Food Chemistry 122 (2010) 920–925

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Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Analytical Methods

Optimisation and validation of an HPLC method for determination

of polycyclic aromatic hydrocarbons (PAHs) in mussels
Francesco Paolo Serpe *, Mauro Esposito, Pasquale Gallo, Luigi Serpe
Istituto Zooprofilattico Sperimentale del Mezzogiorno, Via Salute, 2, 80055 Portici (Napoli), Italy

a r t i c l e i n f o a b s t r a c t

Article history: A method for determination of 11 polycyclic aromatic hydrocarbons (PAHs) in mussels (Mytilus gallopro-
Received 8 September 2009 vincialis), using HPLC coupled to a fluorescence detector, has been optimised and validated, according to
Received in revised form 15 March 2010 European Community rules. Sample preparation involves alkaline digestion of mussel tissue, liquid–
Accepted 16 March 2010
liquid extraction of organic compounds and solid phase clean-up. Accuracy and precision of the method
were determined by a validation study, carried out to demonstrate that the method is useful for both
screening purposes and confirmation. Commission Regulation (EC) No. 2007/333/EC stated the perfor-
Keywords:
mance criteria for the analysis of the only benzo[a]pyrene (BaP) in food products of animal origin, since
Polycyclic aromatic hydrocarbons
Mussels
BaP is the most studied PAH. We extended the BaP analysis performance criteria to other 10 toxic PAHs
HPLC (listed in Commission Recommendation (EC) No. 2005/108/EC) and the validation study was performed
also in agreement with Commission Decision (EC) No. 2002/657/EC. The method was applied to investi-
gate 27 mussel samples from actively producing shellfish plants located in Campania (Italy) and variable
levels of PAHs were detected ranging from <0.2 to 16 lg/kg wet weight.
! 2010 Elsevier Ltd. All rights reserved.

1. Introduction
Polycyclic aromatic hydrocarbons (PAHs) are persistent chemi-
cals with two or more fused aromatic rings. PAHs are ubiquitous
environmental pollutants that have been identified worldwide in
various matrices, such as water, soil or dust particles. PAHs have
been also found as contaminants in various foods, such as dairy
products, vegetables, fruits, oils, and roasted and smoked meat
(Camargo & Toledo, 2003; De Vos, Van Dokkum, Schouten, &
Jong-Berkhout, 1990; Kazerouni, Sinha, Hsu, Greenberg, & Rothman,
2001; Kruijf, Schouten, & Van Der Stegen 1987; Lin, Tu, & Zhu,
2005; Simko 2002).
The movement of PAHs in the environment depends on their
properties; they easily evaporate and disperse into the air and de-
posit in soil, and accumulate in sediments and biota because of
their lipophilic properties. For the most part, PAHs enter the envi-
ronment, and then food chain, through the release from forest fires,
residential wood burning or from geochemical processes, such as
volcanism or petroleum seepages. Moreover, they can enter water-
courses through industrial discharges and waste water treatment
plants and can be found in soils as a result of their escape from
storage containers close to hazardous waste sites (WHO, 1998,
2005). PAHs can also contaminate foods during heating and drying
* Corresponding author.
E-mail address: francesco.serpe@izsmportici.it (F.P. Serpe).
processes that allow combustion products to come into direct con-
tact with food.
The International Agency for Research on Cancer (IARC, 2004)
and the Scientific Committee on Food (Regulation 2006/1881/EC)
concluded that a number of PAHs are genotoxic carcinogens.
Benzo[a]anthracene, benzo[a]pyrene (BaP) and dibenzo[a,h]
anthracene were classified by IARC as probably carcinogenic to hu-
mans (group 2A) while 5-methylchrysene, benzo[b]fluoranthene,
benzo[k]fluoranthene, dibenzo[a,i]pyrene and indeno[1,2,3-cd]pyr-
ene are possibly carcinogenic to humans (group 2B). However, those
PAHs that have not been demonstrated to be carcinogenic, such as
chrysene (IARC group 3), may act as synergists (Hermann, 1981).
In order to protect public health, the EU legislation has fixed maxi-
mum levels for benzo[a]pyrene in certain foods containing fats
and oils, and in smoked and dried foods; smoking and drying pro-
cesses might cause high levels of contamination. According to the
Scientific Committee on Food, benzo[a]pyrene can be used as a mar-
ker for the occurrence and effect of carcinogenic PAH in food, but
Commission Recommendation (EC) No. 2005/108/EC required anal-
ysis of multiple PAHs (15 EU priority PAHs) to get further informa-
tion on levels of contamination from different PAHs in food.
The present paper focused on 11 PAHs included in EC
Recommendation 108/2005 – benzo[a]anthracene (BaA), chrysene
(CHR), 5-methylchrysene (5MC), benzo[b]fluoranthene (BbF),
benzo[k]fluoranthene (BkF), benzo[a]pyrene (BaP), dibenzo[a,l]-
pyrene (DlP), dibenzo[a,h]anthracene (DhA), indeno[1,2,3-cd]pyre-
nen (IcP), dibenzo[a,i]pyrene (DiP), dibenzo[a,h]pyrene (DhP) – in

