) between exposure to black
first exposed during the years 1946–1955: coal tar and to bitumen was
OR = 2.57 (1.27–5.18) not possible
first exposed during the years since 1956:
OR = 1.66 (1.01–2.73)
last year of exposure 1976:
OR = 2.07 (1.17–3.67)
?? 7886 workers (Germany) all tumours: SMR = 1.2 (0.9–1.6) no Frentzel-
exposed to bitumen and lung: SMR = 2.0 (1.2–3.2) part of the IARC study Beyme et

not to tar al. 2000
?? 8964 asphalt workers lung cancer: SIR = 1.3 (1.0-1.7) no Randem et
(Norway) part of the IARC study al. 2000
PMR proportional mortality ratio

SIR standardized incidence ratio
SIR* standardized incidence ratio adjusted for social status
SMR standardized mortality ratio or standardized morbidity ratio
OR odds ratio
CI confidence interval
83
84
Table 8. Case-control studies of the carcinogenicity of bitumen
Collective studied Exposure conditions, Results Included in IARC meta-analysis/ References

exposed groups (95 % confidence interval, comments

unless otherwise specified)
? roofers and slaters no NIOSH

1977a
265 cases with primary liver tumours, road building hepatocellular carcinomas: no Stemhagen
530 controls; OR = 2.6 (0.8–8.2) only liver tumours et al. 1983
New Jersey
212 cases of bladder cancer, work with kerosine OR = 3.1 (0.9–11.0) no Mommsen
259 controls; or asphalt not clear whether persons were et al. 1983,
Denmark exposed to asphalt Mommsen
and
Aagaard
1984
80 cases of hepatocellular carcinoma, highway and street no Austin et
146 hospital controls; construction only liver tumours al. 1987
multicentre study, USA
asphalt 7 cases, 5 controls
OR = 3.2 (0.9–11)
763 cases of lung cancer (white males), roofers and slaters OR = 1.7 (0.7–4.4) yes Schoenberg
900 population controls; New Jersey adjusted for smoking included in Vineis et al. (1988) et al. 1987
petroleum workers collective included in the meta-
analysis of Vineis et al. (1988)
(see below)
96 cases of cancer of the renal pelvis asphalt, tar (men) OR = 5.5 (1.6–19.6) yes Jensen et
and urethra (benign and malignant), 294 adjusted for smoking al. 1988
Volume 17
hospital controls;
eastern Denmark
Volume 17
Table 8. continued

826 cases of bladder cancer tar, asphalt OR = 1.4 (0.8–2.7) yes Risch et al.
792 population controls at least 6 months adjusted for smoking 1988
Canada exposure
at least 6 months OR = 3.1 (1.2–9.7)
exposure and adjusted for smoking
employed 8 to 28
years before
diagnosis
trend with duration OR = 2.0 (1.1–5.0)
of exposure (10 year adjusted for smoking
intervals)
2973 cases of lung cancer roofing and asphalt 45 cases, 37 controls yes Vineis et

3210 controls workers OR = 1.4 (0.9–2.3) meta-analysis of 5 case-control studies al. 1988
USA including Schoenberg et al. 1987 (see
above); OR adjusted for age and
smoking habits; for 2/5 sub-collectives
data only from next of kin interviews
4431 cases of lung cancer (white males) roofers 6 cases, 7 controls yes Zahm et al.
11326 cancer controls from Missouri; OR = 2.1 (0.6–8.2) only the job at the time of diagnosis 1989
Cancer Registry recorded
pavers, surfacers, 32 cases, 64 controls yes
material moving OR = 0.9 (0.6–1.5)
equipment operators
85
86
Table 8. continued


3730 cases of cancer (type specified) asphalt stomach cancer: listed are only the data for cancers Siemiatycki
among men aged 35 to 70 years any exposure OR = 1.0 (90 % CI = 0.5–1.8) with statistically significant (p ≤ 0.10, 1991
533 population controls cancer of the large intestine: single sided) associations in at least
Canada (region Montreal) OR = 1.6 (90 % CI = 1.1–2.5) one exposure range
prostate cancer:
OR = 1.1 (90 % CI = 0.6–2.0)
bladder cancer:
OR = 0.9 (90 % CI = 0.5–1.6)
non-Hodgkin’s lymphoma:
OR = 2.0 (90 % CI = 1.0–4.0)
substantial exposure stomach cancer:
OR = 2.0 (90 % CI = 1.0–4.1)
cancer of the large intestine:
OR = 1.0 (90 % CI = 0.5–2.1)
prostate cancer:
OR = 3.0 (90 % CI = 1.0–9.0)
bladder cancer:
OR = 2.2 (90 % CI = 1.0–4.9)
non-Hodgkin’s lymphoma:
OR = 1.5 (90 % CI = 0.4–5.1)
1793 cases of lung cancer roofers and slaters 7 cases, 6 controls yes Morabia et
3228 controls OR = 2.1 (0.7–6.2) OR adjusted for age, ethnic group, al. 1992
multicentre study, USA region and smoking habits
Volume 17
Volume 17
Table 8. continued

622 cases of non-Hodgkin’s lymphoma, paving occupations 3 cases, 2 controls no Blair et al.
1245 age-matched controls OR = 3.4 (0.6–20.8) OR adjusted for factors including 1993
Iowa and Minnesota smoking and family history of
lymphoproliferative disorders,
adjusted OR for exposure to asphalt
and creosote: 1.0 (95 % CI = 0.7–1.5)
with 53 cases and 105 controls; no
dependence on exposure level
144 cases of lung cancer asphalt fumes 60 cases, 122 controls; adjusted no Chiazze et
260 controls; ≥ 0.01 mg/m3 OR = 1.1 (0.5–2.7) presumably published too late al. 1993
Owens Corning Fiberglas Co., Ohio
OR odds ratio

CI confidence interval
87
88
Table 9. Effects of bitumen on DNA or nucleosides in vitro
Test material Test system Results Comments References

bitumen “obtained from the PM2 DNA to test for DNA single strand breaks: none the authors conclude from their results Hong and
distillation of crude oil from single strand breaks double strand breaks: none that bitumen is directly responsible for Lee 1999
Hanwha Energy (South Korea)”, λDNA linearized with the production of reactive oxygen
dissolved in tetrahydrofuran; hydroxylation of deoxy- species
Hind III to test for double guanosine (8-OH-dG/105 dG):
tested alone or in combination strand breaks It is a deficit of the publication that for
with UVA control: 1.7
8-hydroxylation of tetrahydrofuran: 13.0 neither single nor total effects were
deoxyguanosine bitumen (100 µg/ml): 14.6 errors given.
in all tests: bitumen
concentration 100 µg/ml; UVA (6 mJ/cm2): 14.2
1–24 h, ± UVA in various doses UVA (12.0 mJ/cm2): 15.5
UVA (24 mJ/cm2): 18.4
bitumen (100 µg/ml) plus UVA
(6 mJ/cm2): 22.7
(12 mJ/cm2): 24.0
(24 mJ/cm2): 33.7
condensates of “bitumen of incubation of DNA with adducts detectable after DNA adducts only detectable after Genevois et
45/60 pen hardness derived from and without microsomes metabolism of the condensates metabolism absence of the AH-receptor al. 1998
a heavy Venezuela crude oil”, from wild type mice or by microsomes from wild type caused only a 30 % reduction in the
obtained at 160°C and 200°C, mice deficient in AH mice and from mice deficient in adduct level (compensation by CYP3A4,
dissolved in acetone receptor and CYP1A2 AH-receptor and CYP1A2 in 2C and prostaglandin H synthase?)
(C57BL/6) and microsomes the presence and absence of Condensates from heated bitumen and
from yeast which expressed NADPH2 adducts formed after from coal tar (for comparison) yielded
human CYP1A1, 1A2, 3A4 metabolism by microsomes very different adduct patterns.
and 2C9; DNA adducts from yeasts expressing Temperature at which bitumen
Volume 17
detected by CYP1A1, 3A4, 2C9 and 1A2 condensates were produced did not
32
P-postlabelling affect the adduct pattern and had little
effect on the yield.
Volume 17
Table 9. continued
Test material Test system Results Comments References
condensates from two samples of calf thymus DNA, Aroclor- numerous adducts with all four bitumen condensates more De Méo et
industrial bitumen (penetration 1254-induced rat liver kinds of adducts were formed than with al. 1996
factors 45/60 and 160–210) microsomes, condensate of black coal tar
32
obtained by heating each sample P-postlabelling
to 160°C and 200°C
PBFC1: “Paving bitumen fume calf thymus DNA, Aroclor- numerous adducts, highest adduct levels were at least two orders of Akkineni et
condensate no. 1 sampled from a 1254-induced rat liver yield with the sample RBFC magnitude lower than with black coal al. 2001
hot storage tank (156°C) microsomes, tar; parallel experiments with crude oil
32
containing bitumen P-postlabelling distillates yielded comparable adduct
(PG64-S22/ca. Pen 65)” patterns after metabolism with rat and
PBFC2: “Paving bitumen fume with human microsomes.
condensate no. 2 sampled from a
hot storage tank (163°C)
containing road bitumen
(AC-20/ca. Pen 40)”

