Jürgen Jacob

Profile of the polycyclic aromatic compounds from crude oils

Zusammenfassung8 Rohöle unterschiedlicher Provenienz wurden auf ihren Gehalt an polycyclischen aromatischen Kohlenwasserstoffen, deren isosteren Sauerstoff- (Furane) und Schwefelanaloga (Thiophene) sowie an Carbazolen mit Hilfe der Glas-Capillar-Gas-Chromatographie quantitativ untersucht. Am Beispiel eines Rohöls (Qatar) wurden 226 verschiedene Verbindungen entweder durch Vergleich der Retentionszeiten und Massenspektren mit denen von Referenzsubstanzen identifiziert oder, wo diese nicht vorhanden waren, massenspektrometrisch als polycyclische aromatische Verbindungen (PAC) charakterisiert. Der PAC-Gehalt (>3 Ringe) der untersuchten Öle schwankt innerhalb einer Zehnerpotenz. Der relative Gehalt an schwefelhaltigen PAC (S-PAC) ist mit dem Gesamtschwefelgehalt linear korrelierbar.SummaryEight crude oils of different origin were quantitatively analysed in respect to the content of polycyclic aromatic hydrocarbons, isosteric oxygen (furans) and sulphur (thiophenes) analogues, and carbazoles by means of glass-capillary gas-chromatography. From a crude oil (Qatar) 226 different compounds were either identified by comparing their retention times and mass spectra with those of reference materials, or where these are lacking characterized as polycyclic aromatic compounds (PAC) by mass spectrometry. The PAC-content (>3 rings) of these crude oils investigated ranged within one order of magnitude. The relative content of sulphur containing PAC can be correlated to the total sulphur content.

Profile of the polycyclic aromatic hydrocarbons from lubricating oils inventory by GCGC/MS — PAH in environmental materials, part 1

ZusammenfassungEine Schmierölprobe wurde auf ihren Gehalt an polycyclischen aromatischen Kohlenwasserstoffen (= PAH), Thiophen- und Furanhaltigen polycyclischen aromatischen Verbindungen (S-PAC und O-PAC) untersucht. Diejenigen Verbindungen, deren Gehalt über 0,02 mg/kg lag, wurden durch Vergleich mit Originalsubstanzen identifiziert oder — wenn Referenzsubstanzen nicht zur Verfügung standen — massenspektrometrisch charakterisiert. Diese Bestandsaufnahme wurde mit einer GC-MS-Kombination durchgeführt. Die Massenspektren von typischen Vertretern einiger Stoffklassen (S-PAC, O-PAC und Methylderivate) wurden diskutiert.Die Profile der im Mineralöl enthaltenen polycyclischen aromatischen Verbindungen (= PAC) sind von den PAC-Profilen der durch unvollständige Verbrennung oder Pyrolyse entstandenen deutlich verschieden.Um einen Überblick über den Konzentrationsbereich der PAC in verschiedenen nicht gebrauchten Motoren-Schmierölen zu bekommen, wurden 22 Handelsmuster auf ihren Gehalt an 14 ausgewählten PAC untersucht.SummaryPolycyclic aromatic hydrocarbons (= PAH), sulphur containing and oxygen containing polycyclic aromatic compounds (= S-resp. O-PAC) which occur in concentrations higher than 0.02 mg/kg in a lubricating oil sample were identified by comparison with reference substances (34) or, if reference compounds are lacking, characterized by mass spectrometry (58 compounds). Glass capillary gas chromatography combined with mass spectrometry was used for this inventory. Mass spectra of typical individuals from several classes (S-PAC, O-PAC, and methyl-derivatives) are discussed in detail.The profile of polycyclic aromatic compounds (= PAC) contained in mineral oils is significantly different from that obtained by incomplete combustion or pyrolysis of organic materials.To get a survey of the range of PAC-concentration in different engine oils, 22 commercial samples have been examined for a selected number of PAC.

