Assessment of dietary exposure to polycyclic aromatic hydrocarbons from smoked meat products produced in Latvia

Food Control 54 (2015) 16e22

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Assessment of dietary exposure to polycyclic aromatic hydrocarbons from smoked meat products produced in Latvia le, Ilze Stumpe-Vıksna*, Dzintars Za Irina Rozenta cs, Inese Siksna, Aija Melngaile, Vadims Bartkevi cs Institute of Food Safety, Animal Health and Environment “BIOR”, Lejupes Street 3, Riga, LV, 1076, Latvia

a r t i c l e i n f o

a b s t r a c t

Article history: Received 25 September 2014 Received in revised form 6 January 2015 Accepted 19 January 2015 Available online 28 January 2015

A total of 128 samples of smoked meat and meat products produced in Latvia were analysed using gas chromatography coupled to tandem mass spectrometry (GC-MS/MS) for the content of four priority polycyclic aromatic hydrocarbons (PAHs), including benzo[a]pyrene (BaP), chrysene (Chr), benzo[a] anthracene (BaA) and benzo[b]fluoranthene (BbF). The median content of BaP was 0.21 mg kg
1, significantly below the current maximum level (ML) of 5.0 mg kg
1. The highest content was observed for BaA and Chr with median values of 0.76 and 0.82 mg kg
1, respectively. All median values of individual PAH content and the combined mean levels of four PAHs (PAH4) were higher in smoked chicken samples (7.96 mg kg
1). The highest level of PAH4 was found in smoked pork speck (34.65 mg kg
1). The results showed that almost 14% of Latvian origin smoked meat products will be non-compliant to the new permitted level of BaP expected to be introduced in the European Union in September 2014 (2.0 mg kg
1). To estimate the exposure of Latvian population to polycyclic aromatic hydrocarbons, national individual consumption data and contamination data were analysed. The margin of exposure (MOE) approach indicates that exposure to PAH through smoked meat does not suggest significant toxicological concern for Latvian consumers. © 2015 Elsevier Ltd. All rights reserved.

Keywords: Polycyclic aromatic hydrocarbons (PAHs) Risk assessment Meat products Food safety

1. Introduction Smoking is one of the oldest technologies for the conservation of meat and meat products. During smoking phenolic substances are generated, which have considerable importance in organoleptic properties of smoked meat products. Besides that, phenolic compounds show antimicrobial and antioxidant properties. Nowadays, smoking technology uses mainly the specific effects of various sensory active compounds contained in smoke aromatization of meat products with suitable organoleptic profile, widely demanded on the market (Lorenzo, Purrinos, Garcia Fontan, & Franco, 2010). As undesirable consequence of smoking, PAHs are generated during the incomplete combustion of wood, comprising a large group of organic compounds with two or more fused aromatic rings, and represent a class of organic pollutants that are carcinogenic, teratogenic, and mutagenic (Conde, Ayala, Afonso, & Gonzalez, 2005; Janoszka, Warzecha, & Bodzek, 2004; Yoon, Park, Lee, Yang, &

* Corresponding author. E-mail address: [email protected] (I. Stumpe-Vıksna). http://dx.doi.org/10.1016/j.foodcont.2015.01.017 0956-7135/© 2015 Elsevier Ltd. All rights reserved.

Lee, 2007). The risk from exposure to PAHs depends on the type of diet, eating habits, cooking and smoking practices, which often are linked to regional traditions (Reinik et al., 2007). The composition and concentration of PAHs contaminating smoked meat products depends on multiple factors: type of wood, its moisture content, as well as the temperature achieved during  n, & Simal-Ga ndara, 2005; Guille n, smoke generation (García-Falco Sopelan, & Partearroyo, 2000). As PAHs represent an important class of carcinogens, their presence and amount in food has been intensively studied. The main attention has been paid to the benzo[a]pyrene. The EU Scientific Committee on Food (SCF) has identified 15 PAHs as genotoxic carcinogens, namely: benz[a]anthracene, benzo[b] fluoranthene, benzo[j]fluoranthene, benzo[k]fluoranthene, benzo [a]pyrene, benzo[g,h,i]perylene, chrysene, cyclopenta[c,d]pyrene, dibenz[a,h]anthracene, dibenzo[a,e]pyrene, dibenzo[a,h]pyrene, dibenzo[a,i]pyrene, dibenzo[a,l]pyrene, indeno[1,2,3-cd]pyrene, and 5-methylchrysene. Due to the carcinogenic properties, the PAH content in food should be as low as reasonably achievable (European Commission recommendations, 2005).