PBFC3: “Paving bitumen fume
condensate no. 3 sampled from a
hot storage tank (146°C)
containing road bitumen
(PG64-22/ca. Pen 65)”
RBFC: “Roofing bitumen fume
condensate sampled from a hot
storage tank (227°C) containing
roofing bitumen (type IV)”
89
90
Table 10. Mutagenicity tests with Salmonella typhimurium
Sample tested Test system Metabolic Results a Comments References

activation

4 “petroleum asphalt paints” (solid petroleum TA98, TA100, TA1535, without/with –/– low levels of xylene (≤ 3 %) Robinson
asphalt material cut back to 64 % solid with TA1537, TA1538; et al. 1984
mineral spirits), dissolved in DMSO Aroclor-induced rat liver
microsomes
“vacuum residuum (D6), boiling range > 1070°F”, TA98; 8-fold increased with – Blackburn
dissolved in cyclohexane and extracted with concentration of hamster et al. 1986
DMSO microsomes, increased
NADPH concentration
(8 mM)
asphalt for road construction (“petroleum TA98, TA100 without/with –/– negative results also with Pasquini et
derivative, 80–100 penetration grade”) separated 50 % S9 (instead of 10 %); al. 1989b
from aggregate with benzene, after precipitation in contrast, positive results
of asphaltenes with n-heptane, extracted with obtained under the same test
DMSO to concentrate the PAH, residue dissolved conditions with “petroleum
in cyclohexane pitch”
“5 whole asphalt samples from actual road paving TA98, TA100; without/with DMSO weakly positive result b in Gage et al.
operations in Oklahoma”, extracted with DMSO commercial rat liver S9 extract: TA100 with metabolic 1991
or destilled water (pH 5.0) –/+ activation with 1 of 5 DMSO
H2O extract: extracts; H2O extract of a
–/+ a different sample doubled the
number of colonies of TA98
in the presence of S9
(1) “naphthenic AC-20 asphalt produced from a TA98; without/with –/– abstract McGowan
mix of Nigerian light, Maya and Alaska North modified Salmonella et al. 1992
Slope crudes”; mutagenicity test without/with
Volume 17
(2) “naphthenic coastal residuum”; (Blackburn et al. 1986) –/–
in both cases the sample was dissolved in
cyclohexane and extracted with DMSO
Volume 17
Table 10. continued

activation
“bitumen (vacuum residue; CAS 64741-56-6), TA98; with + (?) Booth et al.
ref. no. TMC21565”, DMSO extract prepared modified Salmonella (mutagenicity 1998
according to the method IP 346 mutagenicity test index: 0.5,
(Blackburn et al. 1986) no other data)
(1) solid bitumen extracted with benzene, TA98, TA100 without/with –/– no positive results even Monarca et
asphaltenes precipitated with n-heptane, heptane- with higher S9 al. 1987
soluble substances extracted with DMSO without/with concentrations (50 %
(2) two bitumen aerosol samples from workplaces, TA98, TA100 –/– instead of 10 %);
filter from the sampling tubes extracted with preparation and use of S9
diethyl ether and then with acetone, extracted not described in detail
material dissolved in DMSO
bitumen B-80, bituminous concrete 0.8; 7 strains (no other details) ? – inadequately documented Bittighofer
in both cases the dichloromethane-soluble fraction et al. 1983
and the distillate, vapour and fumes obtained at

250°C were tested
condensates from: other tester strains were Sonntag
(1) welded polymer bitumen sheeting TA97c, TA98, TA100 without/with –/– mentioned but the results and
(PYE-PV200S5, DIN 52133) not presented Erdinger
(2) welded polymer bitumen sheeting TA98, TA100, TA1535c without/with –/– 1989
(PYE-G200S4, DIN 52133)
(3) welded bitumen sheeting TA98, TA100 without/with –/–
(G200S4, DIN 52131)
(4) bitumen sheeting for roofing TA98, TA100, TA1535c without/with –/–
(G200DD, DIN 52130)
(5) bitumen sheeting for roofing TA98, TA100 without/with –/–
(PV200DD, DIN 52130):
in each case fumes produced at 80°C and trapped
with a cold finger or adsorbed on XAD resin,
then extracted with acetone and pentane c
91
92
Table 10. continued

activation

condensates from: Sonntag
(1) bitumen E (Venezuela) – TA98, TA100 without/with (+)/(+) (1) maximum effect and
oxidized bitumen 85/25 doubling of the number of Erdinger
revertants 1992
(2) bitumen S (“Mittel-Ost”) - TA98, TA100 without/with –/– (2) with TA100 and no
oxidized bitumen 85/25 metabolic activation, an
in each case the samples were heated to 190°C, increasing trend with
the fumes transported in an air stream to a increasing dose is evident
condenser at –38°C and the condensate extracted
with acetone, concentrated and then dissolved in
DMSO or in a 3 % aqueous Tween solution
condensates from: Machado et
(1) 18 “paving asphalts representing 14 different TA98; with + (1) and (2): no good al. 1993
crude oil sources and various process conditions”, modified Ames test with correlation of mutagenicity
obtained at 163°C, extracted with DMSO; Aroclor-induced hamster index with levels of B[a]P
S9 (Blackburn et al. 1986) or selected neutral PAH
(2) 2 “asphalts conforming to ASTM type III TA98; with + (2) numbers of revertants
roofing specifications” modified Salmonella not increased with the
a) “air-blown without the use of a catalyst” mutagenicity test with temperature used to
b) “air-blown using ferric chloride as a catalyst” Aroclor-induced hamster produce the fumes, not
condensates obtained at 232°C and 316°C, S9 (Blackburn et al. 1986) affected by the asphalt
extracted with DMSO production methods or the
origin of the oil
Volume 17
Volume 17
Table 10. continued

activation
condensates from two samples of industrial TA98, TA100, YG1041, without/with –/+ mutagenic activity seen De Méo et
bitumen (45/60 pen and 160–210 pen), YG1042; (tested with the only after metabolic al. 1996
obtained by heating to either 160°C or 200°C modified method of De activation, effect greatest
Méo et al. (1996)); rat in TA100 (3175–1748
liver microsomes induced revertants) with all 4 fume
with Aroclor 1254 condensates
a
standard criterion for a positive result: with at least one concentration of the test substance double the number of revertants in the solvent control
b
positive result defined as double the control number of revertants with at least two concentrations of the test substance
c
only tested without metabolic activation
DMSO dimethylsulfoxide
AHH arylhydrocarbon hydroxylase
B[a]P benzo[a]pyrene
PAH polycyclic aromatic hydrocarbons

