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ISSN: 2157-7110
Journal of Food Processing & Technology
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Safety Assessment of Polycyclic Aromatic Hydrocarbons (PAHs) in Cold Smoked Fish (Mugil Cephalus) Using GC-MS

Hafez NE1*, Awad AM1, Ibrahim SM2 and Mohamed HR2

1Department of Food Science and Technology, Faculty of Agriculture, El-Fayoum University, Egypt

2Fish Processing and Technology Laboratory, Fisheries Division, National Institute of Oceanography and Fisheries, Egypt

*Corresponding Author:
Hafez NE
Department of Food Science
and Technology, Faculty of Agriculture
El-Fayoum University, Egypt
Tel: 084- 6356731
E-mail: [email protected]

Received Date: July 17, 2017 Accepted Date: August 04, 2017 Published Date: August 11, 2017

Citation: Hafez NE, Awad AM, Ibrahim SM, Mohamed HR (2017) Safety Assessment of Polycyclic Aromatic Hydrocarbons (PAHs) in Cold Smoked Fish (Mugil Cephalus) Using GC-MS. J Food Process Technol 8: 688. doi: 10.4172/2157-7110.1000688

Copyright: © 2017 Hafez NE, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Abstract

This work was planned to determine the safety level of polycyclic aromatic hydrocarbons (PAHs) in cold smoked mullet fish obtained from two fish farms (A and B) localized at El-Fayoum Governorate, Egypt during August 2015. PAHs components were determined by GC-MS. Results showed that total of PAHs components was 28.8 and 5 ppb in both smoked fish products (A and B), respectively. Also, low molecular weight (LMW) of PAHs was found especially in smoked fish (A), followed by medium molecular weight (MMW) and however high molecular weight (HMW) was not detectable. Levels of Benzo [a] Pyrene (B {a} P) equivalent were 0.051 and 0.005 in products (A and B), respectively. However, PAH4 (BaP+CHR+BaA+BbF) and PAH8 (Benzo (a) anthracene, Chrysene, Dibenzo (a, h) anthracene, Benzo (g, h, i) perylene, Benzo (b) Fluoranthene, Benzo (k) fluoranthene, Benzo (a) Pyrene, Indeno (1, 2, 3-c, d) Pyrene) were not detectable. Also, categories of PAHs concentration are considered a minimally contaminated (10 to 99 μg/kg) and not contaminated (˂10 ppb) compared with the maximum recommended levels. Based on our results, it could be concluded that Benzo (a) pyrene compound was not detectable in all smoked samples which are considered as a safe product for human consumption.

Keywords

Fish; Smoking; PAHs; GC-MS

Introduction

Contamination of food by polycyclic aromatic hydrocarbons (PAHs) can be resulted from food processing at high temperature such as frying, smoking or roasting Codex [1-3]. There are many parameters limited the amounts of PAHs in food such as composition of the smoke, technology used in smoking, combustion temperature, type of wood, and exposure of the edible parts to the smoke [3-5]. The ranges of MW could be divided into three classes; 152 g/mol to 178 g/mol, 202 and 228 g/mol to 278 g/mol of low, medium and high MW, respectively [6,7]. Seven of the PAHs have been considered human carcnogens; benzo (a) anthracene, benzo (b) fluoranthene, benzo (k) fluoranthene, chrysene, benzo (a) pyrene, dibenzo (a, h) anthracene, and indeno (1, 2, 3-c, d) pyrene. Therefore, this study was designed to determine the safety level of PAHs in cold smoked mullet fish samples that were obtained during August 2015 from two fish farms localized at El-Fayoum governorate, Egypt (Figure 1).

food-processing-technology-chromatogram

Figure 1: GC-MS chromatogram of smoked mullet fish (farm A).

Materials and Methods

Fish samples

Mullet fish (Mugil cephalus) samples were obtained after directly catch from two fish farms (A and B). The main resources of irrigation water were industrial for A and agricultural discharge for B during August 2015 at El-Fayoum governorate. They were transported within two hours using ice box to Fish Processing and Technology Lab, Shakshouk Station for Water Resource, National Institute of Oceanography and Fisheries (NIOF), Egypt. Average of weight 453.3 gm ± 51.7 gm and length 35 gm ± 3 cm for raw samples from Farm A (Industrial discharge) also, average weight and length of raw mullet samples from Farm B (Agricultural discharge) are 526.6 gm ± 18 gm and 38 cm ± 2 cm respectively.

Smoking process and source of PAHs

After that, chilled whole fish samples were washed carefully with tap water, soaked in 10% brined solution for 2 hrs, rinsed with tap water for 1 min, semi-dried at 250°C for two hrs, traditional cold smoked at 35°C to 45°C for 8 to 10 hrs, using sawdust and finally cooled under ambient temperature.

Analytical methods

The edible of smoked mullet fish products was manually separated, homogenized, packed in polyethylene bags and then stored in a freezer at -20°C till analysis.

