Polycyclic aromatic hydrocarbons at fire stations: firefighters’exposure monitoring and biomonitoring, and assessment of the contribution to total internal dose

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ContentslistsavailableatScienceDirect

Journal

of

Hazardous

Materials

j ou rn a l h om epa ge :w w w . e l s e v i e r . c o m / l o c a t e / j h a z m a t

Polycyclic

aromatic

hydrocarbons

at

ﬁre

stations:

ﬁreﬁghters’

exposure

monitoring

and

biomonitoring,

and

assessment

of

the

contribution

to

total

internal

dose

Marta

Oliveira

a,b

_,

_Klara

_Slezakova

a,b

_,

_Maria

_José

_Alves

c

_,

_Adília

_Fernandes

c

_,

João

Paulo

Teixeira

d,e

_,

_Cristina

_{Delerue-Matos}

a

_,

_Maria

_do

_Carmo

_Pereira

b

_,

Simone

Morais

a,∗

a_{REQUIMTE-LAQV,}_Instituto_Superior_de_Engenharia,_Instituto_Politécnico_do_Porto,_R._Dr._António_Bernardino_de_Almeida_431,_4200-072_Porto,_Portugal b_LEPABE,_Departamento_de_Engenharia_Química,_Faculdade_de_Engenharia,_Universidade_do_Porto,_R._Dr._Roberto_Frias,_4200-465_Porto,_Portugal c_Escola_Superior_de_Saúde,_Instituto_Politécnico_de_Braganc¸_a,_Avenida_D._Afonso_V,_5300-121,_Braganc¸_a,_Portugal

d_Instituto_Nacional_de_Saúde_Pública,_Departamento_de_Saúde_Ambiental,_Rua_Alexandre_Herculano_321,_4000-055_Porto,_Portugal e_Universidade_do_Porto,_Instituto_de_Saúde_Pública,_Rua_das_Taipas_135,_4050-600_Porto,_Portugal

h

i

g

h

l

i

g

h

t

s

•FireﬁghtersexposuretoPAHswasassessedbypersonalmonitoringandbiomonitoring.

•AirbornePAHswith2–3ringswerethemostabundantatallﬁrestations.

•1-hydroxynaphthaleneand1-hydroxyacenaphthenewerethepredominantmetabolites.

•NaphthalenecontributedthemosttocarcinogenicPAHsinmajorityofﬁrehouses.

•SigniﬁcantcorrelationswerefoundamongurinaryOH-PAHexcretionandinhaledPAHs.

a

r

t

i

c

l

e

i

n

f

o

Articlehistory:

Received30December2015 Receivedinrevisedform1March2016 Accepted5March2016

Availableonline8March2016 Keywords:

Fireﬁghters Exposure

Polycyclicaromatichydrocarbons(PAHs) Monohydroxyl-PAHs(OH-PAHs) 1-hydroxypyrene

a

b

s

t

r

a

c

t

Thisworkcharacterizeslevelsofeighteenpolycyclicaromatichydrocarbons(PAHs)inthebreathingair zoneoffirefightersduringtheirregularworkshiftateightPortuguesefirestations,andthefirefighters’ totalinternaldosebysixurinarymonohydroxylmetabolites(OH-PAHs).TotalPAHs(PAHs) concentra-tionsvariedwidely(46.4–428ng/m3_),_mainly_due_to_site_specificity_{(urban/rural)}_and_{characteristics}_(age

andlayout)ofbuildings.AirbornePAHswith2–3ringswerethemostabundant(63.9–95.7%PAHs). Similarly,urinary1-hydroxynaphthaleneand1-hydroxyacenaphthenewerethepredominant metabo-lites(66–96%OH-PAHs).NaphthalenecontributedthemosttocarcinogenicPAHs(39.4–78.1%)in majorityoffirehouses;benzo[a]pyrene,themarkerofcarcinogenicPAHs,accountedwith1.5–10%. Statis-ticallypositivesignificantcorrelations(r≥0.733,p≤0.025)wereobservedbetweenPAHsandurinary OH-PAHsforfirefightersoffourfirestationssuggestingthat,atthesesites,indoorairwastheirmajor exposuresourceofPAHs.Firefighter’spersonalexposuretoPAHsatPortuguesefirestationswerewell belowtheexistentoccupationalexposurelimits.Also,thequantifiedconcentrationsofpost-shift uri-nary1-hydroxypyreneinallfirefighterswereclearlylowerthanthebenchmarklevel(0.5␮mol/mol) recommendedbytheAmericanConferenceofGovernmentalIndustrialHygienists.

1. Introduction

Fireﬁghting,alongwithconstruction,mining,andagriculture,

ranksamong themost dangerousprofessions, withits

occupa-tionalexposurebeingregardedaspossiblecarcinogentohumans

∗ Correspondingauthor.

E-mailaddress:sbm@isep.ipp.pt(S.Morais).

bytheInternationalAgencyforResearchonCancer(IARC)andthe

USNationalInstituteforOccupationalSafetyandHealth(NIOSH)

[1,2].Fireﬁghtingisamongthemosthazardousyettheleast

stud-iedoccupationsinterms ofexposures andtheirrelationshipto

occupationaldiseases.

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous

compounds that are released during the incomplete

combus-tion or pyrolysis of organic material. PAHs are well known

for their cytotoxic and mutagenic properties [3,4], with some

http://dx.doi.org/10.1016/j.jhazmat.2016.03.012

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of them being recognized as endocrine disrupting

chemi-cals [5]; USEPA listed 16 priority PAHs [6]. PAHs possess an

undeniable role in the induction of human carcinogenesis [7],

especially if benzo[a]pyrene (known human carcinogenic) and

benz[a]anthracene, benzo[b]ﬂuoranthene, benzo[j]ﬂuoranthene,

benzo[k]ﬂuoranthene, chrysene, indeno[1,2,3-cd]pyrene, and

naphthalene (possible human carcinogens; [4,8] are present.

Dibenzo[a,l]pyrene and dibenz[a,h]anthracene have been also

under scrutinybecause theyare regarded as probable

carcino-gens tohumans due totheir highercarcinogenic potency than

benzo[a]pyrene [9–12]. Electrophilic compounds such as PAHs

playakeyroleinenvironmentalcancerandsomerecentevidences

associate their role in cardiovascular disease [13]. Fireﬁghters’

occupationalexposurehasbeenassociatedwithexcessmorbidity

and mortality withcardiovascular disease being considered as

theleading causeof deathinapproximately 45%of ﬁreﬁghters

andamajorcauseoftheirmorbidity[14–16].Asaconsequence

ﬁreﬁghters’ occupational exposure to PAHs may promote the

developmentoraggravationofcardiovascularillnesses.Firesare

themajorcontributortooccupationalexposuretoPAHs[17–20].