93
94
Table 11. Effects of bitumen on cultured mammalian cells or tissues
Sample tested Test system Results Comments References

bitumen “obtained from the distillation of HL60 cells a; 24 h incu- cell proliferation % DNA cross-linked to protein: Hong and
crude oil from Hanwha Energy (South bation with 10 µg/ml; slightly inhibited; control 4.8 % Lee 1999
Korea)”, dissolved in tetrahydrofuran cell proliferation, DNA- increase in the bitumen [10 µg/ml] 6.3 %
protein cross-links proportion of DNA
assayed (after DNA pre- cross-linked to
cipitation, treatment with protein
proteinase K; result is
% DNA cross-linked)
“black bitumen paint (containing 57 % v/v normal skin from foetal skin: + significance < 0.005; Schoket et
bitumen), purchased locally”, 4 % or 20 % mastectomy samples and adducts: al. 1988b
v/v in tetrahydrofuran; foetuses; 24 h incubation; breast skin from test 0.16–0.22 fmol/µg DNA; control
concentrations tested: 3 mg and 15 mg assay of DNA adducts by mastectomy 0.06–0.1 fmol/µg DNA (1 fmol/µg DNA
32
bitumen in 150 µl tetrahydrofuran P-postlabelling patients: + corresponds to 33 adducts/108 nucleotides)
conclusion: “.. it would seem prudent to
regard preparations of bitumen as
representing a hazard to man comparable
to that of coal-tar and creosote.”
“vacuum residuum (petroleum)” samples test for forward muta- with metabolic mutation frequency increased 2 to 3 times API 1984c,
81-13 und 18-14 (CAS 64741-56-6), taken tions in L5178Y mouse activation: + at high concentrations with metabolic 1984d
up in acetone, because of poor solubility lymphoma cells, meta- without metabolic activation
exact concentration cannot be given bolic activation with S9 activation: –
from rat liver induced
with Aroclor
3 condensates of “straight run, vacuum- test for chromosomal with metabolic and no trend towards an increase with Reinke et al.
distilled 85/100 grade asphalt cement aberrations in CHO cells activation: – increasing concentration 2000
derived from a blend of heavy Canadian (a cell line from Chinese without metabolic
Volume 17
sour crude oil supplied by Koch Materials” hamster ovary) activation: –
obtained in the laboratory at 149°C and
316ºC and from the gas phase of a hot tank
(147–157ºC)
Volume 17
Table 11. continued
Sample tested Test system Results Comments References
“fume condensates of type I (for flat roofs) micronucleus test with both condensates: + 4 concentrations of each tested (63– Qian et al.
and type III (for steep roofs) roofing V79 cells (Chinese 250 µg/ml); significant increase at all 1996
asphalts, .. from local supplier”, obtained at hamster lung fibroblasts) concentrations; 70 % of micronucleated
316°C by the method of NIOSH 1989, cells contained kinetochor-positive
taken up in cyclohexane/acetone (50/50), micronuclei
material dissolved in DMSO “ .. condensates are aneuploidogens and
possess some clastogenic activity ..”; “..
aneuploidy action of .. fume condensates
by blocking spindle assembly via
interaction with tubulin.”
condensate of “type III roofing asphalt”, micronucleus test with dose-response curve steepest with fractions Qian et al.
obtained at 316°C b by the method of V79 cells B and C, 1999
NIOSH 1989, dissolved in (reveals chromosomal dose-dependent increase in the incidence
cyclohexane/acetone (50/50), after drying breaks and aneuploidy) of binucleated cells with the whole
taken up in DMSO; HPLC fractionation condensate and fractions B and C

whole condensate, whole condensate: + “.. roofing asphalt fumes contain chemicals
Fraction A (constituents include C19–C35 Fraction A: – .. which can induce micronuclei and inhibit
alkanes, alkylated benzenes), cytokinesis by damaging spindle fibers and
Fraction B (constituents include Fraction B c: + microfilaments ..”
benzothiophenes and dibenzothiophenes),
Fraction C (constituents include Fraction C c: +
phenylethanones and dihydrofuranones),
Fraction D (constituents include phenols Fraction D: +
and carbazoles),
Fraction E Fraction E: +
a
human leukaemia cell line
b
“This temperature is similar to the high overheat temperature reached in asphalt roofing kettles in the field.”
c
tumourigenic on mouse skin
95
96
Table 12. Genotoxic effects of bitumen in experimental animals
Sample tested Experimental Test system; administration route Results Comments References
animals

“black bitumen paint, ..  assay of DNA adducts by means 1) DNA adducts (detection topical application results Schoket et
purchased ... (containing mouse of 32P-postlabelling after limit 0.09 fmol/µg DNA = in uptake and relocation of al. 1988a
57 % v/v bitumen)”, (Parkes ) 1) single applications of 26.7 or 3 adducts per the substances into the
dissolved in tetrahydrofuran 5.3 mg bitumen paint (diluted to 108 nucleotides) found in lungs where DNA-binding
150 µl with tetrahydrofuran) to skin tissue only after the takes place
the shaved dorsal skin and killing high dose treatment
animals after 24 h
2) a maximum of five weeks 2) time-dependent increase
dermal treatment with 5.3 mg in the levels of adducts in
doses of bitumen paint (diluted the skin and lungs
to 150 µl with tetrahydrofuran)
on days 1 and 4 of every week
and killing the animals on day 5
of the week.
“bitumen (vacuum residue;  detection of adducts in mouse 0.24 fmol PAH-DNA- adduct level only slightly Booth et al.
CAS 64741-56-6), ref. no. mouse (CD) skin by means of 32P-post- adducts/µg DNA and mg above background 1998
TMC21565”, DMSO extract labelling 24 h after a single substance (one adduct per
prepared according to the dermal application of 1 mg 1.4 × 107 nucleotides)
method IP 346, residue extract in 25 µl tetrahydrofuran
dissolved in tetrahydrofuran
“fume condensates of type I (for  DNA adducts determined by both test samples pro- DNA adducts in white Qian et al.
flat roofs) and type III (for steep rat means of 32P-postlabelling after duced a dose-dependent blood cells are not a 1998
roofs) roofing asphalts, ... from (CD) intratracheal instillation at 8 h increase in the amounts of marker for exposure to
local supplier”, produced at intervals of 3 doses of 250–2000 DNA adducts in lung cells condensate of asphalt
316°C mg/kg body weight dissolved in (up to 71 adducts/108 fumes
DMSO; animals killed 6 h after nucleotides); no adducts data for the doses used in
Volume 17
the last dose detectable in white blood the tables do not agree
cells after intratracheal with those in the text
instillation
Volume 17
Table 12. continued
animals
condensate of “bitumens of two rat DNA adducts determined by 32P- DNA adducts produced PAH on the EPA list are Genevois et
viscosities provided by industry (BD4) postlabelling after application of with all four test samples not the only substances al. 1996
(45/60 pen, 160–210 pen)”, 3 per group 100 µg of the undiluted in all the tissues examinedforming adducts; the
produced at 160°C and 200°C; condensate to the shaved dorsal (skin, lung, lymphocytes) authors suggest that
captured on glass fibre filters skin twice in three days; animals systemic effect, maximum thioarenes, carbazoles,
and XAD-2 resin, filters eluted killed 24 h after last treatment of about 8 adducts/108 acridines and alkylated
with benzene, the resin with nucleotides; aromatics make a
diethylether, and the extraction significant contribution;
solvents removed by excretion of 1-hydroxypy- adduct pattern seen after
evaporation rene in the urine not cor- application of bitumen
related with the amounts condensates different from
of adducts in the indi- that produced with coal tar
vidual organs condensates
“bitumen .. (penetration grade one  DNA strand breaks in nuclear relative elution rates: effects were obtained with Pasquini et

80–100), collected during Sprague- DNA of liver cells determined by control (12 assays): dimethynitrosamine as al. 1989b
paving operations, .. extracted Dawley rat alkaline elution, also DNA- 2.37 ± 0.51; bitumen: positive control
with benzene”; asphaltenes per test unwinding 72 h after single 2.87 ± 0.55; difference not
precipitated with n-heptane, intraperitoneal injections of the significant
heptane-soluble material concentrate in doses of 50 mg/kg
extracted with DMSO, body weight DNA-unwinding: no
concentrate taken up in difference
cyclohexane
bitumen for road construction rat mutagenic activity of the urine no increase in the numbers time-dependent but not Pasquini et
(“petroleum derivative, 80–100 (Sprague- determined with Salmonella of revertants after 24 h or dose-dependent increases al. 1992
penetration grade”), extracted Dawley) typhimurium TA98, 24 h after 3 days (not even with in the activities of AHH
with benzene, after precipita-  (4 per single intraperitoneal injection of urine concentrates) in the and glutathione transferase
tion of the asphaltenes with group) 10, 50 or 100 mg/kg body weight presence or absence of rat in the liver but not in the
n-heptane, extraction with in DMSO liver S9 and lung; significant increase
DMSO and drying β-glucuronidase in glucaric acid in urine
97
98
Table 12. continued

animals
“vacuum residuum rat detection of structural no significant increases in API 1984c,

