organic-geochemical differentiation of petroleum-type pollutants and study of their fate in danube...

Organic-geochemical Differentiation of Petroleum-typePollutants and Study of Their Fate in DanubeAlluvial Sediments and Corresponding Water(Pančevo Oil Refinery, Serbia)

Branimir Jovančićević & Miroslav Vrvić &

Jan Schwarzbauer & Hermann Wehner &

Georg Scheeder & Dragomir Vitorović

Received: 1 August 2006 /Accepted: 15 February 2007 / Published online: 31 March 2007# Springer Science + Business Media B.V. 2007

Abstract A review is given in this paper of the up-to-date results observed in differentiation and trans-formation studies on petroleum-type pollutants inunderground and surface waters. Water and particulatematter derived from the locality of Pančevo PetroleumRefinery, Serbia (River Danube alluvial formations). Itwas shown that distributions of n-alkanes, steranesand triterpanes, and δ13CPDB values of n-alkanes maysuccessfully be used for qualitatively differentiatingthe petroleum-type pollutants from native organicmatter in recent sedimentary formations. In under-ground waters, a petroleum-type pollutant is exposedto microbiological degradation which is manifested

through relatively fast degradation of n-alkanes.Following an almost complete degradation of crudeoil n-alkanes in underground water, the biosynthesisof novel, even carbon-number C16–C30 n-alkanes maybe observed. It is shown that the n-alkane distributionobserved in a petroleum-type pollutant may dependon the intensity of its previous interaction with water.The fate of petroleum-type pollutants in environmen-tal waters may be predicted through laboratorysimulative microbiological degradation experimentsby using microorganism consortiums similar to thoseobserved under relevant natural conditions, as well ason corresponding nutrient base.

Keywords n-Alkanes . Alluvial sediments .

Biodegradation . Identification . δ13CPDB.

Pančevo Petroleum Refinery . Petroleum pollutants .

Steranes . Triterpanes

1 Introduction

In spite of remarkable advancement of petroleumexploitation, transport and refining technologies,petroleum and its refining products continue to beone of the most abundant environmental pollutants.Consequently, studies on the environmental fate ofpetroleum-type pollutants remain to be an actualscientific interdisciplinary problem.

Water Air Soil Pollut (2007) 183:225–238DOI 10.1007/s11270-007-9371-7

B. Jovančićević (*) :M. Vrvić :D. VitorovićDepartment of Chemistry, University of Belgrade,PO Box 158, 11001 Belgrade, Serbiae-mail: [email protected]

B. Jovančićević :M. Vrvić :D. VitorovićCenter of Chemistry, IChTM,Belgrade, Serbia

J. SchwarzbauerInstitute of Geology and Geochemistry of Petroleumand Coal, Aachen University,Aachen, Germany

H. Wehner :G. ScheederFederal Institute for Geosciences and Natural Resources,Hannover, Germany

Transformation processes of petroleum-type pollut-ants in the environment were studied by numerousauthors (e.g., Vaajasaari et al. 2002; Booth et al. 2005).Thus, the extent and the way of petroleum-typepollutants’ biodegradation in coastal marine environ-ment (Readman et al. 1996), in estuarine sediments(Oudot et al. 1998), under arctic marine conditions(Garrett et al. 2003), and on the East Mediterraneancoast (Ezra et al. 2000) were comprehensivelyelucidated.

Since the composition of crude oil or some of itsrefining products in environment is mainly changedby the help of microorganisms (e.g., Prince 1993;Blanke and Wibbe 1999; Brandsch et al. 2001; Kahruet al. 2002; Truu et al. 2002; Hallberg and Trepte2003; Rosolen et al. 2005), a number of experimentswere carried out in order to define optimal conditionsfor the most efficient biodegradation (temperature,humidity, nutrients). Application of biodegradationprocesses focussed primarily on bioremediation of

Fig. 1 Location of PančevoOil Refinery within the al-luvial formations of Serbia(with average lithologicalcomposition)