Otherrelevantsourcesincludemotor-vehicleexhaust(especially

diesel), industrial emissions, residential and commercial

heat-ingwithwood,coal,orotherbiomassfuels,and tobaccosmoke

[21–23].Despitesomeavailableinformationregardingﬁreﬁghters’

occupationalexposuretoPAHsduringliveﬁrecombatactivities

in Australia [20] and USA [17–19,23–27], ﬁreﬁghters may also

be exposed to PAHs when they are at ﬁre stations. Recently

some studies revealed that chemical contaminants from ﬁres

weretrackedbacktoﬁrestationsviaﬁrevehiclesandprincipally

throughﬁreﬁghters’personal equipmentssuchasboots,gloves,

and turnout gear [18,20,28–31]. Only two studies were found

regarding firefighters’ occupational exposure to PAHs at fire

stations [23,32],both performed in USA.No information exists

concerningothercountries,even thoughtheavailableexposure

datamaynotbedirectlyapplicable.

Biologicalmonitoringis animportanttool in theprevention

of occupational diseases related to those exposed chemicals

on a regular basis, particularly when multi-route exposure

(inhalation, dermal, ingestion) or abnormal exposure takes

place. 1-hydroxypyrene (1OHPy) is the most widely used

bio-logical indicator of internal dose of PAHs exposure [33,34];

3-hydroxibenzo[a]pyrene (3OHBaP) is the main metabolite

of the known human carcinogenic benzo[a]pyrene.

Acenaph-thene, ﬂuorene, and phenanthrene are common PAHs in

different matrices [10,35,36] and their major urinary

metabo-lites are1-hydroxylacenaphthene(1OHAce), 2-hydroxylﬂuorene

(2OHFlu), and 1-hydroxyphenanthrene (1OHPhen),

respec-tively. Regarding naphthalene there are more than thirty

Table1

ConcentrationsofPM2.5-boundPAHs(mediana;min-max;ng/m3)inthebreathingairzoneoffirefightersatthefirestations(MirandadoDouro(MRD),TorreDonaChama

(TDC),Sendim(SDM),Mirandela(MDL),TorredeMoncorvo(TMC),Vinhais(VNH),Braganc¸a(BRG),andFreixodeEspadaàCinta(FEC)).

Compound Firestation

MRD TDC SDM MDL TMC VNH BRG FEC Naphthalene 12.2 (8.51–15.1) 6.67 (5.61−6.99) 7.95 (6.04−10.3) 11.5 (10.9−13.5) 4.95 (4.27−8.74) 7.19 (5.95−8.53) 9.21 (4.23−13.0) 35.5 (33.4−38.6) Acenaphthylene 110 (75.6−208) 24.0a _56.4 (27.8−86.9) 125** (24.0−198) 24.0a _43.9*** (24.0−89.1) 27.2*** (24.0−111) 199 (60.3−352) Acenaphthene 88.5 (56.1−111) 2.48** (1.62−4.26) 18.7 (18.4−20.8) 97.3** (1.62−121) 10.3 (9.11−16.2) 1.62a _8.54*** (1.62−18.2) 4.33*** (1.62−8.02) Fluorene 1.05 (0.540−1.49) 0.272*** (0.272−0.588) 0.272a _1.86 (1.62−3.53) 0.446** (0.272−0.528) 0.272a _0.272*** (0.272−0.450) 6.73 (3.13−9.94) Phenanthrene 6.07 (3.74−7.27) 4.08 (3.71−5.97) 2.88 (2.76−3.00) 9.45 (6.83−9.88) 3.99 (3.39−4.42) 4.32 (3.42−5.01) 3.63 (2.97−4.84) 28.5 (21.0−35.9) Anthracene 0.223a _0.223*** (0.223−0.364) 0.223a _0.223*** (0.223−1.59) 0.223a _0.223*** (0.223−0.657) 0.223a _0.636 (0.601−0.675) Fluoranthene 0.351*** (0.351−0.603) 0.351*** (0.351−0.592) 0.351a _1.50 (0.968−2.16) 0.351*** (0.351−0.589) 0.596** (0.351−1.02) 0.351*** (0.351−0.631) 1.88*** (0.351−3.88) Pyrene 1.24** (0.292−1.34) 0.292a _0.292a _2.28 (1.76−2.77) 0.473** (0.292−0.755) 0.582** (0.292−1.09) 0.292a _3.41 (2.01−4.92) Benz[a]anthracene 0.205a _0.205a _0.205a _0.205*** (0.205−0.866) 0.205a _0.205*** (0.205−0.452) 0.205*** (0.205−0.324) 0.446*** (0.205−0.824) Chrysene 0.145*** (0.145−0.358) 0.145a _0.145a _0.145*** (0.145−2.01) 0.365** (0.145−0.475) 0.196*** (0.145−0.743) 0.145a _2.41 (2.01−3.08) Benzo[b+j]ﬂuoranthene 0.844a _0.844a _0.844a _0.844*** (0.844−3.55) 0.844a _2.31** (0.844−3.51) 0.844a _24.5 (15.5−33.9) Benzo[k]ﬂuoranthene 0.238** (0.134−0.642) 0.134a _0.134a _0.134*** (0.134−0.412) 0.134a _0.399** (0.134−0.594) 0.134a _3.84 (2.50−5.15) Benzo[a]pyrene 0.277*** (0.277−1.02) 0.277*** (0.277−0.398) 0.277a _0.277*** (0.277−1.24) 0.277a _1.22** (0.277−2.45) 0.277a _15.1 (9.76−20.5) Dibenzo[a,l]pyrene 0.671a _0.671a _0.671a _0.671a _0.671a _0.671a _0.671a _0.671a Dibenz[a,h]anthracene 3.97** (0.499−13.9) 0.499*** (0.499−1.66) 0.499a _0.767** (0.499−3.63) 0.499a _5.85** (0.499−9.96) 0.499a _51.1 (38.1−65.5) Benzo[ghi]perylene 3.09** (0.355−4.48) 5.08 (2.66−6.78) 1.66 (1.44−1.88) 3.80 (2.86−11.8) 3.17 (1.98−4.00) 4.83 (3.06−8.53) 2.29 (1.55−4.18) 32.9 (27.9−40.8) Indeno[1,2,3-cd]pyrene 0.185*** (0.185−1.18) 0.185a _0.185a _0.185a _0.185a _0.185a _0.185a _17.6 (13.9−23.3) PAHs 229 (200−296) 46.4 (44.0−49.4) 91.7 (64.5−124) 256 (77.6−352) 51.1 (48.8−57.5) 74.6 (44.4−125) 55.0 (49.8−137) 428 (250−631) PAHscarc 20.8 (15.8−26.0) 9.82 (8.57−11.1) 10.9 (9.00−13.3) 16.8 (13.9−23.9) 8.24 (7.45−11.7) 19.6 (9.43−23.6) 12.2 (7.19−16.1) 150 (122−186) Note:Detectionfrequencyofeachcompoundwas100%unlessotherwiseindicate.

*_80%_≤_detection_frequency_<100%. **_60%_≤_detection_frequency_<80%. ***_15%_≤_detection._frequency_<60%.

