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Effects of Smoking Temperature, Smoking Time, and Type of Wood Sawdust on Polycyclic Aromatic Hydrocarbon Accumulation Levels in Directly Smoked Pork Sausages
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Food Safety and Toxicology

Effects of Smoking Temperature, Smoking Time, and Type of Wood Sawdust on Polycyclic Aromatic Hydrocarbon Accumulation Levels in Directly Smoked Pork Sausages
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  • Radu C. Racovita
    Radu C. Racovita
    University “Politehnica” of Bucharest, Faculty of Applied Chemistry and Materials Science, Department of Inorganic Chemistry, Physical Chemistry and Electrochemistry, 1-7 Gh. Polizu Street, Bucharest 011061, Romania
  • Catinca Secuianu*
    Catinca Secuianu
    University “Politehnica” of Bucharest, Faculty of Applied Chemistry and Materials Science, Department of Inorganic Chemistry, Physical Chemistry and Electrochemistry, 1-7 Gh. Polizu Street, Bucharest 011061, Romania
    Imperial College London, Department of Chemical Engineering, South Kensington Campus, London SW7 2AZ, United Kingdom
    *E-mail: [email protected]; [email protected]
  • Maria D. Ciuca
    Maria D. Ciuca
    University “Politehnica” of Bucharest, Faculty of Applied Chemistry and Materials Science, Department of Inorganic Chemistry, Physical Chemistry and Electrochemistry, 1-7 Gh. Polizu Street, Bucharest 011061, Romania
  • Florentina Israel-Roming
    Florentina Israel-Roming
    University “Politehnica” of Bucharest, Faculty of Applied Chemistry and Materials Science, Department of Inorganic Chemistry, Physical Chemistry and Electrochemistry, 1-7 Gh. Polizu Street, Bucharest 011061, Romania
    University of Agronomic Sciences and Veterinary Medicine, Biotechnology Department, 59 Marasti Boulevard, Bucharest 011464, Romania
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Journal of Agricultural and Food Chemistry

Cite this: J. Agric. Food Chem. 2020, 68, 35, 9530–9536
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https://doi.org/10.1021/acs.jafc.0c04116
Published August 7, 2020

Copyright © 2020 American Chemical Society. This publication is licensed under these Terms of Use.

Abstract

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Smoking is an excellent food preservation method but also a source of contamination of foodstuffs with carcinogenic polycyclic aromatic hydrocarbons (PAHs). Herein we investigated the influence of smoking temperature, smoking time, and type of wood sawdust used as smoke source on PAH levels attained through controlled smoking of pork sausages. Four PAHs (benz[a]anthracene, chrysene, benzo[b]fluoranthene, benzo[a]pyrene) were monitored, as required by European Commission Regulation 835/2011. PAH concentrations increased continuously both with higher temperatures (55–95 °C) and with longer smoking periods (2–9 h), although the level of benzo[a]pyrene exhibited a tendency to plateau after 6 h. Among seven types of hardwoods tested, plum, alder, and birch yielded PAH concentrations considerably higher than that of commonly used beech, and oak showed similar levels to beech while apple and, to a lesser extent, walnut caused lower levels of sausage contamination. These findings could guide the establishment of good practices in the smoked meat industry.