0308-8146/$ - see front matter ! 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2010.03.062
 F.P. Serpe et al. / Food Chemistry 122 (2010) 920–925
27 mussel samples. Environmental PAH pollution may cause con-
tamination of food, in particular fish and seafood. Marine organ-
isms can accumulate PAHs and can be a risk source for
consumers but at the same time they are a useful indicator for
assessing the eco-health of aquatic environments (Bihari, Fafandel,
& Piskur, 2007). In Campania, mussels breeding is an important
economic resource and many productive mussel-farming areas
are located along the coast. Unfortunately some of those overlook-
ing urban areas are the sources of PAHs environmental contamina-
tion. A study conducted on surface sediment samples collected
from Naples harbour (Campania) provided some data on levels of
contamination by PAHs (Sprovieri et al., 2007). The occurrence of
PAHs in seafood has been demonstrated by many researchers but
few results are available for farmed seafood in Italy (Perugini
et al., 2007; Storelli & Marcotrigiano, 2001).
The European Parliament and Council established in Regulation
(EC) No. 2004/854/EC that Member States shall ensure that live bi-
valve molluscs undergo to official controls as described in Annex II.
The relevant authority must classify production areas from which
it authorises the harvesting of live bivalve molluscs. Classified
relaying and production areas must be periodically monitored to
check for the presence of chemical contaminants in live bivalve
molluscs. Moreover, the Commission Regulation 2006/1881/EC
has fixed maximum levels for benzo[a]pyrene in various food prod-
ucts of marine origin, such as bivalves, crustaceans, fish and ceph-
alopods. Thus, it was necessary to develop an analytical method to
determine PAH levels in these seafoods. Moreover, the majority of
scientific papers concerning PAHs in food matrices have focused
mainly on BaP. A considerable number tracked the set of 16 US
EPA PAHs, but only a small number studied the 15 EU priority PAHs
(Wenzl, Simon, Kleiner, & Anklam, 2006).
Therefore, the aim of the present study was to develop and val-
idate a sensitive and reliable method for the determination of PAHs
in mussel samples. This method was tested by determining the
presence of 11 PAHs in samples of Mytilus galloprovincialis
collected from Campania (Italy).
2. Material and methods
2.1. Mussel samples
Samples of bivalves (M. galloprovincialis) were collected from
commercial shellfish farms located in different areas of the Campa-
nia coast (Italy), in particular from Domitio Coast, Pozzuoli Gulf, Na-
poli Gulf and Salerno Gulf between January 2008 and March 2009.
All samples were from farms in which mussels are harvested
before commercialisation, in natural beds or in long-line plants.
Samples were kept at !20 "C until analysis.
2.2. Standard preparation
PAHs standard solutions at 10 lg/ml were purchased from
Dr. Ehrenstorfer (Ausburg, Germany) which contained:
benzo[a]anthracene, chrysene, 5-methylchrysene, benzo[b]fluo-
ranthene, benzo[k]fluoranthene, benzo[a]pyrene, dibenzo[a,l]
pyrene, dibenzo[a,h]anthracene, indeno[1,2,3-cd]pyrene, diben-
zo[a,i]pyrene, dibenzo[a,h]pyrene. A standard mix solution of 11
PAHs at 0.1 lg/ml (100 lg/kg) for each PAH was prepared from
the 10 lg/ml standard solutions by dilution 1:100 with acetonitrile.
2.3. Sample preparation
2.3.1. Reagents and materials
Cyclohexane, acetonitrile (ACN, HPLC grade), potassium
hydroxide, ethanol and sodium sulphate anhydrous were pur-
921