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Table 2 Monohydroxyl-PAH concentrations (median #; min-max; ␮ mol/mol creatinine) measured in firefighters urine samples from different fire stations (Miranda do Douro (MRD), Torre Dona Chama (TDC), Sendim (SDM), Mirandela (MDL), Torre de Moncorvo (TMC), Vinhais (VNH), Braganc ¸a (BRG), and Freixo de Espada à Cinta (FEC)). Compound * Fire station MRD TDC SDM MDL TMC VNH BRG FEC 1OHNaph + 1OHAce 1.39 0.823–1.60 0.138 # 0.138 − 1.50 1.48 0.731 − 2.25 0.887 0.722 − 2.31 0.707 0.231 − 5.32 1.63 # 0.138 − 8.06 0.650 0.458 − 1.90 3.59 3.26 − 4.14 2OHFlu 0.147 4.40 × 10 − 2− 0.150 5.23 × 10 − 2 2.42 × 10 − 2− 0.152 1.39 × 10 − 2 9.58 × 10 − 3− 2.01 × 10 − 2 0.182 6.95 × 10 − 2− 0.212 4.40 × 10 − 2 3.40 × 10 − 2− 8.75 × 10 − 2 3.58 × 10 − 2 # 5.66 × 10 − 4− 0.134 9.35 × 10 − 2 4.71 × 10 − 2− 0.170 5.86 × 10 − 2 5.32 × 10 − 2− 6.61 × 10 − 2 1OHPhen 4.10 × 10 − 2 6.71 × 10 − 3–0.211 4.17 × 10 − 2 1.65 × 10 − 2− 7.62 × 10 − 2 1.21 × 10 − 2 1.04 × 10 − 2− 1.28 × 10 − 2 7.44 × 10 − 2 4.22 × 10 − 2− 0.103 1.41 × 10 − 2 9.67 × 10 − 3− 2.86 × 10 − 2 3.93 × 10 − 2 3.05 × 10 − 2− 4.54 × 10 − 2 8.38 × 10 − 2 7.07 × 10 − 2− 0.120 3.10 × 10 − 2 1.87 × 10 − 2− 4.33 × 10 − 2 1OHPy 3.59 × 10 − 2 9.23 × 10 − 3− 0.131 2.67 × 10 − 2 1.12 × 10 − 2− 4.10 × 10 − 2 1.35 × 10 − 2 # 1.06 × 10 − 3− 2.79 × 10 − 2 0.146 1.99 × 10 − 2− 0.166 2.10 × 10 − 2 6.07 × 10 − 3− 2.68 × 10 − 2 1.73 × 10 − 2 7.67 × 10 − 3− 4.36 × 10 − 2 5.18 × 10 − 2 3.09 × 10 − 2− 9.91 × 10 − 2 2.70 × 10 − 2 2.35 × 10 − 2− 3.13 × 10 − 2 OH-PAHs 1.61 0.889 − 1.88 0.259 0.133 − 1.56 1.52 0.780 − 2.28 1.29 0.979 − 2.62 0.786 0.281 − 5.42 1.79 0.198 − 8.13 0.879 0.737 − 2.20 3.71 3.37 − 4.27 *1OHNaph + 1OHAce: 1-hydroxynaphthalene + 1-hydroxyacenaphthene; 2OHFlu: 2-hydroxyfluorene; 1OHPHen: 1-hydroxyphenanthrene; 1OHPy: 1-hydroxypyrene. # When the urine concentration was below the LOD, the value of the respective LOD/ √ 2 was used [50] .

identiﬁed metabolites, being 1-hydroxynaphthalene(1OHNaph)

and2-hydroxynaphthalenethemainbiomarkersofoccupational

exposuretonaphthalene [37].Only fourstudieswere foundin

the literature concerning ﬁreﬁghters’ biological monitoring to

PAHs.Amongthem,twocharacterizedexclusively1OHPylevels

[27,38],whiletheothertwoassessedtheurinarylevelsof1OHPy

and1OHNaph[31],andtheconcentrations of1OHPyand three

hydroxyphenanthrenemetabolites[39],respectively.In orderto

evaluateﬁreﬁghters’workplaceconditionsonacontinuousbasis,

emphasisshouldbeplaced onenvironmentalmonitoring,

com-plementedbybiological monitoring.No studywasfoundabout

ﬁreﬁghters’occupational exposuretoPAHs thatsimultaneously

assessedenvironmentalexposurelevelsat ﬁrestations andthe

urinary levels of one or more PAH metabolites. Furthermore,

urinary 1OHAce, 2OHFlu, and 3OHB[a]P were never measured

beforeinﬁreﬁghters.

Thus,thepresentstudyaimstoassessthelevelsof18PAHs(16

USEPApriorityPAHs,dibenzo[a,l]pyrene,andbenzo[j]ﬂuoranthene

recommended by EU [40]) in the breathing air zone of

firefighters’duringtheirworkshiftatPortuguesefirestations,and

theﬁreﬁghters’totalinternaldosebysixbiomarkersofexposure

(1OHNaph,1OHAce,2OHFlu,1OHPhen,1OHPy,and3OHB[a]P)in

urine samples.Thecontributionof personalairborneindividual

(naphthalene, acenaphthene, naphthalene+acenaphthene,

ﬂuo-rene,phenanthrene,pyrene)andPAHsexposuretorespective

individual (1OHNaph+1OHAce, 2OHFlu, 1OHPhen, and 1OHPy)

andOH-PAHsexcretioninurinewasexploredfortheﬁrsttime

inﬁreﬁghters.

2. MaterialsandMethods

2.1. Characterizationofthesamplingsites

PortugalisoneoftheﬁveSouthernEuropeancountriesthatare

themostaffectedbyforestﬁreseveryyear[41],beingtheNorth

andCentreregionsthemostdisturbedareas[42].In2014the

dis-trictofBraganc¸a,inthesub-regionofAltoTrás-os-Montes,(north

ofPortugal),wasthethirdPortuguesedistrictwithahigher

inci-denceofburntarea[42].Thisregionischaracterizedbytypically

hotandverydrysummerswithabsolutemaximumtemperatures

exceedingthe30◦C;wintersareusuallylongandcoldwithabsolute

minimumtemperaturesreachingvaluesbelow0◦C.Samplingwas

performedateightdifferentﬁrestations:MirandadoDouro(MRD),

TorreDonaChama(TDC),Sendim(SDM),Mirandela(MDL),Torre

deMoncorvo(TMC),Vinhais(VNH),Braganc¸a(BRG),andFreixode

EspadaàCinta(FEC)(Fig.1).Twopairsofﬁrestations,namelyMDL

andTDC,aswellasMRDandSDMwere22–23kmawayfromeach

other.TheﬁrestationsFEC,MDL,and MRDwerelocatedinthe

urbancenterofthecity,nearcommercialshopsandbusyroads,

whiletheﬁrehousesTMCandTDCweresituatedinvillages.FEC,

MDL,andMRDwereconstructedbefore1984whiletheotherﬁre

stationswereconstructedand/orrestoredafter2007.AtFEC,MDL

andMRD,thetruckbaywhereﬁreﬁghters’andemergencyvehicles

wereparkedonhaddirectaccesstotheoperationalcontrolcenter

andsocialroomwhereﬁreﬁghtersspendmostoftheirtime(Fig.1S

(a),SupplementaryMaterial).Thedoorthatseparatedthesetwo

microenvironmentswasfrequentlyopened.AtﬁrestationsTDC,

SDM,TMC,VNH,andBRG,thecontrolcenterandsocialroomwere

indirectly(throughcorridorsandwithsomedivisions;Fig.1S(b),

SupplementaryMaterial)connectedwiththetruckbay.