Copyright © 2020 American Chemical Society

1. Introduction

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In addition to drying and salting, smoking is a third preservation method commonly used for meat and fish, among other foods, and dates as far back as 90 000 years ago. (1) The significant extension of a food product’s shelf life through smoking is the result of the combined action of several phenolics native to smoke that are transferred to the foodstuff. (1) Many of these phenolics were proven, in experiments replicating smoke compositions, to have antioxidant, (2) antifungal, (3) and antimicrobial properties. (4) Furthermore, smoking is associated with additional organoleptic benefits: it enhances the visual appearance of meats, through browning reactions between meat amino acids and carbonyl compounds derived from thermal transformations of wood carbohydrates, and imparts a characteristic smoky smell and flavor, for which a combination of phenolics, carbonyls, and lactones have been shown to be responsible. (1) On the other hand, smoking also causes significant contamination of foodstuffs with known or suspected carcinogens and mutagens, such as formaldehyde, (1) heterocyclic amines, (5) and especially polycyclic aromatic hydrocarbons (PAHs). (6−10) The latter are formed through incomplete pyrolysis of the organic fuel (typically wood) used to generate smoke and can raise serious health concerns at high levels of ingestion. (11)
Consequently, in 2008, the Panel on Contaminants in the Food Chain (CONTAM Panel) of the European Food Safety Authority (EFSA) made a recommendation that the cumulative concentration of benzo[a]pyrene (BaP), chrysene (CHR), benz[a]anthracene (BaA), and benzo[b]fluoranthene (BbF) denoted as PAH4, in addition to the individual concentration of BaP, be used as best indicator of PAH contamination levels of food products. (12) In response, the European Commission (EC) adopted Regulation No. 835/2011 that sets the upper limits acceptable for BaP and PAH4 concentrations in various categories of retail foodstuffs marketed within the European Union (EU). (13)
Efforts geared toward correlating PAH accumulation in smoked foods with technical aspects of the smoking process have been made in various laboratories around the world. Among factors that have been found to influence to a higher or lesser degree the levels of PAHs reached in the smoked end-product are the method of smoke generation, such as wood smoldering, thermostated plates, friction, or steam smoking, (1,14) the way of combustion of wood material, whether by ignition from a flame or by direct heating, (15) the specific smoking technique employed, i.e., industrial or traditional, (16) direct or indirect, (17−20) hot or cold, (18,21) classic or using liquid smoke, (14,21,22) smoke generation temperature, (6,8,14) the position of food material in the smokehouse, (10,19,23) the duration of smoking, (7,18,24,25) and the specific type of fuel used to generate smoke, including various kinds of wood, (6,8,17,18,26) charcoal, (20,26) or sugar. (24)
Aside from smoked plant and animal foods, (27) several other studies were published in recent years focusing on a wide range of food products contaminated with PAHs either from environmental sources or through certain processing technologies. These included, among others, bread that was toasted in various ways, (28) plant foods harvested from the vicinity of a recent chemical company fire, (29) animal feed ingredients, (30) instant coffee, (31) alcoholic drinks, (32) canned bivalves in pickle and natural sauce, (33) fish oils, (34,35) and grilled pork marinated with beer marinades. (36)
In the present work, we investigated the influence of smokehouse temperature, duration of smoking, and type of wood sawdust used as fuel for smoke generation on BaP and total PAH4 contamination levels of pork sausages obtained by direct smoking. In the context of food safety and quality assurance, the findings of this study serve, along with the relatively few other studies on this topic mentioned above, as valuable reference data and scientific basis for defining best practices for the food smoking industry.

2. Materials and Methods

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2.1. Chemicals

Acetonitrile (≥99.9%, HPLC grade) was acquired from Scharlab S.L. (Spain) and used as-received. Water from a Milli-Q Reference Water Purification System (18 MΩ cm resistivity) was used in all experiments. Helium (≥99.9999%) and nitrogen (≥99.996%) were purchased from Linde GmbH (Germany). QuEChERS (Quick-Easy-Cheap-Effective-Rugged-Safe) kits for PAH extraction from the food matrix were from Thermo Fisher Scientific Inc. (USA) and, in essence, were 50 mL conical tubes containing 6 g of anhydrous magnesium sulfate and 1.5 g of sodium acetate. QuEChERS kits for purification of PAHs by dispersive solid-phase extraction (dSPE) came from Agilent Technologies Inc. (USA) and were 15 mL conical tubes with 400 mg of PSA (primary-secondary amine), 400 mg of octadecyl-substituted silica, and 1200 mg of anhydrous magnesium sulfate. For gas chromatography–mass spectrometry (GC-MS) method development and validation, a certified reference material (CRM) from Ultra Scientific Inc. (now Agilent Technologies Inc.), JTB-0005, consisting of a 16 PAH mixture including PAH4, was used. Another CRM, BCR-092, i.e., neat 10-azabenzo[a]pyrene (ABaP) (≥99.6%), acquired from LGC Limited (United Kingdom), served as internal standard for GC measurements.

