Food Chemistry 141 (2013) 1–9

Contents lists available at SciVerse ScienceDirect

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

Polycyclic aromatic hydrocarbons in the bakery chain

M. Ciecierska ⇑, M.W. Obiedziński
Division of Food Quality Evaluation, Department of Food Biotechnology, Microbiology and Food Evaluation, Faculty of Food Sciences, Warsaw University of Life Sciences,
Nowoursynowska 159, 02-787 Warsaw, Poland

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

Article history: The level of polycyclic aromatic hydrocarbons occurrence and the possibility of their formation in the
Received 23 September 2012 bakery chain, its raw materials and final products, were examined. Experimental bread baking, with dif-
Received in revised form 21 January 2013 ferent baking temperatures, was performed in the Warsaw bakery, using cyclothermic deck ovens. PAHs
Accepted 4 March 2013
determination was performed by high-pressure liquid chromatography with fluorescent and diode array
Available online 14 March 2013
detectors (HPLC–FLD/DAD) and confirmed by gas chromatography coupled with mass spectrometry (GC/
MS). Total content of 19 PAHs in the grain, flour and bran varied from 1.07 to 3.65 lg/kg and, in bread,
Keywords:
from 1.59 to 13.6 lg/kg depending on the part of bread and baking temperature. Based on the dough’s
PAHs
Bakery chain
contamination level and the influence of the baking temperature on the bread’s PAHs content, it was con-
HPLC-FLD/DAD firmed that PAHs are formed during baking. Considering the results of the average dietary exposure to
GC/MS PAHs and the MOE (Margin of Exposure) analysis, it could be concluded that analysed bread and cereal
Risk assessment products constitute little concern for consumer health.
Ó 2013 Elsevier Ltd. All rights reserved.

1. Introduction
Polycyclic aromatic hydrocarbons (PAHs) are a diverse and
ubiquitous class of chemical contaminants, present throughout
the environment (CCFAC, 2005; EFSA, 2008). The Scientific
Committee on Food of the European Union, in its opinion dated 4
December 2002, concluded that 15 heavy PAHs are genotoxic
carcinogens (SCF, 2002). Since 2005, in accordance with the Euro-
pean Commission Recommendation 2005/108/EC, further analyses
of benzo[a]pyrene and other genotoxic compounds from the 15
PAHs mentioned above are necessary in foodstuffs (Commission
of the European Communities, 2005). However, referring to the
EFSA opinion from 2008 and Commission Regulation (EU) No.
835/2011 of 19 August, 2011, benzo[a]pyrene is not a suitable mar-
ker for the occurrence of the other PAHs in food and therefore a
system of four specific PAHs (benzo[a]pyrene, benzo[a]anthracene,
benzo[b]fluoranthene and chrysene) would be the most suitable
indicators of PAHs in food (Commission of the European Commu-
nities, 2011a; EFSA, 2008). These changes have been introduced
in Commission Regulation (EU) No. 835/2011 of 19 August 2011
amending Regulation (EC) No. 1881/2006 in regard to maximum
levels for polycyclic aromatic hydrocarbons in foodstuffs (Commis-
sion of the European Communities, 2011a).
PAHs may be formed and released as a result of incomplete
combustion or pyrolysis of organic matter, during industrial pro-
cesses and other human activities (CCFAC, 2005; Chang, Fang,
⇑ Corresponding author. Tel.: +48 22 593 76 86; fax: +48 22 593 76 82.
E-mail address: marta_ciecierska@sggw.pl (M. Ciecierska).
Chen, & Wu, 2006; SCF, 2002). Many researchers have detected
their occurrence in air, water, soil and sediments (Amoako, Ansa-
Asare, Karikari, & Dartey, 2011; Chang et al., 2006; Zhang, Song,
& Yuan, 2009) and therefore also in food (Camargo, Antoniolli, &
Vicente, 2012; Ciecierska & Obiedziński, 2013; White, Fernandes,
& Rose, 2008). Occurrence of PAHs in food can be a consequence
of environmental deposition, but also the thermal treatment pro-
cesses used in the preparation and manufacture of foods. Process-
ing procedures, such as smoking, roasting, grilling, baking and
frying, are recognised as a major cause of a potentially high level
of food contamination (Alomirah et al., 2011; Houessou, Goujot,
Heyd, & Camel, 2008; Orecchio & Papuzza, 2009; Rey-Salgueiro,
García-Falcón, Martínez-Carballo, & Simal-Gándara, 2008).
According to the studies on PAHs exposure, food is the main
source of human exposure to PAHs, and cereals constitute one of
the major contributing sources (Ibanez et al., 2005; Martí-Cid,
Llobet, Castell, & Domingo, 2008; Orecchio & Papuzza, 2009). In Po-
land, bread is a major component of people’s diet and the average
monthly per capita consumption of bread and cereals is 7.01 kg, of
which bread is 4.67 kg, that is 56 kg per year (Central Statistical Of-
fice, 2011). Bread’s contamination by PAHs can be dependent on
both the contamination of bakery raw materials, primarily flour,
and the baking process. An important issue is also the temperature
of this thermal treatment and its influence on bread’s contamina-
tion level.
With the background provided, the essential aim of this
research was to determine the level of PAHs occurrence and the
possibility of their formation in the bakery chain, its raw materials
and final products. These include the grain of wheat and rye, three

