jelena avdalovic et al - university of belgradesrdjan/ref/2016_emec_17_javdalovic.pdf · 1institute...
TRANSCRIPT
BIOREMEDIATION BIOREMEDIATION OF OF COMPLEX CONTAMINANT FROM PETROLEUM INDUSTRYCOMPLEX CONTAMINANT FROM PETROLEUM INDUSTRY
IN IN AN ARTIFICIAL SOIL AN ARTIFICIAL SOIL SUBSTRATESUBSTRATE
Jelena AvdalovićJelena Avdalović1,*1,*, Tatjana Šolević Knudsen, Tatjana Šolević Knudsen11, Srđan Miletić, Srđan Miletić11, Mila Ilić, Mila Ilić11, Jelena Milić, Jelena Milić11, Vladimir Beškoski, Vladimir Beškoski22, Miroslav M. Vrvić, Miroslav M. Vrvić22
11Institute of Chemistry, Technology and Metallurgy, University of Belgrade, Institute of Chemistry, Technology and Metallurgy, University of Belgrade, NjegoševaNjegoševa 12, P.O. Box 473, 11001 Belgrade, Serbia12, P.O. Box 473, 11001 Belgrade, Serbia22Faculty of Chemistry, University of Belgrade, Faculty of Chemistry, University of Belgrade, StudentskiStudentski trgtrg 1212--16, P.O. Box 158, 11001 Belgrade, Serbia16, P.O. Box 158, 11001 Belgrade, Serbia
(* corresponding author: [email protected] )(* corresponding author: [email protected] )
BIOREMEDIATION BIOREMEDIATION OF OF COMPLEX CONTAMINANT FROM PETROLEUM INDUSTRYCOMPLEX CONTAMINANT FROM PETROLEUM INDUSTRY
IN IN AN ARTIFICIAL SOIL AN ARTIFICIAL SOIL SUBSTRATESUBSTRATE
Jelena AvdalovićJelena Avdalović1,*1,*, Tatjana Šolević Knudsen, Tatjana Šolević Knudsen11, Srđan Miletić, Srđan Miletić11, Mila Ilić, Mila Ilić11, Jelena Milić, Jelena Milić11, Vladimir Beškoski, Vladimir Beškoski22, Miroslav M. Vrvić, Miroslav M. Vrvić22
11Institute of Chemistry, Technology and Metallurgy, University of Belgrade, Institute of Chemistry, Technology and Metallurgy, University of Belgrade, NjegoševaNjegoševa 12, P.O. Box 473, 11001 Belgrade, Serbia12, P.O. Box 473, 11001 Belgrade, Serbia22Faculty of Chemistry, University of Belgrade, Faculty of Chemistry, University of Belgrade, StudentskiStudentski trgtrg 1212--16, P.O. Box 158, 11001 Belgrade, Serbia16, P.O. Box 158, 11001 Belgrade, Serbia
(* corresponding author: [email protected] )(* corresponding author: [email protected] )
INTRODUCTIONINTRODUCTION
BioremediationBioremediation isis aa processprocess basedbased onon thethe naturalnatural capacitycapacity ofof microorganismsmicroorganisms toto
degradedegrade oror transformtransform toxictoxic substancessubstances intointo lessless toxictoxic oror harmlessharmless productsproducts..
TheThe objectiveobjective ofof thisthis researchresearch waswas toto investigateinvestigate thethe efficiencyefficiency ofof bioremediationbioremediation
ofof complexcomplex contaminantcontaminant fromfrom petroleumpetroleum industryindustry inin anan artificialartificial soilsoil substratesubstrate..
INTRODUCTIONINTRODUCTION
BioremediationBioremediation isis aa processprocess basedbased onon thethe naturalnatural capacitycapacity ofof microorganismsmicroorganisms toto
degradedegrade oror transformtransform toxictoxic substancessubstances intointo lessless toxictoxic oror harmlessharmless productsproducts..
TheThe objectiveobjective ofof thisthis researchresearch waswas toto investigateinvestigate thethe efficiencyefficiency ofof bioremediationbioremediation
ofof complexcomplex contaminantcontaminant fromfrom petroleumpetroleum industryindustry inin anan artificialartificial soilsoil substratesubstrate..
EXPERIMENTALEXPERIMENTAL
TheThe artificialartificial soilsoil substratesubstrate waswas preparedprepared asas aa mixturemixture ofof sandsand andand sawdustsawdust.. AA
complexcomplex contaminantcontaminant fromfrom petroleumpetroleum industryindustry waswas addedadded toto makemake thethe finalfinal massmass ofof
thethe experimentalexperimental biopilebiopile ofof 4545 kgkg.. DuringDuring thethe experiment,experiment, thethe biopilebiopile waswas inoculatedinoculated
withwith multipliedmultiplied consortiaconsortia ofof microorganisms,microorganisms, isolatedisolated fromfrom thethe complexcomplex contaminantcontaminant..