PAHs determination: PAHs were determined at Central Laboratory of Residue Analysis of Pesticides and Heavy Metals in Food (QCAP), Agricultural Research Centre. Cairo, Egypt as described by Forsberg et al. [8], Smoker et al. [9] and Khorshid et al. [10].

B{a}P equivalent: The B{a}Peq was calculated as the Σ B{a}Peqi value for individual PAHs. The B {a}Peqi calculated as the following equation [11]:

equation

CPAHi: Concentration of each PAH in the sample,

TEFPAHi: Toxic equivalency factor for each individual PAH.

Statistical analysis: The results obtained were analyzed statistically using the least significant difference test (LSD) at (P ≤ 0.05) and were expressed as Mean ± SD using SPSS 16 for windows.

Results and Discussion

As previous mentioned that raw mullet samples were obtained from different fish farms based on irrigation main resources; farm (A) was dependent on industrial discharge and localized at eastern Fayoum governorate farms whereas farm (B) was dependent on agriculture discharge and localized at Qarun lake farms (Figure 2).

food-processing-technology-mullet-fish

Figure 2: GC-MS chromatogram of mullet fish ( farm B).

Polycyclic aromatic hydrocarbons (PAHs)

Table 1 shows the PAHs concentration of cold smoked mullet fish flesh. 16 components of PAHs were detected in edible part of investigated products. Concentrations of ANT, FLA, PYR, FLU and PHE were 2.8, 3.3, 4.6, 6.6 and 11.5 ppb, respectively in smoked fish farm (A).

Compound Abbrev. Mw Rings Concentration (ppb)
Farm (A) Farm (B)
Chrysene CHR 228 4 ND ND
Anthracene ANT 178 3 2.8 ND
Acenaphthene ACE 154 3 ND ND
Benzo(b)fluoranthene BbF 252 5 ND ND
Benzo(k)fluoranthene BkF 252 5 ND ND
Dibenzo(a,h)anthracene DahA 278 5 ND ND
Fluorene FLU 166 3 6.6 ND
Naphthalene NA 128 3 ND ND
Benzo(a)pyrene BaP 252 5 ND ND
Benzo(g,h,i)perylene BghiP 276 6 ND ND
Indeno(1,2,3,cd)pyrene IcdP 276 6 ND ND
Acenaphthylene ACY 152 3 ND ND
Fluoranthene FLA 202 4 3.3 2.1
Pyrene PYR 202 4 4.6 2.9
Benzo(a)anthracene BaA 228 4 ND ND
Phenanthrene PHE 178 3 11.5 ND
Σ 16PAHs 28.8 5

Table 1: Molecular weight (MW), number of rings and concentration of PAHs in smoked mullet samples.

Farm (A): Industrial discharge. Farm (B): Agricultural discharge. Mw: Molecular weight.

In the other batch, FLA and PYR levels were 2.1 and 2.9 ppb, respectively in smoked fish farms (B). The PAHs have been grouped according to its molecular weights, we found that the high MW components of PAHs (228 g/mol to 278 g/mol), were Benzo (a) anthracene, Benzo (b) fluoranthene, Benzo (k) fluoranthene, Benzo (j) fluoranthene, Benzo (e) pyrene, Benzo (a) Pyrene, Benzo (ghi) perylene, dibenzo (h) anthracene, chrysene, cyclopenta (cd) pyrene, indo (1, 2, 3-c, d) Pyrene, and anthranthrene. The medium MW components (202 g/mol) were fluoranthene and pyrene. The low molecular weights (152 g/mol to 178 g/mol) included naphthalene, Acenaphthylene, acenaphthene, fluorene, anthracene and phenanthrene. Total of PAHs were 28.8 and 5 ppb in both smoked fish farms (A) and (B), respectively. In addition, rings number of PAHs in smoked fish farm (A) ranged from 3 to 4 rings and its MW ranged from 166 to 178 whereas in case of farm (B) only 4 rings and 202 as MW were found. Comparison of fish species smoked as fillets and as whole fish illustrated that the level of PAHs was higher for smoked fillets in comparison with the same fish species smoked as a whole fish.

Category of PAH concentration

Categories of concentration of PAH are considered a minimally contaminated (10 to 99 ppb) and not contaminated (<10 ppb) compared with recommended levels as set by Soares-Gomes et al. [12]. Concentrations of PAH were 28.8 and 5.0 μg/kg in smoked fish farms (A) and (B), respectively. Based on these results, Farm A (Industrial discharge) and B (Agricultural discharge) classified as minimally contaminated and Not contaminated respectively.

Molecular weight of PAHs in smoked fish

Also, Table 2 exhibits the molecular weight (MW) of PAHs in smoked mullet fish. The total concentration of the low molecular weights (LWM) of PAHs was higher than the medium molecular weights (MMW) in smoked fish farm (A). The high concentration of LWM in smoked fish farm (A) was 20.9 KDa. In other side, total concentration of medium molecular weights of PAHs was 5 KDa. Lipophilic nature of the PAHs and fish’s skin may be make better protection from the HMW-PAHs than LMW as reported by Mohammadi et al. [13]. In addition, results showed that there is no found a HMW-PAHs in different smoked fish farms either (A) or (B). In all products, HMWPAHs were below the limit of quantification or not detectable.