0308-8146/$ - see front matter Ó 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.foodchem.2013.03.006
2 M. Ciecierska, M.W. Obiedziński / Food Chemistry 141 (2013) 1–9
types of flour derived from these grains and their bran, as well as
selected kinds of bread and their doughs obtained from the
above-mentioned raw materials. The scope of the study included
experimental bread baking with different baking temperatures
applied. Moreover, different parts of loaves were analysed in order
to assess the level of their contamination by PAHs. From the group
of 19 PAHs compounds, including 4 light PAHs from the US EPA list
(phenanthrene, anthracene, fluoranthene, pyrene) and 15 heavy
PAHs from the SCF list, were determined by high-pressure liquid
chromatography with fluorescent and diode array detectors. In
our studies and Polish monitoring studies, concerning food con-
tamination by 16 EPA PAHs, it was noted that these 4 light PAHs
were always predominant in the quality profiles of PAHs (Cie-
cierska & Obiedziński, 2010, 2013; Jankowski & Obiedziński,
1998). Therefore, an important issue of this research was also to
define the percentage distribution of the above-mentioned light
and heavy PAHs in the contamination profile of analysed products.
Moreover, the objective of this study was also an attempt to esti-
mate the level of adult exposure to PAHs from bread and cereal
products under investigation.
2. Materials and methods
2.1. Samples
2.1.1. General
The materials investigated were the grain of wheat (winter
wheat, Cobra cultivar) and rye (winter rye, Dańkowskie Nowe cul-
tivar) obtained from a farm, where they were harvested (Miastków
Kościelny, a village located in Garwolin County, Masovian Voivode-
ship, Poland), three types of flour (wheat flour type 850, rye flour
type 720, wholemeal rye flour type 2000) derived exactly from
these grains and their bran (wheat bran, rye bran and bran from
rye grinding to wholemeal rye flour). Furthermore, three selected
kinds of bread (wheat-rye bread – Baltonowski bread, rye bread,
wholemeal rye bread) and their doughs, obtained from the
above-mentioned raw materials, were investigated. Moreover, in
the case of bread, different parts of loaves (crumb and crust) and
the whole of loaves were analysed in order to assess the level of
their contamination by PAHs. Nine samples of every product (three
samples – each one from three different batches) were used for the
study.
2.1.2. Bread baking
Apart from PAHs determination in the above-mentioned raw
materials and final products of the bakery chain, the scope of the
study also included experimental bread baking from these raw
materials, which was performed in the Warsaw bakery, using
cyclothermic deck ovens (heated by fuel oil).
In order to assess whether the higher baking temperature led to
a higher level of bread contamination by PAHs, bread baking was
performed at three temperatures (Table 1). The standard baking
temperature, at which every kind of bread was baked, was chosen
for the lowest temperature of baking. They were equal to 230, 235
and 240 °C, respectively, as for the wheat-rye bread, rye bread, and
wholemeal rye bread. In the case of all analysed kinds of bread,
Table 1
Bread baking conditions.
Kind of bread Baking time (min) Baking temperature (°C)
Wheat-rye bread 40 230 240
Rye bread 40 235 245
Wholemeal rye bread 40 240 250
subsequent baking temperatures were, respectively 10 and 20 °C
higher than their standard baking temperatures.
2.2. Chemicals and materials
The standard mixture of 15 PAHs from the list of SCF (PAH-
Mix 183, Dr. Ehrenstorfer): cyclopenta[c,d]pyrene (C[cd]P),
benzo[a]anthracene (B[a]A), chrysene (Chr), 5-metylchrysene
(5-MChr), benzo[j]fluoranthene (B[j]F), benzo[b] fluoranthene
(B[b]F) and benzo[k]fluoranthene (B[k]F), benzo[a]pyrene (B[a]P),
dibenzo[a,h]anthracene (D[ah]A), benzo[g,h,i]perylene (B[ghi]P),
indeno[c,d]pyrene (I[cd]P), dibenzo[a,l]pyrene (D[al]P), diben-
zo[a,e]pyrene (D[ae]P), dibenzo[a,i]pyrene (D[ai]P) and diben-
zo[a,h]pyrene (D[ah]P) and 16 PAHs by EPA (PAH-Mix 9, Dr.
Ehrenstorfer) were purchased from Witko (Łódź, Poland). The mix-
ture of 16 PAHs by EPA served for the determination of 4 light PAHs:
phenanthrene (Phen), anthracene (Anthr), fluoranthene (F) and pyr-
ene (Pyr). Hexane, acetone, cyclohexane, ethyl acetate, acetonitrile
and toluene (all of analytical grade for residue analysis, HPLC gradi-
ent grade) and anhydrous sodium sulphate (analytical purity
>99.0%) were supplied by POCH (Gliwice, Poland). Deionised water
was obtained from a Millipore Milli-Q water purification system.
The polytetrafluoroethylene (PTFE) filters (1 lm pore size, 25 mm
i.d.) were purchased from Bio Analytic (Gdańsk, Poland).
2.3. Extraction and clean-up
For extraction and clean-up procedures, the method described
by Ciecierska and Obiedziński (2010), with some modifications,
was used. Briefly, homogenised sample (15 g), dried with anhy-
drous sodium sulphate (20 g) in a grinding mortar, was transferred
in an Erlenmeyer flask with 100 ml extraction solvent (hexane/ace-
tone, 60:40, v/v). The flask was covered with aluminium foil to
avoid daylight and placed for 30 min into an ultrasonic bath. Sub-
sequently, the extract was filtered through a layer of anhydrous so-
dium sulphate. The obtained filtrate, after solvent evaporation in a
rotary vacuum evaporator at 35 °C (Büchi No. 141397), was dis-
solved in 4 ml of cyclohexane/ethyl acetate (50:50, v/v) and filtered
through the polytetrafluoroethylene (PTFE) filter with a pore size
of 1 lm. In the evaporation step the most crucial problem was to
avoid evaporation to dryness, as well as to avoid splashes on the
walls of the flask.
For the PAHs isolation step, a clean-up procedure, carried out by
gel permeation chromatography (GPC) and TSK Gel G1000HXL col-
umn (300  7.8 mm, 5 lm), which was purchased from Polygen
(Gliwice, Poland), was used. For this purpose, 1 ml of the extract
obtained previously, was injected onto the column. Chromato-
graphic separation was performed by an isocratic method with a
mixture of cyclohexane/ethyl acetate (50:50, v/v) used as mobile
phase. Samples were eluted at a flow rate, of the mobile phase,
of 0.8 ml/min, with dump time 0–19 min and collect time
19–34 min. A UV–VIS detector with a wavelength of 254 nm was
applied. The purified extract was then subjected to concentration
by rotary vacuum evaporator at 35 °C, subsequently dissolved in
0.5 ml of acetonitrile and transferred to the injector vial.
2.4. HPLC-FLD/DAD
The HPLC-FLD/DAD determination was carried out using a Shi-
madzu HPLC, and consisted of a liquid chromatograph LC-10ATVP,
diode array detector SPD-M10AVP, fluorescence detector RF-10A
XL, degasser DGU-14A, auto injector SIL-10ADVP and system con-
troller SCL-10AVP, co-operated with computer programme LabSo-
250 lution 2.1. A Baker’s chromatographic column named BAKERBOND
255
PAH-16 Plus 250  3 mm, 5 lm (Witko, Poland) was used for
260
chromatographic analyses. Column temperature was isothermal
 M. Ciecierska, M.W. Obiedziński / Food Chemistry 141 (2013) 1–9 3

at 30 °C. Injection volume was 20 ll. A gradient method, with flow
rate of 0.5 ml/min and mobile phase of acetonitrile/water, 50:50
(A) and acetonitrile (B), was applied. The following gradient elution
programme was used: 0–25 min 30% B, 25–50 min 30% B to 100%
B, 50–68.5 min 100% B. After 68.5 min of analysis, the column
was returned to the initial conditions and equilibrated for
20 min. For the PAHs determination by fluorescence detector
(FLD), the following excitation and emission wavelengths
(Ex/Em) were used: 256/370 nm (Phen, Anthr), 270/420 nm (F,
Pyr, B[a]A, Chr, 5-MChr, B[b]F, B[k]F, B[a]P, D[ah]A, D[al]P, B[ghi]P,
D[ae]P), 270/500 nm (B[j]F, I[cd]P), 270/470 nm (D[ai]P, D[ah]P).
For the determination of C[cd]P, a diode array detector (DAD) with
a wavelength of 254 nm was applied.
2.5. Quantification and validation of method
Qualitative–quantitative (19 PAHs) determination was
performed by an external standard method, using the standard
mixture of 15 PAHs from the list of SCF (PAH-Mix 183, Dr. Ehren-
storfer) and 16 PAHs by EPA (PAH-Mix 9, Dr. Ehrenstorfer), which
served for the determination of 4 light PAHs. External standards
mentioned above were used for the preparation of six different
working standard solutions (which cover the concentration range
1–50 lg/l) in order to calibrate and validate the method for the
determination of 19 PAHs by HPLC-FLD/DAD (including 4 light
PAHs from the EPA list and 15 PAHs from the list of SCF).
Triplicate injections of 20 ll of every PAHs working standard
solution were used to construct calibration curves for all analysed
PAHs. The linear relationship of the chromatogram peak area to the
concentration, as well as the squared correlation coefficient (r2) for
different calibration curves and instrument linearity range, are
shown in Table 2. The high values obtained of the squared correla-
tion coefficients for every PAH proved method linearity in the con-
centration range, for which working standard solutions were
prepared, in order to construct calibration curves. The method
linearity was therefore confirmed for the majority of analysed
PAHs in the concentration range 1–50 lg/l, except for C[cd]P,
B[j]F and D[al]P where a linearity range from 2 to 50 lg/l was
confirmed.
With the aim of demonstrating that the method applied is
under analytical control, the limit of detection (LOD), the limit of
quantification (LOQ) as well as recovery experiments, were per-
formed. The LOD and the LOQ, which are defined, respectively, as
numerically equal to three and six times the standard deviation
of the mean of blank determinations (n > 20) (Commission of the
European Communities, 2007) for all of 19 PAHs are presented in
Table 2. Recovery experiments were performed by spiking samples
of wheat-rye bread (Baltonowski bread baked at standard temper-
ature) with three different concentrations of PAHs standard solu-
tion: 0.5, 2 and 10 lg/kg. The fortified samples, as well as
unfortified ones were analysed in triplicate. Recoveries were calcu-
lated as differences in PAHs content in spiked and un-spiked sam-
ples relative to the spiked level. For these recovery analyses, the
repeatability of the method was expressed as the relative standard
deviation (RSD) in %, and therefore as the coefficient of variation
(CV). In accordance with Commission Regulation (EU) No.
836/2011 the HORRATR, values are regarded as a measurement of
the method precision (Commission of the European Communities,
2011b). For all 19 PAHs analysed at each of three different concen-
trations the HORRATR values were calculated, dividing RSD by
RSDR obtained from the Horwitz equation (RSDR = 2(1 0,5log C),
where C = the concentration expressed as a dimensionless frac-
tion). In Table 2, mean recovery, as well as RSD and the HORRATR
values, of these three different concentrations, are presented.
Fig. 1 shows a chromatogram of the standard mixture of 15
PAHs from the list of SCF and the standard mixture of 16 PAHs
by EPA, which served for the determination of 4 light PAHs and
the rye bread (its crust) from the standard baking temperature
(235 °C).
2.6. Confirmation of results by GC/MS
For the comparison of the results of high-pressure liquid chro-
matography with selective detectors, gas chromatography coupled
with mass spectrometry was used. A Shimadzu GCMS-QP 2010,
equipped with a gas chromatograph GC-2010 and mass spectrom-
eter GCMS-QP 2010, was employed for the analysis. Operating con-
ditions were as follows: 30 m  0.25 mm i.d. capillary column with
film thickness of 0.25 lm (ZB-5 ms Zebron, Phenomenex); helium
carrier gas 0.74 ml/min; injector temperature 265 °C, splitless
injection mode, injection volume 1 ll; the ion source and interface
temperature were 230 and 270 °C, respectively; the GC oven tem-
perature programme: 92 °C (1.5 min), 92–140 °C (15 °C/min),
140 °C (1 min), 140–315 °C (5 °C/min) and 315 °C (5 min). The

