Određivanje PAH4 jedinjenja u dimljenom mesu i dimljenim proizvodima od mesa - razrada metode

Smoked meat and smoked meat products represent a significant part of the human diet in Serbia. The process of smoking, i.e. wood combustion, is one of the most important sources of polycyclic aromatic hydrocarcarbons (PAHs). Considering the carcinogenic and mutagenic properties of some PAHs, these compounds have been in the spotlight of scientific interest. The EFSA (European Food Safety Authority) Panel on Contaminants in the Food Chain (CONTAM Panel) reviewed the available data on occurrence and toxicity of PAHs in food. In 2008, the CONTAM Panel concluded that benzo[a]pyrene is not a suitable indicator for the occurrence of PAHs in food. Based on the currently available data relating to occurrence and toxicity, the CONTAM Panel concluded that the sum of benzo[a]pyrene, chrysene, benz[a]anthracene and benzo[b]fluoranthene (PAH4) are the most suitable indicators of PAHs in food. These proposals have become part of the legislation both of EU and Serbia. From 1st September 2014, maximum residue limits (MRL) both for benzo[a]pyrene (2 μg/kg) and sum of PAH4 compounds (12 μg/kg), in smoked meat and meat products, were defined by the legislation of Serbia, which is in accordance with EU regulation. In this paper, the method has been developed for the determination of benzo[a]pyrene, chrysene, benz[a]anthracene and benzo[b]fluoranthene (PAH4 compounds) in smoked meat and smoked meat products. Accelerated solvent extraction (ASE) was used for extraction of lipids and lipophilic compounds. Solid Phase Extraction (SPE) was used in order to remove lipids from analysed samples. High-performance liquid chromatographic with fluorescence detection (HPLC-FL) was applied for identification and quantification of benzo[a]pyrene, chrysene, benz[a]anthracene and benzo[b]fluoranthene. Different conditions of HPLC-FL analysed were applied (mobile phase, HPLC column, oven temperature, flow) in order to achieve optimal conditions for qualitative and quantitative analysis of PAH4 compounds.Dimljeno meso i proizvodi od mesa zauzimaju važno mesto u ishrani stanovništva u Srbiji. S obzirom, da tokom dimljenja tj. procesom sagorevanja drveta nastaju policiklični aromatični ugljovodonici (Polycyclic Aromatic Hydrocarbons, PAH), koji su klasifikovani kao karcinogena i mutagena jedinjenja, proučavanje ovih jedinjenja uvek zaokuplja pažnju javnosti. Na osnovu podataka, koji su rezultat novih bioloških i toksikoloških naučnih istraživanja, evropska naučna komisija o zagađivačima u lancu ishrane je u junu 2008. godine predložila da se suma sadržaja benzo[a]pirena, benzo[a]antracena, benzo[b]fluorantena i hrizena, tj. PAH4 jedinjenja koristi kao marker prisustva drugih PAH jedinjenja u različitoj hrani. Ovi predlozi postali su deo zakonske regulative, kako Evropske unije, tako i Srbije. Zakonska regulativa Srbije, koja je u saglasnosti sa propisima EU, od 1. septembra 2014. godine definiše maksimalno dozvoljenu količinu (MDK) za sumu sadržaja PAH4 jedinjenja (12 μg/kg), kao i za sadržaj benzo[a]pirena (2 μg/ kg) u dimljenom mesu i proizvodima od mesa. U ovom radu razvijena je metoda za određivanje benzo[a]pirena, benzo[a]antracena, benzo[b]fluorantena i hrizena tj. PAH4 jedinjenja u dimljenom mesu i proizvodima od mesa. Za ekstrakciju lipida i lipofilnih jedinjenja iz uzoraka korišćena je ubrzana ekstrakcija pomoću rastvarača. Ekstrakcija na čvrstoj fazi je korišćena kao postupak za uklanjanje molekula lipida iz ispitanih uzoraka. Identifikacija i kvantifikacija benzo[a]pirena, benzo[a]antracena, benzo[b]fluorantena i hrizena rađena je korišćenjem visokoefikasne tečne hromatografije sa fluorescentnim detektorom (HPLC-FL). Primenjeni su različiti uslovi HPLC analize (mobilna faza, HPLC kolona, temperatura peći, protok mobilne faze) u cilju postizanja optimalnih uslova za kvalitativnu i kvantitativnu analizu PAH4 jedinjenja