226 Water Air Soil Pollut (2007) 183:225–238

Fig. 2 Distributions ofn-alkanes and δ13CPDB

values of individualn-alkanes of recentsediments’ bitumenfractions and one oil typepollutant (sample I1) fromPančevo Oil Refinerylocality (Jovančićevićet al. 1997)

Fig. 3 Gas chromatogramsof saturated hydrocarbonfractions isolated fromground water samples fromboreholes I–VII (PančevoOil Refinery locality;Jovančićević et al. 2001a)

Water Air Soil Pollut (2007) 183:225–238 227

different sections of the environmental compartments.For example, it was shown that remarkably higherproportions of petroleum-type pollutant (up to 81%)were removed from fertilized soil than from soildeficient in nutritional additives (up to 56%)(Chaĭneau et al. 2003). Similar effects were observedin marine environments. In the presence of fertilizers,the achieved efficiency of petroleum pollutants’biodegradation was found to be three to five timeshigher (Atlas 1995). On the other hand, laboratoryexperiments have shown Pseudomonas aeruginosa tobe much more efficient in biodegrading the dominanthydrocarbons, i.e., C14–C22 n-alkanes, in the presenceof molasses than in the presence of mineral fertilizers(Al-Hadhrami et al. 1997).

Recent results will be reviewed in this paperobserved by investigating microbial transformationprocesses of petroleum-type pollutants in Danubealluvial sediments and underground and surface watersfrom the locality of Pančevo Oil Refinery, Serbia(Fig. 1), as a potential new contribution for betterunderstanding the fate of oil type pollutant in theenvironment and conditions for its bioremediation.

2 Differentiation of Petroleum-type Pollutants

Identification of petroleum-type pollutants in recentsediments, soil, underground or surface waters,requires reliable and precise differentiation ofnative and anthropogenically released substances(Jovančićević 2002). Organic matter in recent sedi-ments was shown to be composed mainly of fulvo-and humic acids and humin (Tissot and Welte 1984).The soluble sedimentary organic substance, calledbitumen, was generally found in very small amount.Its higher content in sediments may therefore indicatethe presence of anthropogenic contaminants. However,the likely maximal amount of soluble nativeorganic substance in recent sediments was neverstrictly defined. Wide ranges of concentrations wereobserved, starting from the limit of determination upto 10 or 15%. However, considering the phenomenonof “seeping crude oils”, 100% of soluble organicsubstance in recent sediments may also, practically, beexpected (Tissot and Welte 1984). Consequently,observation of a larger amount of bitumen in asediment sample, is just a presumption of thepresence of a petroleum-type pollutant. Hence, a

reliable proof requires a detailed analysis of theextract.

Fortunately, substantial differences between thechemical compositions of the recent sediments,

bitumen and crude oils, involving almost all classesof organic compounds, make the interpretation ofsuch analyses easier. The differences result from thefact that petroleum is a relatively mature form oforganic substance in the geosphere, in contrast tobitumen of recent sediments which are of rather lowdegree of thermal maturity.

The mentioned compositional differences aremainly pronounced with n-alkanes, one of the mostabundant fractions in soluble sedimentary organicmatter. An example of typical petroleum n-alkanedistribution (in the form of a gas chromatogram ofsample I1 representing a petroleum-type pollutantfound in underground water from the area of PančevoOil Refinery), and several examples of distributionscharacteristic of organic substance derived fromrecent sediments, typical for all other samplesoriginating from different localities of the RiverDanube alluvial formations, are shown in Fig. 2(Jovančićević et al. 1997). n-Alkane fraction insample I1 was characterized by uniform distributionof odd and even homologues (CPI around 1) and amaximum at a lower member (n-C19). On the otherhand, the distribution of n-alkanes in nonpollutedsediments was characterized by domination of oddhomologues (CPI considerably above 1) and amaximum at some of the higher homologue members(n-C29 or n-C31). Bearing in mind that identificationof n-alkanes in petroleum or bitumen is relativelysimple (by gas chromatographic analysis of the alkanefraction), saturated hydrocarbons may be considered