Informationonoutdoormeteorologicalconditions,namely

tem-perature,relativehumidity,windspeed,precipitation,andsolar

radiationduringthesamplingcampaignswereretrievedfromthe

localmeteorologicalstation(Table1S,Supplementarymaterial).

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aero-Fig.1.GeographicallocationofthemonitoredﬁrestationsinthedistrictofBraganc¸a.

dynamicdiameterof10␮m(PM10)and2.5␮m(PM2.5),nitrogen

monoxide(NO),dioxide(NO2)andoxides(NOx),sulphurdioxide

(SO2),andozone(O3)werealsomonitoredfromthe

meteorologi-calstationduringthesamplingcampaign(Table1S,Supplementary

material).

2.2. Personalairandurinesampling

Thestudypopulationconsistedofeightunitsofprofessional

ﬁreﬁghters servingat six municipalcities(Fig.1). Astructured

questionnaireadaptedfromvalidatedquestionnaire[43]wasﬁlled

by each ﬁreﬁghter. The questionnaire collectedinformation on

gender,age,numberofyearsasﬁreﬁghter(Table2S,

Supplemen-tary material),and factors reported tobe associated with PAH

exposures,namelysmokinghabits,andthemostconsumedmeals

(boiled,roasted,andgrilled)duringtheﬁvedaysbeforeurine

col-lection.Subjectswereexcludedfromthestudyiftheyparticipated

infirefightingactivitieswithinfivedayspriorthesampling

cam-paignsandiftheywereexposedtotobaccosmoke.Basedonthe

informationcollectedthroughtheindividualquestionnaires,PAHs

intakethroughfoodingestionwasconsiderednotsigniﬁcant.All

participantsreadandsignedinformedconsentformsapprovedby

theEthicCommitteeofUniversityofPorto.

Personalexposuresweremonitoredduringa periodof

fifty-fourdays(JuneandJuly2014)infirefightersthatwerenotdirectly

involvedinfirefightingactivities.Ateachfirestation,airsampling

campaignsof18selectedPAHsinPM2.5wereperformedonasingle

day(6individualsatSDMandFEC;9atMRD,MDLandTMC;12at

TDC,VNHandBRG)andoveracontinuous4-hperiodofaregular

workshiftinsidetheﬁrestation;duplicatesamplesper

individ-ualwerecollected.Duringthesamplingcampaignﬁreﬁghterswere

encouragedtomovefreelyattheﬁrestationasinanormal

work-ingday.Airsamplingwasdonebypersonalconstantﬂowsamplers

(Gilian,modelsGilAir3andProValue3;Sensidyne,USA)thatwere

placedatthewaistofeachﬁreﬁghterinawaythatcouldnotdisturb

theregularactivities;anairﬂowrateof2L/minwasused.Theinlets

werepositionedatthebreadingzoneofﬁreﬁghters.Airsamples

werecollectedonpolytetraﬂuoroethylenemembraneﬁlterswith

polymethylpentenesupportring(2␮mporosity,Ø47mm,SKCLtd.,

UnitedKingdom).PM2.5massesweredeterminedgravimetrically

accordingto[44].Afterthesampling,ﬁlterswerestoredinafreezer

(–20◦C)beforechemicalanalysis.

Aspoturinesamplewascollectedattheendoftheﬁreﬁghter

workshiftinsterilized50mLpolycarbonatecontainers.Samples

werefrozenat −20◦_C_until _analysis._Sampling_campaigns _were

performedintriplicate.

Duringsamplecollection,aresearcherwaspresenttokeepa

recordofpotentialsourceactivitiesandventilationsystemstatus

(doorand windowpositions).Potentialsources ofPAHsineach

ﬁrestationwerealsocollected.Itwasobservedthatduring

sam-plingcampaigns,therewerealwaysvehicles(arriving,parking,or

leaving)onthetruckbay;smokingisnotallowedatﬁrestations.

2.3. PAHsandOH-PAHschromatographicanalysis

Extractionandquantiﬁcation ofPAHsfromPM2.5 ﬁlters,and

OH-PAHsfromurinesampleswereperformedbypreviously

vali-datedanalyticalprocedures[45–47].PAHsandOH-PAHsextracts

wereanalysedusingaShimadzuLCsystem(Shimadzu

Corpora-tion, Kyoto,Japan) equipped withphotodiode array (PAD) and

ﬂuorescence(FLD)detectorsonline.Separationofthecompounds

wasperformedin aC18 column(CC150/4Nucleosil 100–5C18

PAH, 150×4.0mm;5␮m particlesize; Macherey–Nagel,Duren,

Germany) maintained at room temperature (20±1◦C).

Fluo-rescencewavelength programmingwas usedtoperform better

sensitivityandminimalinterference.Eachcompoundwasdetected

atitsoptimumexcitation/emissionwavelengthpair:260/315nm

(naphthalene, acenaphthene and ﬂuorene), 260/366nm

(phenanthrene), 260/430nm (anthracene, ﬂuoranthene,

pyrene, benz[a]anthracene, chrysene, benzo[b+j]ﬂuoranthene,

benzo[k]ﬂuoranthene, benzo[a]pyrene, dibenz[a,h]anthracene,

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0% 20% 40% 60% 80% 100% PAHs distrib utio n MRD TDC SDM MDL TMC VNH BRG FEC Fire station

5 and more rings 4-rings 2-3 rings

Fig.2.PAHsdistributionaccordingtothenumberofaromaticringsatdifferentﬁrestations(MirandadoDouro(MRD),TorreDonaChama(TDC),Sendim(SDM),Mirandela (MDL),TorredeMoncorvo(TMC),Vinhais(VNH),Braganc¸a(BRG),andFreixodeEspadaàCinta(FEC)).

(indeno[1,2,3–cd]pyrene) for PAHs, and 232/337nm (1OHNapt

and 1OHAce), 265/335nm (2OHFlu), 263/363nm (1OHPhen),

242/388nm(1OHPy), and308/432nm(3OHB[a]P)for OH-PAHs.

ThePAHacenaphthylene,whichshowslimitedﬂuorescence,was

analysedat254nminPAD.

CalibrationswithPAHsandOH-PAHsmixedstandards,using

atleast6calibrationpoints,wereperformedinacetonitrileand

methanol, respectively. Calibration curves were linearly ﬁtted

with correlation coefﬁcients always higher than 0.9979. RSD

values ranged from 1.8% (dibenzo[a,l]pyrene) to 9.1%

(naph-thalene) for PAHs extraction from ﬁlters. Regarding urinary

OH-PAHs,RSD values varied between1.3% (1OHNaph+1OHAce

and2OHFlu) to8.1% (1OHNaph+1OHAce,2OHFlu, and1OHPy).

Limits of detection (LODs) and quantiﬁcation (LOQs) were

cal-culated as the minimum detectable amount of analyte with a

signal-to-noiseratioof3:1and10:1,respectively[48].ForPAHs,

LODsbetween1.0pg/m3 _(for_anthracene, _{benzo[k]ﬂuoranthene,}

chrysene, benz[a]anthracene, phenanthrene and

indeno[1,2,3-cd]pyrene) and 148pg/m3 _(for_{acenaphthylene)} _were _obtained,

withcorrespondingLOQsintherange3.4–492pg/m3_._Regarding

OH-PAHs,LODsrangedbetween0.0008␮g/Lurine(for2OHFlu)

to0.195␮g/Lurine(for1OHNaph+1OHAce),whiletherespective

LOQsvariedfrom0.0028␮g/Lurineto0.650␮g/Lurine.