2.2. Wood Sawdust

For studying the effect of smoking temperature and time on PAH4 accumulation in pork sausages, beech wood sawdust purchased from RIA DO Trading S.R.L. (Romania) was used in all experiments. To assess the effect of the type of sawdust used to generate smoke, five other types of wood sawdust (apple, plum, birch, oak, walnut) were acquired from Axtschlag GmbH (Germany) and an additional one (alder) from Jaxon Sp. z o.o. (Poland).

2.3. Pork Sausages

The raw sausages used for smoking under controlled conditions consisted of ground pork in natural hog casing and were purchased from a major retailer in Romania. All had the same batch number and were frozen immediately after purchase. They were stored in frozen state at −18 °C (255 K) for no more than 1 month. For smoking experiments, a sufficient number of sausages were removed from the freezer and left to thaw in a fridge at 4 °C (277 K) for about 24 h prior to use.

2.4. Smoking of Sausages

A Direca Depo Professional Smokehouse with four racks (Direca Depo S.R.L., Romania) and double walls with mineral wool insulation was used in all sausage smoking experiments. It is equipped with temperature control for the smoking chamber, timer, a metal pan for sawdust heated by an electric heating coil (i.e., smoke generator), a lateral sawdust feeder for replenishing sawdust when needed, and a smoke exhaust at the top with adjustable opening. In all experiments, raw sausages were placed at the same height level as the temperature sensor of the smokehouse, namely on the uppermost rack, i.e., farthest from the smoke generator. The smoke exhaust was kept in the completely open position, and new portions of 15 g of sawdust, moistened with 5 mL of water, were added every time the flow rate of the exhaust decreased substantially, which was typically every 20–30 min. Sausages were smoked with smoldering beech sawdust (i) for a fixed duration of 3 h at five different temperatures (55 °C/328 K, 65 °C/338 K, 75 °C/348 K, 85 °C/358 K, and 95 °C/368 K) and (ii) for various durations (2, 3, 5, 6, 8, and 9 h) at a fixed temperature of 75 °C, the latter temperature being recommended for smoking sausages in the smokehouse user’s manual. With the other types of sawdust, smoking was performed at the recommended temperature (75 °C) and for fixed amounts of time (2, 3, 5, 6, 8, and 9 h, respectively). After the desired smoking time, smoked sausages were removed from the smokehouse using clean metal tongs and were allowed to cool to room temperature before being processed as described in section 2.5. Each smoking experiment (at a certain temperature, for a certain amount of time, and using a certain type of sawdust) was repeated three times on different days using roughly equal-sized raw sausages.

2.5. Preparation of Smoked Sausage Samples for GC-MS Analysis

In a typical experiment, smoked sausages were minced and homogenized in a Z-1111-O bowl chopper (Zephyr S.L.G., Germany). About 10 g (weighed to a precision of ±0.1 mg using a KERN ADB 200-4 analytical balance) of finely minced sample was transferred into the 50 mL QuEChERS extraction tube along with 10 mL of water and 10 mL of acetonitrile. Samples were intensely agitated at 2500 rpm using a DLAB MX-S vortex (Dragon Lab, China) for 1 min and then centrifuged at 2720g for 5 min in a Medibas+ centrifuge (Auxilab S.L., Spain). Five milliliters from the acetonitrile layer was transferred to a 15 mL QuEChERS tube for purification by dSPE. Stirring and centrifugation steps were repeated. After settling, 1.5 mL of sample was transferred into a 2 mL GC amber vial (ISOLAB GmbH, Germany), 10 μL of an acetonitrile solution of internal standard (ABaP) of exact concentration was added, and then the sample was evaporated to dryness under a gentle stream of N2 at 50 °C and redissolved with fresh acetonitrile up to a volume of 100 μL in 150 μL vial inserts (Agilent Technologies Inc., Lexington, MA). For GC-MS analysis, a 1 μL aliquot of the resulting solution was injected into the GC inlet.