Fig. 4 Distributions of steranes (SIM, m/z 217; (a) andtriterpanes (SIM, m/z 191, (b) in the alkane fractions isolatedfrom samples I–VII (Pančevo Oil Refinery locality; Jovančićevićet al. 2001a). 1: C27-13β(H),17α(H) diasterane (20S); 2:C27-14α(H),17α(H) sterane (20R); 3: C28-14α(H),17α(H)sterane (20R); 4: C29-14α(H),17α(H) sterane (20S); 5:C29-14β(H),17β(H) sterane (20R); 6: C29-14α(H),17α(H)sterane (20R); a: C27-18α(H)-22,29,30-trisnorhopane (Ts);b : C2 7 -17α (H) -22 ,29 ,30 - t r i sno rhopane (Tm) ; c :C30-17α(H),21β(H)-hopane; d: C30-17β (H),21α(H)-moretane;e: C31-17α(H),21β(H)-homohopanes (22S and 22R); O:oleanane; G: gammacerane. (A detailed identification of thecorresponding peaks based on published data: e.g., Peters et al.2005)

b


to be an effective tool for differentiating petroleum-type pollutants in the environment.

However, exceptionally, n-alkanes in recent sedi-ments may have a similar or even the samedistribution as the crude oil n-alkanes. Namely, recentsediments containing organic substance of exclusivelymarine origin will have a uniform distribution of oddand even n-alkane homologues and a maximum atsome of the lower members, independently of a lowerdegree of thermal maturity (Tissot and Welte 1984).In such a case n-alkanes cannot be reliable means fordifferentiation of native and anthropogenic organicsubstances in recent sediments.

An alternative way for solving the problem may bethe δ13C analysis.

Being a mature organic substance in geosphere,crude oil contains the greatest amount of heaviercarbon isotope, 13C. Consequently, the PDB-normal-ised ratio of 12C and 13C carbon isotopes (δ13CPDB) inpetroleum is less negative compared to the recentsediments, organic matter. For example, the sample I1presented in Fig. 2 (Jovančićević et al. 1997) showedless negative δ13CPDB with C25, C27, C29 and C31 n-alkanes compared to all other samples. As mentionedabove, this particular sample was characterized by atypical petroleum n-alkane distribution, in contrast toall other samples demonstrating distributions typicalfor recent sediments. Hence, it is suggested thatenvironmental native and anthropogenic organic sub-stances may be differentiated based on comparison oftheir carbon isotopes ratios, in other words, thatparameter δ13CPDB may be used as a tool forrevealing petroleum-type pollutants in the environ-ment. For this purpose gas chromatograpic-isotoperatio mass spectrometric (GC-IRMS) analysis ofsingle compounds is required, or the analysis of itscorresponding fractions.

However, since isotopic data (δ13CPDB) do notdepend exclusively on the thermal maturity of theinvestigated organic substance, but also on itsdepositional environment, length of migration path,etc., it may happen that petroleum and its individualhydrocarbon components, in the first place n-alkanes,do have identical δ13CPDB values as recent sediments,

organic substance. In such a case, carbon isotope ratiocan not be a reliable parameter to indicate the presenceof petroleum-type pollutant in the environment.

A further additional step in solving the problemmay be the analysis of polycyclic alkanes of sterane

and triterpane types (Jovančićević et al. 2001a).Namely, these types of petroleum biomarkers arecharacterized by easy recognizable distribution ofbiolipid or geolipid isomers, and it can be used as acrude oil “fingerprint” (Tissot and Welte 1984; Peterset al. 2005). Therefore, in the case when differentia-tion of native and anthropogenic origin of an organicsubstance based on distribution of n-alkanes or carbonisotope analysis becomes questionable (differentillustrative examples of n-alkane distributions inPančevo Oil Refinery underground water extractsare shown in Fig. 3, Jovančićević et al. 2001a),analysis of steranes and triterpanes may be helpful

Fig. 5 Gas chromatograms of alkanes isolated from oilpolluted alluvial ground waters (Pančevo Oil Refinery locality).Samples were taken in November 1997 (a), May 1998 (b), inSeptember 1998 (c), September 1999 (d) and in February 2000(sample e) (Jovančićević et al. 2001b; Wehner et al. 2001)


(corresponding fragmentograms are shown in Fig. 4,Jovančićević et al. 2001a).