Dietary Exposure and Risk Assessment of Aflatoxin M1 for Children Aged 1 to 9 Years Old in Serbia

The present study was conducted to estimate the exposure and characterize the risk for the child population of Serbia to Aflatoxin M1 (AFM1) from milk and milk-based food. A total of 3404 samples comprising milk and different milk-based food samples were collected from various regions of Serbia from 2017 to 2019. Evaluation of AFM1 exposure was carried out using the deterministic method, whereas risk characterization was evaluated using the margin of exposure (MOE) and the risk of hepatocellular carcinoma (HCC). Detection rates for AFM1 in milk and milk-based food samples ranged between 2% and 79%, with the highest incidence (79%) and mean level (22.34 ± 0.018 ng kg−1) of AFM1 being detected in pasteurized and UHT milk. According to the three consumption estimates, the values of estimated daily intake (EDI) were higher for toddlers as compared with children aged 3–9 years. Children aged 1–3 years had the highest risk of exposure to AFM1 in milk, with an estimated daily intake of 0.164 and 0.193 ng kg−1 bw day−1 using lower bound (LB) and upper bound (UB) exposure scenarios, respectively. Such difference could result from the higher consumption to weight in younger children. Based on the estimated daily intake (EDI) found in this study, the risk of AFM1 exposure due to consumption of milk and milk-based food was low since the MOE values obtained were >10,000. In addition, the risk of HCC cases/year/105 individuals of different age groups showed that the value of HCC, using potency estimates of 0.0017 (mean), was maximum (0.00034) in the age group 1–3 years, which indicates no health risk for the evaluated groups. The present study revealed the importance of controlling and preventing AFM1 contamination in milk through continuous monitoring and regular inspection to reduce the risk of AFM1 exposure, especially in children

Konzervirana morska riba na srpskome tržištu: razine cinka, bakra i željeza i njihov doprinos dnevnom unosu ovih esencijalnih metala

The aim of this study was to determine the levels of Zn, Cu, and Fe in three canned fish species marketed Serbia to see if they meet recommended daily intake requirements or exceed safety limits. We collected a total of 207 samples of canned tuna, sardine, and mackerel, in oil or tomato sauce and analysed them with inductively coupled plasma mass spectrometry (ICP-MS) after acid digestion. The highest levels were obtained for Zn (15.1 mg kg-1) and Cu (1.37 mg kg-1) in sardine in oil and tomato sauce, respectively, and for Fe (18.98 mg kg-1) in mackerel in tomato sauce. Our results keep within the ranges reported by several national food databases and available literature data, with a few exceptions. Our findings also single out canned sardines as the richest source of the three essential elements combined. The estimated daily intake (EDI) of the three essential elements, however, was subpar, and ranged between 0.14 % and 0.72 % of the recommended dietary allowance (RDA) for Zn, Cu, and Fe.Cilj ovoga rada bio je utvrditi razinu cinka, bakra i željeza u trima vrstama ribljih konzervi sa srpskoga tržišta kako bi se ustanovilo zadovoljavaju li zahtjeve za preporučenim dnevnim unosom, ili pak prekoračuju sigurnosne granice. Sadržaj esencijalnih elemenata utvrđen je u ukupno 207 uzoraka konzerve tune, sardine i skuše primjenom masene spektrometrije s induktivno spregnutom plazmom (ICP-MS) nakon kisele digestije uzoraka. Najveći sadržaj cinka (15,1 mg kg-1) utvrđen je u sardini u ulju, bakra (1,37 mg kg-1) u sardini u umaku od rajčice, a željeza (18,98 mg kg-1) u skuši u umaku od rajčice. Dobiveni rezultati su u opsegu vrijednosti za analizirane elemente koje je objavilo nekoliko nacionalnih baza podataka o hrani, kao i u opsegu dostupnih literaturnih podataka, s nekoliko iznimaka. Rezultati istraživanja izdvajaju sardinu u konzervi kao najbogatiji izvor proučavanih esencijalnih elementa. Procijenjeni dnevni unos (EDI) bio je između 0,14 % i 0,72 % preporučenih dnevnih unosa cinka, bakra i željeza. Iako sardina u konzervi, u odnosu na tunu i skušu, doprinosi oko 1,5 puta više preporučenom dnevnom unosu (RDI) esencijalnih elemenata, konzervirana morska riba ne može se smatrati značajnim izvorom cinka, bakra i željeza u prehrani srpskoga stanovništva