chased from Carlo Erba (Milan, Italy) and ultrapure water was ob-
tained from a Milli-Q system (Millipore Corp., Bedford, MA). Sep-
Pak (500 mg/3 ml) Vac Silica cartridges were purchased from
Waters (Milford, MA).
2.3.2. Procedure
The mussel tissue (2 g) was homogenised (Ultra-Turrax dis-
perser T25, Bicasa, Bernareggio, Italy) and saponified with 10 ml
of a solution of potassium hydroxide (2 N in ethanol) in a water
bath at 80 "C for 2 h. The digest was cooled at room temperature
and 10 ml of ultrapure water were added, extracted with 20 ml
of cyclohexane and then centrifuged at 3000 rcf for 5 min at 4 "C
(Megafuge, Heraeus). The extraction was carried out three times.
The supernatants were combined (about 60 ml), filtered with a fil-
ter paper (Millipore) containing anhydrous sodium sulphate and
reduced to small volume (about 0.2 ml) using a Rotovapor at
40 "C (Büchi, Flavil, Switzerland). The residue was dissolved in
3 ml ACN, applied on a Sep-Pak cartridge pre-activated with 3 ml
ACN and eluted with 3 ml ACN. Eluate was dried under nitrogen
flow in a thermoblock at 50 "C; the residue was dissolved in 1 ml
ACN, 0.45 lm filtered (nylon syringe on-line filter, Millipore) and
analysed by HPLC.
2.4. HPLC analysis
A protocol for analysis of 11 PAHs was developed and opti-
mised, using a Hitachi-Merck HPLC apparatus equipped with a
pump (L7100), autosampler (L7250) and fluorescence detector
(L7480). Different excitation and emission wavelengths were used
for the detection of different PAHs. The ideal excitation–emission
couple for each PAH was measured by a fluorimeter (Agilent
1200 Series) and a fluorescence detector (FLD) timetable was built
up: at t0 kexcitation was 294 nm and kemission was 404; at t = 15.5 min,
270 and 420; at t = 18.8 min, 294 and 404; at t = 25.4 min, 240 and
450, until the end of the run. The HPLC column was a
2.1 mm " 50 mm " 1.8 lm C18 column (Rapid Resolution HT, Agi-
lent, Santa Clara, CA) and the mobile phase was acetonitrile/water
in gradient mode at a flow rate of 0.4 ml/min with the column oven
set at 25 "C. The gradient elution programme started with an initial
mobile phase at 60:40 (v/v) ACN:water, changing linearly to 100%
ACN in 14 min and, after 5 min, changing back to initial phase
(60:40). Total run time of one analysis was 35 min. The injection
volume was 20 ll. Sample peaks identification was based on stan-
dard peaks’ retention time. The HPLC elution profile of a mix of 11
PAHs and of a mussel sample spiked with a mix of 11 PAHs are re-
ported in Figs. 1 and 2, respectively.
2.5. Quantification
External standard method was used to determine PAH concen-
tration in the samples. Linearity of method was checked by triple
injection of an 11 PAH standard solution ranging from 0.5 lg/kg
to 15 lg/kg. Linear regression was applied to construct a calibra-
tion curve reporting peak area vs PAH concentration. A calibration
curve was made for every sequence of analysis.
2.6. Method validation according to Regulation 2007/333/EC
The present method was optimised and validated according to
the performance criteria required by EC for the analysis of ben-
zo[a]pyrene and described in Commission Regulation 2007/333/
EC (Table 1). These criteria were extended to all 11 PAHs studied.
Limit of detection (LOD) and limit of quantification (LOQ) were cal-
culated for each PAH, using PAH-free mussels spiked with the stan-
dard mix of 11 PAHs at 100 lg/kg to obtain six samples with PAH
concentration of 0.2 lg/kg and six with PAH concentration of
922 F.P. Serpe et al. / Food Chemistry 122 (2010) 920–925