ReinoculationReinoculation waswas carriedcarried outout everyevery thirtythirty daysdays.. TheThe pilepile waswas mixedmixed onceonce perper monthmonth toto
achieveachieve thethe effecteffect ofof aerationaeration.. CompoundsCompounds ofof nitrogen,nitrogen, phosphorusphosphorus andand potassiumpotassium
werewere usedused forfor biostimulationbiostimulation.. TheThe resultsresults ofof thisthis experimentexperiment werewere comparedcompared withwith thethe
resultsresults ofof analysesanalyses ofof aa controlcontrol pile,pile, whichwhich waswas preparedprepared inin thethe samesame wayway asas thethe biopilebiopile
butbut withoutwithout inoculation,inoculation, aerationaeration andand biostimulationbiostimulation.. TheThe experimentexperiment lastedlasted 110110 daysdays..
TotalTotal PetroleumPetroleum HydrocarbonsHydrocarbons (TPH,(TPH, asas perper methodmethod ISO/TRISO/TR 1104611046::19941994)) inin thethe
biopilebiopile andand thethe controlcontrol pilepile werewere analyzedanalyzed atat thethe beginning,beginning, afterafter 6060 days,days, andand atat thethe
endend ofof thethe experimentexperiment..
InIn orderorder toto investigateinvestigate changeschanges inin distributiondistribution andand abundancesabundances ofof differentdifferent
compoundscompounds duringduring thisthis experiment,experiment, TPHTPH extractsextracts fromfrom thethe biopilebiopile werewere analyzedanalyzed byby
gasgas chromatographychromatography –– massmass spectrometryspectrometry (GC(GC--MS)MS)..
EXPERIMENTALEXPERIMENTAL
TheThe artificialartificial soilsoil substratesubstrate waswas preparedprepared asas aa mixturemixture ofof sandsand andand sawdustsawdust.. AA
complexcomplex contaminantcontaminant fromfrom petroleumpetroleum industryindustry waswas addedadded toto makemake thethe finalfinal massmass ofof
thethe experimentalexperimental biopilebiopile ofof 4545 kgkg.. DuringDuring thethe experiment,experiment, thethe biopilebiopile waswas inoculatedinoculated
withwith multipliedmultiplied consortiaconsortia ofof microorganisms,microorganisms, isolatedisolated fromfrom thethe complexcomplex contaminantcontaminant..
ReinoculationReinoculation waswas carriedcarried outout everyevery thirtythirty daysdays.. TheThe pilepile waswas mixedmixed onceonce perper monthmonth toto
achieveachieve thethe effecteffect ofof aerationaeration.. CompoundsCompounds ofof nitrogen,nitrogen, phosphorusphosphorus andand potassiumpotassium
werewere usedused forfor biostimulationbiostimulation.. TheThe resultsresults ofof thisthis experimentexperiment werewere comparedcompared withwith thethe
resultsresults ofof analysesanalyses ofof aa controlcontrol pile,pile, whichwhich waswas preparedprepared inin thethe samesame wayway asas thethe biopilebiopile
butbut withoutwithout inoculation,inoculation, aerationaeration andand biostimulationbiostimulation.. TheThe experimentexperiment lastedlasted 110110 daysdays..
TotalTotal PetroleumPetroleum HydrocarbonsHydrocarbons (TPH,(TPH, asas perper methodmethod ISO/TRISO/TR 1104611046::19941994)) inin thethe
biopilebiopile andand thethe controlcontrol pilepile werewere analyzedanalyzed atat thethe beginning,beginning, afterafter 6060 days,days, andand atat thethe
endend ofof thethe experimentexperiment..
InIn orderorder toto investigateinvestigate changeschanges inin distributiondistribution andand abundancesabundances ofof differentdifferent
compoundscompounds duringduring thisthis experiment,experiment, TPHTPH extractsextracts fromfrom thethe biopilebiopile werewere analyzedanalyzed byby
gasgas chromatographychromatography –– massmass spectrometryspectrometry (GC(GC--MS)MS)..
Figure 1. Total ion chromatograms of the TPH extracts isolated from the Figure 1. Total ion chromatograms of the TPH extracts isolated from the biopilebiopile during the experiment.during the experiment.