Biomonitoring of polycyclic aromatic hydrocarbons in highly exposed coke plant workers by measurement of urinary phenanthrene and pyrene metabolites (phenols and dihydrodiols)

SummaryA filter combination consisting of an impregnated glass fibre and a control filter was used for the collection of air samples in which gaseous and particulate polycyclic aromatic hydrocarbons (PAHs) were determined. To estimate the loss of lower boiling PAHs, d10-phenanthrene was applied as internal standard. A simple, well-reproducible method for the determination of 1-, 2-, 3-, 4- and 9-hydroxyphenanthrene, 1,2-, 3,4- and 9,10 dihydroxydihydrophenanthrene, 1-hydroxypyrene and 1,2-dihydroxy-1,2-dihydropyrene is described. By means of personal air samplers the exposure to PAHs of four coke plant employees working at different locations was measured over 4 days. Simultaneously the 24-h urine was collected and stored frozen until analysed. The main excretion product of pyrene is a 1-hydroxypyrene conjugate, whereas phenanthrene is excreted predominantly as dihydrodiol conjugate. As expected, workers on the battery topside were exposed the most and accordingly excreted by far the highest amounts of PAHs. Up to 34.0 μg phenanthrol conjugates (total of all isomeric phenols) and 195.5 μg dihydrodiol conjugates (total of all isomeric dihydrodiols) were excreted in the 24-h urine (mean of 4 days). The metabolite profiles of five isomeric phenanthrene phenols and three isomeric dihydrodiols exhibited only small percentage variations within one individual whereas significant interindividual differences were observed. These findings may indicate a genetically determined enzyme pattern responsible for the metabolic conversion of PAHs.

Determination of polycyclic aromatic hydrocarbons, azaarenes, and thiaarenes emitted from coal-fired residential furnaces by gas chromatography/mass spectrometry

ZusammenfassungDa die krebserzeugende Wirkung von Emissionen aus Steinkohlebrikett-beheizten Zimmeröfen nahezu vollständig durch die im Kondensat enthaltenene polycyclischen aromatischen Verbindungen (PAC) mit mehr als 3 Ringen verursacht wird, wurde die Zusammensetzung dieser PAC-Fraktion untersucht.Mehr als 170 neutrale [111 polycyclische aromatische Kohlenwasserstoffe (PAH), 57 Thiaarene, 6 Oxaarene] sowie 70 basische Verbindungen (Azaarene) konnten mit der GC/MS-Kombination als PAC charakterisiert werden (Nachweisgrenze 0,1 mg/kg Brikett). Etwa 80 Verbindungen wurden durch den Vergleich mit Referenzsubstanzen identifiziert. Um in einigen zweifelhaften Fällen die Identität der isolierten mit der synthetischen Verbindung zu bestätigen, wurden bei 11 PAH mit einem Molgewicht von 302 zusätzlich auch UV- und Fluorescenzspektren verglichen [Naphtho(1,2-k)fluoranthen, Dibenz(e,k)acephenanthrylen, Naphth(2,3-e)acephenanthrylen, Naphtho(2,3-k)fluoranthen, Dibenzo(de,qr)naphthacen, Coronen, Dibenzo(fg,op)naphthacen, Naphtho(1,2,3,4-def)chrysen, Benzo(b)perylen, Dibenzo(def,j)chrysen, Benzo(rst)pentaphen]. Diese Verbindungen sind von besonderem Interesse, da PAC mit 6 und mehr Ringen im Tierversuch etwa 50% der carcinogenen Wirkung des Steinkohlenbrikett-Emissionskondensates verursachen.SummarySince the carcinogenic effect of emissions from hard coal briquet-fired furnaces is almost entirely caused by polycyclic aromatic compounds (PAC) with more than three rings, the composition of this fraction has been analyzed in detail.More than 170 neutral [111 polycyclic aromatic hydrocarbons (PAH), 57 thiaarenes, 6 oxaarenes] and 70 basic (azaarenes) compounds were characterized as PAC by means of GC/MS (limit of detection 0.1 mg/kg briquet). About 80 compounds were identified by comparison with reference standards. In order to obtain unequivocal evidence for the identification of 11 PAH with a molecular weight of 302, the UV- and fluorescence spectra were compared with those of the synthesized reference compounds [naphtho(1,2-k) fluoranthene, dibenz(e,k)acephenanthrylene, naphth-(2,3e) acephenanthrylene, naphtho(2,3-k)fluoranthene, dibenzo (de,qr)naphthacene, coronene, dibenzo(fg,op)naphthacene, naphtho(1,2,3,4-def)chrysene, benzo(b)perylene, dibenzo (-def,j)chrysene, benzo(rst)pentaphene]. In case of the emission condensate from hard coal briquets, PAC containing 6 and more rings cause about 50% of the carcinogenic effect in animal experiments.