Fig. 1. A 10 lg/kg PAH standard mixture.

Fig. 2. A PAH-free mussel sample spiked with 10 lg/kg PAHs standard mixture.

Table 1
EC); blank mussel tissue was spiked with the standard mix at
Performance criteria required for analysis of benzo[a]pyrene (Commission Regulation 100 lg/kg to obtain samples at concentrations of 5.0, 10.0 and
2007/333/EC). 15.0 lg/kg in six replicates per level for repeatability and six repli-
Applicability Foods specified in regulation 2006/1881/EC
cates plus another six analysed by a different technician, instru-
ment and on a different day for reproducibility. The mean
LOD Less than 0.3 lg/kg
recovery for all PAHs for the assessment of accuracy was then cal-
LOQ Less than 0.9 lg/kg
Precision rHorrat Less than 2 for all culated. Specificity was evaluated analysing 20 PAHs in blank mus-
Precision RHorrat Less than 2 for all sel samples and checking for any interferences in the region of
Recovery 50–120% for all interest where single PAHs were expected to elute.
Specificity Free from matrix or spectral interferences for all (*)
*
Resulted from 20 injections of a negative sample.
2.7. Method validation according to Decision 2002/657/EC

0.8 lg/kg and analysing them to evaluate the repeatability. Preci- For those contaminants for which exists a maximum limit (LM)
sion of the method was assessed through calculation of Horrat permitted (group B, 96/23/EC), in this case only BaP, the CCa con-
coefficient (2007/333/EC) for each PAH at three concentrations, centration (where a error is the probability of a false compliant re-
in repeatability (r) and reproducibility (R) conditions (2007/333/ sponse) can be calculated by a number of replicates of blank
 F.P. Serpe et al. / Food Chemistry 122 (2010) 920–925 923

Table 2
Repeatability inter-die and reproducibility intra-die. r% means the recovery percentage.

ppb BaA CHR 5MC BbF BkF BaP DlP DhA IcP DiP DhP
Repeatability intra-die (r, n = 6)
5 s 0.4 0.4 0.5 0.3 0.2 0.1 0.3 0.4 0.7 0.2 0.3
RSD 10 12 17 7 6 5 8 12 8 7 8
r% 74 66 54 72 66 62 75 73 166 72 68
10 s 0.2 0.4 0.9 0.3 0.3 0.3 0.3 0.3 0.5 0.7 0.8
RSD 3 7 8 4 4 4 5 5 7 11 12
r% 67 63 107 66 67 69 63 64 65 64 69
15 s 3.8 3.6 1 0.8 0.6 0.6 0.8 1.2 1.3 1 1
RSD 36 21 10 8 6 6 8 12 14 10 12
r% 69 113 68 69 64 61 73 65 65 62 63
Reproducibility inter-die (R, n = 12)
5 s 0.7 1 0.7 0.7 0.6 0.5 0.7 0.8 2.4 0.7 0.7
RSD 17 24 25 18 19 16 21 26 39 21 23
r% 86 82 52 78 66 66 69 64 122 64 63
10 s 0.6 0.6 1.7 1.2 0.5 0.5 0.4 0.6 0.9 1 1
RSD 9 10 16 17 8 7 6 10 14 18 20
r% 70 62 103 69 65 66 63 62 66 57 57
15 s 2.9 5.6 1.6 1.1 1.1 0.8 1.6 1.6 2.0 2.0 2.0
RSD 33 43 18 11 12 9 16 18 25 26 25
r% 58 88 60 67 59 58 64 59 55 52 53