All samples shown in Fig. 3, except the sample I,may be supposed to represent a petroleum-typepollutant, since their steranes and triterpanes werecharacterized by typical crude oil distributions(Fig. 4). In addition to biolipid C27–C29 αα (20R)isomers (peaks 2, 3 and 6, Fig. 4a), geolipid isomers,such as diasteranes and C27–C29 isomers withhydrogen atom at C14 and C17 in β-position andS-configuration at C20 (peaks 1, 4 and 5) were alsoobserved in significant amounts. Moreover, withtriterpanes (Fig. 4b), in addition to oleanane andgammacerane, thermodynamically the most stablehopanes (peaks a, c and e, isomer 22S), the pro-portions of their less stable isomers (peaks b, d and e,isomer 22R) were also characteristic of crude oils.

This use of n-alkanes and polycyclic alkanes of thesterane and triterpane types can be defined as an“organic-geochemical approach” in the identificationof oil type pollutants in the environment.

3 Transformation Processes of Petroleum-typePollutants

As already indicated, petroleum-type pollutants inrecent sediments, soil, underground or surface waters,are exposed to microbial degradation. This happensunder various conditions. The degradation intensitydiffers depending on a great number of biological,chemical and physicochemical parameters. Theprocesses may be followed in different ways. For

Fig. 6 Fragmentograms of steranes (m/z 217) and triterpanes (m/z 191) from oil polluted ground waters (d) and (e) (Jovančićević etal. 2001b). (Peak identification in Fig. 4)


example, the process of biodegradation of petroleum-type pollutants in underground waters from Danubealluvial sediments (the locality of Pančevo OilRefinery) was followed through a period fromNovember 1997 to February 2000 by GC analysesof isolated alkane fractions. The corresponding gaschromatograms are shown in Fig. 5 (Jovančićevićet al. 2001b; Wehner et al. 2001).

In the period from November 1997, when the firstsample was taken, to February 2000, when the fifthsample was taken, important changes of the chemicalcomposition were obvious. Relative contribution ofn-alkanes as compared to pristane and phytane insample Ia indicated changes defined as “initialpetroleum biodegradation”. The abundance of C17

and C18 n-alkanes was somewhat smaller than theabundance of pristane (C19) and phytane (C20). Asknown, gas chromatographic retention times of thesehydrocarbons are very similar. Gas chromatogram of

sample b showed that in the period from November1997 to May next year the amount of n-alkanesrelative to isoprenoids was reduced, a phenomenontypical for biodegradation intensity in geochemicalliterature defined as “minimal biodegradation”(Volkman et al. 1983). Later, in September 1998(sample c), the amount of n-alkanes was still smaller.Finally, during the next year, n-alkanes were almostcompletely degraded (sample d). Pristane and phytaneremained nonbiodegraded.

Comparison of Pr/n-C17 and Phyt/n-C18 ratiosobserved in samples a–c (winter 1997–autumn 1998)suggested that biodegradation was considerablymore intensive during the summer period than duringthe winter or spring periods. On the other hand, bycomparing these degradation intensities with thoseobserved in samples originating from close butdeeper localities, it was concluded that biodegradationof the petroleum-type pollutant was more intensive in

Fig. 7 Chromatograms of GC-MS analyses of fractions of alkanes, alcohols and fatty acids (their methyl-esters), isolated fromextracts (d) and (e) (Jovančićević et al. 2003)


shallow undergroundwaters (2.8 vs. 5.8m; Jovančićevićand Polić 2000).