Detection of carcinogenic aromatic amines in the urine of non-smokers.

Smoking is thought to be one of the most important anthropogenic risk factors involved in the development of urinary bladder cancer in humans. Tobacco smoke contains a complex mixture of chemicals including potent carcinogens such as aromatic amines. In the present study the amounts of several freebase aromatic amines including the potent carcinogens 2-aminonaphthalene and 4-aminobiphenyl have been analyzed in the urine of 48 German urban living smokers and non-smokers. The results indicate that (i) both groups excrete the identical set of four aromatic amines; (ii) smokers excrete approximately twice the total amount of these amines, but similar amounts of 2-aminonaphthalene and 4-aminobiphenyl are found in non-smokers; and (iii) the excreted aromatic amines are decomposed in the urine within a few hours thus, explaining why aromatic amines are difficult to detect in this matrix. Their decomposition could be prevented by adding small amounts of p-toluidine to the freshly collected urine. Unlike smokers the origin of aromatic amines detected in the urine of non-smokers is at present unknown. Based on the cotinine levels found in the urine of non-smokers environmental tobacco smoke can be excluded as a major source of aromatic amines. In addition, neither diesel exhaust-related nitroarenes nor the corresponding amino-derivatives, to which they may be metabolically converted, were found. The detected urinary levels of aromatic amines arising from sources other than tobacco smoke or diesel exhaust may play a role in the bladder cancer etiology of non-smokers.

Relevance of polycyclic aromatic hydrocarbons as environmental carcinogens

SummaryAll emissions from incomplete combustion contain polycyclic aromatic hydrocarbons (PAH) which are a well-known class of carcinogens. The question whether additional carcinogenic compounds do exist in these emissions can be answered by separating the emission condensate into a PAH-containing and a PAH-free part. These parts have been tested in an animal experiment by means of a carcinogen-specific test system such as topical application onto the skin of mice or injection into the lung of rats. The investigation on the contribution of PAH-fractions and of benzo(a)pyrene to the carcinogenic potential of emission condensate from gasoline driven engines, Diesel engines, coal combustion in domestic furnaces and sidestream smoke of cigarettes shows that the carcinogenic effect of the particle phase of these pyrolytic condensates is predominantly caused by polycyclic aromatic compounds such as PAH. In all cases investigated the PAH-fraction containing 4 and more rings accounted for more than 75% of the total carcinogenic effect resulting from the implantation into the lung of rats or from the topical application onto mouse skin. The contribution of benzo(a)pyrene to the carcinogenic potency of various condensates, however, is minor in all cases investigated and accounts for only 0.17% to 4% of the total carcinogenicity as evaluated from implantation into the lung of rats (Table 1).

The metabolism of pyrene by rat liver microsomes and the influence of various mono-oxygenase inducers

1. Pyrene metabolite g.l.c. profiles were recorded and metabolites identified by mass spectrometry. 2. Pyrene is metabolized by liver microsomes of untreated rats to 1-hydroxypyrene, 4,5-dihydroxy-4,5-dihydropyrene, two different diphenols and a triol, tentatively identified as 1,4,5-trihydroxy-4,5-dihydropyrene. 3. Pretreatment with phenobarbital or polychlorinated biphenyls favours oxidation at the K-region, whereas cytochrome P-448 inducers stimulate oxidation at the non-K-region of pyrene. 4. 1-Hydroxypyrene does not inhibit pyrene oxidation. 5. Pyrene diphenols are formed by secondary oxidation of 1-hydroxypyrene. 6. Triols are formed from dihydrodiols by secondary oxidation.