Table 2
Performance of the proposed HPLC–FLD/DAD method for the determination of PAHs in the loaf of wheat-rye bread (Baltonowski bread baked at standard temperature).

PAH Method linearity range Calibration curvea Correlation LOD (lg/ LOQ (lg/ Recoveryb RSDb HORRATR
(lg/l) coefficient r2 kg) kg) (%) (%) valueb
Phen 1–50 y = 149144x + 31879 0.9998 0.06 0.11 75.0 8.4 0.7
Anthr 1–50 y = 117707x + 4272.1 0.9999 0.07 0.14 75.6 7.5 0.7
F 1–50 y = 18392x 2547 0.9999 0.13 0.26 82.4 9.1 0.8
Pyr 1–50 y = 88196x + 1193.6 0.9998 0.08 0.16 86.5 9.9 0.9
C[cd]P 2–50 y = 122.78x + 14.9 0.9993 0.47 0.94 110 5.9 0.5
B[a]A 1–50 y = 159530x 45297 0.9997 0.05 0.10 83.7 5.4 0.5
Chr 1–50 y = 51379x 13012 0.9996 0.15 0.29 81.2 5.3 0.5
5-MChr 1–50 y = 97566x 6394.4 0.9999 0.07 0.15 82.2 6.3 0.6
B[j]F 2–50 y = 1600.7x + 196.91 0.9997 0.32 0.64 79.7 7.8 0.7
B[b]F 1–50 y = 78977x 22630 0.9998 0.10 0.20 80.6 4.5 0.4
B[k]F 1–50 y = 78142x 4724.2 0.9997 0.10 0.19 81.2 5.0 0.4
B[a]P 1–50 y = 35742x 8979.4 0.9999 0.12 0.24 106 5.1 0.5
D[ah]A 1–50 y = 39684x 8533 0.9997 0.13 0.26 90.8 6.7 0.6
D[al]P 2–50 y = 359.96x 154.23 0.9995 0.30 0.60 107 8.3 0.7
B[ghi]P 1–50 y = 14600x 5088.6 0.9997 0.15 0.30 91.7 6.4 0.6
I[cd]P 1–50 y = 12337x 839.5 0.9997 0.28 0.56 77.6 7.3 0.7
D[ae]P 1–50 y = 8144.2x + 456.41 0.9997 0.29 0.59 91.3 8.1 0.7
D[ai]P 1–50 y = 226619x 96869 0.9997 0.13 0.25 94.3 6.9 0.6
D[ah]P 1–50 y = 172110x 68308 0.9997 0.16 0.33 79.2 7.7 0.7
a
y = peak area, x = concentration (lg/l).
b
Mean recovery, RSD and HORRATR value of three different concentrations (three spiking levels).
4 M. Ciecierska, M.W. Obiedziński / Food Chemistry 141 (2013) 1–9

(x100.000)

2.00

1.75

1.50
Phen B[a]A 5-MChr B[k]F D[ah]A B[ghi]P D[ae]P D[ai]P D[ah]P
1.25
Anthr Pyr Chr B[b]F B[a]P D[al]P I[cd]P
1.00
F B[j]F
0.75
A
0.50

0.25

0.00

- 0.25

- 0.50
B

- 0.75

- 1.00

- 1.25
25.0 27.5 30.0 32.5 35.0 37.5 40.0 42.5 45.0 47.5 50.0 52.5 55.0 57.5 60.0 62.5 65.0 67.5 min

Fig. 1. Chromatogram of the target PAHs in: (A) the standard mixture of 15 PAHs from the list of SCF (10 pg/ll, PAH-Mix 183, Dr. Ehrenstorfer; in blue colour) and the
standard mixture of 16 PAHs by EPA (10 pg/ll, PAH-Mix 9, Dr. Ehrenstorfer; in brown colour), which served for the determination of 4 light PAHs and (B) the rye bread (its
crust) from the standard baking temperature (235 °C). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this
article.)