FigureFigure 22.. GCGC--MSMS chromatogramschromatograms ofof ionsions m/zm/z == 128128 (naphthalene(naphthalene –– N),N), m/zm/z == 142142 (methyl(methyl--
naphthalenesnaphthalenes –– MN),MN), m/zm/z == 156156 ((dimethyldimethyl--naphthalenesnaphthalenes –– DMN),DMN), m/zm/z == 170170 ((trimethyltrimethyl--naphthalenesnaphthalenes
–– TMN)TMN) andand m/zm/z == 184184 ((tetramethyltetramethyl--naphthalenesnaphthalenes –– TeMNTeMN andand dibenzotiophenedibenzotiophene –– DBT)DBT) ofof thethe TPHTPH
RESULTSRESULTS
TotalTotal decreasedecrease ofof TPHTPH inin thethe controlcontrol pilepile atat thethe endend ofof thethe experimentexperiment waswas
55..5656 %%.. TheThe analysisanalysis ofof thethe biopilebiopile showedshowed aa totaltotal decreasedecrease inin TPHTPH ofof 5454..7575 %%..
TheThe GCGC--MSMS analysisanalysis ofof TPHTPH extractextract fromfrom thethe biopilebiopile atat thethe beginningbeginning ofof thethe
experimentexperiment showedshowed thatthat dominantdominant compoundscompounds inin thisthis contaminantcontaminant werewere isoprenoidsisoprenoids
andand petroleumpetroleum polycyclicpolycyclic aromaticaromatic hydrocarbonshydrocarbons (PPAH)(PPAH):: naphthalene,naphthalene, phenanthrenephenanthrene
andand dibenzotiophenedibenzotiophene,, andand theirtheir CC11--CC44 alkylatedalkylated homologueshomologues (Figures(Figures 11 -- 44))..
AtAt thethe endend ofof thethe experiment,experiment, mostmost ofof thethe PPAHPPAH werewere reducedreduced inin abundances,abundances,
andand somesome ofof themthem werewere completelycompletely removedremoved (Figures(Figures 11--44))..
RESULTSRESULTS
TotalTotal decreasedecrease ofof TPHTPH inin thethe controlcontrol pilepile atat thethe endend ofof thethe experimentexperiment waswas
55..5656 %%.. TheThe analysisanalysis ofof thethe biopilebiopile showedshowed aa totaltotal decreasedecrease inin TPHTPH ofof 5454..7575 %%..
TheThe GCGC--MSMS analysisanalysis ofof TPHTPH extractextract fromfrom thethe biopilebiopile atat thethe beginningbeginning ofof thethe
experimentexperiment showedshowed thatthat dominantdominant compoundscompounds inin thisthis contaminantcontaminant werewere isoprenoidsisoprenoids
andand petroleumpetroleum polycyclicpolycyclic aromaticaromatic hydrocarbonshydrocarbons (PPAH)(PPAH):: naphthalene,naphthalene, phenanthrenephenanthrene
andand dibenzotiophenedibenzotiophene,, andand theirtheir CC11--CC44 alkylatedalkylated homologueshomologues (Figures(Figures 11 -- 44))..
AtAt thethe endend ofof thethe experiment,experiment, mostmost ofof thethe PPAHPPAH werewere reducedreduced inin abundances,abundances,
andand somesome ofof themthem werewere completelycompletely removedremoved (Figures(Figures 11--44))..
Table 1. Decrease of TPH in the control and Table 1. Decrease of TPH in the control and biopilebiopile during the experiment.during the experiment.
REFERENCESREFERENCES::
AvdalovićAvdalović JelenaJelena,, DoctoralDoctoral Dissertation,Dissertation, GenerationGeneration ofof soilsoil inin thethe processprocess ofof bioremediation,bioremediation, FacultyFaculty ofof
Chemistry,Chemistry, Belgrade,Belgrade, 20152015 (in(in Serbian)Serbian)..
ISO/TRISO/TR 1104611046::19941994 InternationalInternational standardstandard:: soilsoil qualityquality -- DeterminationDetermination ofof mineralmineral oiloil contentcontent -- MethodMethod
byby infraredinfrared spectrometryspectrometry andand gasgas chromatographicchromatographic methodmethod..
ACKNOWLEDGMENTSACKNOWLEDGMENTS::
WeWe thankthank thethe MinistryMinistry ofof Education,Education, ScienceScience andand TechnologicalTechnological DevelopmentDevelopment ofof thethe RepublicRepublic ofof SerbiaSerbia
(Projects(Projects 176006176006 andand IIIIII 4300443004)) forfor supportingsupporting thisthis researchresearch..
REFERENCESREFERENCES::
AvdalovićAvdalović JelenaJelena,, DoctoralDoctoral Dissertation,Dissertation, GenerationGeneration ofof soilsoil inin thethe processprocess ofof bioremediation,bioremediation, FacultyFaculty ofof
Chemistry,Chemistry, Belgrade,Belgrade, 20152015 (in(in Serbian)Serbian)..