samples spiked at 10 lg/kg (LM for BaP). The value of CCa is equal
to the mean value of concentration plus 1.64 " standard deviation
and represents the decision limit at which the probability of a false
negative is 5%. Concentration CCb can be calculated in a similar
way. Error b is the probability of a false non-compliant response
and the CCb value is equal to CCa concentration plus 1.64 " ((aver-
age CV% " CCa)/100), where CV% is the variation coefficient in
reproducibility conditions, thus CCb represents the detection capa-
bility at which the probability of a false positive is 5%. Finally, the
ruggedness of the method was evaluated for each PAH with the
ruggedness minor changes approach: Rotavapor bath was utilised
at 35 "C and 55 "C, two different ACN lots and two different HPLC
column lots were used, 1 g and 2 g of anhydrous Na2SO4 were
tested, 40 "C and 60 "C as temperatures of evaporation and 50 ml
and 70 ml of cyclohexane as volumes of extraction were tested.
2.8. Uncertainty of measurement
Measurement uncertainty was calculated by identifying and
quantifying the uncertainty components of the analytical process,
in particular the relative expanded uncertainties (Uc) were calcu-
lated for each PAH (UNI CEI ENV 13005:2000).
3. Results and discussion
3.1. Sample analysis
creating an FLD timetable. This was possible because of the use
of column oven and a good retention time conservation that al-
lowed us also to use of the simple method of external standardiza-
tion for quantification.
3.2. Method validation
Minimum performance criteria for the analysis of benzo[a]pyr-
ene described in Commission Regulation 2007/333/EC (Table 1),
should be applied to the matrices, including mussels, listed in
Commission Regulation 2006/1881/EC. These criteria were ex-
tended to all 11 PAHs: limit of detection (LOD) and limit of quan-
titation (LOQ) values were 0.2 lg/kg and 0.8 lg/kg, respectively,
for eight PAHs, according to Commission Regulation 2007/333/
EC. For CHR, BbF and IcP, LOD and LOQ were higher than the other
PAHs (0.8 lg/kg and 5 lg/kg, respectively; Table 3). Precision Hor-
rat r (repeatability) ranged from 0.10 to 1.2 for all PAHs and Horrat
R (reproducibility) from 0.19 to 1.9 (<2 as requested), calculated at
5.0, 10.0 and 15.0 lg/kg, and the mean recovery for the 11 PAHs at
concentrations of 5.0, 10.0 and 15.0 lg/kg was in the range of 51–
72%, in agreement with 2007/333/EC. The results of LOD, LOQ, Hor-
rat r and Horrat R (at 10.0 lg/kg) for the 11 PAHs are in Table 3.
Moreover, intralaboratory coefficient of variation (CV) under
repeatability conditions for benzo[a]pyrene was between 4.1%
and 20.0% (Table 2), thus the results of calculation of accuracy of
the method were also in agreement with Commission Decision
(EC) No. 2002/657/EC, in which it is reported that CVs, for

Preparation of samples for chromatographic analyses generally

involves two steps: extraction and clean-up. Complexes matrices, Table 3
LOD, LOQ, rHorrat and RHorrat of 11 PAHs.
such as foods, need additional steps. In the case of mussels a useful
additional step involved an alkaline saponification to improve the PAH LOQ (lg/kg) LOD (lg/kg) r
Horrat (10 lg/kg) R
Horrat (10 lg/kg)
yield of extraction (Dunn, 1976). Sample preparation method was BaA 0.8 0.2 0.22 0.28
tested and optimised as described and a study on ruggedness of CHR 5.0 0.8 0.32 0.32
method was carried out (included in validation study). Determina- 5MC 0.8 0.2 0.26 0.51
tion by HPLC/FLD is the most common type of analysis for PAHs BbF 5.0 0.8 0.44 0.53
BkF 0.8 0.2 0.23 0.24
(Simon, Palme, & Anklam, 2007; Wenzl et al., 2006). One of the tar- BaP 0.8 0.2 0.20 0.23
gets of the present work was to improve the sensitivity of the HPLC DlP 0.8 0.2 0.18 0.19
determination, to detect the maximum number of PAHs in a single DhA 0.8 0.2 0.30 0.32
run and to maximise recovery/minimise the interferences. This IcP 5.0 0.8 0.40 0.43
DiP 0.8 0.2 0.48 0.56
was achieved by creating an HPLC method in which the coupled
DhP 0.8 0.2 0.44 0.61
excitation/fluorescence emission was optimal for each analyte, by
924 F.P. Serpe et al. / Food Chemistry 122 (2010) 920–925