quadrupole analyser measured the abundance of ions of m/z from
100 to 400 and detector voltage was 1.5 kV. Electron ionisation
(70 eV) with selected ion monitoring mode was used, and the
two most abundant ions from the molecular ion cluster were mea-
sured for each analysed compound. PAHs identification was per-
formed both on the basis of GC retention time comparison with
available PAHs standard solutions and using characteristic ions
monitored in analyses.
2.7. Statistical analysis
The obtained results were statistically analysed using the Stat-
istica 7.1 programme. To assess the significance of the differences
between the means of 19 PAHs contents in the analysed products,
the method of multiple comparisons, using Tukey’s test, was
applied, at a significance level a = 0.05.
3. Results and discussion
In accordance with the Commission Regulation (EU) No. 836/
2011, performance criteria for methods of analysis for B[a]P,
B[a]A, B[b]F and Chr are the following: LOD less than 0.3 lg/kg,
LOQ less than 0.9 lg/kg, precision – HORRATR values of less than
2 and recovery from 50% to 120% (Commission of the European
Communities, 2011b). The performance of the presented method
showed that, for all 19 PAHs analysed at each of three different
concentrations, both the HORRATR values were well below than 2
and recovery was in the recommended range. Furthermore, the ob-
tained values of RSD (%) are satisfactory, as they ranged from 4.5%
for B[b]F to 9.9% in the case of the light PAH – pyrene. Thereby it
was proved that HPLC-FLD/DAD method for the determination of
19 PAHs, including 4 light PAHs from the EPA list and 15 PAHs from
the list of SCF (as well as fulfilling all requirements of the European
Union concerning the methods of B[a]P, B[a]A, B[b]F and Chr anal-
ysis in foodstuffs) also demonstrated satisfactory validation
parameters for the remaining PAHs.
Data on mean contents of individual PAHs, as well as a total
sum of 19 PAHs content in analysed raw materials of the bakery
chain and its final products, were obtained. The data included
the grain of wheat and rye, three types of flour derived from these
grains and their bran and selected kinds of bread, as well as their
dough obtained from the HPLC-FLD/DAD method, and are pre-
sented in Tables 3–6, respectively. It should be noted that, for all
analysed products, on the basis of the results obtained from the
HPLC-FLD/DAD and GC/MS method, it was confirmed that the dif-
ferences between those results were statistically insignificant.
Taking into consideration contamination of the bakery chain
raw materials, both in the groups of grain, flour and the bran
obtained by grinding the grain (mentioned above) into flour, sim-
ilar quality profiles of PAHs were apparent. In the majority of cases,
they were characterised by relatively high contents of 4 light PAHs
(Phen, Anthr, F, Pyr). In the grain of wheat and rye, these 4 com-
pounds constituted, respectively, 94% and 69% of the total 19 PAHs
content. In the case of wheat flour, rye flour and wholemeal rye
flour, light PAHs constituted 93%, 80% and 61% of all analysed
polyarenes whereas, in the wheat bran, rye bran (obtained as a
result of grinding the grain of rye to wholemeal rye flour), they
constituted 70%, 63% and 74%, respectively. In the group of 15 hea-
vy PAHs, mainly B[a]A, 5-MChr and B[k]F were detected. The
remaining PAHs, e.g. B[a]P, B[j]F, B[ghi]P, I[cd]P and PAHs from
the group of dibenzopyrenes, were not detected in any of analysed
samples of the bakery chain raw materials.
On the basis of the performed statistical analysis, statistically
higher levels of the total 19 PAHs contamination were found in
the grain of rye and rye bran (respectively 2.93 and 2.65 lg/kg)
compared with the grain of wheat (2.35 lg/kg) and wheat bran
(1.87 lg/kg). Statistically significant differentiation in the 19 PAHs
content between wheat flour, rye flour and wholemeal rye flour
were not apparent, as the sums of 19 PAHs content in these
products were equal to 1.07, 1.25 and 1.35 lg/kg, respectively.
Among the brans studied, statistically the lowest and the highest
levels of total PAHs contamination, being equal to 1.87 and
3.65 lg/kg, were observed for wheat bran and bran obtained as a
 M. Ciecierska, M.W. Obiedziński / Food Chemistry 141 (2013) 1–9 5

Table 3
Mean content of PAHs with standard deviation in selected raw materials of the bakery chain (lg/kg).

PAH Wheat Rye Wheat Rye flour Wholemeal Wheat Rye bran Bran from rye grinding to
flour rye flour bran wholemeal rye flour
Phen 1.14 ± 0.09 1.45 ± 0.23 0.64 ± 0.09 0.59 ± 0.11 0.51 ± 0.11 0.79 ± 0.05 0.76 ± 0.09 1.19 ± 0.11
Anthr 0.08 ± 0.00 0.10 ± 0.00 nd nd nd 0.07 ± 0.01 0.08 ± 0.01 0.14 ± 0.02
F 0.58 ± 0.14 0.19 ± 0.04 0.19 ± 0.01 0.20 ± 0.01 0.21 ± 0.01 0.17 ± 0.04 0.59 ± 0.01 0.55 ± 0.03
Pyr 0.42 ± 0.01 0.27 ± 0.01 0.17 ± 0.04 0.21 ± 0.03 0.11 ± 0.01 0.28 ± 0.04 0.24 ± 0.01 0.83 ± 0.08
C[cd]P nda nd nd nd nd nd nd nd
B[a]A 0.05 ± 0.00 0.06 ± 0.00 nd nd nd 0.05 ± 0.01 0.05 ± 0.01 0.08 ± 0.02
Chr nd nd nd nd nd nd nd 0.16 ± 0.03
5-MChr 0.09 ± 0.01 0.32 ± 0.04 0.07 ± 0.01 0.09 ± 0.00 0.27 ± 0.04 0.08 ± 0.01 0.43 ± 0.02 0.47 ± 0.03
B[j]F nd nd nd nd nd nd nd nd
B[b]F nd nd nd nd nd 0.11 ± 0.01 nd nd
B[k]F nd 0.30 ± 0.04 nd 0.10 ± 0.01 0.25 ± 0.03 0.10 ± 0.01 0.30 ± 0.02 0.23 ± 0.02
B[a]P nd nd nd nd nd nd nd nd
D[ah]A nd 0.23 ± 0.02 nd nd nd 0.22 ± 0.01 0.20 ± 0.04 nd
D[al]P nd nd nd nd nd nd nd nd
B[ghi]P nd nd nd nd nd nd nd nd
I[cd]P nd nd nd nd nd nd nd nd
D[ae]P nd nd nd nd nd nd nd nd
D[ai]P nd nd nd nd nd nd nd nd
D[ah]P nd nd nd nd nd nd nd nd
R 19 PAHs (lg/kg) and theirs 2.35 ± 0.15 2.93 ± 0.22 1.07 ± 0.14 1.25 ± 0.15 1.35 ± 0.20 1.87 ± 0.18 2.65 ± 0.17 3.65 ± 0.23
significance of differencesb a1 b1 a2 a2 a2 a3 b3 c3
B1 B2 A1 A2 A3 B1 B2 B3
a
nd – not detected.
b
Different small or capital letters by the same number (namely within one comparison, where the next number represents a further comparison) below the mean values of
sum of 19 PAHs indicate statistically significant difference between means at a = 0.05 level.