In a relatively short period of time, from Septem-ber 1999 to February 2000, the alkane fraction of thepetroleum-type pollutant suffered an unexpectedchange (sample e, Fig. 5, Jovančićević et al. 2001b;Wehner et al. 2001). Namely, while pristane andphytane were found in the same amounts character-ized by approximately the same ratios, in this fractionof the pollutant new even carbon-number C16–C30 n-alkanes were observed. Such a significant changeraised the question of possible new contaminationduring this period of time, e.g., by a pollutantcharacterized by dominating even carbon-number n-alkanes. However, identical distributions of bothsteranes and triterpanes in sample d and e (Fig. 6,Jovančićević et al. 2001b) were a high evidence of thepresence of the same petroleum-type pollutant in bothsamples. Hence, the question concerning the origin ofthe new, even carbon-number C16–C30 n-alkanesbecame provocative.

It was supposed that these even carbon-numbern-alkanes were biosynthesized by some microorgan-

isms. According to literature data, the followingorganisms are known to synthesize such compounds:Desulfovibrio desulfuricans, Corynebacterium sp.,Escherichia coli, Rhizopus stolonifer, Penicillium sp.or algae of Pyrrophyta type (Grimalt and Albaigés1987; Blaženčić 1988). These are all microorganismswhich have grown on different organic basal medium(for example sewerage waste, Jovančićević et al.1998a). Consequently, confirmation of biosynthesisof even carbon-number C16–C30 n-alkanes was triedby detailed analysis of extracts obtained from samplesd and e (Fig. 7, Jovančićević et al. 2003).

It was found that sample Ie, containing remarkableamounts of even n-alkane homologues, contained, aswell, in the alcoholic fraction, a homologous series ofeven carbon-number alcohols in a C14–C20 range anda relatively significant amount of cholesterol. On theother hand, sample d, which did not contain anysignificant amounts of neither odd nor even n-alkanehomologues, did not contain alcohols nor higher fattyacids.

Even carbon-number alcohols and fatty acidsobserved in sample e were taken as a proof of the

Fig. 8 Gas chromatogramsof saturated hydrocarbonsfrom extracts which wereisolated from ground waters(Pančevo Oil Refinery lo-cality) by decantation (sam-ple a), by shaking in aseparatory funnel for 1 min(b), by shaking in a separa-tory funnel for 5 min (c) andby mechanical shaking for24 h (sample d)(Jovančićević et al. 2005)


presence of particular microorganisms, i.e., of unicel-lular, nonphotosynthetic algae of Pyrrophyta type.These types of microorganisms, also known under apopular name of “fire algae”, are known to be able tosynthesize even n-alkane homologues on a suitablebase such as petroleum or petroleum-type pollutants(Blaženčić 1988). They were shown to be more activein the case of closer interaction of petroleum andwater (Šolević et al. 2003). Biosynthesized even n-alkane homologues were observed only in petroleum-type pollutant isolated from underground water afterprolonged shaking in a separating funnel. On the

other hand, with petroleum-type pollutant preliminaryseparated from the same sample by simple decanting,which thus represented a pollutant fraction which hadbeen in weaker interaction with water, such activity ofmicroorganisms was not observed. Distribution of n-alkanes in surface waters_ petroleum-type pollutantsalso depended on the intensity of their interactionwith water.

Hence, it is suggested that the observed distributionof n-alkanes in a petroleum-type pollutant may dependon the method of its isolation from the polluted surfacewater (Fig. 8; Jovančićević et al. 2005).

Fig. 9 Gas chromatogramsof saturated hydrocarbonfractions isolated from dis-charged heavy fuel oil(R-2a → R-9c, increaseof the depth and distancefrom oil spill) (Jovančićevićet al. 1996)


An experiment shown in Fig. 8 will shortly bedescribed as to illustrate the latter statement. The firsttwo samples, isolated after a weak interaction withwater, showed a typical crude oil n-alkane distribu-tion, i.e., a uniform distribution of odd and evenhomologues, with a maximum at n-C18. Even homo-logues were already observed in extract isolated after5 min shaking (sample c, somewhat stronger interac-tion with water). Finally, in the extract isolated afterintensive 24 h shaking (d), larger amount of higherhomologues was observed, which changed the n-alkane distribution into a bimodal distribution.