concentrations lower than 100 lg/kg, should be as low as possible.
As regards the specificity of the method, the matrix was free from
interferences across the range of retention times of analytes. The
relative expanded uncertainty (Uc), expressed as percent value,
was 1.14% for BaP, thus appropriate for residues quantification in
the range of concentrations considered in the present study. The
majority of experiments carried out for application of 2007/333/
EC covers the application of 2002/657/EC (in particular regarding
accuracy, precision and specificity). The validation study was com-
pleted with some prescriptions reported in 2002/657/EC. For or-
ganic contaminants like PAHs (included in 96/23/EC, Annex I,
group B), having a native fluorescence, a HPLC/FLD method is con-
sidered as confirmatory quantitative method (2002/657/EC) and
some additional parameters are required, in particular the calcula-
tion of decision limit (CCa), detection capability (CCb) and rugged-
ness of the method (2002/657/EC). CCa and CCb concentrations for
BaP, calculated at LM, were 7.72 lg/kg and 9.39 lg/kg, respec-
tively, according to 2002/657/CE. The method was not affected
by slight variations of some critical factors (ruggedness minor
changes), in particular as regard repeatability and reproducibility.
Finally, all response curves from analysing standard solutions
exhibited a linear fit from 0.5 to 15 lg/kg and the determination
coefficients (R2) were higher than 0.995 for all PAHs.
3.3. Level of PAHs in mussel samples
The concentrations of 11 PAHs in 27 samples of mussel taken
from different locations in Campania, are reported in Table 4. The
concentration is expressed as lg/kg (wet weight) and all concen-
trations were corrected for recovery as Commission Regulation
2007/333/EC requires for benzo[a]pyrene. The concentrations be-
low the limit of detection were considered as not detected (ND).
The benzo[a]pyrene concentration ranged from 0.3 to 9.3 lg/kg,
with median values of 1.5 lg/kg, and it was under the established
limit of 10.0 lg/kg in all samples. Most of the other PAHs were
detected: benzo[a]anthracene was found in 18 samples (67%) with
a median value of 2.0 lg/kg, 5-methylchrysene in 19 (70%) with a
median of 2.2 lg/kg, benzo[b]fluoranthene in 23 (85%) with a med-
ian of 2.1 lg/kg, indeno[1,2,3-cd]pyrene in 19 (70%) and
dibenzo[a,h]anthracene in 20 (74%) with median values of 1.4
and 1.2 lg/kg, respectively, and benzo[k]fluoranthene in 27 sam-
ples (100%) with a median of 1.9 lg/kg (Table 4). Deviation stan-
dards were from 0.9 to 4.4 lg/kg (Table 4).
Therefore, PAH contamination in mussels originating from
Campania shellfish farms is under the maximum limit permitted
by European Legislation for benzo[a]pyrene in all samples. How-
ever, an interesting result of the present study was that levels of
benzo[a]pyrene were higher than other findings in mussels from
the Adriatic sea (Italy) that indicate values below the instrumental
detection limit in all samples, whereas, they show detectable levels
of others PAHs (Perugini et al., 2007). Levels of benzo[a]pyrene in
mussels from the Campanian Sea are also higher than the Ionian
Sea (Italy) (Storelli & Marcotrigiano, 2001).
4. Conclusion
Validation results of the HPLC/FLD method were in accordance
with performance criteria required by Commission Regulation
2007/333/EC and Commission Decision (EC) No. 2002/657/EC.
Therefore, this method is a confirmed quantitative method that
can be considered useful for routine analyses to monitor 11 PAHs
in mussel samples and to verify whether the concentration of ben-
zo[a]pyrene in mussels is under the maximum permitted limit. In
addition, this method is applicable to determination of environ-
mental pollution by PAHs, using mussels as pollution indicators.
EC assessed a maximum residue limit in bivalve molluscs for
only BaP, while it is desirable to determine a legal limit for the oth-
ers PAHs, in particular benzo[a]anthracene and dibenzo[a,h]antra-
cene (IARC group 2A, as benzo[a]pyrene). The intrinsic toxicity
could be quantifiable as the sum of PAHs below 10 lg/kg as estab-

Table 4
Concentration of PAHs in mussel samples from Campania (Italy) expressed as lg/kg (wet weight) and corrected for recovery, ND – not detected.

