3. FOOD ChEMISTRy & bIOTEChNOLOGy 3.1. Lectures

Chem. Listy, 102, s265–s1311 (2008) Food Chemistry & Biotechnology
P07 CONTENT OF POLyCyCLIC AROMATIC
hyDROCARbONS IN hONEyS FROM ThE
CZECh REPubLIC
PETRA BATELKOVá, IVAnA BORKOVCOVá, KLáRA
BARTáKOVá and LEnKA VORLOVá
University of Veterinary and Pharmaceutical Science Brno
Palackého 1/3, 612 42 Brno,
pbatelkova@vfu.cz
Introduction
Polycyclic aromatic hydrocarbons (PAHs) belong to the
family of persistent organic pollutants having many properties
with a negative impact on human organism (carcinogenicity,
mutagenicity).
In addition to the inhaled air the main exposition source
of polycyclic aromatic hydrocarbons for the most of population
are foodstuffs. PAHs were found in many foodstuffs both
of animal origin (mainly as a result of culinary treatment) and
plant origin (due to atmospheric deposition and growing in
the contaminated soil). The data about PAHs content in honey
are rare 1 . In the study dealing with the PAHs content in honey
considerably high concentrations are reported 2 , especially
for the honeydew honey, not only for those PAHs with 3–4
aromatic rings (acenaphten 187 μg kg –1 , fluoren 163 μg kg –1 ,
phenanthren 625 μg kg –1 , anthracen 635 μg kg –1 ) predominantly
occurring in foodstuffs ant materials of plant origin 3 ,
but also for the toxic PAHs (benzo-k-fluoranthen 58 μg kg –1 ,
benzo-a-pyren 126 μg kg –1 ).
The aim of this study was to estimate content of PAHs
in Czech honeys whose high quality is known (absence of
antibiotics, minimal content of hydroxymethylfurfural). The
content of 15 priority pollutants according to the US EPA was
monitored, namely naphtalen (nAPT), acenaphten (ACEn-
APT), fluoren (FLU), anthracen (AnT), fluoranthen (FLT),
pyren (PY), benzo-a-anthracen (BaA), chrysen (CHRY)
benzo-b-fluoranthen (BbF), benzo-k-fluoranthen (BkF),
benzo-a-pyren (BaP), dibenzo-a,h-anthracen (DBahA),
benzo-g,h,i-perylen (BghiPE) and indeno-1,2,3-cd-pyren
(IPY).
Experimental
I n s t r u m e n t s
The Waters 2695 Alliance chromatographic system
was equipped with the Waters 2475 fluorescence detector.
A column PAH C18 (250 mm × 4.6 mm I.D. 5 µm (Waters,
Germany) was used. The column was held at 30 °C with
a column heater. Mobile phase A: water, B: acetonitrile,
flow 1.4 ml min –1 , linear gradient were used. PAHs were
detected by the fluorescent detector using an excitation and
emission wavelength program (λ exc ranged from 232 nm to
300 nm and λ em from 330 nm to 500 nm). The HPLC system
was controlled and the data were processed by Waters
Empower 2 software.
s587
C h e m i c a l s a n d R e a g e n t s
Acetonitrile (Merck KGaA), dichloromethane (Merck
KGaA) and hexane (Scharlau), the PAHs standard mixture
(PAH mix 9, Dr. Ehrenstorfer), sodium sulphate anhydrous,
sodium chloride, HPLC water. All solvents were HPLC grade
or for residual analysis, other chemicals p.a. quality at least.
S a m p l e s
10 samples of honey obtained directly from Czech beekeepers
and 10 samples of honey from shops with the Czech
Republic declared as a country of origin were analysed. Each
sample was analysed in at least duplicates, blank samples
in each series were performed.
P r o c e d u r e
Three kinds of procedures of sample preparation were
used.
The first procedure: 10 g sample of honey was dissolved
in 100 ml of deionised water, 1 g naCl and 10 ml hexane
added and thoroughly shaken for 30 min. By means of separatory
adapter an aliquot of organic layer was taken and evaporated
to dryness. The residue was dissolved in 1 ml of acetonitrile,
filtered through nylon membrane filter (0.45 μm)
and analysed by HPLC.
The second procedure: 10 g of honey sample was mixed
with anhydrous sodium sulphate, added 40 ml of dichloromethane
and extracted by means of ultrasonic bath and Ultra
Turrax. After filtration the solvent was evaporated and next
process was the same as in the first procedure.
The third procedure: 10 g of sample was dissolved
in 100 ml of deionised water and extracted by means of solid
phase of SPE cartridges. The next process was as in the first
procedure.
Results
Fig. 1. represents typical chromatogram obtained with
honey sample. The average concentrations of each PAHs
ranged from 0,02 to 2.22 μg kg –1 (Table I). Recoveries,
which were calculated by using observed and spiked concentrations
for PAHs, ranged from 60 to 90 %. Limit of quanti-
Table I
Concentrations of PAHs [μg.kg –1 ] in samples of honey
honey from nAPT ACEnAP FLU PHE
bee-keepers 0.29 0.20 0.66 0.32
shops 0.82 0.45 0.49 2.22
AnT FLT PY BaA
bee-keepers 0.07 0.02 0.15 0.55
shops 0.07 0.07 0.05 0.16
CHRY BbF BkF BaPY
bee-keepers 0.15 0.04 0.08 0.05
shops 0.02 0.02 0.02 0.02
DBahA BghiPE IPY
bee-keepers 0.26 0.10 0.07
shops 0.09 0.0 0.02