le et al. / Food Control 54 (2015) 16e22 I. Rozenta

Currently MLs for benzo[a]pyrene and the sum of four PAHs (BaP, CHR, BaA and BbF) in smoked meat products are established in the European Union (EC, 2011). A 5 mg kg
1 ML for benzo[a]pyrene (BaP) in smoked meats and smoked meat products was introduced in 2006 (EC, 2006). EFSA concluded that BaP is not a sufficient indicator for the occurrence of PAHs in food and assessed that the PAH4 value is the most suitable criterion (Jira, 2010). A 30 mg kg
1 ML for the PAH4 value was introduced in 2011 [EC, 2011], and in September 2014 these MLs will be lowered to 2 mg kg
1 for BaP and to 12 mg kg
1 for PAH4, respectively (Commission Regulation (EC) No 1881/2006 amended by Commission Regulation (EU) No 835/ 2011). Over the last 10 years, several studies determining the content of BaP in smoked meat products from different European countries were performed. The BaP content in smoked meat and meat products reported by these studies indicates a significant influence of the smoking technology used. The meat processing industry is interested in research of PAHs concentration in smoked meat and meat products, in order to improve food safety of products and their compliance to the European Union regulations (Djinovic, Popovic, & Jira, 2008; Lorenzo et al., 2011; Santos, Gomes, & Roseiro, 2011). Latvia has a long tradition of meat smoking. Smoked meat is produced not only by large meat processing facilities, but also at home and by small companies that produce products according to the traditional recipes. The main objective of the present study was to determine the content of the EU regulated PAH compounds in representative samples of smoked meat products, including those from companies applying traditional smoking methods, and to assess the exposure of certain Latvian population groups to polycyclic aromatic hydrocarbons from smoked meat products. 2. Materials and methods 2.1. Food samples and sampling procedures For the study, 128 samples of various smoked meat products, produced by 48 different smoked meat manufacturers were obtained in local markets. Food samples included: meat (pork, pork breast, chop, speck, smoked ham, smoked chicken), and meat products (sausages, small sausages, Semi-dry sausages, roulette and other). Samples were collected in conformity with requirements laid down in Commission Regulation (EC) No 333/2007 of 28 March 2007, specifying the methods of sampling and analysis for the official control of the levels of lead, cadmium, mercury, inorganic tin, 3-MCPD, and polycyclic aromatic hydrocarbons in foodstuffs. Samples were processed immediately upon receiving at the laboratory.

17

deuterated standards benzo[a]pyrene-d12, benzo-[b]fluoranthened12, chrysene-d12, and benz[a]anthracene-d12 were purchased from Dr. Ehrenstrofer. The standard mix of PAHs consisted of a solution in acetonitrile with concentration 50 mg L
1 and the concentration of deuterated benzo[a]-pyrene-d12, benzo-[b]fluoranthene-d12, chrysene-d12, benz[a]anthracene-d12 dissolved in cyclohexane was 1000 ng ml
1. The mixtures were stored at 4  C.

2.4. Sample preparation for the analysis Whole samples were homogenized (including skin and muscle) without bones, minced into smaller pieces, and blended. Each homogenised meat sample (2.75 g) was thoroughly mixed with 15 g of anhydrous sodium sulphate to absorb moisture. An aliquot of 27.5 mL toluene solution of internal standards including benzo(a) pyrene, chrysene, benzo(b)fluoranthene, and benzo(a)anthracene with concentration 0.500 mg mL
1 was added. The PAHs were extracted from meat samples by adding 25 mL of dichlormethane/ hexane (1:1, v/v) mixture and performing sonication of about 20 min. After sonication the supernatant of the extracts were decanted and 15 mL of fresh solvent was added for another 20 min of sonication. To avoid the presence of solid particles, all the extracts were filtered. The combined extracts (~40 mL) were rotary evaporated (30  C, 500-100 mbar) to eliminate the solvents, and the fat residue was reconstituted in 5.5 mL of cyclohexane/ethyl acetate (1:1, v/v) solution for further elimination of high molecular compounds by means of gel permeation chromatography (GPC). The extracts were centrifuged at 3000 rpm for 10 min and the solution was transferred into a glass GPC vial. The sample extracts were injected into LC Tech Freestyle™ GPC system (Dorfen, Germany) consisting of an HPLC pump, autosampler, and fraction collector. High molecular substances were removed on a glass column (500  40 mm, 25 mm ID) filled with 50 g of Bio-Beads SX3 (Bio-Rad, Philadelphia, USA) stationary phase with cyclohexane/ ethyl acetate (1:1, v/v) mobile phase at a flow rate of 5 mL min
1. The automated GPC program was as follows: dump time 0e21 min, collection time 21e45 min. The collected fractions were transferred to round-bottom flasks and then rotary evaporated (30  C, 130 mbar) to dryness. The dry residue was dissolved in 3 mL of cyclohexane. Further clean-up was done using SPE cartridges filled with 500 mg of silica. The sorbent of the SPE cartridges was first conditioned with 5 mL of cyclohexane, after which extracts were loaded onto the cartridges. The analytes of interest were eluted from the column with cyclohexane (3  3 mL), the obtained fraction was evaporated under a stream of nitrogen at 40  C, dissolved in 50 mL of cyclohexane, and transferred into a GCeMS/MS vial for the further PAHs analysis.

2.2. Materials and reagents

2.5. Instrumental analysis

Solvents employed were cyclohexane, hexane, dichloromethane, and ethyl acetate, all of them were of pesticide purity grade. Other reagents and materials used were anhydrous sodium sulphate, and 6 mL (500 mg) Phenomenex Strata SI-1 Silica solidphase extraction (SPE) tubes. All the aforementioned solvents, reagents, and materials were commercially purchased from SigmaeAldrich (Steinheim, Germany), Supelco (Bellefonte, PA), and Merck (Darmstadt, Germany).