Distribucija organohlornih pesticida i polihlorovanih bifenila u dve vrste riba iz Dunava

The distribution and contents of organochlorine pesticides (OCPs) and congeners of polychlorinated biphenyls (PCBs) were analyzed in Bream (Blicca bjoerkna) and Barbel (Barbus barbus) from the Danube River, near Batajnica. The analysed compounds were 16 OCPs (α-Hexachlorocyclohexane (α-HCH), β-Hexachlorocyclohexane (β-HCH), Hexachlorobenzene (HCB), γ-Hexachlorocyclohexane (γ-HCH), δ-Hexachlorocyclohexane (δ-HCH), Heptachlor, Aldrin, cis-Heptachloroepoxide (cis-HCE), trans-Heptachloroepoxide (trans-HCE), γ-Chlordane, p,p'-Dichlorodiphenyldichloroethylene (p,p'-DDE), α-Chlordane, Dieldrin, Endrin, p,p'-Dichlorodiphenyldichloroethane (p,p'-DDD), p,p'-Dichlorodiphenyltri-chloroethane (p,p'-DDT)) and 7 PCB congeners (IUPAC numbers 28, 52, 101, 138, 153, 180, 118). Determination and quantification of OCPs and PCBs were performed by a GC-ECD method. The proximate composition of fish fillets was determined by applying standard SRPS ISO methods. The statistically significant difference (p lt 0.05) was found between the OCPs content in Bream and Barbel, as well as between PCBs content in the analyzed fish filets. Among all analyzed pesticides, both fish contained the highest amounts of ΣDDT (expressed as sum of p,p'-DDT + p,p'-DDE + p,p'-DDD), (13.8 ng/g fish - Bream, 2.6 ng/g fish - Barbel). The sum of PCB congeners in Bream filets (40.8 ng/g fish) was significantly higher than in Barbel filets (7.2 ng/g fish). Results of Student's t-test (p = 0.05) showed the differences in the content of lipids, moisture, ash and calculated energy value between the analyzed fish filets. Four times higher fat content in Bream (4.25%) than in Barbel (1.07%) is one of the main reasons for higher content of OCPs and PCBs in Bream compared to Barbel, which were taken from the same location (The Danube River, Batajnica).U ovom radu prikazana je distribucija i sadržaj organohlornih pesticida (organochlorine pesticides, OCP) i polihlorovanih bifenila (polychlorinated biphenyls, PCB) u ribama krupatici (Blicca bjoerkna) i mreni (Barbus barbus) iz Dunava u blizini Batajnice. Ispitano je 16 organohlornih pesticida i 7 kongenera polihlorovanih bifenila. Anlizirana su sledeća jedinjenja: α-heksahlorocikloheksan (α-HCH), β-heksahlorocikloheksan (β-HCH), heksahlorobenzen (HCB), γ-heksahlorocikloheksan (γ-HCH), δ-heksahlorocikloheksan (δ-HCH), heptahlor, aldrin, cis-heptahloroepoksid (cis-HCE), trans-heptahloroepoksid (trans-HCE), γ-hlordan, α-hlordan, p,p'-dihlorodifenildihloroetilen (p,p'-DDE), dieldrin, endrin, p,p'-dihlorodifenildihloroetan (p,p'-DDD) i p,p'- dihlorodifeniltrihloroetan (p,p'-DDT) i PCB kongeneri označeni IUPAC brojevima 28, 52, 101, 138, 153, 180 i 118. Kvalitativna i kvanitativna ispitivanja ovih jedinjenja rađena su GC-ECD metodom. Sadržaj proteina, lipida, vlage i pepela u filetima riba određen je korišćenjem standardnih SRPS ISO metoda. Utvrđena je statistički značajna razlika (p lt 0,05) između sadržaja organohlornih pesticida u krupatici i mreni, kao i između sadržaja polihlorovanih bifenila u ispitanim ribama. Najveći sadržaj među ispitivanim organohlornim pesticidima, u obe vrste ribe utvrđen je za ΣDDT (pp'-DDT + pp'-DDE + pp'-DDD), (13,8 ng/g ribe - krupatica, 2,6 ng/g ribe - mrena). Ukupan sadržaj PCB jedinjenja u filetima krupatice (40,8 ng/g ribe) bio je značajno veći nego u filetima mrene (7,2 ng/g ribe). Na osnovu rezultata Studentovog t-testa (p = 0,05) utvrđeno je da postoji statistički značajna razlika u sadržaju masti, vlage, pepela i izračunate energetske vrednosti između fileta ispitanih riba. Četiri puta veći sadržaj masti u filetima krupatice (4,25%) u odnosu na filete mrene (1,07%) jedan je od glavih razloga većeg sadržaja organohlornih pesticida i polihlorovanih bifenila u krupatici nego u mreni, koje su uzete sa istog lokaliteta (Dunav, Batajnica)