(petroleum)” samples 81-13 (Sprague- chromosomal aberrations in bone chromosomal aberrations 1984d
and 81-14 (CAS 64741-56-6), Dawley) marrow after oral administration
in corn oil ,  of 5 doses; doses: 0.4, 1.3, 4 g/kg
body weight and day; animals
killed 6 h after the last dose
AHH arylhydrocarbon hydroxylase

DMSO dimethylsulfoxide
PAH polycyclic aromatic hydrocarbons
Volume 17
Volume 17
Table 13. Incidence of skin tumours in experimental animals treated epicutaneously with asphalt or bitumen
Type of asphalt or Species, strain Treatment Duration Number of animals Comments References
bitumen and number with skin tumours
“3 residues after 5 rabbits per samples applied to 7 1 year B: 0/5, 0/5, 0/5; no control data Hieger and
fractionation of crudes group; study different skin areas on L: 0/5, 1/5, 1/5; Woodhouse
using vacuum and carried out each animal, 0.3 ml, in both animals 1952
steam” twice, in twice weekly, fur papillomas
Birmingham (B) removed regularly
and London (L)
“3 residues after 50 mice 0.2 ml applied to the 1 year B: 1/50, 0/50, 0/50; no control data
fractionation of crudes (“laboratory interscapular region L: 0/50, 0/50, 0/50;
using vacuum and outbred”) per twice weekly papilloma
steam” group, 25 , 25
; study carried
out twice, in
Birmingham (B)
and London (L)

“Commercially 68 mice, asphalt liquefied with > 54 among 68 animals tumours classified as malignant on Simmers et
manufactured 32 , 36  benzene (no other weeks 12 epidermoid the basis of microscopic criteria; no al. 1959
petroleum asphalt, (C57 Black) details) applied twice carcinomas; metastases; tumour appearance was
mixture of steam and weekly with a glass 5 (other?) animals preceded by desquamation of the skin
air blown asphalts” rod to the with papillomas the papilloma formation
interscapular skin
criticism: only the total number of
tumours was given, not the number of
animals with a tumour
control with benzene 63 mice, benzene (amount not > 54 0/63 no skin tumours; only slight hair loss,
32 , 36  specified) applied weeks dryness and desquamation of the skin;
(C57 Black) twice weekly with a in one control animal leukocytic
glass rod to the invasion of the salivary glands and
interscapular skin lungs
99
100
Table 13. continued

“4 road asphalts, steam 50 mice for asphalt diluted with 2 years 3/250 in addition 7 animals with leukaemia; Hueper and
distillation products of 4 each sample, acetone, concentration 2 papillomas, the authors suggested that absorbed Payne 1960
different crudes .. from 25 , 25  not specified; 1 carcinoma asphalt components could stimulate
Mississippi, California, C57 Black, application twice the development of leukaemia
Venezuela, Oklahoma” 100 for the weekly on the skin of
Venezuelan the neck
asphalt
control with acetone 200 C57 0/200 1/200 animals with leukaemia
Black mice
“petroleum roofing 50 mice, material liquefied by 2 years 1 questionable no mortality data
asphalt, blown” 25 , 25  heating, concentration carcinoma among
C57 Black not specified; applied 50 animals
twice weekly to skin
of the neck
“4 road asphalts, steam 6 rabbits undiluted heated 2 years 0/6
distillation products of New Zealand asphalt applied twice
4 different crudes .. from White weekly to the inside
Mississippi, California, ear and the shaved
Venezuela, Oklahoma” dorsal skin
mixture (1:1:1) of three 50 mice, 75–100 mg material up to 22 3 of 50 animals because of intercurrent pneumonia, Simmers
“steam-refined asphalts 25 , 25  liquefied in a water months developed epider- after 7 weeks only 27 (54 %) of the 1965a
made from West coast C57 Black; bath, applied directly moid cancer at the animals were still alive, after 52
crude oil” later 8  and with a glass rod (after application site, one weeks only 6; skin reactions were
8  added slight cooling on the (of these or an- seen in all animals, therefore the
rod) 3 × weekly; the other?) originated in decimated group was stocked up with
Volume 17
asphalt hardened on skin appendages; in 13 additional animals (8 , 5 );
the skin which re- addition, 2 papil- 16–240 applications
sulted in poor contact lomas were found no control group
Volume 17
Table 13. continued
mixture (1:1:1) of three 50 mice, 75–100 mg material up to 21 2/50; one animal after 7 weeks only 32 animals (64 %)
“air-refined asphalts 25 , 25  liquefied in a water months developed a had survived an attack of
made from West coast C57 Black bath, applied directly papilloma, another a pneumonitis;
crude oil” with a glass rod three tumour of the skin 22 to 270 applications; chronic
times weekly; the as- appendages at the dermatitis of the application site
phalt hardened on the application site; 1
skin which resulted in lung adenoma no control group
poor contact
mixture of three “air- 20 mice, asphalt dissolved in 2 years 9/50; 9 epidermoid 2 lung adenomas;
refined asphalts” 10 , 10  toluene (10:1); carcinomas at the up to 284 applications; chronic
dissolved in toluene C57 Black applied three times application sites, 2 dermatitis
weekly with a glass of which were
rod infiltrating and
metastasizing

control with toluene 15 mice 20–30 mg toluene 19 months 1 small papilloma chronic dermatitis developed during
C57 Black three times weekly the 220–230 applications; authors
stated toluene had not been reported
to cause cancer or to function as a co-
carcinogen in skin-painting studies
mixture of the fractions 50 mice, applied three times up to 20 13/50; 7 animals skin at the application site dry, Simmers
“saturates” and 25 , 25  weekly to the months with epidermoid scaling, hairless; 4 animals with 1965b
“aromatics” from C57 Black interscapular area carcinomas, 5 with hyperkeratosis; 1 leiomyosarcoma of
a
“steam-refined asphalt” with a glass rod and basal cell carcino- the small intestine; in all 72–242
“rubbed into the fur”, mas, 1 with a applications
on average 33.4 mg sebaceous gland
per application carcinoma;
13 papillomas, one
mixed with a
leiomyosarcoma
101
102
Table 13. continued