result of grinding the grain of rye to wholemeal rye flour. More-
over, significantly higher total 19 PAHs contents in the grain of
wheat and rye compared with respectively wheat flour, rye flour
and wholemeal rye flour, were also confirmed. Furthermore, statis-
tically significant differences among the levels of 19 PAHs contam-
ination in wheat bran, rye bran or bran obtained, as a result of
grinding the grain of rye to wholemeal rye flour and respectively
wheat flour, rye flour and wholemeal rye flour, were also proved.
In the case of the sum of 4 heavy PAHs contents (B[a]P, B[a]A,
B[b]F, Chr), which were concluded by the CONTAM Panel in the
EFSA opinion of June 9, 2008 to be the most suitable indicators
of PAHs in food and listed to be monitored in food according to
the Regulation (EU) No 835/2011), similar relations were apparent.
It could be explained by the fact that the majority of PAHs contam-
inating the grain is concentrated in its external parts directly
exposed to the deposition of PAHs.
Available literature reports that grain contamination is depen-
dent on the place of growing, and especially the level of its industri-
alisation, that is PAHs emission sources and vehicular traffic
(Jankowski & Obiedziński, 1998; SCF, 2002, Yang, Conell, Havker,
& Kayal, 1991). American studies have revealed that there are geo-
graphical differences in the levels of PAHs in the grain and that traf-
fic is the major cause of grain contamination (Kobayashi, Okamoto,
Maddalena, & Kado, 2008). Therefore it could be concluded that the
low level of contamination of analysed grains was a consequence of
the place of their origin. They were collected for testing from a farm
located on Polish agricultural land, away from the nearest highway
and the industry, because the dairy was 25 km away.
Taking into consideration contamination of bran, studies have
also revealed their higher PAHs content compared with other prod-
ucts of grain grinding (Dennis, Cripps, Venn, Howarth, & Lee, 1991;
Lawrence & Das, 1986; Tuominen, Pyssalo, & Sauri, 1988).
In all analysed kinds of bread, similar quality profiles of PAHs
were observed. It was found that 4 light PAHs constituted 88–
92%, 91–94% and 86–88% of all 19 PAHs determined in different
variants of wheat-rye bread, rye bread and wholemeal rye bread,
respectively. Among 15 PAHs from the list of SCF, mainly low levels
of B[a]A, Chr, 5-MChr, B[b]- and B[k]F were found. However, the
presence of the latter two compounds was detected only in bread
baked at elevated temperatures. Furthermore, B[a]P was detected
only in the crust of wholemeal rye bread at the highest tempera-
ture of baking. In none of the variants of bread under investigation
were the remaining heavy PAHs, including dibenzopyrenes
detected. In doughs of wheat-rye bread, rye bread and wholemeal
rye bread, 4 light PAHs constituted, respectively, 100%, 100% and
83% of total 19 PAHs content.
Regardless of the baking temperature of Baltonowski bread, sta-
tistically significant differences between the levels of 19 PAHs con-
tamination of individual parts of the loaf were apparent. As an
example, in the standard baking temperature (230 °C) the mean
19 PAHs contents in the crumb, the loaf and the crust of Baltonow-
ski bread were, respectively, 1.59, 2.36 and 3.84 lg/kg whereas, at
the temperature increased to 250 °C, these were equal to 3.02, 4.33
and 7.37 lg/kg.
The influence of baking temperature on the level of PAHs con-
tamination of analysed parts of Baltonowski bread was also
proved. For the crumb, the loaf of bread and the crust, with increas-
ing baking temperature, the total PAHs content significantly
increased. However, in the case of the crumb of this bread, it was
observed that an increase of only 20 °C, above the standard tem-
perature of baking (230 °C), led to a significantly higher PAHs
content.
In the case of rye bread, for every baking temperature, it was
revealed, that the total 19 PAHs concentration in the loaf of bread
was statistically significantly higher than in the crumb, but signif-
icantly lower than in the crust. For example the mean sums of 19
PAHs content in the crumb, the loaf of bread and the crust baked
at the standard temperature of 235 °C were, respectively, 5.62,
6.86 and 7.83 lg/kg, whereas, at the temperature of 255 °C, they
were 8.73, 10.4 and 13.0 lg/kg.
For the rye bread, it was also statistically confirmed that
increases of 10 and 20 °C above the standard temperature of bak-
ing (235 °C) caused significant increases in contamination level of
its crumb, the loaf of bread as well as the crust.
Describing the level of total 19 PAHs contamination in whole-
meal rye bread, for every baking temperature, statistically signifi-
6 M. Ciecierska, M.W. Obiedziński / Food Chemistry 141 (2013) 1–9

Table 4
Mean content of PAHs with standard deviation in wheat-rye bread (Baltonowski bread) and its dough (lg/kg).

PAH Dough of Baltonowski bread baked at temperature

Baltonowski
230 °C 240 °C 250 °C
bread
Crumb Loaf of Crust Crumb Loaf of Crust Crumb Loaf of Crust
bread bread bread
Phen 0.35 ± 0.03 0.65 ± 0.14 0.99 ± 0.11 1.48 ± 0.13 0.88 ± 0.14 1.23 ± 0.10 2.27 ± 0.10 1.29 ± 0.20 1.73 ± 0.10 2.88 ± 0.16
Anthr nda nd 0.07 ± 0.01 0.10 ± 0.02 nd 0.08 ± 0.01 0.19 ± 0.03 0.08 ± 0.01 0.13 ± 0.02 0.17 ± 0.03
F 0.13 ± 0.02 0.49 ± 0.07 0.76 ± 0.09 1.29 ± 0.14 0.66 ± 0.04 0.85 ± 0.04 1.69 ± 0.23 0.88 ± 0.07 1.23 ± 0.14 1.96 ± 0.13
Pyr 0.09 ± 0.02 0.23 ± 0.03 0.42 ± 0.03 0.64 ± 0.11 0.39 ± 0.04 0.59 ± 0.10 0.97 ± 0.18 0.50 ± 0.03 0.82 ± 0.09 1.51 ± 0.17
C[cd]P nd nd nd nd nd nd nd nd nd nd
B[a]A nd 0.05 ± 0.01 0.06 ± 0.01 0.06 ± 0.02 0.05 ± 0.01 0.06 ± 0.01 0.07 ± 0.02 0.06 ± 0.01 0.07 ± 0.01 0.09 ± 0.01
Chr nd nd nd 0.15 ± 0.04 nd nd 0.17 ± 0.04 nd nd 0.23 ± 0.03
5-MChr nd 0.07 ± 0.01 0.08 ± 0.02 0.12 ± 0.03 0.09 ± 0.02 0.10 ± 0.01 0.13 ± 0.06 0.10 ± 0.01 0.12 ± 0.03 0.16 ± 0.02
B[j]F nd nd nd nd nd nd nd nd nd nd
B[b]F nd nd nd nd nd 0.11 ± 0.01 0.12 ± 0.01 0.10 ± 0.02 0.13 ± 0.02 0.15 ± 0.05
B[k]F nd nd nd nd nd nd 0.11 ± 0.01 nd 0.10 ± 0.02 0.12 ± 0.02
B[a]P nd nd nd nd nd nd nd nd nd nd
D[ah]A nd nd nd nd nd nd nd nd nd nd
D[al]P nd nd nd nd nd nd nd nd nd nd
B[ghi]P nd nd nd nd nd nd nd nd nd nd
I[cd]P nd nd nd nd nd nd nd nd nd nd
D[ae]P nd nd nd nd nd nd nd nd nd nd
D[ai]P nd nd nd nd nd nd nd nd nd nd
D[ah]P nd nd nd nd nd nd nd nd nd nd
R 19 PAHs (lg/kg) and 0.57 ± 0.05 1.59 ± 0.19 2.36 ± 0.14 3.84 ± 0.35 2.05 ± 0.22 3.04 ± 0.15 5.71 ± 0.46 3.02 ± 0.21 4.33 ± 0.33 7.37 ± 0.59
theirs significance of a1 b1 c1 d1 a2 b2 c2 a3 b3 c3
differencesb A1 A2 A3 A1 B2 B3 B1 C2 C3
a
nd – not detected.
b
Different small or capital letters by the same number (namely within one comparison, where the next number represents a further comparison) below the mean values of
sum of 19 PAHs indicate statistically significant difference between means at a = 0.05 level.