Mlađ i konzumna kalifornijska pastrmka (Oncorhynchus mykiss) - hemijski sastav, sadržaj holesterola i masnokiselinski sastav fileta

Rainbow trout (Onchorhynchys mykiss) is well known fish species in the nature. Changes in proximate composition of fish meat are associated with age and size of fish. Cholesterol content in animal tissues is associated with feeding method and quality of food, in spite of regulatory mechanism of cholesterol synthesis and absorption. Objective of this study was determination and comparison of proximate composition, cholesterol content and fatty acid profile of fingerlings and marketable size rainbow trout from aquaculture. Samples of fingerlings (average mass of 99 g and length of 18.6 cm) and marketable size rainbow trout (average mass of 229 g and length of 23.3 cm) were collected in August 2010, in the fishpond 'Ribnik', Mrkonjić city, Republic Srpska - Bosnia and Herzegovina. Fingerlings and marketable size rainbow trout were fed complete mixture of similar composition for both fish categories (fish products, oils and fats, cereal products and oil seeds). Obtained results showed that there was no statistically significant difference (p> 0.05) in the content of total lipids (3.81%, fingerlings and 4.17%, marketable size trout) and ash (1.27%, fingerlings and 1.29%, marketable size trout). Higher protein content was determined in marketable size trout fillets (18.69%), as well as lower water content (75.40%) compared to their content in fingerlings (17.72% proteins and 77.11% water). Cholesterol content was 82.59 mg/100g (fingerlings) and 70.12 mg/100g (marketable size trout). Statistically significant differences (p lt 0.05) in content of total saturated fatty acids was established between fingerlings and marketable size trout (31.81% and 29.14%, respectively), also in polyunsaturated fatty acids-PUFA (33.93% and 36.78%, respectively) and n-3 fatty acids (17.17% and 19.20%, respectively). Content of monounsaturated fatty acids was similar and ranged from 33.30%, in fingerlings, to 33.05% in marketable size trout. There was no statistically significant difference (p > 0.05) in content of total n-6 fatty acids in fingerlings (16.76%) and marketable size fish (17.58%). Higher quantities of n-3 PUFA in fingerlings (17.17%) and commercial trout (19.20%) and lower quantities of n-6 PUFA (16.76% in fingerlings and 17.58% in marketable size trout) resulted in favourable n-3/n-6 ratio (1.02 in fingerlings and 1.09 in commercial trout). Content of eicosapentaenoic acid (EPA, C20:5 n-3) and docosahexaenoic acid (DHA, C22:6 n-3) was 2.78% and 8.21% in fingerlings and 3.36% and 9.29% in marketable size fish, respectively. Content of EPA+DHA in total fatty acids was 10.99% in fingerlings and 12.65% in marketable size rainbow trout. By consumption 200 g of this fish intake of desirable fatty acids, EPA and DHA, is 0.84 g in fingerlings, and 1.06 g in marketable size rainbow trout, which is in accordance with recommendation of the American Heart Association for persons with cardiovascular disease (daily intake: in total 1g EPA and DHA). Trout growth was accompanied by increase of the protein content, decrease of the water content and increase in content of PUFA