2.6. Quantification of PAHs in Smoked Sausages by GC-MS

A Thermo Scientific Focus GC with a Polaris Q ion trap MS detector was used. Sample injections were performed in splitless mode, using a TriPlus Autosampler interfaced with the GC, at an injector temperature of 250 °C (523 K). A TR-5MS column from Thermo Fisher Scientific (30 m long, 0.25 mm i.d., 0.25 μm film thickness) was employed. Helium at a flow rate of 1 mL/min was the mobile phase. The MS ion source temperature was 250 °C (523 K), and the transfer line was set at 280 °C (553 K). The temperature program was as follows: 1 min hold at 70 °C (343 K); ramp 10 °C/min to 300 °C (573 K); 10 min hold. All measurements were performed in single ion monitoring (SIM) mode, for increased sensitivity, using the following m/z characteristic ratios: 228 (BaA), 228 (CHR), 252 (BbF), 252 (BaP), 252 (ABaP).

2.7. GC-MS Method Validation

The method was validated intralaboratory in agreement with EC Regulation No. 836/2011. (37) Accordingly, calibration curves for peak areas as a function of individual PAH concentration were obtained and found to be linear over 4 orders of magnitude (10–4– 1 μg/mL) in the case of BaA, CHR, and BaP, and 3 orders of magnitude (10–3– 1 μg/mL) in the case of BbF, respectively. Analytical sensitivities Si, i.e., the slopes of calibration lines for each analyte i, were, in the following order: 3.73 × 105 (BaA), 5.78 × 105 (CHR), 3.18 × 105 (BbF), and 3.76 × 105 (BaP). The limits of detection, LOD, and quantification, LOQ, were determined as LODi = 3σi/Si and LOQi = 10σi/Si, where σi and Si are the standard deviation of the mean peak area obtained using a series of 20 blanks for analyte i and the sensitivity, respectively. LODs obtained were as follows: 0.093 (BaA), 0.070 (CHR), 0.058 (BbF), and 0.083 μg/kg (BaP), while LOQs were found to be 0.309 (BaA), 0.232 (CHR), 0.195 (BbF), and 0.275 μg/kg (BaP), respectively. All LODs were below 0.3 μg/kg and all LOQs below 0.9 μg/kg as required for in-house validation. (37) As all replicates of raw pork sausages analyzed using the same procedure described in section 2.5 for smoked sausages exhibited undetectable levels (<LOD) for all four PAHs, they were used as matrixes for recovery assessments. In these experiments, the same protocol described in section 2.5 was used, with the only additional step that the 10 g of minced raw sausage was spiked with 200 μL of a PAH solution obtained by diluting 1 mL of JTB-0005 CRM with 9 mL of acetonitrile. Percent recoveries of spiked amounts, given as averages of three replicates, were found to be in the following order: 60.1 (BaA), 58.8 (CHR), 67.6 (BbF), and 65.4% (BaP), and were thus within the 50–120% range stipulated by EC Regulation 836/2011. (37) For precision testing, Horwitz ratios HORRATR were calculated for each EC-regulated PAH by division of relative standard deviations of the mean concentrations determined for each analyte in the same spiked samples with the predicted value of 22% given by the modified Horwitz equation for trace analysis. (38) In order, they were found to be 0.277 (BaA), 0.214 (CHR), 0.948 (BbF), and 0.437 (BaP), and were therefore all well below the upper limit of 2. (37)

2.8. Statistical Analysis

A randomized experimental design was used considering the fixed effect of each type of smoking treatment and random effect of replicate within independent groups of raw sausages as the experimental unit. All PAH concentration data measured were subjected to one-way analysis of variance (ANOVA). Significant pairwise differences resulting from a given treatment variable (temperature, time, or type of wood sawdust) were tested using Student’s two-sided t test. A calculated p value below 0.05 was considered to be statistically significant. All PAH concentrations were reported as mean value ± standard deviation. Statistical analysis procedures were carried out using Microsoft Excel (part of Microsoft Office Suite 365, Redmond WA).

3. Results and Discussion

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Concentrations of the four PAHs in the whole smoked sausages were determined individually and compared (i) as a function of smoking temperature when beech sawdust was used for a fixed smoking time; (ii) as a function of smoking time when beech sawdust was used at a fixed temperature; (iii) as a function of the type of wood sawdust used to produce smoke at a constant temperature for fixed durations.