cant differentiation was observed between individual parts of the
loaf. The mean sums of 19 PAHs contents in the crumb, the loaf
and the crust of this bread were, respectively, 2.67, 5.08 and
8.78 lg/kg for the standard baking temperature (240 °C) and
7.22, 9.56 and 13.6 lg/kg when the temperature increased by
20 °C.
In the case of wholemeal rye bread, it was statistically
confirmed that, for its crumb, an increase of only 20 °C above the
standard temperature of baking (240 °C) resulted in a significantly
higher level of total PAHs contamination. However, for the loaf of
bread as well as its crust it was proved that both increases of 10
and 20 °C above the temperature of 240 °C led to a significant
increase in contamination level.
According to the European Food Safety Authority report dated
29 June, 2007, prepared on the basis of the results of studies per-
formed in 16 Member States dealing with PAHs in food, with
special attention to benzo[a]pyrene, the mean contents of this
compound in bread, flour and grain were 0.22, 0.10 and 0.09 lg/
kg, respectively. Moreover, the percentage of samples exceeding
the limit of detection in the case of bread was only 8%, in flour
31%, and in grain 53%. None of the analysed samples exceeded
the maximum tolerable limit of 1 lg B[a]P/kg for processed
cereal-based foods (EFSA, 2007).
Other research has revealed that, in samples of Italian bread
baked, using wood as fuel, concentrations of B[a]P were, on aver-
age, lower than the limit set in the Commission Regulation (UE)
No. 835/2011 (Orecchio & Papuzza, 2009). In a survey performed
in Spain, dealing with PAHs contamination of toasted bread, simi-
lar results were found. B[b]F, B[k]F and B[a]P were detected in 3 of
24 samples of commercial toasted bread in ranges of 0.25–0.45,
0.059–0.098 and 0.13–0.23 lg/kg, respectively. Nevertheless
B[a]P concentrations in these samples were much lower than the
maximum level regulated by food law requirements and no sam-
ples obtained by electric oven and toaster were polluted by heavy
PAHs (Rey-Salgueiro et al., 2008). In another work, also concerning
PAHs content in toasted bread, B[a]P was not detected in 10 of 18
samples whereas, in the rest of the samples, it varied from 2.83 lg/
kg to even 16.5 lg/kg. Furthermore, the total heavy PAHs concen-
tration was in the range 1.06–44.2 lg/kg (Al-Rashdan, Helaleh,
Nisar, Ibtisam, & Al-Ballam, 2010).
Although data on PAHs content in the group of raw materials
and final products of the bakery chain are scarce, our results corre-
spond well with the data cited above. To summarise, among the
analysed bakery chain’s raw materials, the lowest levels of total
PAHs contamination were found in flour. The mean content of 19
PAHs in flour under investigation was over 2 times lower than
the relatively low level of grain and bran contamination and from
about 2 to 6 times lower in comparison with loaves of bread from
the standard temperature of baking. Moreover, considering the
percentage share of flour in the recipe for analysed kinds of bread,
a relatively low contribution of these raw materials in the contam-
ination of final products was proved. Furthermore, the obtained
results indicated unambiguously low levels of PAHs in analysed
final products of the bakery chain. Taking into consideration the
maximum tolerable limit for B[a]P and the sum of 4 heavy PAHs
(B[a]P, B[a]A, B[b]F, Chr) in processed cereal-based foods stated
in the Commission Regulation (UE) No. 835/2011, it was proved
that analysed products fulfilled the European Union food law
requirements. Only at the highest, unconventional temperature
of baking in the case of wholemeal rye bread crust, was the mean
content of 4 heavy PAHs above the maximum tolerable limit of
1 lg/kg, by about 30%.
The mean sums of 4 heavy PAHs contents were in the range 0.05–
0.47, 0.23–0.45 and 0.21–1.29 lg/kg in the case of wheat-rye bread,
rye bread and wholemeal rye bread, respectively. Moreover, regard-
less of baking temperature, contamination by light PAHs was
predominant. For all analysed kinds of bread statistically significant
differences among levels of total PAHs contamination of individual
parts of loaves were apparent, and the influence of baking tempera-
ture on the sum of 19 PAHs content was confirmed. Furthermore, for
every bread, the levels of both total 19 PAHs content and 4 heavy
PAHs content in their doughs were statistically lower than those
 M. Ciecierska, M.W. Obiedziński / Food Chemistry 141 (2013) 1–9 7

Table 5
Mean content of PAHs with standard deviation in rye bread and its dough (lg/kg).

PAH Dough of Rye bread baked at temperature

rye bread
235 °C 245 °C 255 °C
Crumb Loaf of Crust Crumb Loaf of Crust Crumb Loaf of Crust
bread bread bread
Phen 0.38 ± 0.05 2.57 ± 0.10 3.30 ± 0.28 3.49 ± 0.16 3.18 ± 0.48 3.97 ± 0.33 4.65 ± 0.39 3.89 ± 0.41 4.55 ± 0.49 5.99 ± 0.59
Anthr nda 0.12 ± 0.03 0.15 ± 0.01 0.21 ± 0.02 0.17 ± 0.02 0.20 ± 0.01 0.26 ± 0.07 0.20 ± 0.06 0.36 ± 0.07 0.48 ± 0.05
F 0.13 ± 0.02 1.58 ± 0.10 1.77 ± 0.25 2.02 ± 0.26 1.93 ± 0.30 2.09 ± 0.13 2.29 ± 0.41 2.32 ± 0.31 2.73 ± 0.28 3.21 ± 0.45
Pyr 0.15 ± 0.02 1.01 ± 0.09 1.26 ± 0.01 1.67 ± 0.05 1.53 ± 0.15 1.89 ± 0.17 2.20 ± 0.25 1.82 ± 0.14 2.11 ± 0.06 2.25 ± 0.05
C[cd]P nd nd nd nd nd nd nd nd nd nd
B[a]A nd 0.08 ± 0.01 0.09 ± 0.03 0.11 ± 0.01 0.07 ± 0.02 0.09 ± 0.00 0.12 ± 0.04 0.12 ± 0.05 0.14 ± 0.05 0.18 ± 0.07
Chr nd 0.15 ± 0.02 0.17 ± 0.02 0.19 ± 0.02 0.17 ± 0.05 0.22 ± 0.04 0.23 ± 0.04 0.19 ± 0.03 0.24 ± 0.06 0.27 ± 0.08
5-MChr nd 0.11 ± 0.02 0.12 ± 0.02 0.14 ± 0.01 0.13 ± 0.02 0.17 ± 0.02 0.19 ± 0.04 0.19 ± 0.07 0.22 ± 0.05 0.26 ± 0.05
B[j]F nd nd nd nd nd nd nd nd nd nd
B[b]F nd nd nd nd nd nd nd nd nd nd
B[k]F nd nd nd nd nd nd 0.10 ± 0.00 nd nd 0.40 ± 0.03
B[a]P nd nd nd nd nd nd nd nd nd nd
D[ah]A nd nd nd nd nd nd nd nd nd nd
D[al]P nd nd nd nd nd nd nd nd nd nd
B[ghi]P nd nd nd nd nd nd nd nd nd nd
I[cd]P nd nd nd nd nd nd nd nd nd nd
D[ae]P nd nd nd nd nd nd nd nd nd nd
D[ai]P nd nd nd nd nd nd nd nd nd nd
D[ah]P nd nd nd nd nd nd nd nd nd nd
R 19 PAHs (lg/kg) and 0.66 ± 0.07 5.62 ± 0.26 6.86 ± 0.33 7.83 ± 0.41 7.20 ± 0.63 8.63 ± 0.50 10.04 ± 0.64 8.73 ± 0.72 10.35 ± 0.48 13.04 ± 0.71
theirs significance of a1 b1 c1 d1 a2 b2 c2 a3 b3 c3
differencesb A1 A2 A3 B1 B2 B3 C1 C2 C3
a
nd – not detected.
b
Different small or capital letters by the same number (namely within one comparison, where the next number represents a further comparison) below the mean values of
sum of 19 PAHs indicate statistically significant difference between means at a = 0.05 level.

in the crumb, the loaf of bread and the crust baked at standard tem-
peratures. In the case of the total 4 heavy PAHs content, for every
bread, significant differences between the crumb and the crust were
also apparent and the influence of baking temperature on their
content was also proved. Therefore, on the basis of the performed
study it could be concluded that PAHs can be formed during baking
and that baking conditions applied significantly influence the level
of bread contamination by PAHs; however, they do not cause high
or undesirable PAHs concentrations, especially by heavy ones.
Despite the fact that processed cereal-based foods are usually
characterised by low levels of PAHs contamination, it should be
noted that, due to their high level of consumption, they can be a

Table 6
Mean content of PAHs with standard deviation in wholemeal rye bread and its dough (lg/kg).