The PAHs analysis was carried out by ThermoScientific TSQ Quantum XLS Ultra GCeMS/MS system equipped with a DB-17 capillary column (30 m long  0.25 mm i.d.  0.25 mm film thickness) and operating in a splitless mode. The operating conditions were as follows: helium gas was used as the carrier gas at a constant flow of 1.2 mL/min; inlet temperature 260  C; MS transfer line temperature 280  C; source temperature 250  C. The oven temperature was set initially at 80  C (2 min hold), increased to 265  C at 15  C/min. At 265  C, temperature increased at a rate of 5  C/min to 290  C and then to 320  C at a rate of 20  C/min (20 min hold). The total run time was 45.8 min. The injection volume was 1 mL. The data were acquired by operating the MS in selective reaction monitoring mode (SRM).

2.3. Standards Mixture of 4 PAH standards: benz[a]anthracene (BaA), benzo-[b] fluoranthene (BbF), benzo[a]pyrene (BaP), chrysene (Chr), and

18

le et al. / Food Control 54 (2015) 16e22 I. Rozenta

2.6. Quality assurance/quality control

2.9. Statistical analysis

Identification criteria for the analytes of interest were based on the retention times of native PAHs and deuterated PAH surrogates, and the isotopic peak ratios of the SRM transitions. The acceptable deviation of the isotope ratio of two monitored ions or SRM transitions (target/confirmation) was within 15% of the value obtained for the medium calibration point. A five-point calibration curve was checked with relative response factors (RRFs) over the sample concentration range of 1.00e50.0 ng g
1 and was used for quantifying the analytes of interest in each sample run. Procedural blanks consisting of anhydrous sodium sulphate aliquots typically used for routine samples were taken through all steps of analytical procedure and analysed in each sample sequence. The procedural blanks were found to be uncontaminated with the analytes of interest. The quantitation of analytes of interest was based on stable isotope dilution with the deuterated PAH surrogates and on internal standardization. The LOQ values were evaluated on the basis of noise obtained with the analysis of the blank samples, and were defined as the concentration of analyte that produced a signal-to-noise ratio of 10.

The one-way analysis of variance (ANOVA) and t-test with the Microsoft Excel Data Analysis Toolpack was used to test the differences between PAHs in different food samples. A criterion of p < 0.05 was considered to indicate statistical significance.

2.7. Smoked meat consumption data evaluation The dietary survey was conducted in the year 2012 and almost two thousand participants from the age group of 19e64 were reached. Due to the fact that smoked meat consumption is a specific part of food consumption, focussed research for smoked meat consumption in Latvia in the age group of 19e64 was carried out in 2014. For dietary survey, the food frequency questionnaire and 24 h recall method were used. An additional questionnaire for smoked meat producers about processing technologies was used.

2.8. Risk characterization The actual contribution of smoked meat products in the overall exposure to BaP and PAH4 was studied during this research. The scientific opinion of EFSA on a harmonized approach for risk assessment of substances which are both genotoxic and carcinogenic (EFSA, 2005) was used to characterize the risk related to consumption of smoked meat products. In general, it was assumed that the margin of exposure (MOE) of 10000 or higher would be of low concern from the viewpoint of public health and might be considered low priority for risk management actions. The BMDL10 (benchmark dose lower confidence limit 10%) - an estimate of the lowest dose which is 95% certain to cause no more than a 10% cancer incidence in rodents e was used to obtain the MOE. Taking into account the findings of the EFSA research on PAHs in food, the BMDL10 for BaP was 0.07 mg kg
1 b.w. per day, and the BMDL10 for the PAH4 was 0.34 mg kg
1 b.w. per day, that were used as reference values in the MOE calculations (EFSA, 2008). The MOE values were calculated by dividing the reference BMDL10 values with the mean, median, and 75 percentile estimates of the dietary exposure to BaP and PAH4. To calculate the exposure of the whole population and specific population groups, data on the mean, median, and 75 percentile consumption of smoked meat products were used. Comparison of the MOE indicators within different groups of consumers was carried out to conclude whether consumption of smoked meat products could present a risk to public health, taking into account the characteristic consumption patterns.