3.1. Effect of Smoking Temperature on PAH Concentrations

Aside from some minor fluctuations, all PAH concentrations showed a generally increasing trend as smoking temperature increased (Figure 1). Irrespective of temperature, the highest concentration was always that of CHR, followed by BaA and BaP, BbF being the least abundant. The total concentration of PAH4 showed an almost linear increase with temperature, within error limits. The concentration of BaP alone also increased, albeit less regularly, the most significant increases being from 55 °C to 65 °C and then from 75 °C to 85 °C. Notably, only for sausage samples smoked for 3 h at the lowest temperature, 55 °C, were the concentrations of BaP and PAH4 found below the EC-imposed limits of 2 and 12 μg/kg, respectively. (13)

Figure 1

Figure 1. Variation of individual concentrations of benzo[a]pyrene (BaP), benz[a]anthracene (BaA), chrysene (CHR), and benzo[b]fluoranthene (BbF) and the sum of their concentrations (PAH4) in smoked sausages as a function of smoking temperature (smoke source: beech wood sawdust, smoking time: 3 h). Error bars correspond to standard deviations of the mean (n = 3). Identical letters with the same color indicate there is no significant difference for different smoking temperatures (two-tailed t test, p < 0.05).

Overall, a 40 °C temperature increase (from 55 °C to 95 °C) triggered a more than 3-fold increase of both BaP and PAH4 contents, demonstrating that smokehouse temperature has a major effect on foodstuff contamination during smoking. Increases of PAH concentrations with smoking temperature are not surprising and were expected, given that PAHs are produced through thermal treatment of organic matter, including foodstuffs. (12) Such temperature-induced increases have been reported previously in the literature for smoked fish, (18) as well as for smoked meat. (19,39)

3.2. Effect of Smoking Time on PAH Concentrations

Smoking time also had a strong influence on PAH accumulation in pork sausages (Figure 2). The total concentration of PAH4 and that of the most abundant of all PAHs, i.e. CHR, exhibited continuous increases over the course of the first 8 h of smoking and only in the last hour no longer increased significantly, although it cannot be excluded that the increase would have continued had the smoking process not been ceased after 9 h. The concentrations of BaP and BaA also increased for the first 6 h of smoking, after which they came to a halt. This could be due to substantial loss of the lipid fraction in the sausage through dripping by this time point, as BaP and PAH4 contents in smoked sausages have been previously reported to vary proportionally to fat content. (10,21,40) The increase in BaP concentration was more significant, such that after 5 and 6 h of direct smoking, BaP became roughly equally abundant as BaA. BbF was least abundant of all PAHs at any time, the only notable increase in its concentration being recorded between 3 and 5 h of smoking time, after which it also remained roughly constant. Once again, it was found that even after only 2 h of smoking at 75 °C, the EC limits for BaP and PAH4 were both exceeded.

Figure 2

Figure 2. Variation of individual concentrations of BaP, BaA, CHR, and BbF and the sum of their concentrations (PAH4) in smoked sausages as a function of smoking time (smoke source: beech wood sawdust, smoking temperature: 75 °C). Error bars correspond to standard deviations of the mean (n = 3). Identical letters with the same color indicate that there is no significant difference for different smoking times (two-tailed t test, p < 0.05).

Past investigations in the literature on the effect of increasing smoking time also showed increases of PAH contents of smoked foodstuffs, in some studies with certain time points beyond which further increases were no longer observed, either for BaP alone (7) or for the sum of all PAHs, (18) and in others with no obvious tendency for concentrations to level off even after 5 h of smoking meat. (19) In any case, our data clearly showed that a 3-fold increase in smoking time (from 2 to 6 h) determined a 3-fold increase of BaP concentration and more than 2-fold increase of total PAH4, proving that smoking duration also has an important impact on PAH accumulation in foods.

3.3. Effect of Type of Wood Sawdust on PAH Concentrations

The types of sawdust used in the experiments spanned a wide range of hardwoods, specifically from seven tree species commonly used in the smoking process. These species included members of the Fagaceae family (beech and oak), the Betulaceae family (alder and birch), the Juglandaceae family (walnut), and the Rosaceae family (apple and plum).
For a smoking duration of 3 h at a constant temperature of 75 °C, the concentrations of PAHs varied widely depending on the type of wood sawdust used (Figure 3). Beech wood is the most common hardwood used for smoking meat products, (9) so it was taken as reference for comparisons here.