Increased amount of higher n-alkane homologueswas observed during the migration of a petroleum-type pollutant through aquatic sediments (Fig. 9;Jovančićević et al. 1996). It was then supposed that

polar petroleum components (nitrogen, sulphur, oxy-gen and NSO-compounds) were forming colloidalmicelles with water and that higher homologues of n-alkanes were forming more stable inclusion com-pounds with developed colloidal micelles. In that waythey were “transported” preferentially, compared toshorter-chain n-alkanes (the shift of n-alkane maxi-mum from n-C19 to n-C26 in Fig. 9).

The fate of a petroleum-type pollutant in environ-mental water may be foreseen on the basis oflaboratory simulation experiments of microbiologicaldegradation of petroleum using microorganism con-sortiums similar to those typical for the naturalenvironment, activated on a corresponding nutrientbase (Warren et al. 2002). As an example, Fig. 10shows a gas chromatogram of the alkane fraction of a

Fig. 10 Gas chromato-grams of the alkane frac-tions derived from paraffinictype petroleum of Sirakovoafter 90 days of simulatedbiodegradation with Phor-midium foveolarum,Achanthes minutissima,Nitzschia communis andChlorella communis withKp medium in the light(Sir-1), with Kp medium inthe dark (Sir-2), with Bhmedium in the light (Sir-3),with Bh medium in the dark(Sir-4) together with achromatogram of the alkanefraction typical for the con-trol experiments (Sir-1C)(Antić et al. 2006)


paraffinic-type crude oil originating from Sirakovo,Serbia, oil field (Sir-1C) (Jovančićević et al. 1998b),and gas chromatograms of alkane fractions of thesame crude oil isolated after 90 days of simulatedbiodegradation on an inorganic “Knop” base(Ca(NO3)2×4H2O, K2HPO4, MgSO4×7H2O, KCl,FeCl3, H2SO4, H2O; pH≈8) under daylight (Topa-chevskii 1975) (Sir-1) and in absence of light (Sir-2),as well as on a “Bujon” organic base (tryptone, yeastextract, glucose, distilled water, pH≈7) under daylight(Sir-3) and in darkness (Sir-4). The experiments werecarried out with microorganisms consortium similar tothat one identified as dominant in the investigatedsurface sewage water in the channel of the PančevoOil Refinery (Phormidium foveolarum, Achanthesminutissima, Nitzschia communis, Chlorella commu-nis) (Stewart 1974; Topachevskii 1975; Blaženčić1988; Cvijan and Blaženčić 1996; Antić et al. 2006).

In an experiment on a Knop base, which maximal-ly corresponded to natural conditions, by biodegrada-tion under daylight (Sir-1), n-alkanes were almostcompletely degraded. In darkness, the degradationwas less effective (Sir-2). In experiments on a Bujonbase, n-alkanes were found to be much less degraded(Sir-3 and Sir-4).

On the other hand, in experiments carried out inthe same way, using the same nutrient bases andidentical microorganisms, but with a naphthenic-typecrude oil originating from the Velebit, Serbia, oil fieldas substrate (Šaban et al. 1990), it was possible tofollow the degradation of isoprenoid aliphatic com-pounds (Antić et al. 2006). Comparison of Phyt/C30-

hopane ratios observed in the investigated samplesafter 90 days of simulated biodegradation (Ve-1–Ve-4) with those observed in control tests (Ve-1C–Ve-4C;Table 1), showed the degradation of isoprenoids to bealso most pronounced on the Knop inorganic nutrientbase and under daylight. Under identical experimentalconditions, polycyclic alkanes of sterane and triter-pane types retained their original distributions.