3. Results and discussion 3.1. PAHs in smoked meat products In this study, 128 samples of smoked meat products were analysed and the content of four PAHs was determined. Table 1 shows the mean, median, minimum, and maximum concentrations of single PAH compounds, as well as the total content of PAH4. In some samples the concentration of BaP and BbF was below the limit of quantification (LOQ). The median of BaP contents was 0.21 mg kg
1, being significantly below the ML of 5.0 mg kg
1. The higher content was observed for BaA and Chr with median values of 0.76 and 0.82 mg kg
1, respectively. Results showed (see Table 2) that all median values of individual PAH content and the mean levels of PAH4 were higher in smoked chicken samples (7.96 mg kg
1), although no significant differences (p > 0.05) were observed. The highest level of PAH4 for individual samples was found in smoked pork speck (34.65 mg kg
1) that could be due to the high concentration of Chr in some samples. An important factor for PAH contamination is the surface/mass ratio. General smoked chicken meat being of lesser size and thickness than smoked pork meat, showed a larger surface per unit of volume, which causes the elevated concentration of PAHs. The PAH values observed in smoked chicken and smoked pork meat were higher than those reported for smoked meat products in Italy, Estonia, and Germany (Jira, 2010; Purcaro, Moret, & Conte, 2009; Reinik et al., 2007). However, data from previous studies show that during traditional smoking of meat high levels of BaP appear. Wretling, Eriksson, Eskhult, and Larsson (2010) reported 9 (out of 38) samples with high BaP lavels ranging from 6.6 to 36.9 mg kg
1 in Swedish smoked meat samples exceeding the 5 mg kg
1 level. High levels of BaP were detected in samples where traditional sauna smoking is used. Also, our research shows that the higher concentrations of PAHs appear in samples from small producers, where traditional smoking methods could be used and the intensity of smoke deposition is uncontrolled, thus depends on the environmental conditions (temperature and relative humidity), and the type of wood used. In this case, the foodstuff is in direct contact with all components of the generated smoke and it could be highly contaminated with PAHs. Contrary to traditional smoking methods, smoke production in industrial smoking ovens is closely controlled and the removal of undesirable compounds is facilitated by the smoke generators being separated from the smoking chamber (Simko, 2005).

Table 1 Mean, median, minimum, and maximum concentrations of single PAH4 compounds and sum of PAH4 in smoked meat products (n ¼ 128). Compound

Mean, mg kg
1

Median, mg kg
1

Maximum, mg kg
1

Minimum, mg kg
1

BaA Chr BbF BaP PAH4 sum

2.38 2.44 0.82 0.74 6.36

0.76 0.82 0.32 0.21 2.10

14.21 14.50 4.60 6.03 34.65

0.05 0.10 <0.05 <0.05 0.15

le et al. / Food Control 54 (2015) 16e22 I. Rozenta Table 2 Average and median concentrations and range of individual PAHs and PAH4 sum in each type of meat.

19

Table 4 The occurrence of BaP in smoked meat products. BaP concentration

Concentration (mg kg
1) BaA

Chr

Smoked pork, n ¼ 14 Average 2.73 2.78 Median 1.11 1.25 Min-max 0.05e10.60 0.10e11.07 Samples >0 14 14 Smoked pork breast, n ¼ 18 Average 2.26 2.27 Median 0.74 0.67 Min-max 0.06e9.60 0.10e11.24 Samples >0 18 18 Smoked chop, n ¼ 12 Average 2.37 2.21 Median 1.07 0.88 Min-max 0.08e8.36 0.14e8.30 Samples >0 12 12 Smoked pork speck, n ¼ 10 Average 2.72 3.08 Median 0.72 0.80 Min-max 0.13e12.23 0.25e14.50 Samples >0 10 10 Smoked ham, n ¼ 4 Average 1.88 2.17 Median 0.56 0.69 Min-max 0.12e6.26 0.29e6.99 Samples >0 4 4 Smoked chicken, n ¼ 12 Average 3.30 2.98 Median 2.29 2.01 Min-max 0.30e12.32 0.63e12.08 Samples >0 12 12

<0.05 mg kg
1 2 mg kg
1 >2 mg kg
1 >5 mg kg
1 BbF

BaP

PAH4

1.11 0.37 <0.05e4.60 13

1.05 0.29 <0.05e6.03 13

7.68 3.07 0.15e27.25 14

0.81 0.33 <0.05e4.28 16

0.60 0.23 <0.05e2.99 16

5.92 1.83 0.16e28.10 18

0.93 0.44 <0.05e3.00 11

0.99 0.24 <0.05e4.17 8

6.48 2.95 0.23e20.16 12

1.05 0.34 0.11e4.29 10

0.91 0.31 <0.05e3.62 9

7.75 2.23 0.48e34.65 10

0.81 0.34 0.11e2.46 4

0.66 0.25 <0.05e2.10 3

5.50 1.84 0.52e17.81 4

0.94 0.62 0.20e4.55 12

0.74 0.47 0.08e2.80 12

7.96 5.38 1.21e31.74 12

All analysed sausages were distributed in three groups e smoked sausages, small sausages, and semi-dry sausages. The highest average concentrations of BaA (2.40 mg kg
1), Chr (2.58 mg kg1), BbF (0.77 mg kg
1), BaP (0.84 mg kg
1), and PAH4 sum (6.57 mg kg
1) were found in smoked sausage samples (see Table 3). Significant (p < 0.05) lower concentrations of BaP with an average value of 0.11 mg kg
1 were found in half dried sausage samples. Our results were higher than those obtained for Spanish (Lorenzo et al., 2010), Italian (Purcaro et al., 2009), and Swedish sausages, where Table 3 Average and median concentration and range of individual PAHs and PAH4 sum in smoked sausages and other smoked products. Concentration (mg kg
1) BaA Smoked sausage, n ¼ 21 Average 2.40 Median 0.31 Min-max 0.08e14.21 Samples >0 21 Crackers or small sausages, Average 1.92 Median 0.36 Min-max 0.19e13.13 Samples >0 10 Semi-dry sausage, n ¼ 8 Average 0.43 Median 0.36 Min-max 0.13e0.88 Samples >0 8 Roulette, n ¼ 6 Average 0.35 Median 0.30 Min-max 0.12e0.78 Samples >0 6