Figure 3

Figure 3. Variation of individual concentrations of BaP, BaA, CHR, and BbF and the sum of their concentrations (PAH4) in smoked sausages as a function of the type of wood sawdust used to generate smoke (smoking temperature: 75 °C, smoking time: 3 h). Error bars correspond to standard deviations of the mean (n = 3). Identical letters with the same color indicate that there is no significant difference for different types of wood sawdust (two-tailed t test, p < 0.05).

In terms of relative abundance of the four PAHs, just like in the case of beech discussed in sections 3.1 and 3.2, CHR was the most abundant and BbF was least abundant among PAH4 for the other six types of sawdust as well. BaP was generally third most abundant, however, with levels as low as that of BbF in the cases of apple, walnut, and oak woods. BaA was consistently more abundant than BaP irrespective of wood type and, in the cases of plum and oak woods, it reached concentration levels as high as those of CHR after 3 h of smoking at 75 °C (Figure 3). Supporting Information Tables S1–S6 show, in addition, that the relative distributions of the four PAHs in the PAH4 mixture remained the same for other smoking times too, within error limits.
After 3 h of smoking, the highest concentrations of BaP in smoked sausages were caused, in order, by plum, alder, and birch sawdust, and they were all significantly higher than in the case of beech sawdust (Figure 3). The wood of the other Fagaceae species, oak, caused similar levels of BaP as beech, while apple was always associated with lower levels of BaP in the final food product. Walnut also showed comparatively lower BaP levels not only for the first 3 h, but also up to 6 h of smoking, yet at longer times it reached levels similar to beech, as BaP concentration did not level off as early as it did for beech (Tables S1–S6).
BaA followed a very similar trend, with significantly higher concentrations than for beech in the cases of alder, birch, and especially plum wood. Oak- and walnut-smoked sausages exhibited comparable or only slightly higher levels of BaA to that of beech, while apple sawdust was the only to yield a consistently lower BaA contamination level at all smoking times (Figure 3 and Tables S1–S6).
As far as the most abundant PAH is concerned, CHR, its level was highest for plum, followed by the two Betulaceae species, alder and birch, in this order, while the remainder of wood types investigated had very similar levels of contamination with CHR (Figure 3 and Tables S1–S6).
With respect to the least abundant PAH, i.e. BbF, the same variation as for CHR was observed, except that apple wood-induced levels of BbF were found significantly lower than those caused by beech, but only within the first 3 h (Figure 3 and Tables S1–S6).
Most importantly, total PAH4 concentration varied greatly as a function of sawdust used as fuel. Compared to beech sawdust, sausages smoked with plum sawdust had a total PAH4 level almost five times greater, while alder and birch showed roughly 50% and 25% increases, respectively, within 3 h of smoking (Figure 3). Longer duration augmented these differences (Tables S1–S6). There was no significant difference between beech and oak at any time, and for walnut only a slight decrease (ca. 10%) was recorded for the first 3 h (Figure 3), which disappeared at longer times (Tables S1–S6). Finally, apple wood showed a substantial (ca. 25–30%) decrease in PAH4 level as compared to beech at all smoking times (Figure 3 and Tables S1–S6). Even so, only for the first 2 h of smoking was this level below the EC limit of 12 μg/kg (Table S1).
These data show some degree of correlation between PAH accumulation levels in sausages during direct, hot smoking with wood sawdust and the families to which the trees belong to, in the sense that Betulaceae species (alder and birch) showed consistently higher PAH concentrations than Fagaceae species (beech and oak). However, representatives of the Rosaceae family were at opposite poles, as plum wood yielded the highest levels of PAH contamination and apple the lowest; therefore, such correlations are limited. An alternative correlation could be in terms of lignin content of respective wood types, as lignin is an already aromatic precursor to PAHs, (41) so a higher lignin content may correlate with higher intake of PAHs into smoked meats. Indeed, compared to lignin mass content of beech wood, which is around 22%, (42) apple wood was reported to have about 1% lower lignin content, (43) while plum wood was shown to have a substantially larger lignin portion in its structure, at about 32–33%, (44) which correlates well with the much higher levels of all PAHs observed in sausages smoked with plum sawdust. The correlation also stands for walnut (20–22% lignin) (45) and oak (with highly variable reported lignin content, 17–30%), (46) given that some PAH levels were lower and others equal for these two wood types as compared to beech, but is not as good with respect to alder (ca. 22% lignin) and birch (18–20% lignin). (45)
Somewhat similar trends have been reported in the few past studies on wood type effect on PAHs in smoked meat products, although these used predominantly other kinds of wood. (6,8,26) Malarut and Vangnai (8) compared, among other properties, the PAH contents of sausages smoked with beech woodchips against four other tree species native to Southeast Asia (neem, copper pod, earleaf acacia, and eucalypt), all very different from the species studied in the present work, so comparisons cannot be made. Hitzel et al. (6) investigated PAH accumulation levels in Frankfurter-type sausages (made of pork and beef) and minisalamis (pork) when smoked with a selection of hardwood chips (oak, poplar, hickory, alder, beech) and softwood chips (spruce and fir), respectively. Similar to our results, they also observed an increase of both BaP and PAH4 concentrations when smoking sausages with alder as compared to beech, but they also noted a sensible decrease of BaP and PAH4 levels in both sausages and minisalamis when using oak woodchips. On the contrary, Stumpe-Viksna et al. (26) observed the second lowest BaP and total PAH levels in pork smoked with alder wood in a series of smoking experiments using 10 different types of wood: apple, alder, alder + juniper, spruce, maple, hazel, plum, aspen, bird-cherry, and rowantree. Their species selection did not include beech, but still, in agreement with our findings, they also recorded the lowest BaP and cumulative PAH levels for apple wood and about seven times higher levels for plum, which was the second worst choice of wood in their study, after spruce. Addition of juniper to alder wood during smoking resulted in two to three times higher concentrations of the four PAHs, so, considering that spruce and juniper are both softwoods, known to have higher levels of lignin in their composition in comparison to hardwoods, (45) it follows that there may indeed be a relationship between the lignin content of a certain type of wood and the level of PAHs transferred to meat products during smoking with sawdust from that wood.
As a recommendation for the smoked meat products industry, it follows from this work that cold (15–25 °C) or warm (25–50 °C) smoking techniques are more desirable to avoid high contamination levels with PAH4, because the higher the smoking temperature the higher the concentrations of PAHs will be in the end product. Smoking time should also be kept to a minimum that is still acceptable for sensory properties, because even though some of the PAHs may level off after at a certain time point in the smoking process (6 to 8 h in this work), others may still continue to accumulate in the smoked foodstuff. In terms of choice of wood sawdust for smoking, apple and walnut sawdust seem better options, as they reduce somewhat the PAH content in comparison to the staple beech wood used by the smoking industry. Oak wood appears equally suitable as its Fagaceae relative, and plum and Betulaceae species, like birch and alder, should be avoided, as they consistently tend to yield higher PAH concentrations in smoked foods.