no specific control group no details of controls; only reference
to more than 1000 C57 Black mice
used in the laboratory in several years
“3 sludge asphalts mice 40 % in benzene 19 months animals with BN-5: 1 animal with a keratinizing Kireeva
(straight distillation) SS-57 White (dose not specified) tumours/number of squamous cell carcinoma, 1 with a 1968
from .. the Ukraine and .. for survivors at the time sebaceous gland adenoma; 5/43
the Volga region”; BN-5: 50, once weekly; control of appearance of the (additional?) animals with lung
BN-5 BN-3: 50, benzene alone first tumour: adenomas and adenocarcinomas
BN-3 BN-2: 40, BN-5: 2/43, BN-3: 2/43; 1 animal with a subcu-
BN-2 control: 23 BN-3: 2/43, taneous fibrosarcoma, 1 with papil-
BN-2: 0/30, loma; 1 animal with lung adenoma, 2
control: 0/23; with lymphoreticular sarcomas, 1
4/116 animals with hepatic haemangioma;
developed skin control: 1/23 with lung adenoma;
tumours of which 3 many of the treated animals devel-
were malignant, 1 oped hyperkeratosis, focal hyper-
benign plasia and inflammation of the skin
“2 asphalts from mice 40 % in benzene 19 months animals with BN-5: 5/49 animals with keratinizing
cracking-residue”, SS-57 White (dose not specified) tumours/number of squamous cell carcinomas, 1/49 with
origin: oils from the for: once weekly survivors at the time fibrosarcoma, 3 with papillomas, all
Ukraine and Volga- BN-5: 52, of appearance of the at the application site; also 7 animals
Region (see above); BN-4: 47 first tumour: with lung adenomas and adenocarci-
BN-5 BN-5: 9/49 nomas, 1 with stomach carcinoma,
BN-4 BN-4: 4/42 “systemic lesions” in 2 animals
control: 0/23
BN-4: one animal with a keratinizing,
Volume 17
(see above)
one with non-keratinizning squamous
cell carcinoma, 2 animals with papil-
lomas; all 4 animals with tumours
also had lung adenomas;
survival poorly documented
Volume 17
Table 13. continued
“8 road paving grade 8 groups of 10 % in benzene; > 20 6/218; 5 papillomas, generally “asphalt” induced marked Wallcave et
asphalts, vacuum 12–14  and 2.5 mg “asphalt” in months 1 skin carcinoma skin hyperplasia often with al. 1971
distilled, penetration 12–14  25 µl solution applied infiltration of inflammatory cells into
85/100” (according to Swiss Albino twice weekly to the dermis and occasionally with
b
ASTM)] mice shaved dorsal skin ulcers; the authors considered that the
number of tumours was too low to
demonstrate tumourigenicity of
“asphalt”.
control with benzene 15 , 15  25 µl benzene applied > 20 1/30 animals with
mice twice weekly months skin papilloma
“roofing dust from 20  mice 50 mg of 1:1 (w/w) 30 weeks 12/20; 10 animals survivors at the end of the study: 14 Emmett et
surface of a 25-year-old C3H/HeJ solution of the solid developed al. 1981
coal tar pitch-containing in toluene applied malignant skin
roof” twice weekly to the tumours, 2

interscapular skin; fur papillomas
removed regularly
“1 standard roofing 50  mice 50 mg of 1:1 (w/w) 20 months 0/50 survivors at the end of the study: 26
asphalt” C3H/HeJ solution of the solid
in toluene applied
twice weekly to the
interscapular skin; see
above
control with toluene 50  mice 50 mg toluene, 20 months 0/50 survivors at the end of the study: 37
C3H/HeJ applied twice weekly
to the interscapular
skin; fur removed
regularly
103
104
Table 13. continued

c
“condensed volatiles 50  mice 25 mg of the material 18 months C3H/HeJ : 47/49 C3H/HeJ: in all 76 tumours, NIOSH
from fumes of type I C3H/HeJ ; in 50 µl cyclohexane: animals with including 34 papillomas, 10 1981
roofing asphalt (dead 50  mice acetone (1:1) applied tumours; 24/49 withkeratoacanthomas, 25 squamous cell
level), heated at 232°C, CD-1 Swiss- twice weekly to the papillomas, 22/49 carcinomas;
.. the recommended Albino shaved interscapular with carcinomas with UV-B: 43/48 animals with
application temperature” skin tumours, 14/48 with papillomas,
–/+ UV-B CD-1: 8/45 animals 27/48 with carcinomas
with tumours; 8/45 CD-1: in all 12 papillomas;
with papillomas with UV-B: 4/17 animals with
tumours, 4 with papillomas
c
“condensed volatiles 50  mice 25 mg of the material 18 months C3H/HeJ : 45/47 C3H/HeJ: in all 78 tumours,
from fumes of type I C3H/HeJ in 50 µl cyclohexane: animals with including 27 papillomas, 7
roofing asphalt (dead acetone (1:1) applied tumours; 13/47 with keratoacanthomas, 31 squamous cell
level), heated at 316°C, twice weekly to the papillomas, 31/47 carcinomas;
“.. a temperature in shaved interscapular with carcinomas with UV-B: 44/47 animals with
excess of the recom- 50  mice skin tumours, 18/47 with papillomas,
mended application CD-1 Swiss- CD-1: 21/48 26/47 with carcinomas
d
temperature” Albino animals with CD-1: in all 19 tumours, 18
–/+ UV-B tumours, 20/48 with papillomas, 1 fibrosarcoma
papillomas, 1/48 with UV-B: 7/38 animals with
with a fibrosarcoma tumours, 7 with papillomas
c
from fumes of type III C3H/HeJ in 50 µl cyclohexane: animals with including 32 papillomas,
roofing asphalt (steep), acetone (1:1) applied tumours; 15/47 with 3 keratoacanthomas, 19 squamous
heated at 232°C, .. the twice weekly to the papillomas, 25/47 cell carcinomas, 9 fibrosarcomas;
Volume 17
recommended applic- shaved interscapular with carcinomas with UV-B: 34/42 animals with
d
ation temperature” skin tumours, 11/42 with papillomas,
–/+ UV-B 20/42 with carcinomas
Volume 17
Table 13. continued
50  mice CD-1: 14/48 CD-1: in all 13 tumours,

CD-1 Swiss- animals with 11 papillomas, 1 keratoacanthoma,
Albino tumours, 13/48 with 1 squamous cell carcinoma;
with a carcinoma tumours, 9 with papillomas, 2 with
carcinomas
c
from fumes of type III C3H/HeJ in 50 µl cyclohexane: animals with including 24 papillomas,
roofing asphalt (dead acetone (1:1) applied tumours; 12/45 with 6 keratoacanthomas, 36 squamous
level), heated at 316°C, twice weekly to the papillomas, 28/45 cell carcinomas, 9 fibrosarcomas
.. a temperature in excess shaved interscapular with carcinomas with UV-B: 38/48 animals with
of the recommended skin tumours, 20/48 with papillomas,
application tempera- 18/48 with carcinomas
d
ture” 50  mice CD-1: 20/46 CD-1: in all 20 tumours,

–/+ UV-B CD-1 Swiss- animals with 17 papillomas, 2 fibrosarcomas,
Albino tumours, 17/46 with 1 squamous cell carcinoma
with carcinomas tumours, 5 with papillomas, 1 with a
carcinoma
c
positive control with 50  mice 5 µg in 50 µl 18 months C3H/HeJ : 38/42 C3H/HeJ: in all 53 tumours,
benzo[a]pyrene C3H/HeJ cyclohexane:acetone animals with including 12 papillomas,
–/+ UV-B (1:1) applied twice tumours; 11/42 with 8 keratoacanthomas, 29 squamous
weekly to the shaved papillomas, 27/42 cell carcinomas, 2 fibrosarcomas
interscapular skin with carcinomas with UV-B: 36/45 animals with
tumours, 8/45 with papillomas, 27/45
with carcinomas
105
106
Table 13. continued

control with 50  mice 50 µl cyclohexane: 18 months C3H/HeJ: 0/50 C3H/HeJ with UV-B: 1/50 with 2
cyclohexane:acetone 1:1 C3H/HeJ acetone (1:1) applied with UV-B: 1/50 papillomas
–/+ UV-B 50  mice 2 × weekly to the CD-1: 0/50
CD-1 Swiss- shaved interscapular with UV-B: 0/50
Albino skin
control with 50  mice 50 µl cyclohexane: 18 months C3H/HeJ: 0/50 no additional irradiation with UV-B
cyclohexane:acetone 1:1 C3H/HeJ acetone (1:1) applied CD-1: 1/50 with a
(“cage only”) 50  mice 2 × weekly to the papilloma
CD-1 Swiss- shaved interscapular
Albino skin
no treatment 50  mice 18 months C3H/HeJ: 1/50 one animal with a squamous cell
C3H/HeJ CD-1: 0/50 carcinoma
50  mice
CD-1 Swiss-
Albino
initiation-promotion 8 groups of  single applications of 40 weeks A: 200 µl: 18/40 A: 200 µl: 2 carcinomas/40 Bull et al.
study with 4 “asphalt- Sencar mice, doses of 200 µl or 600 µl: 21/40 600 µl: 2 carcinomas/40 1985;
paints” (A to D) “from 40 per group 600 µl of the 4 B: 200 µl: 17/40 B: 200 µl: no carcinomas/40 Robinson et
solid asphalt, cut back “asphalt paints” to the 600 µl: 20/40 600 µl: 2 carcinomas/40 al. 1984
with mineral spirits to shaved dorsal skin; C: 200 µl: 19/40 C: 200 µl: 5 carcinomas/40
64%” from 2 weeks later 600 µl: 23/40 600 µl: 4 carcinomas/40
topical application of D: 200 µl: 21/40 D: 200 µl: 6 carcinomas/40
d
1 µg TPA in 200 µl 600 µl: 15/40 600 µl: 3 carcinomas/40
acetone 3 × weekly carcinoma yield significantly higher
in exposed groups than in the control
Volume 17
control with acetone 40  Sencar 200 µl acetone con- 40 weeks 4 animals with no carcinomas
mice taining the dissolved papillomas/40
promoter, TPA,
topical 3 × weekly
Volume 17
Table 13. continued
control with petroleum 40  mice 600 µl “mineral 40 weeks 5/40 no carcinomas