ISO/TRISO/TR 1104611046::19941994 InternationalInternational standardstandard:: soilsoil qualityquality -- DeterminationDetermination ofof mineralmineral oiloil contentcontent -- MethodMethod
byby infraredinfrared spectrometryspectrometry andand gasgas chromatographicchromatographic methodmethod..
ACKNOWLEDGMENTSACKNOWLEDGMENTS::
WeWe thankthank thethe MinistryMinistry ofof Education,Education, ScienceScience andand TechnologicalTechnological DevelopmentDevelopment ofof thethe RepublicRepublic ofof SerbiaSerbia
(Projects(Projects 176006176006 andand IIIIII 4300443004)) forfor supportingsupporting thisthis researchresearch..
CONCLUSIONSCONCLUSIONS
TheThe resultsresults ofof thisthis researchresearch showedshowed thatthat bioremediationbioremediation ofof petroleumpetroleum pollutantpollutant
disperseddispersed inin thethe artificialartificial soilsoil substratesubstrate cancan removeremove asas muchmuch asas 5555 %% ofof totaltotal petroleumpetroleum
hydrocarbonshydrocarbons inin 110110 daysdays.. ConsideringConsidering thethe factfact thatthat thethe onlyonly compoundscompounds affectedaffected inin
thisthis researchresearch werewere PPAH,PPAH, itit cancan bebe concludedconcluded thatthat thethe procedureprocedure describeddescribed cancan removeremove
upup toto 5555 %% ofof PPAHPPAH fromfrom petroleumpetroleum pollutantpollutant inin relativelyrelatively shortshort period,period, whichwhich makesmakes
itit especiallyespecially attractiveattractive inin environmentalenvironmental applicationsapplications..
CONCLUSIONSCONCLUSIONS
TheThe resultsresults ofof thisthis researchresearch showedshowed thatthat bioremediationbioremediation ofof petroleumpetroleum pollutantpollutant
disperseddispersed inin thethe artificialartificial soilsoil substratesubstrate cancan removeremove asas muchmuch asas 5555 %% ofof totaltotal petroleumpetroleum
hydrocarbonshydrocarbons inin 110110 daysdays.. ConsideringConsidering thethe factfact thatthat thethe onlyonly compoundscompounds affectedaffected inin
thisthis researchresearch werewere PPAH,PPAH, itit cancan bebe concludedconcluded thatthat thethe procedureprocedure describeddescribed cancan removeremove
upup toto 5555 %% ofof PPAHPPAH fromfrom petroleumpetroleum pollutantpollutant inin relativelyrelatively shortshort period,period, whichwhich makesmakes
itit especiallyespecially attractiveattractive inin environmentalenvironmental applicationsapplications..
extractsextracts isolatedisolated fromfrom thethe biopilebiopile duringduring thethe experimentexperiment..
FigureFigure 33.. GCGC--MSMS chromatogramschromatograms ofof ionsions m/zm/z == 178178 ((phenanthrenephenanthrene –– PP andand anthraceneanthracene –– A),A), m/zm/z ==
192192 (methyl(methyl--phenanthrenesphenanthrenes –– MP),MP), m/zm/z == 206206 (dimethyl(dimethyl--phenanthrenesphenanthrenes –– DMP)DMP) andand m/zm/z == 220220
((trimethyltrimethyl--phenanthrenesphenanthrenes –– TMP)TMP) ofof thethe TPHTPH extractsextracts isolatedisolated fromfrom thethe biopilebiopile duringduring thethe
experimentexperiment..
FigureFigure 44.. GCGC--MSMS chromatogramschromatograms ofof ionsions m/zm/z == 184184 ((dibenzotiophenedibenzotiophene –– DBT),DBT), m/zm/z == 198198 ((methylmethyl--
dibenzotiophenesdibenzotiophenes –– MDBT),MDBT), andand m/zm/z == 212212 ((dimethyldimethyl--dibenzotiophenesdibenzotiophenes –– DMDBT)DMDBT) ofof thethe TPHTPH
extractsextracts isolatedisolated fromfrom thethe biopilebiopile duringduring thethe experimentexperiment..
Day of the Day of the
experimentexperimentTPHTPH Degraded TPH (%)Degraded TPH (%)
Control pileControl pile
0. day0. day 21.0521.05 00
60. day60. day 20.3920.39 3.143.14
110. day110. day 19.8819.88 5,565,56
BiopileBiopile
0. day0. day 18.0118.01 00
60. Day60. Day 13.7413.74 23.7123.71
110. day110. day 8.158.15 54.7554.75