Critical Effects of Smoking Parameters on the Levels of

Research ArticleCritical Effects of Smoking Parameters on the Levels of PolycyclicAromatic Hydrocarbons in Traditionally Smoked Fish and MeatProducts in Finland

Mirja Hokkanen 1 Ulla Luhtasela2 Pirkko Kostamo2 Tiina Ritvanen1

Kimmo Peltonen1 and Marika Jestoi2

1Finnish Food Safety Authority Evira Research and Laboratory Services Department Helsinki FI-00790 Finland2Finnish Food Safety Authority Evira Food Safety Department Helsinki FI-00790 Finland

Correspondence should be addressed to Mirja Hokkanen mirjahokkanenevirafi

Received 8 June 2018 Revised 7 August 2018 Accepted 3 September 2018 Published 10 October 2018

Guest Editor Alberto Fiore

Copyright copy 2018 Mirja Hokkanen et al is is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work isproperly cited

Eighty fish products and 62 meat products were sampled and analysed in Finland in the year of 2012 for four marker polycyclic aromatichydrocarbons (PAH4) with an accredited gas chromatography-tandem mass spectrometry method In general the determined PAH4levels were relatively low and below the maximum levels e mean concentrations of smoked fish samples were 07μgmiddotkgminus1 for benzo[a]pyrene and 39μgmiddotkgminus1 for the PAH4 sum whereas in smokedmeat samples mean benzo[a]pyrene and PAH4 sum levels were 22μgmiddotkgminus1

and 11μgmiddotkgminus1 respectively However PAH4 sum concentrations ranged from not detected to 200microgmiddotkgminus1 particularly among meatproducts underlining the importance of controlling the smoking process In this study the effect of selected smoking parametersie smoking technique (directindirect) smoking time (less than five hoursmore than five hours) smoke generation temperature(optimisednonoptimised) and the distance (less than five metresmore than five metres) between the food and the smoke sourceconfirmed the linkage between the smoking factors and the PAH4 levels formed in fish andmeat products As guidance for a safe smokingprocess it was demonstrated that an indirect smoking technique a shorter smoking time an optimised smoke generation temperature anda longer distance from the smoke source generated lower PAH concentrations in food products However while a shorter smoking timegenerated lower PAH levels in meat products the levels in fish products were unexpectedly higher than in those smoked for a longer timeOther factors such as the smoking type (cold smokingwarm or hot smoking) and the fish size may have affected this result

1 Introduction

Polycyclic aromatic hydrocarbons (PAHs) consist of a ver-satile group of organic compounds that have at least two ormore aromatic rings joint together [1 2]ey are fat solubleand chemically stable compounds that are classified ashuman carcinogens [1] Several metabolic pathways mayresult in reactive intermediates inducing mutagenic orcarcinogenic processes of PAHs [3] e carcinogenic ca-pacity varies between them despite having similar structuralproperties [4] ose with four to six fused rings such asbenzo[a]pyrene (BaP) are effective carcinogens belonging toGroup 1 carcinogens according to the International Agencyfor Research on Cancer (IARC) [5] Additionally PAHs

have teratogenic haematological and immunotoxic effectsand their concentrations in food should therefore be as lowas reasonably achievable (ALARA principle) [6ndash8]

PAHs are formed during the incomplete combustion oforganic matter and they are widely distributed in the envi-ronment via air [6 7] Industry traffic smoking forest firesand volcanic eruptions generate PAHs and humans areconsequently mainly exposed by inhalation skin contact andingestion [6 7] Despite also being environmental contami-nants PAHs are formed in food processing such as dryinggrilling roasting and smoking [6 7 9] For nonsmokers thediet appears to be the main source of PAH exposure [9 10]

Food smoking is one of the oldest preservation methodsand is still widely used [11ndash14] However smoking is

HindawiJournal of ChemistryVolume 2018 Article ID 2160958 14 pageshttpsdoiorg10115520182160958

nowadays mainly used to obtain the desired colour flavouraroma and appearance in the smoked food rather than forpreservation purposes [14 15] Traditional smoking is generallyperformed by the formation of smoke from wood [16 17]Smoke is defined as the result of thermal pyrolysis of woodwhenaccess to oxygen is limited [6] PAHs and other chemicalcompounds occur in smoke particles which canmigrate into thefood product being smoked [2 18] Wood smoke containsa combination of antioxidant and antimicrobial chemicals(eg phenols carboxylic acids aldehydes and acetic acids) butalso someharmful compounds such as PAHs [12 17 19] PAHsare potential health hazards associated with smoked foods inwhich they typically occur as a complex mixture [6 13] InFinland smoking with direct and indirect techniques is widelyused in the processing of meat and fish products For directsmoking smoke is generated from an open fire in the samechamber as the smoked product whereas in indirect smokingthe smoke is generated in an external chamber separated fromthe food and the smoke is led to the product from the externalsmoke generator [16 20] Alongside the smoking technique thetype of process (grilling roasting smoking and drying) thedistance between the food and the smoke source the processtime and temperature impact the formed PAH levels [6]

Commission Regulation (EC) No 18812006 specifiesthe maximum levels (MLs) of BaP in different foodstuffs[21] In 2008 the European Food Safety Authority (EFSA)concluded that BaP alone is not a suitable indicator for theoccurrence and toxicity of PAHs in foods [3] erefore ad-ditional MLs for the sum of four PAHs (PAH4 BaP benz[a]anthracene (BaA) chrysene (CHR) and benzo[b]fluoranthene(BbFA)) were set in Commission Regulation (EU) No8352011 [22] e MLs for smoked meat and fish productssmoked from 192012 to 3182014 are 50 microgmiddotkgminus1 for BaPand 300 microgmiddotkgminus1 for the PAH4 sum e MLs applicablefrom 192014 for smoked meat and fish products are20 microgmiddotkgminus1 for BaP and 120 microgmiddotkgminus1 for the PAH4 sum [22]However in some Member States the lower levels set from192014 could not realistically be reached for traditionallysmoked products erefore Commission Regulation (EU) No13272014 amending Regulation (EC) No 18812006 was setfor three years permitting certain Member States includingFinland to produce and consume traditionally smokedmeat andmeat products as well as fish and fish products in their territorythat comply with the PAH MLs of 50microgmiddotkgminus1 for BaP and300microgmiddotkgminus1 for the PAH4 sum [23] In this study the PAH4levels of Finnish smoked fish and meat products were de-termined In addition particular attention was paid to in-vestigating the effect of selected smoking parameters on thePAH4 levels to develop science-based guidance for a safesmoking procedure Particularly aimed at small andmedium-sized enterprises (SMEs) guidance will strengthentheir expertise and thereby prevent and reduce PAH con-tamination in traditionally smoked fish and meat products

2 Materials and Methods

21 Samples During 2012 the municipal food control au-thorities and the inspection veterinarians of the FinnishFood Safety Authority Evira collected 142 samples (80

fishfishery products and 62 meatmeat products) throughoutFinland from smokehouses and retail shops with their ownsmoking chambers Among the meat and meat productsdifferent types of pork products were the most common(68) (Figure 1(a)) and for the fish and fish products eitherwarm or cold smoked salmon products were predominant(82) (Figure 1(b)) Sampling was conducted accordingto Commission Regulation (EC) No 3332007 [24] andfollowed a risk-based sampling plan (eg the smokingtechnique used and production volumes) Only so-calledworst-case samples from the consumer perspective whichwere most likely to contain PAH compounds were chosenfor chemical analysis erefore liquid-smoked samples forinstance were not included During the sampling detailedinformation on the production process was recordedHowever incompletely reported smoking parameters wereomitted

22 Chemicals and Reagents Organic solvents (dichloro-methane hexane acetone cyclohexane methanol andtoluene) were HPLC or ultra resi-grade and purchased fromJ T Baker (Deventer Netherlands) Ethyl acetate (HPLCgrade) was obtained from VWR Chemicals (Fontenay-sous-Bois France) Ethanol was provided by Altia Oyj (RajamakiFinland) and was of Aa grade (ge995) Celite 545 andflorisil (100ndash200 mesh) used for accelerated solvent extraction(ASE 200reg Dionex Corporation) were obtained fromMerck(Darmstadt Germany) and from Sigma-Aldrich (SteinheimGermany) respectively Supelclean ENVI-Chrom P (6mL500mg) solid-phase extraction cartridges were providedby Supelco (Bellefonte PA USA) e analytical standardmixture (PAH-Mix 183 10 microgmiddotmLminus1) was purchased fromDr Ehrenstorfer GmbH (Augsburg Germany) In additionthe isotopically labelled compounds (13C6 BaA 13C6 CHR13C6 BbFA and 13C4 BaP) containing 100 microgmiddotmLminus1 eachwere supplied by LGC Standards GmbH (Wesel Germany)and used as internal standards e stock solutions of theindividual 13C PAHs were prepared and combined intoluene All the standard solutions were stored at +4degCprotected from the light