especially n-3 essential fatty acids. Due to significant content of proteins and unsaturated fatty acids and lower amounts of fat, rainbow trout can be considered as one of the most valuable food stuffs in human nutrition.Kalifornijska pastrmka (Oncorhynchus mykiss) je jedna od najpoznatijih vrsta ribe u prirodi. Promene u hemijskom sastavu mesa ribe su povezane sa starošću i veličinom ribe. Sadržaj holesterola u tkivima životinja je u vezi sa načinom i kvalitetom ishrane, uprkos regulatornom mehanizmu sinteze i apsorpcije holesterola. Cilj ovih ispitivanja je bio određivanje i poređenje hemijskog sastava, sadržaja holesterola i masnokiselinskog profi la mlađi i konzumne kalifornijske pastrmke iz akvakulture. Uzorci mlađi (prosečne mase 99 g i dužine 18,6 cm) i konzumne kalifornijske pastrmke (prosečne mase 229 g i dužine 23,3 cm) su sakupljeni u avgustu 2010. godine u ribnjaku 'Ribnik', Mrkonjić Grad, Republika Srpska - Bosna i Hercegovina. Mlađ i konzumna pastrmka su hranjene kompletnom hranom za pastrmku, sličnog sastava (riblji proizvodi, ulja i masnoće, proizvodi od žita i semena uljarica). Rezultati ispitivanja su pokazali da nema statistički značajne razlike (p > 0,05) u sadržaju ukupnih lipida (3,81%, mlađ i 4,17%, konzumna pastrmka) i pepela (1,27%, mlađ i 1,29%, konzumna pastrmka). U filetima konzumne pastrmke je utvrđen veći sadržaj proteina (18,69%) i manji sadržaj vode (75,40%) u poređenju sa njihovim sadržajem u filetima mlađi (17,72% proteina i 77,11% vode). Sadržaj holesterola je bio 82,59 mg/100 g (mlađ) i 70,12 mg/100 g (konzumna pastrmka). U filetima mlađi i konzumne pastrmke utvrđena je statistički značajna razlika (p lt 0,05) u sadržaju ukupnih zasićenih masnih kiselina (31,81% i 29,14%, respektivno), polinezasićenih masnih kiselina - PNMK (33,93% i 36,78%, respektivno) i n-3 masnih kiselina (17,17% i 19,20%, respektivno). Sadržaj mononezasićenih masnih kiselina je bio sličan i iznosio je u mlađi 33,30% i u konzumnoj pastrmci 33,05%. Nije postojala statistički značajna razlika (p > 0,05) u sadržaju ukupnih n-6 masnih kiselina u mlađi (16,76%) i konzumnoj ribi (17,58%). Veće količine n-3 PNMK u fi letima mlađi (17,17%) i konzumne pastrmke (19,20%) i manje količine n-6 PNMK (16,76% u mlađi i 17,58% u konzumnoj pastrmci) daju povoljan odnos n-3 i n-6 (1,02 u mlađi i 1,09 u konzumnoj pastrmci). Sadržaj eikozapentaenske (EPA, C20:5 n-3) i dokozaheksaenske kiseline (DHA, C22:6 n-3) bio je 2,78% i 8,21% u mlađi i 3,36% i 9,29% u konzumnoj ribi, respektivno. Sadržaj EPA+DHA u ukupnim masnim kiselinama je bio 10,99% u mlađi i 12,65% u konzumnoj kalifornijskoj pastrmci. Konzumiranjem 200 g ove ribe unos poželjnih masnih kiselina, EPA i DHA, iznosi 0,84 g za mlađ, odnosno 1,06 g za konzumnu kalifornijsku pastrmku, što je u skladu sa preporukom Američkog udruženja za srce za osobe sa kardiovaskularnim oboljenjima (dnevni unos: ukupno 1 g EPA i DHA). Rast pastrmke je bio praćen povećanjem sadržaja proteina, smanjenjem sadržaja vode i povećanjem PNMK, naročito n-3 esencijalnih masnih kiselina. Zbog značajnog sadržaja proteina i nezasićenih masnih kiselina i male količine masti, kalifornijska pastrmka se može svrstati u jednu od nutritivno najvrednijih namirnica u ishrani ljudi