AllOH-PAHconcentrationswerenormalizedbyurinary

creati-nine(mol/mol).Urinarycreatininemeasurementswereperformed

accordingtotheJaffecolorimetricmethod[49].

Analyticalblanksandstandardswereanalyseddailyand

regu-larly.Eachanalysiswasperformedatleastintriplicate.

2.4. Statisticalanalysis

StatisticalanalysiswasperformedusingSPSS(IBMSPSS

Statis-tics20).PAHsandOH-PAHsmedianvalueswerecomparedthrough

thenonparametric Mann−Whitney Utest, since normal

distri-butionwasnotobservedbyKolmogorov-SmirnovwithLilliefors

correctionandShapiro−Wilk’stests.Whentheconcentrationwas

belowtheLOD,thevalueoftherespectiveLOD/√2wasused[50].

Statisticalsigniﬁcancewasdeﬁnedasp≤0.05.Spearman

correla-tioncoefﬁcientswerecalculatedtoexaminetherelationbetween

airborneindividualandtotalPAHconcentrationswithﬁreﬁghters’

urinaryindividualandtotalOH-PAHsforeachﬁrestation.

3. ResultsandDiscussion

3.1. Personalexposuremonitoring

Theconcentrations of individualand total PAHs (PAHs)in

thebreathingairzoneoffirefightersworkingateachfirestation

are presentedin Table 1.Among the18 PAHs considered,

ace-naphthylene wasthe mostabundant compound (exceptin the

environmentalexposuresoftheruralsitesofTMCandTDCwhere

itwasnotdetected)withconcentrationsrepresenting46.4%(at

FEC)to61.5%(atSDM)ofPAHs.Acenaphthyleneisacomponent

ofcrude oil, coaltar, tobaccosmokeand a productof

combus-tion which maybe produced and released tothe environment

during ﬁres [20,30,51]; emissions from wood combustion

con-tainmoreacenaphthylenethanotherPAHs[52].Forworkersat

MRD,SDM,MDL,TMC,andBRGﬁrehouses,theothermost

abun-dantPAHswere,bydescendingorder,acenaphthene,naphthalene,

andphenanthrenewithcontributionsrangingfrom15.5(BRG)to

38.6%(MRD), 4.48(MDL)to16.8% (BRG),and from2.65(MRD)

to7.82%(TMC),respectively.Togetherwithacenaphthylene,these

fourPM2.5-boundPAHsrepresented84.6–95.0%ofPAHsforthose

firestations.InthebreathingairzoneoffirefightersfromFECand

VNH,dibenz[a,h]anthracenewasrespectivelythesecondandthird

predominantPAH, accounting with11.9% and 7.84% ofPAHs.

Dibenz[a,h]anthraceneconcentrationsinworkersfromthe

remain-ingsixﬁrestationsrepresentedonly0.30–1.73%of_PAHs.The

prevalenceof this compound indicatesa higher inﬂuencefrom

light-dutygasolinevehicleemissions[53]inthesurroundingareas

ofFECandVNHﬁrestations.GloballyPM2.5-boundPAHswith2–3

aromaticrings(naphthalene,acenaphthylene,andacenaphthene)

represented 63.9%(FEC) to 95.7% (MDL)of PAHs; 4 aromatic

ringcompounds(anthracene,ﬂuoranthene,andpyrene)accounted

with0.79%(MRD)to2.05%(TMC)and5ormorearomaticringPAHs

correspondedto2.74%(MDL)to34.7%(FEC)ofPAHs(Fig.2).Itis

worthtomentionthatlighterPAHsusuallypredominateinthegas

phase[10,36]suggestingthatthedeterminedpersonalexposures

maybeunderestimated.OverallairbornePAHproﬁlesobtainedat

FECandVNHexhibitedthehighestcontributionsoftheheavier

molecularweightcompounds(themosthazardous),whilethoseat

MRD,SDMandMDLpresentedthelowest.Asageneralrule,PAHs

toxicityincreasesasthenumberofringsincreases(withthe

excep-tionofnaphthalene,2-ringscompoundthatisclassiﬁedaspossible

humancarcinogen;[8]).

GlobalmeanPAHsvariedwidely.Thehighesttotalmedian

content of PAHs was found, by descending order, in the

breathing air zone of ﬁreﬁghters from FEC» MDL>MRD»

SDM>VNH>BRG>TMC≈TDC, withlevels at FEC(428ng/m3₎₂

(256ng/m3_,_MDL)_to₉_(46.4_ng/m3_,_TDC)_times_{signiﬁcantly}_higher

(p≤0.036)thanattheotherﬁrehouses.IndoorPAHlevels,and

per-sonalexposure,dependonactivitiesconducted,occupancyrates,

physicalcharacteristicsof buildings(permeability,particle

size-speciﬁcdifference),sitespeciﬁcity,ventilationhabits,seasonand

meteorology.Theobserveddifferencesmaybemainlyattributedto

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Fig.3. PAHsdistributionofa)totalcarcinogenicPAHs(PAHscar)amongthe18PAHsunderstudy(PAHs)ateachﬁrestation;b)individualcarcinogenicPAHsamongthe

differentﬁrestations(MirandadoDouro(MRD),TorreDonaChama(TDC),Sendim(SDM),Mirandela(MDL),TorredeMoncorvo(TMC),Vinhais(VNH),Braganc¸a(BRG),and FreixodeEspadaàCinta(FEC)).

twofirehouseslocatedatruralareaswithlow trafficinfluences

(TMCandTDC)presentedthelowestlevelsofPAHsinthebreathing

airzoneofﬁreﬁghters(Table1),followedbythosesituatedinurban

sitesbutconstructedand/orrestoredmorerecently(BGR,VNHand

SDM).Vehiculartrafﬁcemissionsareoneofthemajorsourcesof

PAHsinurbanareas[54].Inaddition,theageofabuildingreﬂects

itsconditionand affectspenetrationofoutdoorPAH

concentra-tionstoindoorair.Theolderabuilding,thegreaterwillbethe

impactofoutdoorsourcesowingtohigherairexchangethrough

suchroutesaspoorlyﬁttingwindowsanddoors[55].Theurban

location(nearbusyroads),buildingage(constructedbefore1984)

andlay-outofFEC,MDL,andMRDcontributedsigniﬁcantlytothe

elevatedfirefighters’environmentalexposureatthesefirehouses.

ThetruckbayatFEC,MDLandMRDwasphysicallyseparatedfrom

theoperationalcontrolcenterandsocial room(where

ﬁreﬁght-ersspendmostoftheirtime)onlybyadoor(thatwasfrequently

opened).Moreover,thedeterminedlevelsatthesethreestations

werehigherthanthosereported(30.5–126.5ng/m3₎_for_two_sites

locatedintheoutskirtsofthecityofPortothatisamore

urban-izedareaandhasmuchhighertrafﬁcdensitythanBraganc¸adistrict

(MetropolitanAreaofPortohas2042km2_with_1.7_million_citizens,

whileBraganc¸adistricthasanareaof6608km2_with₁₃₉₃₄₄

citi-zens);Portodistrictcorrespondstothenearestgeographicalregion

(Fig.1)thatwaspreviouslycharacterizedintermsofatmospheric

PAHlevels[56].