23 Sample Preparation PAH4 compounds were deter-mined using a gas chromatography-tandem mass spec-trometry (GC-MSMS) method accredited according to ISO17025 e method was based on the publication of Veyrandet al [25] with minor modifications All samples werepacked in aluminium foil or in a sales package and stored at+5degC prior to homogenisation in a food mixer According tothe manufacturerrsquos information the fish or meat skin wasremoved before homogenisation if the skin was not an ediblepart of the product MLs are only applicable to the edibleparts of smoked productse samples were homogenised ina mixer and approximately 100 g of each sample was takento be freeze-dried Half a gram of the dried sample wasspiked with internal standards (13C-labelled PAHs) followedby the extraction of PAHs using ASE 200reg e extractioncells were filled with celite and florisil and prewashed withdichloromethane before adding the sample and extracting

2 Journal of Chemistry

with hexaneacetone (5050 vv) 13e extract was evapo-rated diluted into cyclohexane and puried in the solid-phase extraction (SPE) cartridge 13e SPE cartridge was rstconditioned (ethyl acetate cyclohexane) and washed withcyclohexaneethanol (7030 vv) after which the sampleextract was introduced into the SPE cartridge 13e PAHcompounds were eluted with cyclohexaneethyl acetate(4060 vv) Finally the solvent was evaporated with ni-trogen and the sample was redissolved in 100 microl toluene andtransferred to a GC vial For the GC-MSMS analysis onemicroliter of the nal sample extract was injected into theGC injector In the case of canned sh samples the sh andoil were separated and analysed separately 13e individualresults were considered when calculating the nal result forthe canned sh product

24 GC-MSMS Analysis 13e nal detection and quanti-cation of the analytes were performed using a gas chro-matograph (Agilent 6890N) coupled to a Micromass QuattroMicro GC triple quadrupole analyser (Waters Micromass)Analysis was carried out using the Select PAH capillary columnfromAgilent JampW (30mtimes 025mmtimes 015microm)Helium (AGAFinland) was applied as the carrier gas at the constant ow rateof 10mLmiddotminminus1 and argon (AGA Finland) as the collision gas13e injection was done in splitless mode by utilising theelectron ionisation (EI) technique (70 eV) Ion transitions weremonitored with the multiple reaction monitoring (MRM)mode and the two most predominant fragments for eachanalyte were utilised Precursor ions and the selectedproduct ions are given in Table 1 13e injector and thetransfer line temperature were held at 300degC and the ionsource at 275degC 13e GC oven temperature was initially keptat 110degC for 07min and then raised at 5degCmiddotminminus1 to 180degC3degCmiddotminminus1 to 230degC for 7min 28degCmiddotminminus1 to 280degC for

15min and 14degCmiddotminminus1 to 350degC for 5min Quanticationwas performed by using the internal standard calibrationcurve with seven standards from 0 to 800 ngmiddotmLminus1 13e datawere processed using MassLynx V41 software supplied byWaters (Manchester United Kingdom)

25 Statistical Analysis 13e statistical analysis was per-formed using IBM SPSS Statistics version 25 (SPSS IncChicago Illinois) Boxplots were performed to visualise thedistribution of the data sets

26 Method Validation and Quality Control 13e GC-MSMSmethod complies with the criteria for the ocentcial controlof PAH4 according to Commission Regulation (EU) No8362011 [26] Method validation was performed for shmeat and oil matrices using Commission Decision (EC)No 6572002 as a guideline [1] 13e following parameterswere successfully tested specicity selectivity linearityrepeatability within-laboratory reproducibility apparentrecovery the limit of detection (LOD) the limit ofquantication (LOQ) and trueness 13e prociency test

68

11

11

5 3 2

PorkSalamiSausage

ReindeerTurkeyOthers

(a)

82

5

52 3 3

SalmonWhitefishCanned fish

HerringVendaceOthers

(b)

Figure 1 Percentage distribution of sampled (a) smoked meat (n 62) and (b) sh (n 80) products

Table 1 Precursor and product ions of PAH4 compounds

Compound and precursor ion (mz) Product ion (mz)Benzo[a]pyrene 2523 2262 2501ISTD benzo[a]pyrene 13C4 2560 2541Benz[a]anthracene 2282 2022 2262ISTD benz[a]anthracene 13C6 2340 2322Chrysene 2282 2021 2261ISTD chrysene 13C6 2340 2541Benzo[b]uoranthene 2523 2260 2501ISTD benzo[b]uoranthene 13C6 2580 2562Underlined product ions are used as quantier ions

Journal of Chemistry 3

organised by FAPASreg was used to assess the trueness of themethod Smoked meat test material was analysed for BaPBaA and BbFA and our z-scores were minus07 01 and minus04respectively

Quality controls of the method were performed bothbefore and during the sample run Each GC-MSMS run waspreceded by a system performance check with the standardsolutions containing 0011 and 0064 microgmiddotmLminus1 of PAH4standards (PAH-Mix 183 Dr Ehrenstorfer GmbH Augs-burg Germany) In addition the standard solutions wereinjected at the beginning in the middle and at the end ofeach sample batch Individual internal standards were usedfor each PAH4 compound

e spiked matrix sample was used to check the func-tionality of the whole method by spiking the blank fish ormeat sample with PAH4 standards at the level of 50microgmiddotkgminus1and analysing in each sample batch e performance criteriaset in Commission Regulation (EU) No 8362011 consider therecovery values 50ndash120 the LODle 030microgmiddotkgminus1 and theLOQle 090microgmiddotkgminus1 for each of the four substances acceptable[26] All our performance values were within these limitsAccording to Commission Decision (EC) No 6572002 [1]the maximum permitted tolerances for relative ion in-tensities were defined during fish and meat validation andapplied as an identification criteria (Table 2) A chro-matographic peak which was eluting at the retention time ofa target PAH was identified if qualifier ion was present andthe ion ratio between quantifier and qualifier was within theacceptable ion ratio As an example benzo[a]pyrene ina meat sample with ion ratio of 53 is illustrated in Figure 2

3 Results

e method used for the determination of PAH4 isroutinely used and is accredited according to ISO 17025[27] e LOD and LOQ of the method were 026 microgmiddotkgminus1and 087 microgmiddotkgminus1 for each PAH4 analyte in fish and meatrespectively Upper bound concentrations were calcu-lated assuming that all values of the individual PAH4compounds less than the LOQ were equal to the LOQFurthermore the concentrations below the LOD werecalculated as zero Measurement uncertainty (MU) wasbased on the validation and proficiency test data [28]Estimated as expanded uncertainty MU varied from 16 to27 in fish and from 10 to 22 in meat depending on thePAH4 compound For the PAH4 sum MU was 44 in fishand 38 in meat

In general the determined PAH4 levels in smoked fishand meat samples were relatively low and below the pre-vailing MLs Regarding the smoked fish products the meanconcentrations were 07 microgmiddotkgminus1 for BaP and 39 microgmiddotkgminus1 forthe PAH4 sum For smoked meat products the mean BaPand PAH4 sum levels were 22 microgmiddotkgminus1 and 11 microgmiddotkgminus1 re-spectively It is noteworthy however that our results dis-played wide variation particularly in meat products forwhich the PAH4 sum concentrations ranged from not de-tected to 200 microgmiddotkgminus1 (Table 3) Moreover median PAHlevels in pork products were higher than in other meatproducts

e smoked large fish products (n 72) contained 79fillets 13 pieces and 8 whole fish samples whereas alleight small fish samples represented the whole fish Whencomparing the data between the products of larger fishspecies (eg salmon and whitefish) and smaller fish(eg herring vendace and roach) those of smaller fishwere found to contain higher median concentrations ofPAHs (Table 3) e minimum and maximum values wereapproximately at the same level in both large and smallfish species

Within the framework of the study four meat productsamples were found noncompliant with the legislation andin all these cases direct smoking was used e competentauthorities enforced the operator of these products tomodify the smoking process After modifications follow-upsamples were taken to ensure the products were compliantwith the legislation before allowing the placement of theproducts on the market

Four canned smoked fish products were analysedcomprising either vendace or roach e final results werecalculated by combining the relative proportions of PAHs inthe fish and vegetable oil in a can In all of the cases thePAH4 contamination was almost five times higher in thevegetable oil than in the fish part A similar oil-fish PAHratio (5 1) was reported by Lawrence and Weber [29] incanned smoked sardine However all the canned fishsamples were compliant with the legislation

31 Effect of the Selected Smoking Parameters on the PAH4Levels e effect of the selected smoking parameters wasstudied using smoked fish and meat samples Additionallysmoked salmon fillets (n 57) were evaluated separately inorder to eliminate the interfering factors such as other fishspecies and effect of the skin Salmon fillets were all smokedwithout the skin Regarding the smoking time more in-vestigation in fish was needed thus cold and warmhotsmoked salmon fillets as well as large and small fish specieswere studied