Supplementary data for the article: Herceg Romanić, S.; Jovanović, G.; Mustać, B.; Stojanović-Đinović, J.; Stojić, A.; Čadež, T.; Popović, A. Fatty Acids, Persistent Organic Pollutants, and Trace Elements in Small Pelagic Fish from the Eastern Mediterranean Sea. Marine Pollution Bulletin 2021, 170, 112654. https://doi.org/10.1016/j.marpolbul.2021.112654.

Supplementary material for: [https://doi.org/10.1016/j.marpolbul.2021.112654]Related to publisshed version: [https://cherry.chem.bg.ac.rs/handle/123456789/4593

Emission of Polycyclic Aromatic Hydrocarbons from Beech Wood Combustion

This study reports results on 16 EU priority polycyclic aromatic hydrocarbons in smoke produced by beech wood combustion from Zlatibor region, Serbia. Smoke samples were collected with two different types of tubes (PUF and XAD-2), at a height of 2 and 5 m away from the firebox; 16 EU priority polycyclic aromatic hydrocarbons [European Commission (2005), Commission Recommendation 2005/108/EC, Official Journal of European Union L34:4345; JECFA (2005), Summary and Conclusion. Paper No. JECFA/64/SC, Joint FAO/WHO Expert Committee on Food Additives, Sixty-Fourth Meeting, Rome, Italy, February 817] were identified and quantified by Fast-GC/HRMS. The analyzed PAHs were: benzo[c]fluorene (BcL), benzo[a]anthracene (BaA), cyclopenta[c,d]pyrene (CPP), chrysene (CHR), 5-methylchrysene (5MC), benzo[b]fluoranthene (BbF), benzo[j]fluoranthene (BjF), benzo[k]fluoranthene (BkF), benzo[a]pyrene (BaP), benzo[g,h,i]perylene (BgP), dibenzo[a,h]anthracene (DhA), indeno[1,2,3-cd]pyrene (IcP), dibenzo[a,e]pyrene (DeP), dibenzo[a,h]pyrene (DhP), dibenzo[a,i]pyrene (DiP) and dibenzo[a,l]pyrene (DlP). Results show that total emission of polycyclic aromatic hydrocarbons at a height of 2 m was 6.7 and 7.4 mg m3 , in PUF and XAD-2 tubes, respectively. At height of 5 m polycyclic aromatic hydrocarbons emission was 2.4 and 4.3 mg m3 , in PUF and XAD-2 tubes, respectively. BaP contribution to total sum of 16 EU priority polycyclic aromatic hydrocarbons was 4% (PUF tubes) and 7.2% (XAD-2 tubes) at a height of 2 m, and 5% (PUF tubes) and 8.3% (XAD-2 tubes) at a height of 5 m. Total benzo[a]pirene equivalent concentrations (BaPeq) in smoke samples were calculated

Non-dioxin like polychlorinated biphenyls in mackerel (Scomber scombrus) available on the Serbian market