PAHs Dough of Wholemeal rye bread baked at temperature

wholemeal
240 °C 250 °C 260 °C
rye bread
Crumb Loaf of Crust Crumb Loaf of Crust Crumb Loaf of Crust
bread bread bread
Phen 0.26 ± 0.04 1.67 ± 0.30 3.22 ± 0.32 4.14 ± 0.28 1.96 ± 0.32 4.14 ± 0.39 5.13 ± 0.52 3.22 ± 0.38 4.34 ± 0.26 5.65 ± 0.16
Anthr nda 0.09 ± 0.02 0.19 ± 0.02 0.31 ± 0.09 0.11 ± 0.06 0.23 ± 0.00 0.36 ± 0.11 0.15 ± 0.02 0.25 ± 0.01 0.40 ± 0.03
F 0.13 ± 0.03 0.49 ± 0.08 1.15 ± 0.10 2.74 ± 0.35 0,92 ± 0.08 1.69 ± 0.29 3.35 ± 0.25 1.99 ± 0.10 2.74 ± 0.50 3.62 ± 0.22
Pyr 0.09 ± 0.02 0.11 ± 0.01 0.14 ± 0.02 1.14 ± 0.10 0.22 ± 0.02 0.44 ± 0.08 1.68 ± 0.34 1.36 ± 0.10 1.41 ± 0.17 1.94 ± 0.17
C[cd]P nd nd nd nd nd nd nd nd nd nd
B[a]A nd 0.06 ± 0.01 0.10 ± 0.02 0.13 ± 0.02 0.07 ± 0.02 0.14 ± 0.02 0.26 ± 0.06 0.09 ± 0.01 0.18 ± 0.00 0.29 ± 0.05
Chr nd 0.15 ± 0.01 0.16 ± 0.02 0.18 ± 0.01 0.18 ± 0.01 0.21 ± 0.07 0.35 ± 0.01 0.23 ± 0.04 0.31 ± 0.05 0.53 ± 0.02
5-MChr 0.10 ± 0.01 0.10 ± 0.02 0.12 ± 0.04 0.14 ± 0.01 0.15 ± 0.07 0.20 ± 0.01 0.25 ± 0.01 0.18 ± 0.01 0.23 ± 0.01 0.27 ± 0.03
B[j]F nd nd nd nd nd nd nd nd nd nd
B[b]F nd nd nd nd nd nd nd nd 0.10 ± 0.05 0.23 ± 0.09
B[k]F nd nd nd nd nd nd nd nd nd 0.15 ± 0.03
B[a]P nd nd nd nd nd nd nd nd nd 0.24 ± 0.07
D[ah]A nd nd nd nd nd nd nd nd nd 0.23 ± 0.05
D[al]P nd nd nd nd nd nd nd nd nd nd
B[ghi]P nd nd nd nd nd nd nd nd nd nd
I[cd]P nd nd nd nd nd nd nd nd nd nd
D[ae]P nd nd nd nd nd nd nd nd nd nd
D[ai]P nd nd nd nd nd nd nd nd nd nd
D[ah]P nd nd nd nd nd nd nd nd nd nd
R PAHs (lg/kg) and 0.58 ± 0.06 2.67 ± 0.27 5.08 ± 0.39 8.78 ± 0.79 3.61 ± 0.31 7.05 ± 0.47 11.38 ± 0.69 7.22 ± 0.35 9.56 ± 0.52 13.55 ± 0.63
theirs significance of a1 b1 c1 d1 a2 b2 c2 a3 b3 c3
differencesb A1 A2 A3 A1 B2 B3 B1 C2 C3
a
nd – not detected.
b
Different small or capital letters by the same number (namely within one comparison, where the next number represents a further comparison) below the mean values of
sum of 19 PAHs indicate statistically significant difference between means at a = 0.05 level.
8 M. Ciecierska, M.W. Obiedziński / Food Chemistry 141 (2013) 1–9
significant cause of exposure to PAHs. The same opinion has been
expressed by many researchers studying the issue of dietary PAHs
intake (Ibanez et al., 2005; Jankowski & Obiedziński, 1998; Orec-
chio & Papuzza, 2009; SCF, 2002, White, Fernandes, & Rose, 2002).
In the EFSA opinion, reported in 2008, the Panel on Contami-
nants in the Food Chain (the CONTAM Panel) presented the MOE
(Margin of Exposure) analysis as a new approach to risk assess-
ment for genotoxic and carcinogenic PAHs (EFSA, 2008). The
MOE is calculated by dividing the toxicity estimate (BMDL – bench
mark dose lower confidence limit) from animal experiments by the
estimated intake of food. Moreover, it should be emphasised that,
in MOE analysis and estimation of PAHs intake, an upper bound
approach to treat the ‘‘not detected’’ data is appropriate for calcu-
lations. According to the Polish Statistical Yearbook of Agriculture
– 2011, average monthly per capita (70 kg body weight) consump-
tion of bread and cereal products in households in 2010 was equal
to 7.01 kg, that is 0.23 kg/day (0.0039 kg/kg b.w./day) (Central Sta-
tistical Office, 2011). Considering the mean B[a]P content, as well
as the sum of 4 heavy PAHs (calculated according to an upper
bound approach) in analysed products (in the case of bread the
loaves of bread from standard baking temperature were taken for
estimation), the average dietary B[a]P and 4 heavy PAHs intakes
(B[a]P, Chr, B[a]A and B[b]F) were 0.5 and 1.7 ng/kg b.w./day,
respectively. Consequently, the MOE for B[a]P was about 213,700
and for 4 PAHs was 198,100. These MOEs, which are about 12
and 11 times higher than the MOE for average adult consumers
in the case of B[a]P and the sum of 4 PAHs, indicated a very low
concern for human health. Moreover, the average dietary exposure
to 19 PAHs estimated in this study (22.1 ng/kg b.w./day) coincides
with the intake of mainly 4 light PAHs. On the basis of the obtained
values of the average dietary exposure to PAHs and the resulting
MOEs, it could be concluded that bread and cereal products under
investigation constitute a low concern for consumer health.
4. Conclusions
This study focused on the determination of 4 light PAHs (listed
by the US EPA) and 15 PAHs (prioritised by the Scientific Commit-
tee on Food of the European Union) in raw materials and final
products of the bakery chain. The obtained validation data showed
the suitability of the applied method for the determination of 19
PAHs in bread. Considering the maximum tolerable limit for
B[a]P and the sum of 4 heavy PAHs (B[a]P, B[a]A, B[b]F, Chr) in pro-
cessed cereal-based foods stated in the Commission Regulation
(UE) No. 835/2011, it was proved that analysed products fulfilled
the European Union food law requirements. Furthermore, the ob-
tained results indicated a low level of total PAHs contamination
with light PAHs being predominant, in both raw materials and final
products of the bakery chain. For all analysed kinds of bread, statis-
tically the highest total 19 PAHs contents were found in the crust,
and the lowest in the crumb. On the basis of contamination level of
the dough of analysed bread and the stated influence of baking
temperature on the bread’s PAHs content, it was confirmed that
PAHs are formed during baking. Moreover, during baking in cyclo-
thermic deck ovens, the temperature significantly influenced the
level of bread contamination by PAHs; however, it did not cause
high and undesirable PAHs concentrations, including by heavy
PAHs. To sum up, taking into consideration the obtained values
of the average dietary exposure to PAHs and MOE analysis, it could
be concluded that bread and cereal products under investigation
constitute a low concern for consumer’s health. However, since
processed cereal-based foods have been shown to be the major
source of PAHs in the diet, further research on PAHs content in
such products is still needed to continuously control the dietary
exposure to these compounds.
Acknowledgement
This research was performed with the financial support of the
Polish State Committee for Scientific Research KBN under Grant
No. 501 0928 00 29.
References
Alomirah, H., Al-Zenki, S., Al-Hooti, S., Zaghloul, S., Sawaya, W., Ahmed, N., et al.
(2011). Concentrations and dietary exposure to polycyclic aromatic
hydrocarbons (PAHs) from grilled and smoked foods. Food Control, 22(12),
2028–2035.
Al-Rashdan, A., Helaleh, M. I. H., Nisar, A., Ibtisam, A., & Al-Ballam, Z. (2010).
Determination of the levels of polycyclic aromatic hydrocarbons in toasted
bread using gas chromatography mass spectrometry. International Journal of