311 Smoking Technique e samples were smoked usingeither a direct or an indirect smoking technique In smokedfish samples direct smoking technique produced only slightlyhigher BaP and PAH4 sum concentrations than indirectsmoking Exceptionally one sample smoked with indirecttechnique contained highest PAH4 concentrations(Figure 3(a)) For smoked salmon fillets the results

Table 2e calculated maximum permitted ion ratio tolerance forfish and meat samples

CompoundMaximum permittedion ratio tolerancefor fish samples

Maximum permittedion ratio tolerancefor meat samples

Benzo[a]pyrene 40ndash75 45ndash84Benz[a]anthracene 40ndash66 39ndash73Chrysene 36ndash59 37ndash61Benzo[b]fluoranthene 62ndash12 50ndash93


illustrated direct smoking producing higher PAH levels thanthe indirect smoking (Figure 3(b)) Moreover directsmoking generated clearly higher PAH levels in smokedmeat samples than indirect smoking (Figure 3(c))

312 Smoking Time 13e ecopyect of smoking time was exam-ined by dividing the PAH results into two categories accordingto whether the smoking process took less or more than vehours Surprisingly the shorter smoking time generated higherPAH concentrations in sh samples particularly observed forthe PAH4 sum (Figures 4(a) and 4(b)) 13e comparison be-tween cold smoked and warm or hot smoked salmon lletsindicated that in cold smoking process only two samplescontained detectable PAH concentrations (Figure 4(c)) Inwarm or hot smoked salmon llets which were mostlysmoked in less than ve hours PAH levels were clearlyhigher than in cold smoked products Moreover the shsize may acopyect the observed PAH levels which was studied

between the large and small sh products All the small shproducts were smoked in less than ve hours and there-fore the comparison between the large and small shproducts was performed in that time range 13e resultsshow the small sh samples containing higher medianPAH levels (Figure 4(d)) In smoked meat samples thePAH concentrations were higher for both BaP andthe PAH4 sum when the smoking time exceeded vehours (Figure 4(e))

313 Smoke Generation Temperature 13e temperaturerequired for smoke generation was dened as optimised ifthe thermal eld was maintained at 400ndash600degC during thewhole process In a nonoptimised case the temperature mayvary during the process and it is not therefore controlled Asexpected both for sh and meat samples lower amounts ofPAHs were formed when the temperature was optimised(Figures 5(a)ndash5(c)) Particularly as shown in Figure 5(b)

3175 3200 3225 3250 3275 3300 3325 3350 3375 3400 3425 3450 3475 3500

()

11

F5 MRM of 3 channels EI+256 gt 25412

14021320 Smooth (Mn 2times3)

2479e + 004

(min)

()

11



7205e + 003

()

2



4510e + 004Benzo[a]pyrene2523 gt 2501

RT = 3262

Benzo[a]pyrene2523 gt 2262

RT = 3261

ISTDBenzo[a]pyrene13C4

2560 gt 2541RT = 3261

Figure 2 Multiple reaction monitoring chromatogram of benzo[a]pyrene at the level of 51 microgmiddotkgminus1 in a meat sample

Table 3 Median and minimum-maximum concentrations (microgmiddotkgminus1) for BaP and PAH4 in products of large and small sh and in pork andother meat products

Product typeBaP PAH4 sum

Median (microgmiddotkgminus1) Minimum-maximum(microgmiddotkgminus1) Median (microgmiddotkgminus1) Minimum-maximum

(microgmiddotkgminus1)Large sh products (n 72) 0 0ndash49 03 0ndash26Small sh products (n 8) 09 0ndash38 8 0ndash21Pork products (n 42) 08 0ndash40 19 0ndash200Other meat products (n 20) 0 0ndash68 04 0ndash38


Direct smoking (n = 35)

(μg

kgndash1

)

30

25

20

15

10

5

0

Indirect smoking (n = 45)Smoking technique

BaP BaPPAH4 PAH4

lowast lowastlowast

lowast

lowast

lowast

(a)

(μg

kgndash1

)

25

20

15

10

5

0

Direct smoking (n = 27) Indirect smoking (n = 30)Smoking technique

BaPBaP PAH4 PAH4

lowastlowast

lowast

(b)

200

150

100

50

0


BaP BaPPAH4 PAH4

(μg

kgndash1

)

lowastlowast

lowast

lowastlowastlowastlowastlowast

(c)

Figure 3 (a) Boxplot diagrams of BaP and PAH4 concentrations in microgmiddotkgminus1 wet weight for fish products divided according to the smokingtechnique used e line in the central box represents the median the whiskers show the distribution outside the central quartiles and thecircles and stars are outliers N is the number of samples (b) Boxplot diagrams of BaP and PAH4 concentrations in microgmiddotkgminus1 wet weight forsalmon fillet products divided according to the smoking technique usede line in the central box represents themedian the whiskers showthe distribution outside the central quartiles and the circles and stars are outliers N is the number of samples (c) Boxplot diagrams of BaPand PAH4 concentrations in microgmiddotkgminus1 wet weight for meat products divided according to the smoking technique used e line in the centralbox represents the median the whiskers show the distribution outside the central quartiles and the circles and stars are outliers N is thenumber of samples


Smoking time gt5h (n = 18)

30

25

20

15

10

5

0 BaP PAH4 PAH4BaPSmoking time lt5h (n = 54)

Smoking time

(μg

kgndash1

)

lowastlowastlowastlowast

lowast

(a)

lowast

30

25

20

15

10

5

0 BaP PAH4 PAH4BaPSmoking time gt5h (n = 16)Smoking time lt5h (n = 36)

Smoking time

(μg

kgndash1

) lowast

lowast

lowast

lowast

(b)

Warm or hot smoked salmon fillet (n = 23)

25

20

15

10

5

0 BaP BaPPAH4 PAH4 BaP BaPPAH4 PAH4

Smoking time Smoking time

Smoking typeCold smoked salmon fillet (n = 13)

lt5h(n = 2) gt5h(n = 11) lt5h(n = 21) gt5h(n = 2)

(μg

kgndash1

)

lowast

lowastlowast

(c)

Figure 4 Continued


PAH levels were clearly higher for smoked salmon llets inthe nonoptimised system

314 Distance between the Food and the Smoke SourceIn this study the reported length between the food andsmoke source was either under or over ve metres 13elonger distance (over ve metres) generated lower concen-trations of PAHs in both sh and meat products(Figures 6(a)ndash6(c)) 13e comparison between PAH levelsin smoked sh (Figure 6(a)) and salmon llets (Figure 6(b))demonstrated that similar results were obtained Furthermore

there were no detectable levels of BaP in meat productswhen the distance from the smoke source was over vemetres (Figure 6(c))

4 Discussion

Food safety and especially the risk management of PAHsmust be the rst priority in food smoking processes al-though smoking extends the food storage period and im-proves the avour composition and preservation of thevitamin content [20] 13e Codex Alimentarius recommendsoptimisation of the smoking process to produce smoked

30

25

20

15

10

5

0

Fish size

BaP BaPPAH4 PAH4Small fish (n = 8)Large fish (n = 46)

lowastlowastlowast


(μg

kgndash1

)

(d)

200

150

100

50

0

Smoking time gt5h (n = 22)Smoking time lt5h (n = 22)Smoking time

BaP BaPPAH4 PAH4

(μg

kgndash1

)

lowastlowastlowast lowast

lowast

lowast

(e)

Figure 4 (a) Boxplot diagrams of BaP and PAH4 concentrations in microgmiddotkgminus1 wet weight for sh products divided according to the smokingtime 13e line in the central box represents the median the whiskers show the distribution outside the central quartiles and the circles andstars are outliers N is the number of samples 72 samples contained the relevant information (b) Boxplot diagrams of BaP and PAH4concentrations in microgmiddotkgminus1 wet weight for salmon llet products divided according to the smoking time13e line in the central box representsthemedian the whiskers show the distribution outside the central quartiles and the circles and stars are outliersN is the number of samples52 samples contained the relevant information (c) Boxplot diagrams of BaP and PAH4 concentrations in microgmiddotkgminus1 wet weight for coldsmoked and warm or hot smoked salmon llets divided according to the smoking time13e line in the central box represents the median thewhiskers show the distribution outside the central quartiles and the circles and stars are outliers N is the number of samples 36 samplescontained the relevant information (d) Boxplot diagrams of BaP and PAH4 concentrations in microgmiddotkgminus1 wet weight for large and small shproducts smoked in less than ve hours 13e line in the central box represents the median the whiskers show the distribution outside thecentral quartiles and the circles and stars are outliers N is the number of samples 54 samples contained the relevant information (e)Boxplot diagrams of BaP and PAH4 concentrations in microgmiddotkgminus1 wet weight for meat products divided according to the smoking time13e linein the central box represents the median the whiskers show the distribution outside the central quartiles and the circles and stars areoutliers N is the number of samples 44 samples contained the relevant information


25

20

15

10

5

0

Temperature optimised (n = 18) Temperature not optimised (n = 17)Temperature optimisation