Chr

BbF

BaP

PAH4

2.58 0.49 0.13e11.19 21 n ¼ 10 1.92 0.58 0.26e12.79 10

0.77 0.25 <0.05e4.09 18

0.84 0.11 <0.05e4.17 14

6.57 1.30 0.22e33.66 21

0.68 0.23 0.13e4.03 10

0.67 0.08 0.05e4.63 10

5.19 1.19 0.63e34.58 10

0.54 0.53 0.21e1.01 8

0.22 0.24 0.10e0.33 8

0.11 0.07 <0.05e0.22 6

1.29 1.23 0.48e2.44 8

0.45 0.37 0.21e0.88 6

0.25 0.26 0.12e0.39 6

0.15 0.12 <0.05e0.36 5

1.19 1.06 0.46e2.40 6

Smoked meat products Smoked chicken Smoked pork Smoked pork breast Smoked chop Smoked pork speck Smoked ham Smoked sausage Small sausages Semi-dry sausage Roulette

n ¼ 128 18.8%

85.9%

14.1%

0.8%

n ¼ 12 n ¼ 14 n ¼ 18

0% 7.1% 11.1%

91.7% 78.6% 88.9%

8.3% 21.4% 11.1%

0% 7.1% 0%

n ¼ 12 n ¼ 10

33.3% 10.0%

83.3% 80.0%

16.7% 20.0%

0% 0%

¼ ¼ ¼ ¼

25.0% 33.3% 0% 20.0%

75.0% 81.0% 90.0% 100%

25.0% 19.0% 10.0% 0%

0% 0% 0% 0%

16.7%

100%

0%

0%

n n n n

4 21 10 8

n¼6

the content of BaP was below the limit of detection. (Wretling et al., 2010). Low BaP content of 0.13e0.16 mg kg
1 was determined in Danish sausages (Duedahl-Olesen, White, & Binderup, 2006) smoked by indirect smoking methods, followed by 0.24e0.33 mg kg
1 in smoked sausages from Serbia (Djinovic et al., 2008), 0.36e0.63 mg kg
1 in Portuguese traditional smoked meat and blood sausages (Santos et al., 2011), and 0.13e0.59 mg kg
1 (Lorenzo et al., 2010) in Spanish traditional smoked sausage varieties “Androlla” and “Botillo”. In a study of Italian traditional smoked sausages “Pitina” the BaP content was found 0.8 mg kg
1 (Purcaro et al., 2009). The higher concentrations of BaP and PAH4 sum were found in Portuguese traditional meat and blood sausages (Roseiro, Gomes, Patarata, & Santos, 2012), where BaP levels in meat products were from 0.36 up to 4.75 mg kg
1 and in blood-derived products from 0.32 to 5.65 mg kg
1. In these products very high BaA and Chr concentrations were found (up to 132.51 and 150.56 mg kg
1 respectively), hence the maximum PAH4 sum was found to be 294.50 mg kg
1, which is several times higher than the 30 mg kg
1 ML for PAH4. The proportion (%) of samples exceeding the maximum permitted level and planned maximum concentration of BaP from September 2014 are shown in Table 4, and for PAH4 sum in Table 5. The frequency of cases exceeding the EU specified limits for PAHs varied from 7% for smoked pork to 0% for other samples, although in the case if the 5 mg kg
1 ML for BaP of will be lowered to 2 mg/kg in September 2014, 14.1% of smoked meat products in Latvia will be non-compliant to the new permitted level of BaP. Regarding the sum of PAH4, currently 3.9% of all smoked meat samples exceed the existing EU limit (30 mg kg
1) and with the introduction of the new EU limits (12 mg kg
1) more than 20% of samples will exceed that limit. The overall results indicate that the production of smoked meat products with BaP contamination levels below 2.0 mg kg
1 and PAH4 concentration below 12.0 mg kg
1 for manufacturers applying traditional smoking methods is problematic, and a greater effort of changing processing practices and regional consumption habits is needed.