Moredetailedanalysiswasperformedconcerningthe

carcino-genicPAHs(PAHscar;Table1).Theinter-ﬁrestationscomparison

of the content of PAHscar in the breathing air zone of

ﬁre-ﬁghterswasFEC»MRD≈VNH>MDL>BRG≈SDM≈TDC≈TMC.In

accordance with thePAH levels,workers from FECexhibited

signiﬁcantly higher (p≤0.002) PAHscar (Fig. 3a). Globally the

compoundthatcontributedthemosttoPAHscar was

naphtha-lene(39.4%atVNHto78.1%atMDL),exceptforﬁreﬁghtersfrom

FECﬁrehouse. For thatﬁre station,dibenz[a,h]anthracene(with

atoxicityequivalentfactor5timeshigherthanbenzo[a]pyrene;

[9]wasthepredominantPAHaccountingwith33.8%ofPAHscar,

followedbynaphthalene(23.5%)(Fig.3b).Inaddition,ﬁreﬁghters

workingatVNHﬁrehousealsopresentedarelevantcontribution

of dibenz[a,h]anthracene (32.1% of PAHscar) in their

breath-ingair zone;for theothersites,values varied from4.1%(BRG)

to 21.1% (MRD) of PAHscar. The maximum concentrations of

benzo[a]pyrene,the PAHsmarker ofcarcinogenicity, accounted

with10%atFEC,6.7%atVNH,1.5–3.4%ofPAHscarfortheother

ﬁrehouses(Fig.3b).Furthermore,itisimportanttomentionthat

the median concentrations of benzo[a]pyrene of FEC

ﬁreﬁght-ersexceeded10to20timestheexistentlimitvalueof1ng/m3

(annualmeantotalcontentinPM10fraction)forambientair[40].

Dibenzo[a,l]pyrene, which carcinogenic potency has been

esti-matedtobeapproximately100timesthatofbenzo[a]pyrene[9],

presented concentrations lowerthan 0.671ng/m3 _(0.44–8.2% _of

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TDC 66% 16% 11% 7% 1OHNapt+1OHAce 2OHFlu 1OHPhen 1OHPy MRD 85% 7% 5% 3% 1OHNapt+1OHAce 2OHFlu 1OHPhen 1OHPy SDM 96% 1% 1% 2% 1OHNapt+1OHAce 2OHFlu 1OHPhen 1OHPy MDL 76% 12% 5% 7% 1OHNapt+1OHAce 2OHFlu 1OHPhen 1OHPy TMC 88% 8% 2% 2% 1OHNapt+1OHAce 2OHFlu 1OHPhen 1OHPy VNH 87% 3% 7% 3% 1OHNapt+1OHAce 2OHFlu 1OHPhen 1OHPy BRG 75% 10% 9% 6% 1OHNapt+1OHAce 2OHFlu 1OHPhen 1OHPy FEC 96% 2% 1% 1% 1OHNapt+1OHAce 2OHFlu 1OHPhen 1OHPy

Fig. 4. Distribution of monohydroxyl-PAHs (1OHNaph+1OHAce: 1-hydroxynaphthalene+1-hydroxyacenaphthene; 2OHFlu: 2-hydroxyfluorene; 1OHPHen: 1-hydroxyphenanthrene;1OHPy:1-hydroxypyrene)intheurinesamplesoffirefightersatdifferentfirestations(MirandadoDouro(MRD),TorreDonaChama(TDC),Sendim (SDM),Mirandela(MDL),TorredeMoncorvo(TMC),Vinhais(VNH),Bragança(BRG),andFreixodeEspadaàCinta(FEC)).

FireﬁghterspersonalexposuretoPAHsatPortugueseﬁre

sta-tionswerewellbelowtheexistentPAHsoccupationalexposure

limits proposed by the American Conference of Governmental

IndustrialHygienists(ACGIH),theNIOSHandtheUSOccupational

SafetyandHealthAdministration(OSHA)(Table3S,Supplementary

material).

Studiesregardingﬁreﬁghters’occupationalexposuretoPAHs

atﬁrestationsare veryscarce[23,32];noneinEuropean

facili-ties.Moreover,comparisonbetweenvariousstudiesishampered

bydifferent designs(analysed particulate fractionor/and

num-berofdeterminedPAHs).Shenandcoworkers[32]collecteddust

samplesfromvacuumcleanerbagsusedtoroutinelycleantwenty

Americanﬁrehouses.MediantotalPAHsconcentrationindustwas

4124ng/g.Amongthe10compoundsunderstudy,pyrenewasthe

predominantPAHmeasuredinthesedustsamples,followed,in

decreasingorder,bybenzo[ghi]perylene,ﬂuorantheneand

chry-sene.Baxterandcolleagues[23]conductedanairmonitoringstudy

inthreedifferentenvironments,includingintheﬁretruckbayand

equipmentstoragearea,thekitchen/commonarea,andthe

sleep-ingquartersoftwoAmericanﬁrestations[23].Surprisingly,out

of16quantiﬁedPAHs,onlynaphthalene(at9.22–9.24␮g/m3₎_was

(8)

truckbayofoneﬁrehouse.Medianconcentrationsofnaphthalene

(4.95atTMCto12.2ng/m3_at_MRD,_Table₁₎_in_the_breathing_air_zone

ofPortugueseﬁreﬁghterswerelowerthanthosereportedlevels

[23].Furthermore,somerecentstudies[20,30]havereportedthat

aquantifiabledepositionofPAHsoutsideandinsidefirefighters’

ensembles(helmets,gloves,hoods,andturnoutgear)occurswhich

maysigniﬁcantlycontributetototalPAHsbodyburden.Further

researchisrequiredtoinvestigatethisaspect.

3.2. Biomonitoring

Biomonitoringofﬁreﬁghtersisaneffectiveassessmenttoolto

understandtotalexposures,especiallytocompoundssuchasPAHs

thatarenotabsorbedexclusivelybyinhalation,butalsothrough

dermalcontactandfoodingestion[18–20].Medianconcentrations

ofindividualandtotalOH-PAHs(OH-PAHs)intheurinesamples

ofﬁreﬁghtersarepresentedinTable2.UrinaryOH-PAHlevelswere

normalizedbythecreatininelevelsofeachﬁreﬁghter.Globally

cre-atinineconcentrationsintheurineofﬁreﬁghtersrangedbetween

0.82 to2.74g/L, beingwithinthe acceptedcreatininerange for

healthypeople(>0.3and<3g/L)[57].SincePAHsareubiquitous

pollutants, each participatingfirefighterwas requestedto fill a

questionnaireregardingtheexistenceofpossibleexposuretoPAHs

outsidetheoccupationalenvironment.Onlynon-smoking

ﬁreﬁght-erswithdietaryexclusiveofbarbecueanddeep-friedfoodswithin

ﬁvedaysbeforeurinesamplescollectionwereconsidered.Thus,

itwasassumedthatfirefightershadsimilar(non-significant)