Fish farm HMW MMW LMW
A - 7.9 20.9
B - 5 -

Table 2: Molecular weight (MW) of PAHs in cold smoked fish.

Toxic equivalent factors (TEFs) and B {a} P equivalent of PAHs

TEF is an estimate of the relative toxicity of individual PAH fraction compared to benzo(a) pyrene. The toxic equivalent factors (TEFs) and B [a] P Equivalent of PAHs in smoked mullet fish are present in Table 3. Concentrations of phenanthrene, flourene, pyrene, fluoranthene and anthracene in smoked fish farm (A) were 11.5, 6.6, 4.6, 3.3 and 2.5 ppb, respectively and total B [a] P Equivalent was 0.051. On the other side, concentrations of pyrene and fluoranthene in smoked fish farm (B) were 2.9 and 2.1 ppb, respectively and total B [a] P Equivalent was 0.005.

Compound TEF Farm (A) Farm (B)
PAHs (µg/kg) BaPeqi PAHs (µg/kg) BaPeqi
Naphthalene 0.001 ND - ND -
Acenaphthylene 0.001 ND - ND -
Acenaphthene 0.001 ND - ND -
Fluorene 0.001 6.6 0.0066 ND -
Phenanthrene 0.001 11.5 0.0115 ND -
Anthracene 0.01 2.5 0.025 ND -
Fluoranthene 0.001 3.3 0.0033 2.1 0.0021
Pyrene 0.001 4.6 0.0046 2.9 0.0029
Benzo(a)anthracene 0.1 ND - ND -
Chrysene 0.01 ND - ND -
Benzo(b)fluoranthene 0.1 ND - ND -
Benzo(k)fluoranthene 0.1 ND - ND -
Benzo(a)pyrene 1 ND - ND -
Indeno(1,2,3,c)pyrene 0.1 ND - ND -
Dibenzo(a,h)anthracene 1 ND - ND -
Benzo(g,h,i)perylene 0.01 ND - ND -
∑ (BaPeqi )     0.051   0.005

Table 3: Toxic Equivalent factors (TEFs) and B [a] P Equivalent of PAHs in smoked mullet fish.

Benzo [a] pyrene (BaP), PAH4 and PAH8

In this study the PAH4 (the sum of BaP, chrysene, benz [a] anthracene and benzo [b] fluoranthene) and PAH8 {Benzo (a) anthracene, Chrysene, Dibenzo (a, h) anthracene, Benzo (g, h, i) perylene, Benzo (b) Fluoranthene, Benzo (k) fluoranthene, Benzo (a) Pyrene, Indeno (1, 2, 3-c, d) Pyrene} were not detected in smoked fish samples too. So, in the current study, cold smoked products are safety for human consumption because BaP did not detect comparing to the maximum permissible limit 2 ppb as set by the European Commission Regulation (OJEU, 835/2011) [14]. These results are in accordance to the findings by El-Lahamy et al. [15] who reported that BaP not detected in cold and hot smoked catfish fillets.

Sources and assessment of PAHs

Table 4 shows that source characterization and assessment of PAHs comparing with the previous references. It is well known that the sawdust wood was used as source of PAHs in this work as mentioned above (material and methods part). Results showed that the ratio of anthrancene to anthracene plus phenanthrene [An/(An+Phen)] was 0.19 in case of product obtained from farm A while it was not detected in farm B. This indicates that the wood combustion is the main source of PAHs compared with the mass 178. [An/(An+Phen)] ratio <0.10 usually is referred to petroleum while a ratio >0.10 indicates dominance of combustion [16-18]. The [Fl/(Fl+Py)] ratio also ranged from 0.41 to 0.42 in farm A and B products, respectively. The ratios ranged between 0.4 and 0.5 the refers to some amount of fossil fuel combustion sources (vehicular, fat and crude oil) of PAHs [19-22].

Source of PAHs PAH Ratios
[An/
(An + Phen)] 178
[Fl /
(Fl + Py)] 202
Wood combustion >0.10 >0.5
Petroleum <0.10 0.40
Saw dust Farm A 0.19 0.41
Farm B ND 0.42

Table 4: Sources and assessment of PAHs comparing with the previous reference

Conclusion

In conclusion, the safety of smoked fish has been controlled by measuring benzo (a) pyrene level, which is one of the most carcinogenic PAHs. European Commission has limited the maximum acceptable concentrations of benzo (a) pyrene at 2 ppb for smoked fish and smoked fishery products, excluding bivalve molluscs. In addition, the categories of concentration of PAH are considered a minimally contaminated and not contaminated compared with international recommended levels.

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