Analytical Chemistry. http://dx.doi.org/10.1155/2010/821216.
Amoako, J., Ansa-Asare, O. D., Karikari, A. Y., & Dartey, G. (2011). Levels of polycyclic
aromatic hydrocarbons (PAHs) in the Densu River Basin of Ghana.
Environmental Monitoring and Assessment, 174, 471–480.
Central Statistical Office. (2011). Statistical Yearbook of Agriculture – 2011
(pp. 317). Warsaw: Statistical Publishing Establishment.
Camargo, M. C. R., Antoniolli, P. R., & Vicente, E. (2012). Evaluation of polycyclic
aromatic hydrocarbons content in different stages of soybean oils processing.
Food Chemistry, 135(3), 937–942.
Codex Committee on Food Additives and Contaminants, CCFAC. (2005). Discussion
paper on polycyclic aromatic hydrocarbons (PAHs) contamination. Thirty-seventh
Session, 25–29 April 2005. The Hague, the Netherlands.
Chang, K. F., Fang, G. C., Chen, J. C., & Wu, Y. S. (2006). Atmospheric polycyclic
aromatic hydrocarbons (PAHs) in Asia: A review from 1999 to 2004.
Environmental Pollution, 142, 388–396.
Ciecierska, M., & Obiedziński, M. W. (2010). Polycyclic aromatic hydrocarbons in
infant formulae, follow-on formulae and baby foods available in the Polish
market. Food Control, 21(8), 1166–1172.
Ciecierska, M., & Obiedziński, M. W. (2013). Polycyclic aromatic hydrocarbons in
vegetable oils from unconventional sources. Food Control, 30(2), 556–562.
Commission of the European Communities. (2005). Commission Recommendation
(EC) No. 108/2005 of 4 February 2005 on the further investigation into the
levels of polycyclic aromatic hydrocarbons in certain foods. Official Journal of the
European Union, L 34/3.
Commission of the European Communities. (2007). Commission Regulation (EC) No.
333/2007 of 28 March 2007 laying down the methods of sampling and analysis
for the official control of the levels of lead, cadmium, mercury, inorganic tin, 3-
MCPD and benzo[a]pyrene in foodstuffs. Official Journal of the European Union, L
88/29.
Commission of the European Communities. (2011a). Commission Regulation (EU)
No. 835/2011 of 19 August 2011 amending Regulation (EC) No. 1881/2006 as
regards maximum levels for polycyclic aromatic hydrocarbons in foodstuffs.
Official Journal of the European Union, L 215/4.
Commission of the European Communities. (2011b). Commission Regulation (EU)
No. 836/2011 of 19 August 2011 amending Regulation (EC) No. 333/2007 laying
down the methods of sampling and analysis for the official control of the levels
of lead, cadmium, mercury, inorganic tin, 3-MCPD and benzo(a)pyrene in
foodstuffs. Official Journal of the European Union, L 215/9.
Dennis, M. J., Cripps, G., Venn, I., Howarth, N., & Lee, G. (1991). Factors affecting the
polycyclic aromatic hydrocarbon content of cereals, fats, and other food
products. Food Additives and Contaminants, 8, 517–530.
European Food Safety Authority, EFSA. (2008). Polycyclic aromatic hydrocarbonsin
food. Scientific opinion of the panel on contaminants in the food chain adopted
on 9 June 2008. The EFSA Journal, 724, 1–114.
European Food Safety Authority, EFSA. (2007). Findings of the EFSA data collection on
polycyclic aromatic hydrocarbons (PAHs) in food. A report from the Unit of Data
Collection and Exposure on Request from the European Commission, 29 June
2007, EFSA/DATEX/002. Available from <http://www.efsa.europa.eu/EFSA/
Scientific_Document/datex_report_pah.pdf>.
Houessou, J. K., Goujot, D., Heyd, B., & Camel, V. (2008). Modeling the formation of
some polycyclic aromatic hydrocarbons during the roasting of Arabica coffee
samples. Journal of Agricultural and Food Chemistry, 56, 3648–3656.
Ibanez, R., Agudo, A., Berenguer, A., Jakszyn, P., Tormo, M. J., Sanchez, M. J., et al.
(2005). Dietary intake of polycyclic aromatic hydrocarbons in a Spanish
population. Journal of Food Protection, 68(10), 2190–2195.
Jankowski, P.S., & Obiedziński, M.W. (1998). The occurrence of polycyclic aromatic
hydrocarbons in agricultural products and foodstuffs. An attempt to assess the
risk of Polish consumer by PAHs [in Polish]. In The Polish National Scientific
Conference in Olsztyn, 3rd Session, 23 September 1998. Olsztyn, Poland.
Kobayashi, R., Okamoto, R. A., Maddalena, R. L., & Kado, N. Y. (2008). Polycyclic
aromatic hydrocarbons in edible grain: A pilot study of agricultural crops as a
human exposure pathway for environmental contaminants using wheat as a
model crop. Environmental Research, 107(2), 145–151.
Lawrence, J. F., & Das, B. J. (1986). Determination of nanogram/kilogram levels of
polycyclic aromatic hydrocarbons in foods by HPLC with fluorescence detection.
International Journal of Environmental Analytical Chemistry, 24(2), 113–131.
Martí-Cid, R., Llobet, J. M., Castell, V., & Domingo, J. L. (2008). Evolution of the
dietary exposure to polycyclic aromatic hydrocarbons in Catalonia, Spain. Food
and Chemical Toxicology, 46(9), 3163–3171.
 M. Ciecierska, M.W. Obiedziński / Food Chemistry 141 (2013) 1–9
Orecchio, S., & Papuzza, V. (2009). Levels, fingerprint and daily intake of polycyclic
aromatic hydrocarbons (PAHs) in bread baked using wood as fuel. Journal of
Hazardous Materials, 164(2–3), 876–883.
Rey-Salgueiro, L., García-Falcón, M. S., Martínez-Carballo, E., & Simal-Gándara, J.
(2008). Effects of toasting procedures on the levels of polycyclic aromatic
hydrocarbons in toasted bread. Food Chemistry, 108(2), 607–615.
Scientific Committee on Food, SCF. (2002). Polycyclic aromatic hydrocarbons –
Occurrence in foods, dietary exposure and health effects. Report No. SCF/CS/CNTM/
PAH/29 Add1 Final, 4 December 2002. Available from <http://ec.europa.eu.int/
comm/food/fs/sc/scf/index_en.html> Retrieved 01.10.04.
Tuominen, J. P., Pyssalo, H. S., & Sauri, M. (1988). Cereal products as a source of
polycyclic aromatic hydrocarbons. Journal of Agriculture and Food Chemistry, 36,
118–120.
9
White, S., Fernandes, A., & Rose, M. (2002). Polycyclic aromatic hydrocarbons (PAHs)
in the UK 2000 total diet samples. Report No. FD 01/42, December 2002. York,
United Kingdom: Central Science Laboratory.
White, S., Fernandes, A., & Rose, M. (2008). Investigation of the formation of PAHs in
foods prepared in the home and from catering outlets to determine the effects of
frying, grilling, barbecuing, toasting and roasting. Report No. FD 06/13.1, May
2008. York, United Kingdom: Central Science Laboratory.
Yang, S. Y. N., Conell, D. W., Havker, D. W., & Kayal, S. I. (1991). Polycyclic aromatic
hydrocarbons in air, soil, and vegetation in the vicinity of urban roadway.
Science of the Total Environment, 102, 229–240.
Zhang, P., Song, J., & Yuan, H. (2009). Persistent organic pollutant residues in the
sediments and mollusks from the Bohai Sea coastal areas, North China: An
overview. Environment International, 35, 632–646.