BaPBaP PAH4 PAH4

(μg

kgndash1

)

lowast

lowastlowast

(a)

25

20

15

10

5

0


lowastlowastlowast

(μg

kgndash1

)

BaPBaP PAH4 PAH4

(b)

200

150

100

50

0


(μg

kgndash1

)

lowastlowastlowast

lowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(c)

Figure 5 (a) Boxplot diagrams of BaP and PAH4 concentrations in microgmiddotkgminus1 wet weight for sh products divided according to thetemperature optimisation13e line in the central box represents the median the whiskers show the distribution outside the central quartilesand the circles and stars are outliers N is the number of samples 35 samples contained the relevant information (b) Boxplot diagrams ofBaP and PAH4 concentrations in microgmiddotkgminus1 wet weight for salmon llet products divided according to the temperature optimisation 13e linein the central box represents the median the whiskers show the distribution outside the central quartiles and the circles and stars areoutliers N is the number of samples 24 samples contained the relevant information (c) Boxplot diagrams of BaP and PAH4 in microgmiddotkgminus1 wetweight for meat products divided according to the temperature optimisation13e line in the central box represents the median the whiskersshow the distribution outside the central quartiles and the circles and stars are outliers N is the number of samples 26 samples containedthe relevant information


30

25

20

15

10

5

0

Distance lt5m (n = 60) Distance gt5m (n = 9)Distance

(μg

kgndash1

)

lowastlowastlowast

lowast

lowastlowast

lowast

lowast

BaPBaP PAH4 PAH4

(a)

25

20

15

10

5

0


(μg

kgndash1

)

lowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(b)

200

150

100

50

0


(μg

kgndash1

)

lowastlowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(c)

Figure 6 (a) Boxplot diagrams of BaP and PAH4 concentrations in microgmiddotkgminus1 wet weight for sh products divided according to the distance13e line in the central box represents the median the whiskers show the distribution outside the central quartiles and the circles and starsare outliers N is the number of samples 69 samples contained the relevant information (b) Boxplot diagrams of BaP and PAH4concentrations in microgmiddotkgminus1 wet weight for salmon llet products divided according to the distance 13e line in the central box represents themedian the whiskers show the distribution outside the central quartiles and the circles and stars are outliersN is the number of samples 50samples contained the relevant information (c) Boxplot diagrams of BaP and PAH4 concentrations in microgmiddotkgminus1 wet weight for meat productsdivided according to the distance 13e line in the central box represents the median the whiskers show the distribution outside the centralquartiles and the circles and stars are outliers N is the number of samples 43 samples contained the relevant information


products that have good microbiological quality and sensoryproperties with low PAH levels [20] In order to control thePAH levels in smoked products it is important to identifythe critical parameters

Overall due to the risk-based ie selective sampling inthis study the detected PAH4 levels in fish and meat wereprobably higher than would have been obtained using ob-jective sampling ey were nevertheless relatively low andcomparable to other studies [18 30 31] with some ex-ceptions However the large variation in PAH levels be-tween the samples demonstrated how variable the smokingprocess can be in Finnish smokehouses e smokingequipment used are rather self-made than commercial thuseg the length of smoking tube and the model of the filtermay vary from one device to another In 1999 the levels of 11PAHs in different food categories were determined inFinland Among smoked meat products the highest BaPconcentrations were observed in smoked pork products(56ndash132 microgmiddotkgminus1) whereas in the smoked fish productcategory a smoked herring sample contained a sum con-centration of 11 PAHs of 270 microgmiddotkgminus1 but a relatively lowBaP concentration of 07 microgmiddotkgminus1 [32] Due to the differentcombinations of PAHs analysed comparison with our re-sults is challenging but we observed the same trend of higherPAH levels in smoked pork products compared to othermeat products e data analysis did not reveal any par-ticular smoking parameter alone to be the reason for thehigher PAH levels in pork but some factors were foundrelated to long smoking time up to 36 hours no distancebetween the food and smoke source or irregular cleaning ofthe smoking equipment Altogether the high variationamong the results demonstrated the smoking process eitherto be controlled or not Products of herring and also othersmall fish species such as vendace roach and mackerelcontained higher PAH levels than those of larger fish species(salmon and whitefish) is may be attributed to the as-sumption that herring is often more heavily smoked thansalmon due to the smaller surface area-to-volume ratio[16 29]

Additionally small fish species are consumed with theskin which contains more PAHs than the other parts of thefish According to previous studies [18 33] the skin acts asa barrier to penetration by smoke particles thereby pre-venting PAH contamination e uptake of PAHs into thefish tissue is greater if the skin has been removed beforesmoking When reaching the product surface PAHs accu-mulate on the skin surface and may migrate into the un-derlying fatty tissue due to their solubility in fat e wateractivity and fat content have a significant role in the diffusionprocess [15] For smoked meat similar conclusions weremade by Ciecierska and Obiedzinski [34] who demon-strated lower PAH levels in the interior than the exterior ofthe same products In canned fish products PAHs canmigrate from the fish to the oil or remain in the fish lipid[35] However the oil mainly acts as a preservative andmany consumers pour away most of it before food con-sumption [35]

As expected the smoking technique had a clear impacton the PAH levels For both meat and fish products direct

smoking generated higher concentrations of BaP and PAH4than indirect smoking e European Commission reportedin 2004 an average BaP concentration of 53 microgmiddotkgminus1 fordirectly smoked fish whereas for indirectly smoked fish theBaP concentrations ranged from 01 microgmiddotkgminus1 to 20 microgmiddotkgminus1[36] e direct contact of the product with combustiongases is definitely an important source of PAH contami-nation [3 6 15] In the direct smoking process the com-bustion temperature is often very high and challenging tocontrol [16 18] e replacement of direct smoking byindirect smoking may reduce PAH contamination [7 34]However 100 of the directly smoked fish products and82 of the directly smoked meat products were compliantwith the legislation so the direct technique is applicablewhen properly controlled Similar findings were observed inSwedish meat and fish products that had been traditionallydirectly ldquosaunardquo smoked in which the smoked meatproducts contained higher levels of BaP than the smoked fishproducts e ldquosaunardquo smoking produced increased BaPlevels in nine meat samples (66ndash369 microgmiddotkgminus1) and in six fishsamples (84ndash144 microgmiddotkgminus1) [18]

e smoking time is one of the critical factors to considerin the smoking process and it should be kept as short aspossible taking into account food safety and the productshelf life because a prolonged smoking time increases thePAH exposure of the product [19 20 37]is was especiallynoticeable in the meat sample results whereas for fishsamples a shorter smoking time generated unexpectedlyhigher PAH concentrations than a longer smoking timeFish results were divided into different groups to be able toevaluate them better Similar findings were done in salmonfillets which in turn were divided into cold smoked andwarm or hot smoked fillets e temperature differs gen-erally from 12 to 25degC in cold smoking from 25 to 45degC inwarm smoking and from 40 to 100degC in hot smoking [13]Warm or hot smoking procedure generated more PAHsthan the cold smoking with some exceptions Most of thecold smoked salmons (85) were smoked more than fivehours whereas 91 of the warm or hot smoked productswere smoked less than five hours which explains thehigher PAH levels in salmon fillets within shorter smokingtime e results are in agreement with Danish studieswhich indicated hot smoking producing higher PAHlevels than cold smoking [16] Moreover all the small fishspecies such as herring and vendace were smoked in lessthan five hours with higher median PAH levels comparedto larger fish species (eg salmon and whitefish) Asmaller product size may result in the formation of higherPAH levels [16]

e most significant parameter influencing the PAHconcentrations in smoked products is the smoke generationtemperature [13] e composition of smoke is dependenton the temperature which needs to be regulated to reduce theformation of PAHs [20] According to Gobel [37] PAHs easilyform in the temperature range between 660 and 740degC Whentemperature is increased within the range 400ndash1000degC there isa linear increase in the formation of PAHs in smoke [18 38]Based on our results smaller amounts of PAHs were formedwhen the smoke generation temperature was optimised to


400ndash600degC than in nonoptimised conditions e difference inPAH levels between the optimised and nonoptimised processeswas obvious in both fish and meat samples which emphasisesthe significance of temperature optimisation Typically thesmoke generation temperature varies between 500 and 800degCand is controlled by the air supply [33]

Besides the temperature the distance between thefood and the smoke source is among the important factorsto observe PAHs are bound to smoke particles anda longer distance from the smoke source to the smokedfood may therefore lead to lower PAH levels in the food[20] e analysed fish and meat samples smoked ata distance over five metres from the smoke source con-tained lower levels of PAHs In indirect smoking thelength of the smoking tube varied between 02 and23 metres Naturally all the samples smoked with thedirect technique were placed in the same chamber as thesmoke source and the distance was therefore alwaysunder five metres It can be expected that PAH levels willthen be higher because as previously mentioned directsmoking leads to higher concentrations of PAHs in theproducts To be able to reduce the PAH levels in the finalproduct an external smoke generator combined witha long tube could be a good option