3.2. Consumption of smoked and grilled products in Latvia Food consumption database is the significant information source for risk assessment, since it contains information on food consumption habits in Latvia. Due to the fact that smoked meat

le et al. / Food Control 54 (2015) 16e22 I. Rozenta

20 Table 5 The occurrence of PAH4 in smoked meat products.

PAH4 sum concentrations

Smoked meat products Smoked chicken Smoked pork Smoked pork breast Smoked chop Smoked pork speck Smoked ham Smoked sausage Small sausages Semi-dry sausage Roulette

n n n n n n n n n n n

¼ ¼ ¼ ¼ ¼ ¼ ¼ ¼ ¼ ¼ ¼

128 12 12 18 12 10 4 21 10 8 6

12 mg kg
1

30 mg kg
1

>30 mg kg
1

78.9% 75.0% 71.4% 77.8% 75.0% 80% 75% 76.2% 90.0% 100% 100%

96.1% 91.7% 100% 100% 100% 90.0% 100% 95.2% 90.0% 100% 100%

3.9% 8.3% 0% 0% 0% 10.0% 0% 4.8% 10.0% 0% 0%

Table 6 Consumption of meat and fish during a year (margins of error reported at 95% level of confidence). N ¼ 1811

Median 75th Mean and Std. 25th Percentile margin of Dev. Percentile error

Meat, kg Fish, kg Meat and Fish, kg Smoked meat and fish, kg Grilled/barbequed meat and fish, kg Smoked, grilled, barbequed meat and fish, kg Smoked meat and fish, % from all meat and fish consumed Grilled/barbequed meat and fish, % from all meat and fish consumed Smoked, grilled, barbequed meat and fish, % from all meat and fish consumed

81.8 13.0 94.8 14.6 13.9

±3.0 ±0.8 ±3.4 ±0.9 ±1.0

65.9 34.9 18.2 2.6 74.1 42.1 18.9 3.3 22.2 1.2

64.1 6.7 73.7 8.3 5.3

110.8 15.4 125.2 18.2 17.3

28.6 ±1.5 31.5

7.3

17.6

37.0

15.0 ±0.6 12.2

6.0

12.2

20.9

14.0 ±0.7 15.3

2.1

8.2

21.6

29.0 ±0.8 17.8 14.7

27.3

41.2

consumption is a specific part of food consumption, focussed research for smoked meat consumption in Latvia in the age group of 19e64 years was performed in 2014. The data for smoked meat and fish consumption is presented in Table 6. Differences in meat consumption between men and women are statistically significant at exceptionally high levels (p < 0.0001 independent sample Student t-test, Mann-Whitney-Wilcoxon U-test). Women consume 171 g of meat per day, while men consume significantly more e 280 g per day. 3.3. Assessment of consumer preferences in relation to smoked products Respondents were asked to note products they use every day and those that are usually consumed on festive occasions. Semi-dry, dried sausages, and smoked chicken legs are consumed more on every day basis, but smoked whole chicken, pork chop, ham and chicken roulette are usually eaten on festive occasions. On average, 33% of consumers purchase smoked products in supermarkets, 23% buy at farmers markets, and 15% get them directly from farmers. Few families make smoked meat and fish products by themselves. Almost 15% do it on their own, while 10% mentioned that they have friends, relatives or neighbours who smoke meat for them. It is possible to slightly reduce the consumed amounts of BaP and PAH4 if the skin or dark rind of meat is removed. Almost half of consumers (47.6%) replied that they remove skin of smoked chicken. These habits are more common among women (62.2%) than men (33.1%). The differences are statistically significant at high level (Pearson c2 test p < 0.001). The consumers who remove skin from smoked products also consume less smoked products than those who do not. An average of 22% respondents have planned or have already reduced their smoked meat consumption to reduce intake of BaP and PAH4, but 23% mentioned that they would not change anything in their eating habits to reduce BaP and PAH and are worried that there will be changes in the taste of traditionally smoked products if levels of BaP and PAH4 have to be lowered. To detect what type and what colour of smoked meat products consumers prefer, they were offered photos of 4 groups of products and indicated their preferences. All the pictures shown in Fig. 1 contain products that are easily available on the market.

Fig. 1. Colour scale for assessment of surface colour of smoked chicken. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Fig. 2. Colour scale for the assessment of surface colour of smoked pork fat. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

le et al. / Food Control 54 (2015) 16e22 I. Rozenta

21

Fig. 3. Colour scale for the assessment of home-made sausage colour. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Table 7 The calculation of consumer exposure to BaP and PAH4, and calculation of MOE indicator for all consumers of smoked meat products (the mean calculated values of BaP concentration and PAH4 sum are used). Consumption of smoked meat products (kg day
1)

BaP PAH4 sum

Mean

Median

75 %tile

0.036 0.036

0.022 0.022

0.047 0.047

Mean BaP/SPAH4 conc. (mg kg
1)

0.73 6.36

Consumer exposure (ng kg
1 b.w. day
1)

BMDL10 (ng kg
1 b.w. day
1)

Mean

Median

75 %tile

0.33 2.91

0.20 1.78

0.44 3.80

70000 340000

MOE Mean

Median

75 %tile

209627 116868

343026 191238

160565 89516

Table 8 The calculation of consumer exposure to BaP and PAH4, and calculation of MOE indicator for all consumers of smoked meat products in the case of worst possible scenario (the maximum allowed values of BaP and PAH4 sum used). Consumption of smoked meat products (kg day
1)

BaP PAH4 sum

Mean

Median

75 %tile

0.036 0.036

0.022 0.022

0.047 0.047

Mean BaP/SPAH4 (mg kg
1)