ether Sencar spirits” containing the
dissolved initiator
applied topically 3 ×
weekly
“1 petroleum asphalt 40  mice 200 µl “asphalt paint” 30 weeks 1 papilloma/40 no other carcinomas; all animals Robinson et
paint, from solid asphalt, Sencar applied to the shaved killed after 52 weeks; the authors al. 1984
cut back with mineral dorsal skin, 60% in suggested that the lack of tumours
spirits to 64 %” petroleum ether, once was the result of the low doses and
weekly the only once weekly application
control 40  mice 200 µl petroleum 30 weeks 3 papillomas/40 no other carcinomas; all animals
Sencar ether applied once killed after 52 weeks
weekly

vacuum residue D6 C3H/HeJ 50 mg in toluene (?) about 2 1/43 (3/350) positive control (50 mg Blackburn
applied 2 × weekly years benzo[a]pyrene): 179/184; et al. 1986
documentation inadequate
“bitumen from 50  mice 25 µl, 70 % in for life 2/50; 1 animal with average survival 19 months McKee et
Athabasca tar sands” C3H/HeJ toluene, applied to a a benign tumour, 1 al. 1986
regularly shaved area with a malignant
of interscapular skin tumour
3 × weekly
control with toluene 50  mice 25 µl toluene applied for life 1 skin papilloma/50 average survival 19 months
C3H/HeJ 3 × weekly to a
shaved area of skin
107
108
Table 13. continued

“bitumen derived from 50  25 µl aliquots of a 2 years in all 8 malignant average survival 88 weeks; tumour McKee and
the Cold Lake Oil Sands C3H/HeJ mice 75 % suspension in and 5 benign skin yield significantly different from the Lewis 1987
deposit, .. located in toluene applied to the tumours (in 26 % of control value;
Northeast Alberta, interscapular region; the animals) the authors concluded that Cold Lake
Canada”, obtained by weekly dose 56 mg “asphalt” is a moderately strong skin
steam injection in situ; carcinogen; no differences from the
material used directly in usual types of “asphalt”
road construction
control with toluene 50  25 µl toluene 2 years 0/50 average survival 83 weeks
C3H/HeJ mice
“naphthenic AC-20 50  37.5 µl aliquots of test ? 0/50 no other data McGowan
asphalt from a mix of C3H mice material (30 % w/v in et al. 1992
Nigerian light (4 %), “USP” mineral oil)
Maya (36%) and Alaska applied 2 × weekly to
North Slope (60%) the shaved dorsal skin
crudes”
“naphthenic coastal 50  37.5 µl aliquots of test ? 0/50 no other data
residuum, a vacuum C3H mice material (30 % w/v in
bottom used as a USP mineral oil)
blendstock for asphalt” applied 2 × weekly to
the shaved dorsal skin
control with mineral oil 50  37.5 µl aliquots of ? 0/50 application not described separately
C3H mice (?) 100% mineral oil ap- but certain to be like that for the test
plied 2 × weekly (?) material
Volume 17
Volume 17
Table 13. continued
1) “raw asphalt” 30  mice 25 mg in 50 µl 2 years 4/30; 1 animal with average survival 610 days; 15 animals Sivak et al.
specification: “standard C3H/HeJ cyclohexane:acetone a papilloma, 3 with died prematurely; number of tumours 1997
commercial type III, (1:1) applied 2 × a carcinoma per tumour-bearing animal: 1.0
roofing, steep”, weekly to the shaved
“produced by distillation dorsal skin
and air-blowing of
Arabian crude” (only
dissolved)
control with 30  mice 50 µl cyclohexane: 2 years 0/30 19 animals died prematurely
cyclohexane:acetone 1:1 C3H/HeJ acetone (1:1) applied
2 × weekly to the
shaved dorsal skin
2) “condensed fumes 30  mice 25 mg in 50 µl 2 years 21/30; 21 animals 28 animals died before the end of the

generated from type III C3H/HeJ cyclohexane:acetone had 25 carcinomas study, mainly because of large
asphalt at temperature of (1:1) applied 2 × and 12 papillomas tumours; number of tumours per
e
316°C , which is in weekly to the shaved tumour-bearing animal: 1.8
excess of recommended dorsal skin
application
temperature”;
specification: “roofing,
steep”
“.. asphalt obtained by
distillation and air
blowing of Arabian
crude”
109
110
Table 13. continued

3) “condensed fumes 30  mice 2.3 mg of fraction B; 2 years 11/30; 11 animals 18 animals died before the end of the
generated from type C3H/HeJ for details see 2) had 10 carcinomas study, some because of tumours;
III roofing asphalt at and 2 papillomas number of tumours per tumour-
e
316°C , which is in bearing animal: 1.1
excess of recommended
temperature”;
specification, see 2)
above,
f
HPLC-Fraktion B
4) “condensed roofing 30  mice 2.6 mg of fraction C; 2 years 20/30; 20 animals 24 animals died before the end of the
asphalt fumes, generated C3H/HeJ for details see 2) had 18 carcinomas study, some because of tumours;
at 316°C”; specification, and 4 papillomas number of tumours per tumour-
see 2) bearing animal: 1.1
f
HPLC fraction C
5) “condensed roofing 30  mice 24.8 mg of the 2 years 25/30 average survival 555 days; 29 animals
asphalt fumes, generated C3H/HeJ fractions mixed in the 23 carcinomas, died prematurely; number of tumours
at 316°C”; specification: proportions w/w: 30 papillomas per tumour-bearing animal: 1.0
see 2) 64.1 % A, 8.3 % B,
HPLC fractions 10.5 % C, 11.5 % D,
f
A+B+C+D+E 5.6 % E; for details
see 2)
as given in 4), 30  mice 18.3 mg of the 2 years 13/30 average survival 608 days; 22 animals
HPLC fractions C3H/HeJ fractions; for details 8 carcinomas, died prematurely; number of tumours
A+B see 2) 10 papillomas per tumour-bearing animal: 1.4
Volume 17
A+C see 2) 12 papillomas per tumour-bearing animal: 1.9
Volume 17
Table 13. continued
B+C+D+E see 2) 9 papillomas per tumour-bearing animal: 1.4
A+B+C+D see 2) 17 papillomas per tumour-bearing animal: 1.6
A+B+C+D see 2) 26 papillomas per tumour-bearing animal: 2.1
B+C+D see 2) 15 papillomas per tumour-bearing animal: 1.8