Evaluating the significance of smoked meat and fish inthe average diets of several European countries the ScientificCommittee on Food (SCF) estimated the intake of BaP fromsmoked fish and meat to form only a small part of the totaldietary intake [13] However in local communities or incertain countries eg in Finland or Sweden [18] wheresmoked fish and meat traditionally constitute a signifi-cant proportion of the diet the intake of BaP and otherPAH4 compounds from these sources is significantlyhigher [13] Regarding the European Commissionrsquos es-timation in 2004 the average BaP intake in Finland is upto 270 ngpersonday whereas elsewhere in Europe theintake varies between 14 and 320 ngpersonday [36] eCodex Alimentarius code of practice advises that insmoking attention should be paid to choosing wood witha low lignin content filtering or cooling the smokereplacing direct with indirect smoking when possibleoptimising the smoking time temperature and airflowavoiding fat dripping onto the heat source regularlycleaning and maintaining the equipment increasing thedistance of the product from the smoking source andwashing the product with water after smoking [20] Inorder to precisely determine the influence of the smokingmethod and other processing parameters on the PAHcontamination of food the Codex Alimentarius and EUregulators require further scientific information [20]

5 Conclusions

Based on our results to obtain safe fish and meat productsregarding PAHs in traditional smoking indirect smokingwith distance more than five meters between food andsmoke source as well as smoking time less than five hoursand an optimised temperature (without jeopardising mi-crobiological food safety) are recommended

Data Availability

e data used to support the findings of this study areavailable from the corresponding author upon request

Conflicts of Interest

e authors declare that there are no conflicts of interestregarding the publication of this paper

Acknowledgments

We are very grateful for the sampling performed by themunicipal food control authorities and the inspection vet-erinarians of Evira We thank Kristian Alho for performingthe PAH analysis and Dr Jukka Ranta for the statisticaladvice In addition we would like to thank the whole samplepretreatment team

References

[1] Commission of the European Communities ldquoCommissiondecision of 12 August 2002 implementing Council Directive9623EC concerning the performance of analytical methodsand the interpretation of resultsrdquo Official Journal of theEuropean Union vol L221 pp 8ndash36 2002

[2] P Simko ldquoDetermination of polycyclic aromatic hydrocar-bons in smoked meat products and smoke flavouring foodadditivesrdquo Journal of Chromatography B vol 770 no 1-2pp 3ndash18 2002

[3] European Food Safety Authority ldquoPolycyclic aromatic hy-drocarbons in food Scientific opinion of the panel on con-taminants in the food chain adopted on 9 June 2008rdquo EFSAJournal vol 724 pp 1ndash114 2008

[4] M-K Song M Song H-S Choi Y-J Kim Y-K Park andJ-C Ryu ldquoIdentification of molecular signatures predictingthe carcinogenicity of polycyclic aromatic hydrocarbons(PAHs)rdquo Toxicology Letters vol 212 no 1 pp 18ndash28 2012

[5] International Agency for Research on Cancer ldquoChemicalagents and related occupations A review of human carcin-ogens IARC Monographs on the evaluation of carcinogenicrisks to humansrdquo December 2016 httpsmonographsiarcfrwp-contentuploads201806mono100Fpdf

[6] G Purcaro S Moret and L S Conte ldquoOverview on polycyclicaromatic hydrocarbons occurrence legislation and innova-tive determination in foodsrdquo Talanta vol 105 pp 292ndash3052013

[7] Scientific Committee on Food ldquoOpinion of the ScientificCommittee on Food on the risks to human health of poly-cyclic aromatic hydrocarbons in food expressed on 4 De-cember 2002rdquo Report No SCFCSCNTMPAH29 FinalScientific Committee on Food Brussels Belgium October2016 httpseceuropaeufoodsitesfoodfilessafetydocssci-com_scf_out153_enpdf

[8] World Health Organization ldquoEvaluation of certain foodadditives and contaminants in WHO food additive reportseriesrdquo vol 28 October 2016 httpwwwinchemorgdocumentsjecfajecmonov28je18htm

[9] M Rose J Holland A Dowding et al ldquoInvestigation into theformation of PAHs in foods prepared in the home to de-termine the effects of frying grilling barbecuing toasting androastingrdquo Food and Chemical Toxicology vol 78 pp 1ndash92015


[10] V Bansal and K Kim ldquoReview of PAH contamination in foodproducts and their health hazardsrdquo Environment Interna-tional vol 84 pp 26ndash38 2015

[11] E Ledesma M Rendueles and M Dıaz ldquoSpanish smokedmeat products benzo(a)pyrene (BaP) contamination andmoisturerdquo Journal of Food Composition and Analysis vol 37pp 87ndash94 2015

[12] J M Lingbeck P Cordero C A OrsquoBryan M G JohnsonS C Ricke and P G Crandall ldquoFunctionality of liquid smokeas an all-natural antimicrobial in food preservationrdquo MeatScience vol 97 no 2 pp 197ndash206 2014

[13] A Stołyhwo and Z E Sikorski ldquoPolycyclic aromatic hydro-carbons in smoked fishmdasha critical reviewrdquo Food Chemistryvol 91 no 2 pp 303ndash311 2005

[14] A eobald D Arcella A Carere et al ldquoSafety assessment ofsmoke flavouring primary products by the European FoodSafety Authorityrdquo Trends in Food Science amp Technologyvol 27 no 2 pp 97ndash108 2012

[15] E Fasano I Yebra-Pimentel E Martınez-Carballo andJ Simal-Gandara ldquoProfiling distribution and levels of car-cinogenic polycyclic aromatic hydrocarbons in traditionalsmoked plant and animal foodsrdquo Food Control vol 59pp 581ndash590 2016

[16] L Duedahl-Olesen J H Christensen A Hoslashjgard K GranbyandM Timm-Heinrich ldquoInfluence of smoking parameters onthe concentration of polycyclic aromatic hydrocarbons(PAHs) in Danish smoked fishrdquo Food Additives amp Contam-inants Part A vol 27 no 9 pp 1294ndash1305 2010

[17] P Visciano M Perugini F Conte and M AmorenaldquoPolycyclic aromatic hydrocarbons in farmed rainbow trout(Oncorhynchus mykiss) processed by traditional flue gassmoking and by liquid smoke flavouringsrdquo Food andChemical Toxicology vol 46 no 5 pp 1409ndash1413 2008

[18] S Wretling A Eriksson G A Eskhult and B LarssonldquoPolycyclic aromatic hydrocarbons (PAHs) in Swedishsmoked meat and fishrdquo Journal of Food Composition andAnalysis vol 23 no 3 pp 264ndash272 2010

[19] D K Essumang D K Dodoo and J K Adjei ldquoEffect ofsmoke generation sources and smoke curing duration on thelevels of polycyclic aromatic hydrocarbon (PAH) in differentsuites of fishrdquo Food and Chemical Toxicology vol 58pp 86ndash94 2013

[20] Codex Alimentarius Commission ldquoCode of practice forthe reduction of contamination of food with polycyclicaromatic hydrocarbons (PAH) from smoking and directdrying processes CACRCPrdquo vol 68 July 2017 httpwwwcodexalimentariusorgdownloadstandards11257CXP_068epdf

[21] Commission of the European Communities ldquoCommissionregulation (EC) no 18812006 of 19 December 2006 settingmaximum levels for certain contaminants in foodstuffsrdquo OfficialJournal of the European Union vol L364 pp 5ndash24 2006

[22] European Commission ldquoCommission regulation (EU) no8352011 of 19 August 2011 amending regulation (EC) no18812006 as regards maximum levels for polycyclic aromatichydrocarbons in foodstuffsrdquo Official Journal of the EuropeanUnion vol L215 pp 4ndash8 2011

[23] European Commission ldquoCommission regulation (EU) no13272014 of 12 December 2014 amending regulation (EC)no 18812006 as regards maximum levels of polycyclicaromatic hydrocarbons (PAHs) in traditionally smokedmeat and meat products and traditionally smoked fish andfishery productsrdquo Official Journal of the European Unionvol L358 pp 13-14 2014

[24] Commission of the European Communities ldquoCommissionregulation (EC) no 3332007 of 28 March 2007 laying downthe methods of sampling and analysis for the official control ofthe levels of lead cadmium mercury inorganic tin 3-MCPDand benzo(a)pyrene in foodstuffsrdquo Official Journal of theEuropean Union vol L88 pp 29ndash38 2007

[25] B Veyrand A Brosseaud L Sarcher et al ldquoInnovativemethod for determination of 19 polycyclic aromatic hydro-carbons in food and oil samples using gas chromatographycoupled to tandem mass spectrometry based on an isotopedilution approachrdquo Journal of Chromatography A vol 1149no 2 pp 333ndash344 2007

[26] e European Commission ldquoCommission regulation (EU) no8362011of 19 August 2011 amending regulation (EC) no 3332007 laying down the methods of sampling and analysis forthe official control of the levels of lead cadmium mercuryinorganic tin 3-MCPD and benzo(a)pyrene in foodstuffsrdquoOfficial Journal of the European Union vol L215 pp 9ndash162011