5.00 30.00

Consumer exposure (ng kg
1 b.w. day
1) Mean

Median

75 %tile

2.29 13.72

1.40 8.39

2.99 17.92

Most consumers prefer chicken that is prepared and looks like in Fig. 1 sample a e 31%, but 26% rate d as the favourite. Our concern is that the product in Fig. 1, sample d has such high level because this is one of most easily available products in all supermarkets and the only one available on market that is packed in convenient vacuum packaging. The same situation is with smoked pork fat, where 38.8% of consumers prefer products which look like sample a in Fig. 2, while only 7% prefer the one in Fig. 2 sample c. In assessing the situation with home-made smoked sausages, we also concluded that most consumers (52.3%) prefer sausage with darker surface colour (sample a in Fig. 3). This is a product typically eaten with bread and mainly chosen by seniors, as it may have a strong taste of garlic and other spices.

3.4. Risk characterization

BMDL10 (ng kg
1 b.w. day
1)

MOE Mean

Median

75 %tile

70000 340000

30606 24776

50082 40542

23443 18977

Two different scenarios were developed for evaluating the possible impact of smoked meat products on public health. For the 1st scenario, the data on average BaP concentration and average PAH4 sum were used, taking into account the research findings. For the 2nd scenario, the worst possible scenario was simulated, and the data on the maximum allowed BaP concentration and the maximum allowed PAH4 sum were used (5 mg kg
1 and 30 mg kg
1, respectively, according to Regulation 1881/2006). Calculations of both the consumer exposure to BaP and PAH4, and the MOE indicators for all consumers of smoked meat products are demonstrated in Tables 7 and 8. The calculation of dietary exposures and MOEs suggests that MOE indicators for all consumer groups studied within this research far exceeds 10 000 e the MOE value derived on the base of precautionary approach and recommended by EFSA (Tables 9 and 10). The comparison of MOE indicators for different consumer groups with regard to BaP exposure revealed that a relatively higher dietary exposure and thus a comparatively lower MOE is

To characterize the risks due to PAHs, the margins of exposure (MOEs) were calculated for different groups of respondents. Table 9 Comparison of MOE indicators within different groups of consumers (dietary exposure to BaP). Groups of consumers

MOE (in case of mean BaP)

MOE (in the case of maximum allowed BaP/ worst possible scenario)

Mean

Median

75 %tile

Mean

Median

75 %tile

All consumers 19e35 years old 36e50 years old 51e64 years old Men Women

209627 212702 198174 217697 172803 261486

343026 350959 297260 383562 259204 424914

160565 171199 148630 167808 127608 206019

30606 31055 28933 31784 25229 38177

50082 51240 43400 56000 37844 62038

23443 24995 21700 24500 18631 30079

Table 10 Comparison of MOE indicators within different groups of consumers (dietary exposure to PAH4 sum). Groups of consumers

MOE (in the case of mean PAH4 sum)

MOE (in the case of maximum allowed PAH4 sum/worst possible scenario)

Mean

Median

75 %tile

Mean

Median

75 %tile

All consumers 19e35 years old 36e50 years old 51e64 years old Men Women

116868 118582 110482 121367 96338 180939

191238 195660 165723 213836 144507 294025

89516 95444 82862 93553 71142 142558

24776 25139 23422 25730 20424 30905

40542 41480 35133 45333 30635 50221

18977 20234 17567 19833 15082 24349

22

le et al. / Food Control 54 (2015) 16e22 I. Rozenta

characteristic for men and middle-age consumers; nevertheless, the MOE values were significantly higher than 10 000. The comparison of MOE indicators for different consumer groups with regard to the exposure to PAH4 sum indicated that a relatively higher dietary exposure and thus a comparatively lower MOE is characteristic for men, rather for women. It should be emphasized that even in the case of the worst possible scenarios e that is, if the consumers most at risk would always consume only meat products which are smoked according to the traditional methods, and if these smoked meat products would always contain the highest allowed concentration of BaP (5 mg kg
1) and PAH4 sum (30 mg kg
1), MOE indicators would still remain significantly above the reference value of 10 000. Thus, the MOE indicators calculated during the research demonstrate a low concern for consumer health at the mean, median, and 75 percentile exposures even in the case of worst possible scenarios.

4. Conclusions The content of BaP in the analysed smoked meat and smoked meat product samples varied from levels below the LOQ to 6.03 mg kg
1, while the content of PAH4 sum varied from 0.15 to 34.65 mg kg
1. The overall results indicate that almost 14% of Latvian origin smoked meat products will be non-compliant to the new permitted level of BaP expected to be introduced in the European Union in September 2014 (2.0 mg kg
1) and therefore the continued production of smoked meat products for manufacturers applying traditional smoking methods presents a challenge, requiring a greater effort towards improving the processing practices. In addition, consumption data in Latvia showed that many consumers purchase smoked meat products directly from small farmers or smoke meat and fish products by themselves, and most of consumers prefer products with darker surface colour. However, the risk characterization results using MOE approach have revealed a low risk to consumers' health. Even for high volume consumers and assuming BaP concentration and PAH4 sum equal to the current legal limit, the calculated MOE values for all groups of consumers are higher than the critical limit of 10 000 proposed by EFSA. The overall risk to public health, contribution of other sources of BaP and PAHs in foods, as well as the impact of food production technologies or preparation practices should be further studied.