lev-elsofPAHsexposurethroughfoodconsumptionandthattobacco

smokecontributionwasnegligible.Still,theassessmentoftobacco

biomarkerdata,andtheuseofpre-exposureurinesamples,would

beprecioustovalidatetheseassumptions.Knowledgeofthe

elim-inationkineticsoftheurinaryOH-PAHsisessentialtodeﬁnethe

urine sampling strategy. Limited studies regarding elimination

kineticsofOH-PAHsinhumansareavailable:half-lifeofurinary

1OHPyexcretionratesvariesbetween6to35hafterinhalation

exposure[58,59]and 4.4[60]to12h[61]afteringestion

expo-sure;half-liferangingfrom3.3to6.2hfor1OHNaph,2.3to4.0h

for2OHFlu,and4.3to6.1hfor1-OHPhenwerealsoreportedfor

ingestionexposure[37].1OHNaph+1OHAceweredetectedinmore

than87%ofsamples,2-OHFluand1OHPypresenteddetectionrates

above96%and1OHPhenwasdetectedinallurinesamples.In

agree-mentwithsomepreviousstudies,urinary3OHB[a]P(metabolite

ofthePAHmarkerofcarcinogenicity)presentednullorverylow

detectionrates[37,62,63].Somestudieswithanimalsrevealedthat

urinaryconcentrationsof3OHB[a]Prepresentonly0.1–0.2%ofthe

benzo[a]pyrenedose,duetoacomplexmetabolismthatproduces

severaldifferentmetaboliteswhicharemainlyexcretedinfeces

[64].

Overalltheinter-comparisonofurinaryOH-PAHsof

firefight-ersfromfirestations:FEC»VNH>MRD≈SDM>MDL>BRG>TMC»

TDC,followedthesametrendasthecontentofairbornePAHs

withtheexceptionofworkersfromVNHandMDLthatexchanged

position.Accordingly,urinaryOH-PAHsinindividualsfromFEC

weresigniﬁcantlyelevated(p≤0.002)(2(MRD)to14(TDC)times

higher)than at other ﬁrehouses.The concentrations of urinary

OH-PAHsinﬁreﬁghterswereinverselyrelatedtothesizeof

com-pounds,i.e.,thehigherthemolecularweightthelowerthedetected

OH-PAH concentrations (Table 2). Urinary 1OHNaph+1OHAce

wereby far thepredominant compounds accounting with66%

(TDC)to96%ofOH-PAHs(FEC,SDM),beingfollowedby2OHFlu

(1%atSDMto16%atTDC),1OHPhen(1%atSDMandFECto11%

atTDC),and1OHPy(1%atFECto7%atTDC)(Fig.4).This

distri-butionproﬁlefollowsthesamepatternasthePAHsdistribution

inthebreathingairzoneoffirefighterswhileworkingatfire

sta-tions(Table1);airbornePAHswithtwoandthreearomaticrings

werethemostabundantonesinallﬁrestations(63.9%atFECto

95.7%ofPAHsatMDL,Fig.2).Urinary1OHNaph+1OHAce

con-centrationsweresigniﬁcantlydifferent(p≤0.002)fromtheother

metabolitesfor all groups of individuals (Table2). Themedian

concentrationsofurinary1OHPyinPortugueseﬁreﬁghtersranged

between1.36×10−2 (SDM)to0.146(MDL)␮mol/molcreatinine

(Table2).Nowadays,measurementsofurinary1OHPyareroutinely

appliedtocontrolindustrialexposuretoPAHsincokeovensand

primaryaluminumproductionandtocontrolexposureof

profes-sionalswhen handlingcoal tarderived products[34]. Although

noreferencestandardguidelinesareestablishedforurinary

OH-PAHs,Jongeneelen[33,34]proposedano-biologicaleffectlevelof

1.4␮mol/molcreatinineof1-hydroxypyreneinurineofexposed

workers,i.e.thelowestreportedlevelatwhichnogenotoxiceffects

werefound.InadditiontheBiologicalExposureIndexCommittee

ofACGIHstatedthatthepresenceofurinary1OHPyabovea

bench-marklevelofabout1␮g/L(0.5␮mol/mol)indicatesoccupational

exposure to PAHs, since very few non-occupationally exposed

persons, smokers or non-smokers, will excrete this amount of

1OHPy[65].ACGIHalsorecommendsthatthebenchmarkshould

beconsideredasapost-shiftlevel.Thequantiﬁedconcentrations

of post-shifturinary 1OHPy infirefighters fromallfire stations

werewellbelowthoserecommendedvalues.Thisobservationis

importantsincefirefightersparticipatingatfirefightingactivities

areheavilyexposedtoPAHs[18–20]andthusmaypresenthigh

backgroundlevelsof1OHPy.

Regardingcharacterizationofﬁreﬁghters’exposure,1OHPyand

1OHNaph are the more investigated metabolites [27,31,38,39].

The comparison ofthedetected levelswithpreviousreports is

extremelydifﬁcultsince,inthemajorityofthestudies,

concen-trationsarenotadequatelynormalizedwiththepersonalurinary

creatininelevels.Creatinineiseliminatedfromthehumanbodyat

aconstantrate,andthusiswidelyusedtominimizethevariability

ofparameterssuchasindividualﬂuidintake,bodytemperature,

physicalexerciseandambienttemperature,whichchangesfrom

persontoperson.Still,urinary1OHPyconcentrationsinPortuguese

ﬁreﬁghters(overallrange:0.078–3.28nmol/L)wereslightlyhigher

thanlevelsfoundinﬁreﬁghtersbeforetheirparticipationingas

simulators(0.6–1.2nmol/L)butlowerthanconcentrationsat

div-ingsimulatorﬁres(0.6–9.2nmol/L)[32].Inaddition1OHPylevels

weresimilarwiththelevelsreportedinﬁreﬁghtersbeforetheir

participationinprescribedpileburns(<0.01–0.56␮g/L[27])but

slightly higher than theconcentrations observedin thecontrol

groupofﬁreﬁghtersthatwerepresentattheWorldTradeCenter

Collapsein2001(157ng/Linthisstudyversus62.5ng/L[39]).Only

twostudiesreportedconcentrationsofothermetabolites,namely

severalOHPhen[39]and1OHNaph[31]compoundsamong

ﬁre-ﬁghter’surine.Urinary1OHPhenlevelsweresimilarwiththose

observedinthecontrolgroupofﬁreﬁghters(170ng/Linthisstudy

versus158ng/L[39]).Inﬁreﬁghters,urinary1OHAce,2OHFlu,and

3OHB[a]Pwereneverassessedbefore.UrinaryOH-PAHsexcretion

infirefightersduringtheirregularworkatfirestationswaslower

thanlevelsreportedforotherindustrialworkerswithknownPAHs

exposure[63,66–70].

3.3. CorrelationsbetweenairbornePAHsandurinaryOH-PAHs

The possible contribution of airborne individual

(naph-thalene, acenaphthene, naphthalene+acenaphthene, ﬂuorene,

phenanthrene,and pyrene)andPAHs torespective individual

(1OHNaph+1OHAce, 2OHFlu,1OHPhen, and 1OHPy)and

OH-PAHsexcretioninthepost-shifturine wasestimated.Moderate

tostrongcorrelationswereobservedbetweenPAHsandurinary

OH-PAHs(ranging fromr=0.367,p=0.332atTMCtor=0.886,

p=0.019at FEC)for firefightersfromsixfire stations(FEC, BRG,

MDL, MRD, TDC, TMC); statistical signiﬁcance was reachedfor

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BRG(r=0.839,p=0.001)andFEC(r=0.886,p=0.019)whichwere

threeoftheﬁrehouseswherethehighestenvironmentalexposure

weredetected(Table1).