[27] International Organisation for Standardization ldquoGeneralrequirements for the competence of testing and calibrationlaboratories ISOIEC 17025 2005rdquo July 2018 httpwwwisoorgisohomehtm

[28] Nordic Committee on Food Analysis Procedure No 5 Es-timation and Expression of Measurement Uncertainty inChemical Analysis Nordic Committee on Food AnalysisOslo Norway 2003

[29] J F Lawrence and D F Weber ldquoDetermination of polycyclicaromatic hydrocarbons in some Canadian commercial fishshellfish and meat products by liquid chromatography withconfirmation by capillary gas chromatography-mass spec-trometryrdquo Journal of Agricultural and Food Chemistry vol 32pp 789ndash794 1984

[30] L Duedahl-Olesen S White and M L Binderup ldquoPolycyclicaromatic hydrocarbons (PAH) in Danish smoked fish andmeat productsrdquo Polycyclic Aromatic Compounds vol 26no 3 pp 163ndash184 2006

[31] M Reinik T Tamme M Roasto K Juhkam T Tenno andA Kiis ldquoPolycyclic aromatic hydrocarbons (PAHs) in meatproducts and estimated PAH intake by children and thegeneral population in Estoniardquo Food Additives and Con-taminants vol 24 no 4 pp 429ndash437 2007

[32] V Hietaniemi M Ovaskainen and A HallikainenldquoPAH-yhdisteet ja niiden saanti markkinoilla olevistaelintarvikkeistardquo Elintarvikevirasto Survey vol 6 pp 1ndash271999 in Finnish

[33] M Pohlmann A Hitzel F Schwagele K Speer and W JiraldquoInfluence of different smoke generation methods on thecontents of polycyclic aromatic hydrocarbons (PAH) andphenolic substances in Frankfurter-type sausagesrdquo FoodControl vol 34 no 2 pp 347ndash355 2013

[34] M Ciecierska and M Obiedzinski ldquoInfluence of smokingprocess on polycyclic aromatic hydrocarbonsrsquo content in meatproductsrdquo Acta Scientiarum Polonorum Technologia Ali-mentaria vol 6 pp 17ndash28 2007

[35] S Moret G Purcaro and L S Conte ldquoPolycyclic aromatichydrocarbons in vegetable oils from canned foodsrdquo EuropeanJournal of Lipid Science and Technology vol 107 no 7-8pp 488ndash496 2005

[36] Directorate-General Health and Consumer ProtectionldquoCollection of occurrence data on polycyclic aromatic hy-drocarbons in foodrdquo in Reports on Tasks for Scientific Co-operation Report of Experts Participating in Task 3212October 2004 pp 1ndash36 EFSA Publication Parma Italy 2004


[37] A Gobel ldquoSachstandsbericht polycyclische aromatischekohlenwasserstoffe (PAH)-ermittlung von herstellungs- undverarbeitungsverfahren mit dem ziel der reduzierung andvermeidung von PAH in lebensmittelnrdquo Journal fur Ver-braucherschutz und Lebensmittelsicherheit vol 1 no 4pp 271ndash284 2006 in German

[38] M Jagerstad and K Skog ldquoGenotoxicity of heat-processedfoodsrdquo Mutation ResearchFundamental and MolecularMechanisms of Mutagenesis vol 574 no 1-2 pp 156ndash1722005


TribologyAdvances in

Hindawiwwwhindawicom Volume 2018


International Journal ofInternational Journal ofPhotoenergy


Journal of

Chemistry


Advances inPhysical Chemistry

Hindawiwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2018

Bioinorganic Chemistry and ApplicationsHindawiwwwhindawicom Volume 2018

SpectroscopyInternational Journal of


Hindawi Publishing Corporation httpwwwhindawicom Volume 2013Hindawiwwwhindawicom

The Scientific World Journal

Volume 2018

Medicinal ChemistryInternational Journal of


NanotechnologyHindawiwwwhindawicom Volume 2018

Journal of

Applied ChemistryJournal of



Biochemistry Research International


Enzyme Research


Journal of

SpectroscopyAnalytical ChemistryInternational Journal of


MaterialsJournal of



BioMed Research International Electrochemistry

International Journal of


Na

nom

ate

ria

ls


Journal ofNanomaterials

Submit your manuscripts atwwwhindawicom

nowadays mainly used to obtain the desired colour flavouraroma and appearance in the smoked food rather than forpreservation purposes [14 15] Traditional smoking is generallyperformed by the formation of smoke from wood [16 17]Smoke is defined as the result of thermal pyrolysis of woodwhenaccess to oxygen is limited [6] PAHs and other chemicalcompounds occur in smoke particles which canmigrate into thefood product being smoked [2 18] Wood smoke containsa combination of antioxidant and antimicrobial chemicals(eg phenols carboxylic acids aldehydes and acetic acids) butalso someharmful compounds such as PAHs [12 17 19] PAHsare potential health hazards associated with smoked foods inwhich they typically occur as a complex mixture [6 13] InFinland smoking with direct and indirect techniques is widelyused in the processing of meat and fish products For directsmoking smoke is generated from an open fire in the samechamber as the smoked product whereas in indirect smokingthe smoke is generated in an external chamber separated fromthe food and the smoke is led to the product from the externalsmoke generator [16 20] Alongside the smoking technique thetype of process (grilling roasting smoking and drying) thedistance between the food and the smoke source the processtime and temperature impact the formed PAH levels [6]

Commission Regulation (EC) No 18812006 specifiesthe maximum levels (MLs) of BaP in different foodstuffs[21] In 2008 the European Food Safety Authority (EFSA)concluded that BaP alone is not a suitable indicator for theoccurrence and toxicity of PAHs in foods [3] erefore ad-ditional MLs for the sum of four PAHs (PAH4 BaP benz[a]anthracene (BaA) chrysene (CHR) and benzo[b]fluoranthene(BbFA)) were set in Commission Regulation (EU) No8352011 [22] e MLs for smoked meat and fish productssmoked from 192012 to 3182014 are 50 microgmiddotkgminus1 for BaPand 300 microgmiddotkgminus1 for the PAH4 sum e MLs applicablefrom 192014 for smoked meat and fish products are20 microgmiddotkgminus1 for BaP and 120 microgmiddotkgminus1 for the PAH4 sum [22]However in some Member States the lower levels set from192014 could not realistically be reached for traditionallysmoked products erefore Commission Regulation (EU) No13272014 amending Regulation (EC) No 18812006 was setfor three years permitting certain Member States includingFinland to produce and consume traditionally smokedmeat andmeat products as well as fish and fish products in their territorythat comply with the PAH MLs of 50microgmiddotkgminus1 for BaP and300microgmiddotkgminus1 for the PAH4 sum [23] In this study the PAH4levels of Finnish smoked fish and meat products were de-termined In addition particular attention was paid to in-vestigating the effect of selected smoking parameters on thePAH4 levels to develop science-based guidance for a safesmoking procedure Particularly aimed at small andmedium-sized enterprises (SMEs) guidance will strengthentheir expertise and thereby prevent and reduce PAH con-tamination in traditionally smoked fish and meat products

2 Materials and Methods

21 Samples During 2012 the municipal food control au-thorities and the inspection veterinarians of the FinnishFood Safety Authority Evira collected 142 samples (80

fishfishery products and 62 meatmeat products) throughoutFinland from smokehouses and retail shops with their ownsmoking chambers Among the meat and meat productsdifferent types of pork products were the most common(68) (Figure 1(a)) and for the fish and fish products eitherwarm or cold smoked salmon products were predominant(82) (Figure 1(b)) Sampling was conducted accordingto Commission Regulation (EC) No 3332007 [24] andfollowed a risk-based sampling plan (eg the smokingtechnique used and production volumes) Only so-calledworst-case samples from the consumer perspective whichwere most likely to contain PAH compounds were chosenfor chemical analysis erefore liquid-smoked samples forinstance were not included During the sampling detailedinformation on the production process was recordedHowever incompletely reported smoking parameters wereomitted

22 Chemicals and Reagents Organic solvents (dichloro-methane hexane acetone cyclohexane methanol andtoluene) were HPLC or ultra resi-grade and purchased fromJ T Baker (Deventer Netherlands) Ethyl acetate (HPLCgrade) was obtained from VWR Chemicals (Fontenay-sous-Bois France) Ethanol was provided by Altia Oyj (RajamakiFinland) and was of Aa grade (ge995) Celite 545 andflorisil (100ndash200 mesh) used for accelerated solvent extraction(ASE 200reg Dionex Corporation) were obtained fromMerck(Darmstadt Germany) and from Sigma-Aldrich (SteinheimGermany) respectively Supelclean ENVI-Chrom P (6mL500mg) solid-phase extraction cartridges were providedby Supelco (Bellefonte PA USA) e analytical standardmixture (PAH-Mix 183 10 microgmiddotmLminus1) was purchased fromDr Ehrenstorfer GmbH (Augsburg Germany) In additionthe isotopically labelled compounds (13C6 BaA 13C6 CHR13C6 BbFA and 13C4 BaP) containing 100 microgmiddotmLminus1 eachwere supplied by LGC Standards GmbH (Wesel Germany)and used as internal standards e stock solutions of theindividual 13C PAHs were prepared and combined intoluene All the standard solutions were stored at +4degCprotected from the light