Supporting Information

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The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acs.jafc.0c04116.

  • Tables S1–S6 showing comparative concentrations of BaP, BaA, CHR, BbF, and total PAH4 for every type of wood sawdust used that were attained after 2, 3, 5, 6, 8, and 9 h, respectively, of controlled smoking at 75 °C. Figure S1 showing an example of a typical gas chromatogram (PDF)

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Author Information

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  • Corresponding Author
    • Catinca Secuianu - University “Politehnica” of Bucharest, Faculty of Applied Chemistry and Materials Science, Department of Inorganic Chemistry, Physical Chemistry and Electrochemistry, 1-7 Gh. Polizu Street, Bucharest 011061, RomaniaImperial College London, Department of Chemical Engineering, South Kensington Campus, London SW7 2AZ, United KingdomOrcidhttp://orcid.org/0000-0001-5779-6415 Email: [email protected] [email protected]
  • Authors
    • Radu C. Racovita - University “Politehnica” of Bucharest, Faculty of Applied Chemistry and Materials Science, Department of Inorganic Chemistry, Physical Chemistry and Electrochemistry, 1-7 Gh. Polizu Street, Bucharest 011061, RomaniaOrcidhttp://orcid.org/0000-0002-6396-9869
    • Maria D. Ciuca - University “Politehnica” of Bucharest, Faculty of Applied Chemistry and Materials Science, Department of Inorganic Chemistry, Physical Chemistry and Electrochemistry, 1-7 Gh. Polizu Street, Bucharest 011061, RomaniaOrcidhttp://orcid.org/0000-0001-5530-0439
    • Florentina Israel-Roming - University “Politehnica” of Bucharest, Faculty of Applied Chemistry and Materials Science, Department of Inorganic Chemistry, Physical Chemistry and Electrochemistry, 1-7 Gh. Polizu Street, Bucharest 011061, RomaniaUniversity of Agronomic Sciences and Veterinary Medicine, Biotechnology Department, 59 Marasti Boulevard, Bucharest 011464, Romania
  • Funding

