Determination of PAH4 compounds in smoked meat and meat products: Elaboration of the method

Abstract

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.