B+C see 2) 12 papillomas per tumour-bearing animal: 1.5
A+C+D+E see 2) 5 papillomas per tumour-bearing animal: 1.1
A+B+D+E see 2) 5 papillomas per tumour-bearing animal: 1.3
HPLC fractions C3H/HeJ fractions; for details 2 papillomas died prematurely; number of tumours
A+D+E see 2) per tumour-bearing animal: 1.0
111
Table 13. continued
112

positive controls: 30  Sencar 50 µl benzo[a]pyrene 2 years 0.01 %: 27/30 average survival period:
benzo[a]pyrene: 0.01 % mice per in cyclohexane: 1 papilloma, 0.01 %: 449 days
benzo[a]pyrene: group acetone (1:1) applied 28 carcinomas; 0.001 %: 666 days
0.001 % twice weekly to the 0.001 %: 5/30 0.0001 %: 630 days
benzo[a]pyrene: shaved dorsal skin 2 papillomas,
0.0001 % 3 carcinomas;
0.0001 %: 0/30
“condensed roofing 30  Sencar unfractionated 2 years 20/30; 20 animals average survival period: 571 days; 25
asphalt fumes, generated mice condensate in 50 µl had 18 carcinomas animals died before the end of the
at 316°C”; specification: cyclohexane:acetone and 21 papillomas study, some because of tumours;
“standard commercial 1:1 number of tumours per tumour-
type III, steep, produced bearing animal: 2.0
by distillation and air-
blowing of Arabian
crude”
control with 30  Sencar 50 µl cyclohexane: 2 years 0/30 average survival period: 672 days; 12
cyclohexane:acetone 1:1 mice acetone (1:1) applied animals died prematurely
twice weekly to the
shaved dorsal skin
a
fractionation of “steam-refined asphalt” into 4 fractions: “asphaltenes, aromatics, saturates, resins”. The fractions containing the saturated and
aromatic compounds were chosen because they contained fluorescent polycyclic substances and mixed, 1:1, and applied to the skin of the animals.
b
ASTM American Society for Testing and Materials
c
number of animals in a group still alive at the time of appearance of the first tumour in the group
d
TPA: 12-O-tetradecanoylphorbol-13-acetate
e
the authors state: “Heating in roofing kettle operations is often poorly controlled, and the materials may be heated as high as 638 °F [337°C].”
f
Volume 17
HPLC fraction A contains: alkanes, alkenes, cycloalkanes, alkylated benzene, naphthalenes, etc. (fraction A induces no tumours). Fraction B
(tumourigenic) contains: alkylated benzothiophenes, dibenzothiophenes and naphthothiophenes, anthracenes, phenanthrenes, fluorenes, pyrenes,
fluoranthenes. Fraction C (tumourigenic) contains: cycloalkenones, alkadienones, dihydrofuranones, dihydroindenones, hydroxyphenylthiols,
pyrenes, fluoranthenes, chrysenes, thiophenes with fused rings. Fraction D contains: alkanones, cycloalkenones, alkadienones, alkanoic acids,
alkylated carbazoles, dihydrofuranones, furanones, naphthols, phenols; fraction D induces no tumours;
Fraction E contains: alkanones, alkanoic acids, alkylated benzoic acids; fraction E induces no tumours.
Volume 17
Table 13a. Average concentrations [µg/ml] of polycyclic aromatic hydrocarbons in condensed fumes from two American roofing asphalts, “type I
asphalt” and “type III asphalt” (from: Niemeier et al. 1988: A comparison of the skin carcinogenicity of condensed roofing asphalt and coal tar pitch
fumes)
Substance Ion masses detected by Condensed fumes from Condensed fumes from Condensed fumes from Condensed fumes from
GC/MS “asphalt type I” heated “asphalt type I” heated “asphalt type III” “asphalt type III”
to 232°C to 316°C heated to 232°C heated to 316°C
naphthalene 128 22 4 17 49
fluorene 166 36 22 39 28
carbazole 167 20 1 6 –
anthracene/phenanthrene 178 180 53 300 69
fluoranthene 202 86 10 97 7
pyrene 202 70 9 63 8
benz[a]anthracene 228 11 10 8 6
chrysene/triphenylene 228 25 19 13 14

benzofluoranthene 252 2 4 5 –
benzo[e]pyrene 252 6 8 4 1
benzo[a]pyrene 252 2 2 3 –
indeno[c,d]pyrene 276 3 3 2 –
benzo[ghi]perylene 276 1 2 1 –
various dibenzanthracenes 278 2 – 2 –
various dibenzopyrenes 302 – – – –
Total analysed substances
[µg/ml] 466 147 560 182
113
114
Table 14. Incidence of lung tumours in experimental animals exposed to asphalt by inhalation
Type of asphalt Species Treatment Duration Tumour Comments References

or bitumen (strain) incidence

a
“air-blown rat, 65 , 0.7–1.0 kg asphalt heated in an 2 years 0/65 3 liver tumours, 2 sarcomas, 1 uterus Hueper and
roofing asphalt” (Bethesda evaporation vessel in the ventilated tumour (“All cancers seen in rats .. are Payne 1960
(“from a Black), exposure chamber to 121–135°C; evidently of ‘spontaneous’ origin.”);
commercial 2 months “brownish fumes”; concentration in Results of lung examination: marked
source”; flash old the chamber not determined; 2–10 g chronic fibrotic pneumonitis with
point: 200°C) asphalt evaporated daily, animals peribronchial adenomatosis, epithelial
exposed 5 h/day, 4 days/week metaplasia of the bronchial mucosa and
no control bronchiectasis.
“air-blown guinea pig, 0.7–1.0 kg asphalt heated in an 2 years 0/30 no other tumours;
roofing asphalt” 30 evaporation vessel in the ventilated results of lung examination: marked
(“from a (strain 13), exposure chamber to 121–135°C; chronic fibrotic pneumonitis with
commercial 2 months “brownish fumes”; concentration in peribronchial adenomatosis, epithelial
source”; flash old, sex not the chamber not determined; 2–10 g metaplasia of the bronchial mucosa and
point: 200°C) specified asphalt evaporated daily, animals bronchiectasis.
exposed 5 h/day, 4 days/week
no control
bitumen-water mouse, 4 15, 30, 45 – examination of the distribution of the Simmers
aerosol labelled (C57 Black) and 60 material in the lung; after inhalation 1964
with 131I min trachea and lung tissue were separated
and assayed: “satisfactory evidence that
material was getting in the
tracheobronchial tree.”
Volume 17
Volume 17
Table 14. continued
Type of asphalt Species Treatment Duration Tumour Comments References

or bitumen (strain) incidence
asphalt mixture mouse, 20 bitumen-water aerosol (produced by 16.5 1/20 3/20 animals survived until the end of the Simmers
from 6 “steam (C57 Black) stirring and aerosolization of hot months papillary study; 280–410 exposures; findings 1964
and air blown 10 , 10  asphalt in hot water); concentration lung included occasionally severe localized
samples from not specified; during exposure the adenoma bronchitis, dilation of the bronchioles,
California animals were fixed in plastic tubes “patchy regions of emphysema”
refineries” with their noses in an opening in the 17 of the 20 animals were subjected to
exposure chamber; exposure: 30 autopsy
min/day; 5 days/week
control mouse, 6  0/6
(C57 Black)
asphalt mixture mouse, 30 animals exposed in a whole animal 21 months 1 2/30 animals survived until the end of the
from 6 “steam (C57 Black) exposure chamber containing 6 cages, bronchial study; the animals ate contaminated food
and air blown sex not each with 5 mice; exposure 6–7.5 adenoma but suffered no weight loss; no stomach

samples from specified h/day, 5 days/week; yellowish brown (only 21 or intestinal tumours; histological
California asphalt fumes produced by heating animals findings: bronchitis with peribronchial
refineries” about 300 g asphalt each time to examined) infiltration of large round cells,
121°C; concentration in the chamber hyperplasia, loss of cilia, flattening and
not determined; enormous losses by necrosis of the bronchial epithelium,
deposition of the material on chamber emphysema, loss of respiratory
walls and in the tubing; during 21 epithelium; findings like those produced
months 2.24 kg asphalt was blown by inhalation of cigarette smoke
through the exposure chamber
control mouse, 6 21 months 0/6 no data
(C57 Black)
a
For comparison: after exposure of 75 rats in the same way to hard coal tar no lung tumours were detected but 6 liver tumours, 1 sarcoma and 1
animal with leukaemia. In a parallel study, 42 guinea pigs developed no lung tumours.
115
116
Table 15. Incidence of sarcomas at the injection site after subcutaneous or intramuscular injection of asphalt or bitumen
Type of asphalt or bitumen Species Treatment Duration Sarcoma Comments References