Contribution of polycyclic aromatic hydrocarbons and nitro-derivatives to the carcinogenic impact of diesel engine exhaust condensate evaluated by implantation into the lungs of rats

Diesel exhaust condensate was separated by a liquid-liquid distribution into a hydrophilic (I; about 25% by weight of the total condensate) and a hydrophobic part (II; about 75%-wt.). To evaluate the carcinogenicity, the proportionately dosed fractions have been implanted into the lungs of Osborne Mendel rats and compared with several doses of benzo[a]pyrene and the vehicle, a mixture of trioctanoin plus beeswax. Only the hydrophobic part which contained polycyclic aromatic compounds (PAC) resulted in 5 malignant tumors in a group of 35 animals. In addition, the hydrophobic part was separated by column chromatography on Sephadex LH 20 and subsequently on silica gel into several fractions, such as non-aromatic compounds plus PAC with 2 and 3 rings (IIa; 72%-wt of the total condensate), polycyclic aromatic compounds (PAH) with 4 and more rings (IIb; 0.8%-wt), polar PAC (IIc; 1.1%-wt) and nitro-PAH (IId; 0.7%-wt). PAH consisting of 4 and more rings (IIb) were found to be the most potent subfraction and provoked when proportionately dosed 6 carcinomas in a group of 35 rats. Only a low contribution to the carcinogenicity was observed by the subfraction of nitro-PAH (IId) which produced 1 carcinoma/35 rats. The polar PAC (IIc) and the fraction of non-aromatics plus PAC with 2 and 3 rings (IIa), although the main subfraction (72%-wt of the total condensate) did not provoke any tumors. The reconstitution of all hydrophobic subfractions (IIa-d) resulted in the same carcinogenic potency as the unfractionated hydrophobics (II), provoking 7 carcinoma in 35 rats. It may be concluded from these findings that most of the carcinogenicity of diesel exhaust originates from the PAH consisting of 4 or more rings.

Profile of urinary phenanthrene metabolites in smokers and non-smokers

Phenanthrene metabolites (phenols and dihydrodiols) and 1-hydroxypyrene excreted in the 24-h urine of smokers, non-smokers and lung cancer patients, who after heavy smoking became light smokers, were determined and compared. In contrast to 1- hydroxypyrene, no significant differences of the absolute amounts of phenanthrene metabolites were found between smokers and non-smokers. A ratio phenanthrene metabolites/l-hydroxypyrene of 10.4 was observed for non-smokers and 9.9 for lung cancer patients, but 4.2 for smokers. Significantly different ratios for the regiospecific oxidation of phenanthrene were found for smokers when compared with non-smokers (1,2-oxidation vs 3,4-oxidation was 1.45 in the case of smokers, but 2.34 in the case of non-smokers) indicating a cigarette smoke - but not PAH - caused induction of CYP 1A2 in smokers. As a consequence of the degree of PAH exposure the ratio dihydrodiols/phenols depends on the total amount of metabolites excreted. Phenols predominate, equally in smokers and non-smokers after low exposure, while dihydrodiols become more prominent in highly exposed persons (coke plant workers). Both (i) the regiospecific oxidation of PAH and (ii) the ratio of dihydrodiol vs phenol formation may be recognized from the urinary phenanthrene metabolite profile. This pattern mirrors the enzymatic status (balance of the CYP isoforms and epoxide hydrolase) in individuals. Accordingly, more detailed information may be obtained from the urinary metabolite pattern than from 1- hydroxypyrene, commonly used in PAH biomonitoring.

Urinary and faecal excretion of pyrene and hydroxypyrene by rats after oral, intraperitoneal, intratracheal or intrapulmonary application

The urinary and faecal excretion of pyrene and 1-hydroxypyrene after oral (53.4%), intraperitoneal (3.1%), intratracheal (30-37%) and intrapulmonary application (0.003%) to rats has been determined by means of gas chromatography/mass spectrometry and the excretion rates were found to depend on the mode of application. With regard to the low urinary excretion rates, 1-hydroxypyrene seems not to be very suitable as a biological marker for PAH exposure to man.