Samples location Sample BaA CHR 5MC BbF BkF BaP DlP DhA IcP DiP DhP
Bacoli 1 1.6 ND 14.8 8.6 5.7 9.3 4.3 ND 3.9 4.1 0.8
2 ND ND 6.4 4.3 1.4 1.5 ND 1.2 ND ND ND
3 0.9 0.6 1.3 1.1 3.9 3.7 ND 0.8 ND 0.9 ND
4 1.9 1.1 1.8 1.8 4.6 1.5 ND 1.0 0.3 ND ND
5 ND ND ND 4.1 1.7 0.5 ND 0.8 8.0 ND ND
6 ND ND 2.9 4.4 1.4 0.3 ND ND ND ND ND
7 2.3 ND ND 0.8 3.2 0.7 ND ND 5.0 ND ND
8 2.0 1.2 3.1 3.1 9.2 3.6 ND 2.2 1.8 0.6 ND
9 ND ND 5.2 2.8 1.2 0.4 ND 0.3 0.7 ND ND
10 2.2 0.8 2.5 0.8 2.3 0.8 ND 0.5 ND ND ND
11 7.0 8.4 4.0 8.8 4.6 6.4 2.6 4.3 1.8 1.8 ND
12 4.0 ND 5.0 4.8 1.6 1.2 1.1 ND 0.4 ND ND
Ercolano 1 1.2 ND ND ND 0.4 0.5 ND 2.0 ND ND ND
Giugliano 1 ND 6.0 ND ND 1.3 2.0 ND ND 6.3 2.6 2.0
Napoli 1 4.2 4.1 7.4 6.5 16.2 5.0 1.8 1.2 1.4 ND ND
2 2.3 0.7 2.2 ND 5.6 3.3 0.7 0.7 1.1 ND ND
3 ND ND ND ND 1.7 0.7 ND ND ND ND ND
4 ND ND ND 2.1 2.6 1.1 ND ND 5.5 ND ND
Pozzuoli 1 6.0 ND 3.0 2.7 2.3 2.6 1.9 3.0 1.2 ND ND
2 1.9 ND 2.9 2.0 1.3 1.4 1.5 1.4 5.4 ND 2.4
3 ND 12.3 2.9 1.9 1.8 2.7 1.9 2.4 9.4 2.9 3.5
Torre del Greco 1 2.4 ND 2.0 2.4 0.9 0.9 0.7 1.8 ND ND 1.1
2 2.0 ND ND 1.9 1.5 1.5 1.6 1.7 7.0 ND 2.6
3 2.4 3.8 4.5 2.2 1.9 2.0 1.5 1.3 1.3 3.7 ND
4 3.2 ND ND 2.1 2.1 1.9 1.5 1.3 1.4 12.6 3.0
5 2.3 ND ND 2.5 1.5 1.3 ND 1.8 ND 2.3 2.1
Battipaglia 1 ND ND ND 1.4 2.2 1.9 1.2 ND 3.7 17.1 2.2
Mean 2.1 1.7 2.1 2.5 3.0 1.8 0.8 1.2 2.7 2.0 0.8
Median 2.0 0.0 2.2 2.1 1.9 1.5 0.7 1.2 1.4 0.0 0.0
Standard dev. 1.9 3.3 2.1 2.1 3.4 1.5 0.9 1.1 2.9 4.4 1.2
 F.P. Serpe et al. / Food Chemistry 122 (2010) 920–925
lished for BaP and in agreement with that ‘‘precautionary princi-
ple” assessed by Regulation 2002/178/EC, in order to protect the
public health. Moreover, the constant presence of BaP and others
PAHs in Italian seas suggest the need to include the determination
of PAHs as an additional parameter for classification and for official
controls of the areas where mussels are harvested.
References
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