(strain) incidence

“Commercially manufac- 62 mice, interscapular s.c. injection of > 36 8/62 all sarcomas at the injection site; Simmers et
tured petroleum asphalt, 33 , 29  0.2 ml of a 1 % suspension of weeks 1 rhabdomyosarcoma, al. 1959
mixture of 6 different steam (C57 Black) asphalt in olive oil, twice 7 fibrosarcomas, no distant
and air blown asphalts weekly, from week 41 once metastases;
supplied by Southern weekly one sarcoma was successfully
California refineries” transplanted into a C57 Black mouse
control with olive oil 60 mice, interscapular s.c. injection of > 36 0/60 also no tumours at sites distant from
32 , 28  0.2 ml olive oil, twice weekly, weeks the injection site
(C57 Black) from week 41 once weekly
“4 road asphalts of different groups of 50 50 % in tricaprylin, 0.1 ml 2 years Ven: 1/50 Ven: 1 × leukaemia Hueper and
manufacture”; from crudes: mice for intramuscular (thigh), 6 × at MS: 1/50 MS: 1 × leukaemia Payne 1960
“3 steam-distilled products” each type of two-week intervals OK: 0/50 OK: 1 tumour in a liver lymph node
from Venezuela (Ven), asphalt, CA: 1/50 CA: 1 skin carcinoma; 1 tumour in a
Mississippi (MS), Oklahoma (C57 Black) liver lymph node
(OK) and 1 steam-vacuum-
distilled product from
California (CA)
control with tricaprylin mouse, 144 50 % tricaprylin in acetone, 2 years 0/144
(C57 Black) 0.1 ml intramuscular (thigh),
6 × at two-week intervals
“4 road asphalts of different groups of 30 50 % tricaprylin in acetone, 2 years Ven: 2/30 Ven: 2 tumours in a liver lymph node
manufacture”; from crudes: rats for each 0.2 ml intramuscular (thigh), MS: 1/30 MS: 3 tumours in a liver lymph node,
“3 steam-distilled products” type of 12 × at two-week intervals 3 lymph node sarcomas, 1 uterus
(from Venezuela, asphalt, carcinoma
Mississippi [MS], Oklahoma (Bethesda OK: 4/30 OK: 1 uterus carcinoma
Volume 17
[OK]) and 1 steam-vacuum- Black) CA: 6/30 CA: 4 tumours in a liver lymph node,
distilled product (from 3 lymph node sarcomas, 1 uterus
California [CA]) carcinoma
Table 15. continued
Volume 17
(strain) incidence
control with tricaprylin 60 animals 50 % in tricaprylin, 0/60

0.2 ml intramuscular
(thigh), 12 × at two-
week intervals
mixture of three “steam- 50 mice, 1–2 interscapular s.c. up to 23 0/32 only in 12 animals was asphalt found at the Simmers
refined asphalt samples 25 , 25  injections of 200 mg months injection site; one mouse had asphalt in the 1965a
from West coast crude (C57 Black) heated material (about abdominal cavity, another near the kidney,
petroleums” 60°C) another near the liver, in two others asphalt
was found in the thorax;
1 lung adenoma;
only 4 animals survived a pneumonia
epidemic; autopsy of 32 animals
mixture of three “air- 50 mice, 1–2 interscapular s.c. up to 23 5/50 1 animal developed a large rhabdomyo-
refined asphalt samples 25 , 25  injections of 200 mg months sarcoma (enclosing the injected material)

from West coast crude (C57 Black) heated material (about which had formed adhesions with the muscles
petroleums” 60°C) of the thigh and back; in another animal a
rhabdomyosarcoma of the abdominal wall
enclosed the injected asphalt; another animal
had a tumour at the injection site originating
from a hair follicle or sebaceous gland, and at
the same time the left lung had been replaced
by histologically identical tumour tissue;
in addition a total of 5 lung adenomas
mixture of the aromatic 126 mice in (see below) 15.6 ? summary of the 3 experiments described Simmers
and saturated fractions of 3 groups months below: 25.7 % of the animals subjected to 1966
petroleum asphalt (see below) autopsy had treatment-related tumours; of
(C57 Black) these, 9 % had lung tumours, 2 % lymphatic
(autopsy of leukaemia, 7.1 % adenocarcinomas of the
117
98 animals) skin appendages (c.f. control (see below):
0.16 %)
118
Table 15. continued

(strain) incidence

Experiment 1 47 mice one single interscapular ≤ 17 ? 5/36 animals with lung adenomas (1
(autopsy of subcutaneous injection of months animal also adenocarcinoma of a skin
36) 0.5 ml “oil” appendage);
1/36 animals with lymphatic leukaemia;
2/36 with adenocarcinomas of the skin
appendages:
of 36 animals subjected to autopsy, 22 %
had tumours
Experiment 2 49 mice 0.25 ml “of asphaltic ≤ 16.5 ? 3/39 with lung adenomas;
(autopsy of material” every second months 1/39 infiltrating lymphoma in the lung
39) week, a total of 8 (?); 2/39 leiomyosarcomas; 1/39
injections fibrosarcoma; 1/39 adenocarcinomas of
the lung; 1/39 adenocarcinoma of the skin
appendages; of 39 animals subjected to
autopsy 18.4 % had tumours
Experiment 3 27 mice 1 ml “of aromatic-saturate ≤ 13.5 ? 1/19 with lymphatic leukaemia;
(autopsy of fractions” fractionated so months 2/19 with adenocarcinoma of the skin
19) that 8 animals received a appendages;
maximum of 11 injections 1/19 epidermoid carcinoma of the nose;
2/19 adenocarcinoma of the lung, in
addition in one animal adenoma of the
bile duct; 1/19 adenomas of the bladder;
of 19 animals subjected to autopsy, 7
(36.8%) had tumours
no specific control group 587 non- 2 animals with lymphatic leukaemia
Volume 17
exposed C57 (0.3%); 1 with an adenocarcinoma of the
Black mice skin appendages (0.16%)
subjected to
autopsy

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Bitumen (vapour and aerosol)

* n-octanol/water distribution coefficient

1 Composition and Analysis

1.1 Definition and terms

1.2 Analysis of PAH in bitumen and its vapour and aerosols

1.3 Bitumen vapour and aerosol levels during hot processing

2 Toxic Effects and Mode of Action

3 Toxicokinetics and Metabolism

4.1 Repeated exposures

4.2 Local effects on skin and mucous membranes

4.3 Allergenic effects

The possibility that sensitization may be caused by individual components of bitumen

In collectives of roof construction workers, an increased incidence of DNA adducts in

In 1994, the IARC published a meta-analysis of the results of carcinogenicity studies

5 Animal Experiments and in vitro Studies

5.1 Acute toxicity

5.2 Subacute, subchronic and chronic toxicity

5.2.3 Dermal absorption

5.3 Local effects on skin and mucous membranes

5.4 Allergenic effects

In experimental animals given doses of bitumen solutions, extracts or condensates, local

5.6.1 Dermal application

Characterization of the animal studies

2) The combination of condensate with UV-B irradiation (for 30 to 45 minutes after

Fractions A, D and E had no detectable carcinogenic effects. Combinations of frac-

5.6.3 Subcutaneous or intramuscular injection

5.7 Other effects

6 Manifesto (MAK value/classification)

ACGIH (American Conference of Governmental and Industrial Hygienists) (2000) Asphalt

Bittighofer PM, Peleschka M, Carbonell F, Fliedner TM, Reichenbach K (1984) Untersuchungen

De Méo M, Genevois C, Brandt H, Laget M, Bartsch H, Castegnaro M (1996) In vitro studies of

HVBG (Hauptverband der gewerblichen Berufsgenossenschaften) (1997) Von den Berufsgenos-

Appendix: Tables 1–15

Table 1. Use of bitumen in Germany in 1998 (Bau-Berufsgenossenschaft 2001)

Use Tonnage per year (1000 t) %

rolled asphalt 2500 74.5

Job Concentration (mg/m3)

production of bitumen 2.6

Level of PAH (mg/kg)

Bitumen product HB 90/100 B 45 B 65 B 80 B 200 B 85/25 B 95/35