References Conde, F. J., Ayala, J. H., Afonso, A. M., & Gonzalez, V. (2005). Polycyclic aromatic hydrocarbons in smoke used to smoke cheese produced by the combustion of rock rose (Cistus monspeliensis) and tree heather (Ericaarborea) wood. Journal of Agricultural and Food Chemistry, 53(1), 176e182.

Djinovic, J., Popovic, A., & Jira, W. (2008). Polycyclic aromatic hydrocarbons (PAHs) in different types of smoked meat products from Serbia. Meat Science, 80, 449e456. Duedahl-Olesen, L., White, S., & Binderup, M. L. (2006). Polycyclic aromatic hydrocarbons (PAH) in Danish smoked fish and meat products. Polycyclic Aromatic Compounds, 26, 163e184. EC. (2005). Commission Recommendation of 4 February 2005 (2005/108/EC) on the further investigation into the levels of polycyclic aromatic hydrocarbons in certain foods. Official Journal of the European Union, L34, 43e45. EC. (2006). Commission regulation 1881/2006/EC of 19 December 2006 setting maximum levels for certain contaminants in foodstuffs. Official Journal of the European Union, L364, 5e24. EC. (2011). European Commission Regulation 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, L215, 4e8. EFSA. (2005). Opinion of the Scientific Committee on a request from EFSA related to a harmonised approach for risk assessment of substances which are both genotoxic and carcinogenic. EFSA Journal, 282, 1e31. EFSA. (2008). Scientific opinion of the panel on contaminants in the food chain on a request from the European Commission on polycyclic aromatic hydrocarbons in food. EFSA Journal, 724, 1e114. n, M. S., & Simal-Ga ndara, J. (2005). Polycyclic aromatic hydrocarbons García-Falco in smoke from different woods and their transfer during traditional smoking into chorizo sausages with collagen and tripe casing. Food Additives and Contaminants, 22(1), 1e8. n, M. D., Sopelan, P., & Partearroyo, M. A. (2000). Polycyclic aromatic hydroGuille carbons in liquid smoke flavourings obtained from direct types of wood. effect of storage in polyethylene flasks on their concentrations. Journal of Agricultural and Food Chemistry, 48, 5083e5087. Janoszka, B., Warzecha, U., & Bodzek, D. (2004). Organic compounds formed in thermally treated high-protein food. Part I: polycyclic aromatic hydrocarbons. Acta Chromatographica, 14, 115e118. Jira, W. (2010). Polycyclic aromatic hydrocarbons in German smoked meat products. European Food Research and Technology, 230, 447e455. Lorenzo, J. M., Purrinos, L., Bermudez, R., Cobas, N., Figueiredo, M., & GarcíaFont an, M. C. (2011). Polycyclic aromatic hydrocarbons (PAHs) in two Spanish traditional smoked sausage varieties: “Chorizo gallego” and “Chorizo de cebolla”. Meat Science, 89, 105e109. Lorenzo, J. M., Purrinos, L., Garcia Fontan, M. C., & Franco, D. (2010). Polycyclic aromatic hydrocarbons (PAHs) in two Spanish traditional smoked sausage varieties: „Androlla ” and „Botillo”. Meat Science, 86, 660e664. Purcaro, G., Moret, S., & Conte, L. S. (2009). Optimisation of microwave assisted extraction (MAE) for polycyclic aromatic hydrocarbon (PAH) determination in smoked meat. Meat Science, 81, 275e280. Reinik, M., Tamme, T., Roasto, M., Juhkam, K., Tenno, T., & Kiis, A. (2007). Polycyclic aromatic hydrocarbons (PAHs) in meat products and estimated PAH intake by children and the general population in Estonia. Food Additives and Contaminants, 24(4), 429e437. Roseiro, L. C., Gomes, A., Patarata, L., & Santos, C. (2012). Comparative survey of PAHs incidence in Portuguese traditional meat and blood sausages. Food and Chemical Toxicology, 50, 1891e1896. Santos, C., Gomes, A., & Roseiro, L. C. (2011). Polycyclic aromatic hydrocarbons incidence in Portuguese traditional smoked meat products. Food and Chemical Toxicology, 49, 2343e2347. Simko, P. (2005). Factors affecting elimination of polycyclic aromatic hydrocarbons in smoked meat foods and liquid smoke flavorings. Molecular Nutrition and Food Research, 49, 637e647. Wretling, S., Eriksson, A., Eskhult, G. A., & Larsson, B. (2010). Polycyclic aromatic hydrocarbons (PAHs) in Swedish smoked meat and fish. Journal of Food Composition and Analysis, 23, 264e272. Yoon, E., Park, K., Lee, H., Yang, J. H., & Lee, C. (2007). Estimation of excess cancer risk on time-weighted lifetime average daily intake of PAHs from food ingestion. Human and Ecological Risk Assessment, 13, 669e680.