Mackerel is rich in omega-3 fatty acids, but this fish may be contaminated with environmental pollutants. The aim of this study was to evaluate concentrations of non-dioxin like polychlorinated biphenyls (ndl-PCBs) in 160 samples of mackerel available on the Serbian market. Average and maximum ndl-PCB concentrations were 7.9 ng/g and 74 ng/g, respectively. A higher average level of ndl-PCBs was found in mackerel from Spain, but this was still three times lower than the permitted level. We conclude that levels of ndl-PCBs in mackerel available on the Serbian market are acceptable

Cadmium in pheasant tissues as a bioindicator of environmental pollution in 23 Serbian districts

Concentrations of cadmium (Cd) were measured in leg muscle and liver of pheasants (n=316) from 23 different Serbian regions under the Serbian National Residue Monitoring Program during 2013-2015. Levels of Cd in pheasant tissues were determined by inductively-coupled plasma mass spectrometry (ICP-MS). Most of the pheasants examined had Cd in their tissues. In the third year of the residue monitoring program, the Cd concentrations detected in leg muscles were nearly double the levels detected in the first year. The highest Cd levels in muscles were measured in pheasants from Podunavlje, Macva and Zlatibor districts. All leg muscle samples had Cd concentrations below permitted maximum residue levels (MRL=0.05 m kg-1). However, fifteen liver samples had Cd levels that exceeded the permitted MRL (0.50 mg kg-1). The number of non-compliant pheasant liver samples increased over the years. The lowest mean Cd level in pheasant livers was measured in birds from Morava district (0.035 mg kg-1), while the highest was in birds from Branicevo district (0.574 mg kg-1). This monitoring program shows that the Cd levels measured in pheasants, which are suitable bioindicators of environmental pollution, indicate that environmental pollution with Cd is increasing

Benzo[a]pyrene, benz[a]anthracene, benzo[b]fluoranthene and chrysene in smoked meat and smoked meat products: Validation of the method

Polycyclic aromatic hydrocarbons (PAHs) are products of the incomplete combustion or pyrolysis of organic material and they are among the most harmful compounds. During process of meat smoking, wood combustion is one of the most important sources of PAH compounds, which can be adsorbed by the surface of meat. The EFSA (European Food Safety Authority) Panel on Contaminants in the Food Chain (CONTAM Panel) recommended to the member states of European Union to use the sum of benzo[a]pyrene (BaP), benz[a]anthracene (BaA), benzo[b]fluoranthene (BbF) and chrysene (CHR), (PAH4 compounds), as a marker for the occurrence and impact of carcinogenic PAHs in food, instead of benzo[a]pyrene. The maximum content of BaP and sum of all four compounds (PAH4) has been established by European Commission Regulation No. 835/2011. For smoked foods, from 1st September 2014, the maximum BaP content was lowered to 2 μg/kg, while the content of PAH4 is allowed to 12 μg/kg. The new maximum residue limits (MRL) both for BaP and sum of PAH4 compounds in smoked meat and meat products were defined by the legislation of Serbia, as well, and it is in accordance with EU regulation. The aim of this paper was the validation of the method for identification and determination of benzo[a]pyrene, chrysene, benz[a]anthracene and benzo[b]fluoranthene in smoked meat and smoked meat products. Accelerated solvent extraction (ASE) was used for extraction of lipids and lipophilic compounds. Solid Phase Extraction (SPE) was used in order to remove lipids from analysed samples. High-performance liquid chromatographic with fluorescence detection (HPLC-FL) was applied for identification and quantification of PAH4 compounds. Fluorescence detector operated at excitation/emission wavelength 275/385 nm for BaA and CHR, 256/446 nm for BbF and 260/410 nm for BaP, respectively. On the base of experimental results it is possible to conclude as follows: The in-house validation procedure of the method meets all criteria (applicability, specificity, repeatability, reproducibility, recovery, LOD and LOQ) set out by EU Regulation No. 836/2011. The method is suitable for operative control of PAH4 content in smoked meat products. [Projekat Ministarstva nauke Republike Srbije, br. III 46009