4 Conclusions

A review is given in this paper of the up-to-dateresults observed in differentiation and transformationstudies of petroleum-type pollutants in undergroundand surface waters. Water and particulate matterderived from the locality of Pančevo PetroleumRefinery, Serbia, which belongs to the River Danubealluvial formations. A few conclusions are proposedon both the differentiation of petroleum-type pollu-tants, as the first stage of any similar study, as well ason the subsequent study of the pollutant transforma-tion processes.

Thanks to different thermal maturity of petroleum,on the one hand, and the recent sediments, bitumen,on the other, n-alkanes may successfully be used fordifferentiating the petroleum-type pollutants, as an-thropogenic organic substances, from native organicmatter in recent sedimentary formations.

Furthermore, being a more mature organic sub-stance compared to the recent sediments, bitumen,petroleum contains a higher amount of heavier carbonisotope, i.e., it is characterized by a less negativeδ13CPDB value. The δ13CPDB is thus shown to beanother parameter for successful differentiation ofpetroleum-type pollutants.

Finally, the distribution of polycyclic alkanes ofsterane and triterpane types is typical for crude oils.Therefore, these classes of compounds, i.e., theirdistributions, may also efficiently be used for reveal-ing the presence of petroleum-type pollutants inrecent formations.

In underground waters, a petroleum-type pollutantis exposed to microbiological degradation which ismanifested through relatively fast degradation of n-alkanes. Their degradation is more efficient during thesummer period as well as in shallow water.

Following an almost complete degradation ofcrude oil n-alkanes, the biosynthesis of novel, even

Table 1 Parameters based on gas chromatograms of Velebitcrude oil alkane fractions (Antić et al. 2006)

Pr/Phyt Phyt/C30-hopane

Ve-1C 0.21 1.68Ve-1 ND NDa

Ve-2C 0.37 3.81Ve-2 ND NDa

Ve-3C 0.46 4.00Ve-3 0.52 1.67Ve-4C 0.10 0.50Ve-4 0.30 0.32

ND – parameter was not calculated due to total degradation ofpristane and phytane.

NDa – parameter was not calculated due to total degradation ofphytane.


carbon-number C16–C30 n-alkanes may be observed.For example, unicellular nonphotosynthetical algae ofPyrrophyta type are able to biosynthesize even n-alkane homologues on a suitable base such aspetroleum or petroleum-type pollutant. Hence, thistype of organisms may be the source of newly formedhydrocarbons.

It was shown that the n-alkane distribution ob-served in a petroleum-type pollutant may depend onthe intensity of its previous interaction with water,and therefore it may not be the same in all pollutantfragments. In other words, the distribution of thepollutants, n-alkanes will depend on the extractionprocedure. The petroleum pollutant fraction separatedfrom the polluted water by simple decanting or byextraction after a short shaking in a separating funnel,shall contain n-alkanes of characteristic petroleumdistribution. On the other hand, even n-alkanehomologues, as well as higher homologues, will berevealed only by thorough extraction.

The fate of petroleum-type pollutants in environ-mental waters may be predicted through laboratorysimulative microbiological degradation experimentsby using microorganism consortiums similar to thoseobserved under relevant natural conditions, as well ason corresponding nutrient base. For example, inillustrative simulation experiments with carefullychosen microorganisms consortium, on an inorganicbase (Ca(NO3)2×4H2O, K2HPO4, MgSO4×7H2O,KCl, FeCl3, H2SO4, H2O) which corresponded tonatural conditions in the waste water channel of thePančevo Oil Refinery, under daylight, n-alkanes of aparaffinic-type petroleum were almost completelydegraded. In contrast, on an organic base, consistingof tryptone, yeast extract, glucose and distilled water,these hydrocarbons were much less degraded.

Acknowledgments We thank the Alexander von HumboldtFoundation and the Ministry of Science and EnvironmentProtection of the Republic of Serbia for supporting thisresearch.

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organic-geochemical differentiation of petroleum-type pollutants and study of their fate in danube...

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