23 Sample Preparation PAH4 compounds were deter-mined using a gas chromatography-tandem mass spec-trometry (GC-MSMS) method accredited according to ISO17025 e method was based on the publication of Veyrandet al [25] with minor modifications All samples werepacked in aluminium foil or in a sales package and stored at+5degC prior to homogenisation in a food mixer According tothe manufacturerrsquos information the fish or meat skin wasremoved before homogenisation if the skin was not an ediblepart of the product MLs are only applicable to the edibleparts of smoked productse samples were homogenised ina mixer and approximately 100 g of each sample was takento be freeze-dried Half a gram of the dried sample wasspiked with internal standards (13C-labelled PAHs) followedby the extraction of PAHs using ASE 200reg e extractioncells were filled with celite and florisil and prewashed withdichloromethane before adding the sample and extracting







68

11

11

5 3 2

PorkSalamiSausage


(a)

82

5

52 3 3



(b)








3 Results

















3175 3200 3225 3250 3275 3300 3325 3350 3375 3400 3425 3450 3475 3500

()

11



2479e + 004

(min)

()

11



7205e + 003

()

2




RT = 3262


RT = 3261


2560 gt 2541RT = 3261








(μg

kgndash1

)

30

25

20

15

10

5

0


BaP BaPPAH4 PAH4

lowast lowastlowast

lowast

lowast

lowast

(a)

(μg

kgndash1

)

25

20

15

10

5

0


BaPBaP PAH4 PAH4

lowastlowast

lowast

(b)

200

150

100

50

0


BaP BaPPAH4 PAH4

(μg

kgndash1

)

lowastlowast

lowast


(c)




30

25

20

15

10

5


Smoking time

(μg

kgndash1

)


lowast

(a)

lowast

30

25

20

15

10

5


Smoking time

(μg

kgndash1

) lowast

lowast

lowast

lowast

(b)


25

20

15

10

5





(μg

kgndash1

)

lowast

lowastlowast

(c)

Figure 4 Continued





4 Discussion


30

25

20

15

10

5

0

Fish size


lowastlowastlowast


(μg

kgndash1

)

(d)

200

150

100

50

0


BaP BaPPAH4 PAH4

(μg

kgndash1

)


lowast

lowast

(e)



25

20

15

10

5

0


BaPBaP PAH4 PAH4

(μg

kgndash1

)

lowast

lowastlowast

(a)

25

20

15

10

5

0


lowastlowastlowast

(μg

kgndash1

)

BaPBaP PAH4 PAH4

(b)

200

150

100

50

0


(μg

kgndash1

)

lowastlowastlowast

lowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(c)



30

25

20

15

10

5

0


(μg

kgndash1

)

lowastlowastlowast

lowast

lowastlowast

lowast

lowast

BaPBaP PAH4 PAH4

(a)

25

20

15

10

5

0


(μg

kgndash1

)

lowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(b)

200

150

100

50

0


(μg

kgndash1

)

lowastlowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(c)














5 Conclusions


Data Availability




Acknowledgments


References















































Journal of

Chemistry





Journal of

Volume 2018






Volume 2018




Journal of






Enzyme Research


Journal of



MaterialsJournal of






Na

nom

ate

ria

ls









68

11

11

5 3 2

PorkSalamiSausage


(a)

82

5

52 3 3



(b)








3 Results

















3175 3200 3225 3250 3275 3300 3325 3350 3375 3400 3425 3450 3475 3500

()

11



2479e + 004

(min)

()

11



7205e + 003

()

2




RT = 3262


RT = 3261


2560 gt 2541RT = 3261








(μg

kgndash1

)

30

25

20

15

10

5

0


BaP BaPPAH4 PAH4

lowast lowastlowast

lowast

lowast

lowast

(a)

(μg

kgndash1

)

25

20

15

10

5

0


BaPBaP PAH4 PAH4

lowastlowast

lowast

(b)

200

150

100

50

0


BaP BaPPAH4 PAH4

(μg

kgndash1

)

lowastlowast

lowast


(c)




30

25

20

15

10

5


Smoking time

(μg

kgndash1

)


lowast

(a)

lowast

30

25

20

15

10

5


Smoking time

(μg

kgndash1

) lowast

lowast

lowast

lowast

(b)


25

20

15

10

5





(μg

kgndash1

)

lowast

lowastlowast

(c)

Figure 4 Continued





4 Discussion


30

25

20

15

10

5

0

Fish size


lowastlowastlowast


(μg

kgndash1

)

(d)

200

150

100

50

0


BaP BaPPAH4 PAH4

(μg

kgndash1

)


lowast

lowast

(e)



25

20

15

10

5

0


BaPBaP PAH4 PAH4

(μg

kgndash1

)

lowast

lowastlowast

(a)

25

20

15

10

5

0


lowastlowastlowast

(μg

kgndash1

)

BaPBaP PAH4 PAH4

(b)

200

150

100

50

0


(μg

kgndash1

)

lowastlowastlowast

lowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(c)



30

25

20

15

10

5

0


(μg

kgndash1

)

lowastlowastlowast

lowast

lowastlowast

lowast

lowast

BaPBaP PAH4 PAH4

(a)

25

20

15

10

5

0


(μg

kgndash1

)

lowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(b)

200

150

100

50

0


(μg

kgndash1

)

lowastlowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(c)














5 Conclusions


Data Availability




Acknowledgments


References















































Journal of

Chemistry





Journal of

Volume 2018






Volume 2018




Journal of






Enzyme Research


Journal of



MaterialsJournal of






Na

nom

ate

ria

ls







3 Results

















3175 3200 3225 3250 3275 3300 3325 3350 3375 3400 3425 3450 3475 3500

()

11



2479e + 004

(min)

()

11



7205e + 003

()

2




RT = 3262


RT = 3261


2560 gt 2541RT = 3261








(μg

kgndash1

)

30

25

20

15

10

5

0


BaP BaPPAH4 PAH4

lowast lowastlowast

lowast

lowast

lowast

(a)

(μg

kgndash1

)

25

20

15

10

5

0


BaPBaP PAH4 PAH4

lowastlowast

lowast

(b)

200

150

100

50

0


BaP BaPPAH4 PAH4

(μg

kgndash1

)

lowastlowast

lowast


(c)




30

25

20

15

10

5


Smoking time

(μg

kgndash1

)


lowast

(a)

lowast

30

25

20

15

10

5


Smoking time

(μg

kgndash1

) lowast

lowast

lowast

lowast

(b)


25

20

15

10

5





(μg

kgndash1

)

lowast

lowastlowast

(c)

Figure 4 Continued





4 Discussion


30

25

20

15

10

5

0

Fish size


lowastlowastlowast


(μg

kgndash1

)

(d)

200

150

100

50

0


BaP BaPPAH4 PAH4

(μg

kgndash1

)


lowast

lowast

(e)



25

20

15

10

5

0


BaPBaP PAH4 PAH4

(μg

kgndash1

)

lowast

lowastlowast

(a)

25

20

15

10

5

0


lowastlowastlowast

(μg

kgndash1

)

BaPBaP PAH4 PAH4

(b)

200

150

100

50

0


(μg

kgndash1

)

lowastlowastlowast

lowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(c)



30

25

20

15

10

5

0


(μg

kgndash1

)

lowastlowastlowast

lowast

lowastlowast

lowast

lowast

BaPBaP PAH4 PAH4

(a)

25

20

15

10

5

0


(μg

kgndash1

)

lowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(b)

200

150

100

50

0


(μg

kgndash1

)

lowastlowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(c)














5 Conclusions


Data Availability




Acknowledgments


References















































Journal of

Chemistry





Journal of

Volume 2018






Volume 2018




Journal of






Enzyme Research


Journal of



MaterialsJournal of






Na

nom

ate

ria

ls








3175 3200 3225 3250 3275 3300 3325 3350 3375 3400 3425 3450 3475 3500

()

11



2479e + 004

(min)

()

11



7205e + 003

()

2




RT = 3262


RT = 3261


2560 gt 2541RT = 3261








(μg

kgndash1

)

30

25

20

15

10

5

0


BaP BaPPAH4 PAH4

lowast lowastlowast

lowast

lowast

lowast

(a)

(μg

kgndash1

)

25

20

15

10

5

0


BaPBaP PAH4 PAH4

lowastlowast

lowast

(b)

200

150

100

50

0


BaP BaPPAH4 PAH4

(μg

kgndash1

)

lowastlowast

lowast


(c)