Theseﬁndingsareinagreementwiththosedescribedforcoke

ovenworkers(r=0.680,p≤0.01betweenPAHsof15compounds

and OH-PAHs of 10 metabolites) [61]. The positive

correla-tionsconﬁrmthatOH-PAHexcretionincreaseswithanincreasing

exposureof PAHs at ﬁre stations. It seems that, atthese sites,

firefighters had fire stationindoor air as theirmajor exposure

source of PAHs. Also, PAHs correlated well with 1OHPy, the

biomarkerofPAHsexposure,forindividualsfromMRD(r=0.703,

p=0.035),SDM(r=0.941,p=0.005),MDL(r=0.262,p=0.531)and

VNH(r=0.944,p≤0.001).Theseurbansitesareinthelistofthe

5ﬁrehouseswhereenvironmentalPAHsexposurewerethe

high-est(FEC»MDL>MRD»SDM>VNH>BRG>TMC≈TDC;Table1);the

onlyobservedexceptionwasFEC.Theseresultssuggestthat1OHPy

maybeasuitable biological indicatorofinternal doseof

expo-suretothosePAHsthatareemittedbythepredominantsource.

Thesecorrelations arehigher thanthose reportedincoke oven

workers(r=0.456,p<0.01;[63])butsimilarwiththerelationships

described in workersemployed in coke production, production

ofgraphite electrodes, special carbon products, and production

ofrefractorymaterials(r=0.717,p<0.01)[70].Othersigniﬁcant

relationshipswerereachedbetweennaphthalene+acenaphthene

withurinary1OHNaph+1OHAceforﬁreﬁghtersatMDL(r=0.833,

p=0.010),ﬂuoreneand2OHFluatFEC(r=0.771,p=0.072)andBRG

(r=0.736,p=0.006),phenanthreneand1OHPhenatFEC(r=0.971,

p=0.001);andpyrenewith1OHPyatTMC(r=0.878,p=0.002)and

VNH(r=0.745,p=0.005).Yamanoetal.[63]andRossbachetal.

[70]alsoreportedmoderatetostrongcorrelationsbetweenurinary

metaboliteconcentrationandpersonalexposuretotherespective

PAH.

Globallytheseresultssuggest thatairbornePAHsatﬁre

sta-tionsmaycontributetoﬁreﬁghters’totalPAHsbodyburden;the

influenceismoreevidentinfirefightersexposedtothehighest

lev-els.Stilltheexistenceofothersigniﬁcantcommonsources(food,

homeairand/oroutdoorair)cannotbediscarded.Naphthaleneand

acenaphthenearethemorevolatilecompoundsandmostoftheir

environmentallevelsenterinthehumanbodymainlythroughair.

Fluorene,phenanthreneandpyreneareabsorbedthroughthe

res-piratorytractbutalsobythegastrointestinaltracts, andbythe

skin.

4. Conclusions

Thisworkcharacterizedﬁreﬁghters´ıexposuretoPAHsduring

theirworkshiftateightPortugueseﬁrestationsbypersonal

expo-suremonitoringandbiomonitoring.Overall,PM2.5-boundPAH

concentrationsrangedfrom46.4–428ng/m3_,_with_2–3_aromatic

ringsaccountingwith63.9–95.7%ofPAHs.PAHswith4–5ormore

aromaticrings correspondedto0.789–2.05% and 2.54–34.7% of

PAHs,respectively.Theobtaineddatahighlightedtheimportance

ofsitespeciﬁcityandpoorbuildingconstruction/conservation,as

wellasinappropriatebuildinglayoutonenvironmentalPAH

lev-elsatthestudiedfirestations.Still,firefighter’spersonalexposure

toPAHsatPortugueseﬁrestationswaswellbelowtheexistent

occupationalexposurelimits.

In accordance with the airborne PAHs proﬁle, urinary

1-hydroxynaphthalene and 1-hydroxyacenaphthene were the

predominant metabolites (66–96% OH-PAHs). Thus, it is

rec-ommendedthattotalbodyburdenofPAHsshouldnotbebased

exclusivelyon1OHPybiomonitoring,asithasbeenperformedin

thelargemajorityofstudies.Inaddition,thecontributionofsome

selectedairborneindividualcompoundsandPAHstorespective

individualandOH-PAHs excretioninthepost-shift urinewas

estimatedforthefirsttimeinfirefighters.Theattainedsignificant

positivecorrelationsforfirefightersfromfourfirestationsindicated

theinﬂuenceofoccupationalexposureonmetabolitelevels.Results

foralargernumberofﬁrestations,participantsandnumberof(air

andurine)samplesareneededtodrawmoremeaningfuland

sta-tisticallymoresigniﬁcantconclusions.Also,amorecomprehensive

monitoringthatincludesallpossiblesources(food,homeand

out-doorair,etc.)wouldbeprecioustogivemorequantitativesupport

totheimpactofoccupationalexposureonﬁreﬁghter’stotalPAHs

internaldose.

Conﬂictofinterest

Theauthorsdeclarethattherearenoconﬂictsofinterest.

Statementofnovelty

Thisworkcharacterizedﬁreﬁghters’exposuretoPAHsduring

theirworkshiftateightﬁrestationsbypersonalmonitoring(18

PAHs)andbiomonitoring(sixurinarymetabolites).Onlytwo

stud-ieswerefoundregardingﬁreﬁghters’environmentalexposureto

PAHsatﬁrestations,bothperformedinUSA.Noinformationexists

concerningothercountries. Moreover,nostudywasfoundthat

simultaneouslyassessedairbornePAHexposurelevelsatﬁre

sta-tionsand urinary levelsof oneor more PAH metabolites. Also,

contributionofpersonalairborneindividualandtotalPAHs

expo-suretorespectiveindividualandtotalOH-PAHsexcretioninurine

wasexploredforthefirsttimeinfirefighters.

Acknowledgments

TheauthorsarethankfulforthecooperationofEscolaSuperior

deSaúdefromInstitutoPolitécnicodeBragançaandtoallfirefighters

involvedinthestudy.ThisworkwassupportedbyEuropeanUnion

(FEDERfundsthroughCOMPETE)andNationalFunds(FCT)through

projectsUID/QUI/50006/2013andUID/EQU/00511/2013-LEPABE,

bytheFCT/MECwithnationalfundsandco-fundedbyFEDERin

thescopeofthe<gn1>P2020</gn1>PartnershipAgreement.

Addi-tionalﬁnancialsupportwasprovidedbyFCTthroughfellowships

SFRH/BD/80113/2011andSFRH/BPD/65722/2009.

AppendixA. Supplementarydata

Supplementarydataassociatedwiththisarticlecanbefound,in

theonlineversion,athttp://dx.doi.org/10.1016/j.jhazmat.2016.03.

012.

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