    This work was supported by a grant of Ministry of Research and Innovation, CNCS - UEFISCDI, project number PN-III-P1-1.1-PD-2016-0934, within PNCDI III.

  • Notes
    The authors declare no competing financial interest.

Acknowledgments

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The authors are grateful to Dr. Maria Mihaly (University “Politehnica” of Bucharest) for access to instrumentation, CRMs provided, and many fruitful discussions.

Abbreviations

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ABaP

10-azabenzo[a]pyrene

ANOVA

analysis of variance

BaA

benz[a]anthracene

BaP

benzo[a]pyrene

BbF

benzo[b]fluoranthene

CHR

chrysene

CRM

certified reference material

dSPE

dispersive solid-phase extraction

EC

European Commission

EFSA

European Food Safety Authority

EU

European Union

g

gravitational acceleration

GC-MS

gas chromatography–mass spectrometry

HORRATR

Horwitz ratio (for reproducibility)

HPLC

high performance liquid chromatography

LOD

limit of detection

LOQ

limit of quantification

QuEChERS

Quick-Easy-Cheap-Effective-Rugged-Safe

PAH

polycyclic aromatic hydrocarbon

PAH4

sum of benz[a]anthracene, benzo[a]pyrene, benzo[b]fluoranthene, chrysene

PSA

primary-secondary amine

SIM

single ion monitoring

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Journal of Agricultural and Food Chemistry

Cite this: J. Agric. Food Chem. 2020, 68, 35, 9530–9536
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https://doi.org/10.1021/acs.jafc.0c04116
Published August 7, 2020

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  • Abstract

    Figure 1

    Figure 1. Variation of individual concentrations of benzo[a]pyrene (BaP), benz[a]anthracene (BaA), chrysene (CHR), and benzo[b]fluoranthene (BbF) and the sum of their concentrations (PAH4) in smoked sausages as a function of smoking temperature (smoke source: beech wood sawdust, smoking time: 3 h). Error bars correspond to standard deviations of the mean (n = 3). Identical letters with the same color indicate there is no significant difference for different smoking temperatures (two-tailed t test, p < 0.05).

    Figure 2

    Figure 2. Variation of individual concentrations of BaP, BaA, CHR, and BbF and the sum of their concentrations (PAH4) in smoked sausages as a function of smoking time (smoke source: beech wood sawdust, smoking temperature: 75 °C). Error bars correspond to standard deviations of the mean (n = 3). Identical letters with the same color indicate that there is no significant difference for different smoking times (two-tailed t test, p < 0.05).

    Figure 3

    Figure 3. Variation of individual concentrations of BaP, BaA, CHR, and BbF and the sum of their concentrations (PAH4) in smoked sausages as a function of the type of wood sawdust used to generate smoke (smoking temperature: 75 °C, smoking time: 3 h). Error bars correspond to standard deviations of the mean (n = 3). Identical letters with the same color indicate that there is no significant difference for different types of wood sawdust (two-tailed t test, p < 0.05).

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  • Supporting Information

    Supporting Information


    The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acs.jafc.0c04116.

    • Tables S1–S6 showing comparative concentrations of BaP, BaA, CHR, BbF, and total PAH4 for every type of wood sawdust used that were attained after 2, 3, 5, 6, 8, and 9 h, respectively, of controlled smoking at 75 °C. Figure S1 showing an example of a typical gas chromatogram (PDF)


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