30

25

20

15

10

5


Smoking time

(μg

kgndash1

)


lowast

(a)

lowast

30

25

20

15

10

5


Smoking time

(μg

kgndash1

) lowast

lowast

lowast

lowast

(b)


25

20

15

10

5





(μg

kgndash1

)

lowast

lowastlowast

(c)

Figure 4 Continued





4 Discussion


30

25

20

15

10

5

0

Fish size


lowastlowastlowast


(μg

kgndash1

)

(d)

200

150

100

50

0


BaP BaPPAH4 PAH4

(μg

kgndash1

)


lowast

lowast

(e)



25

20

15

10

5

0


BaPBaP PAH4 PAH4

(μg

kgndash1

)

lowast

lowastlowast

(a)

25

20

15

10

5

0


lowastlowastlowast

(μg

kgndash1

)

BaPBaP PAH4 PAH4

(b)

200

150

100

50

0


(μg

kgndash1

)

lowastlowastlowast

lowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(c)



30

25

20

15

10

5

0


(μg

kgndash1

)

lowastlowastlowast

lowast

lowastlowast

lowast

lowast

BaPBaP PAH4 PAH4

(a)

25

20

15

10

5

0


(μg

kgndash1

)

lowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(b)

200

150

100

50

0


(μg

kgndash1

)

lowastlowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(c)














5 Conclusions


Data Availability




Acknowledgments


References















































Journal of

Chemistry





Journal of

Volume 2018






Volume 2018




Journal of






Enzyme Research


Journal of



MaterialsJournal of






Na

nom

ate

ria

ls





(μg

kgndash1

)

30

25

20

15

10

5

0


BaP BaPPAH4 PAH4

lowast lowastlowast

lowast

lowast

lowast

(a)

(μg

kgndash1

)

25

20

15

10

5

0


BaPBaP PAH4 PAH4

lowastlowast

lowast

(b)

200

150

100

50

0


BaP BaPPAH4 PAH4

(μg

kgndash1

)

lowastlowast

lowast


(c)




30

25

20

15

10

5


Smoking time

(μg

kgndash1

)


lowast

(a)

lowast

30

25

20

15

10

5


Smoking time

(μg

kgndash1

) lowast

lowast

lowast

lowast

(b)


25

20

15

10

5





(μg

kgndash1

)

lowast

lowastlowast

(c)

Figure 4 Continued





4 Discussion


30

25

20

15

10

5

0

Fish size


lowastlowastlowast


(μg

kgndash1

)

(d)

200

150

100

50

0


BaP BaPPAH4 PAH4

(μg

kgndash1

)


lowast

lowast

(e)



25

20

15

10

5

0


BaPBaP PAH4 PAH4

(μg

kgndash1

)

lowast

lowastlowast

(a)

25

20

15

10

5

0


lowastlowastlowast

(μg

kgndash1

)

BaPBaP PAH4 PAH4

(b)

200

150

100

50

0


(μg

kgndash1

)

lowastlowastlowast

lowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(c)



30

25

20

15

10

5

0


(μg

kgndash1

)

lowastlowastlowast

lowast

lowastlowast

lowast

lowast

BaPBaP PAH4 PAH4

(a)

25

20

15

10

5

0


(μg

kgndash1

)

lowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(b)

200

150

100

50

0


(μg

kgndash1

)

lowastlowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(c)














5 Conclusions


Data Availability




Acknowledgments


References















































Journal of

Chemistry





Journal of

Volume 2018






Volume 2018




Journal of






Enzyme Research


Journal of



MaterialsJournal of






Na

nom

ate

ria

ls





30

25

20

15

10

5


Smoking time

(μg

kgndash1

)


lowast

(a)

lowast

30

25

20

15

10

5


Smoking time

(μg

kgndash1

) lowast

lowast

lowast

lowast

(b)


25

20

15

10

5





(μg

kgndash1

)

lowast

lowastlowast

(c)

Figure 4 Continued





4 Discussion


30

25

20

15

10

5

0

Fish size


lowastlowastlowast


(μg

kgndash1

)

(d)

200

150

100

50

0


BaP BaPPAH4 PAH4

(μg

kgndash1

)


lowast

lowast

(e)



25

20

15

10

5

0


BaPBaP PAH4 PAH4

(μg

kgndash1

)

lowast

lowastlowast

(a)

25

20

15

10

5

0


lowastlowastlowast

(μg

kgndash1

)

BaPBaP PAH4 PAH4

(b)

200

150

100

50

0


(μg

kgndash1

)

lowastlowastlowast

lowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(c)



30

25

20

15

10

5

0


(μg

kgndash1

)

lowastlowastlowast

lowast

lowastlowast

lowast

lowast

BaPBaP PAH4 PAH4

(a)

25

20

15

10

5

0


(μg

kgndash1

)

lowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(b)

200

150

100

50

0


(μg

kgndash1

)

lowastlowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(c)














5 Conclusions


Data Availability




Acknowledgments


References















































Journal of

Chemistry





Journal of

Volume 2018






Volume 2018




Journal of






Enzyme Research


Journal of



MaterialsJournal of






Na

nom

ate

ria

ls







4 Discussion


30

25

20

15

10

5

0

Fish size


lowastlowastlowast


(μg

kgndash1

)

(d)

200

150

100

50

0


BaP BaPPAH4 PAH4

(μg

kgndash1

)


lowast

lowast

(e)



25

20

15

10

5

0


BaPBaP PAH4 PAH4

(μg

kgndash1

)

lowast

lowastlowast

(a)

25

20

15

10

5

0


lowastlowastlowast

(μg

kgndash1

)

BaPBaP PAH4 PAH4

(b)

200

150

100

50

0


(μg

kgndash1

)

lowastlowastlowast

lowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(c)



30

25

20

15

10

5

0


(μg

kgndash1

)

lowastlowastlowast

lowast

lowastlowast

lowast

lowast

BaPBaP PAH4 PAH4

(a)

25

20

15

10

5

0


(μg

kgndash1

)

lowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(b)

200

150

100

50

0


(μg

kgndash1

)

lowastlowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(c)














5 Conclusions


Data Availability




Acknowledgments


References















































Journal of

Chemistry





Journal of

Volume 2018






Volume 2018




Journal of






Enzyme Research


Journal of



MaterialsJournal of






Na

nom

ate

ria

ls




25

20

15

10

5

0


BaPBaP PAH4 PAH4

(μg

kgndash1

)

lowast

lowastlowast

(a)

25

20

15

10

5

0


lowastlowastlowast

(μg

kgndash1

)

BaPBaP PAH4 PAH4

(b)

200

150

100

50

0


(μg

kgndash1

)

lowastlowastlowast

lowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(c)



30

25

20

15

10

5

0


(μg

kgndash1

)

lowastlowastlowast

lowast

lowastlowast

lowast

lowast

BaPBaP PAH4 PAH4

(a)

25

20

15

10

5

0


(μg

kgndash1

)

lowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(b)

200

150

100

50

0


(μg

kgndash1

)

lowastlowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(c)














5 Conclusions


Data Availability




Acknowledgments


References















































Journal of

Chemistry





Journal of

Volume 2018






Volume 2018




Journal of






Enzyme Research


Journal of



MaterialsJournal of






Na

nom

ate

ria

ls




30

25

20

15

10

5

0


(μg

kgndash1

)

lowastlowastlowast

lowast

lowastlowast

lowast

lowast

BaPBaP PAH4 PAH4

(a)

25

20

15

10

5

0


(μg

kgndash1

)

lowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(b)

200

150

100

50

0


(μg

kgndash1

)

lowastlowast

lowast

lowast

lowast

BaPBaP PAH4 PAH4

(c)














5 Conclusions


Data Availability




Acknowledgments


References















































Journal of

Chemistry





Journal of

Volume 2018






Volume 2018




Journal of






Enzyme Research


Journal of



MaterialsJournal of






Na

nom

ate

ria

ls















5 Conclusions


Data Availability




Acknowledgments


References















































Journal of

Chemistry





Journal of

Volume 2018






Volume 2018




Journal of






Enzyme Research


Journal of



MaterialsJournal of






Na

nom

ate

ria

ls







5 Conclusions


Data Availability




Acknowledgments


References















































Journal of

Chemistry





Journal of

Volume 2018






Volume 2018




Journal of






Enzyme Research


Journal of



MaterialsJournal of






Na

nom

ate

ria

ls








































Journal of

Chemistry





Journal of

Volume 2018






Volume 2018




Journal of






Enzyme Research


Journal of



MaterialsJournal of






Na

nom

ate

ria

ls












Journal of

Chemistry





Journal of

Volume 2018






Volume 2018




Journal of






Enzyme Research


Journal of



MaterialsJournal of






Na

nom

ate

ria

ls









Journal of

Chemistry





Journal of

Volume 2018






Volume 2018




Journal of






Enzyme Research


Journal of



MaterialsJournal of






Na

nom

ate

ria

ls




Documents

Critical Effects of Smoking Parameters on the Levels of