Kurdistan Regional Government-Iraq Ministry of Higher Education & Scientific Research Salahaddin University-Erbil College of Science Department of Chemistry
Review: determination and characterization of Heterocyclic Amines and Polycyclic Aromatic Hydrocarbons in food Products
A Review Article in Analytical Chemistry In Partial Fulfillment of the Requirements for the Degree of Ph. D
Prepared by: Yousif Taha Maaroof, [email protected] Supervised by: Asst. Prof. Dr. Media Noori Abdullah, [email protected]
(2020 – 2021)
Review: determination and characterization of Heterocyclic Amines and Polycyclic Aromatic Hydrocarbons in food Products
Abstract This review article was studies to produce, effects, and analyzing of heterocyclic amines, and polycyclic aromatic hydrocarbons in cooked food. Heterocyclic amine compounds are produce in the resulting high temperatures of cooking of amino acids reacts with creatine. As well as, polycyclic aromatic hydrocarbons are produce in cooking of meat, when fat pyrolysis and form intermediate products when, cooking at high temperatures. As a results of cooking methods Heterocyclic amines and polycyclic aromatic hydrocarbons will be form especially foods can be contaminated at high temperatures cooking methods such as, at grilling, smoking of chicken, barbecuing meat, and smoking steak. When fish and/or meat is smoked, or grilled, or barbecued in open fire at high temperatures higher polycyclic aromatic hydrocarbons production. Heterocyclic amines are generally carcinogenic compounds and causes to colon cancer, breast cancer, with prostate cancer, but polycyclic aromatic hydrocarbons do not effect to cancer once, although different polycyclic aromatic hydrocarbons as a mixture linked together effects to cancer. The main four polycyclic aromatic hydrocarbons which more effects on the carcinogenic in human body, due tobenzo[a]pyrene, and bennz[a]anthracene, and benzo[b]fluoranthene, and chrysene. Some of the organizations working to evaluation occurrence polycyclic aromatic hydrocarbons as well as, to evaluation toxicity of polycyclic aromatic hydrocarbons in different foods, and beverages.
Contents 1-Introduction ...... 3 2. Carcinogenic effect of PAHs with HCAs ...... 6 3. Formation of HCAs and polycyclic aromatic hydrocarbons in cooking foods ...... 6 4. Reducing HCA and PAH ...... 8 5. Analysis of PAHs with HCAs in foods...... 8 5.1. HPLC analysis of PAHs with HCAs in foods ...... 9 5.1.1. Determination of PAHs...... 10 5.1.2. Determination of HCAs ...... 10 5.2. Gas chromatography ...... 11 6. Conclusions ...... 11 7. References ...... 12
Keywords: Heterocyclic Amines, Polycyclic Aromatic Hydrocarbons, smoke, meat
1-Introduction In recent years, the publication most discussed and focused on the risks from food hazards (Adams, Moss et al. 1995, Amos-Tautua, Inengite et al. 2013). Nowadays, Food safety is a global phenomenon which continuously shown that is an increase levels of chemical contaminants like Polycyclic Aromatic Hydrocarbons (PAHs) and Heterocyclic amines (HCAs) in cooking food processing. Heterocyclic amines detected in cooked meats by a group of mutagenic compounds form, like meats. Heterocyclic amine compounds are produce in the resulting high temperatures of cooking of amino acids reacts with creatine (Knize and Felton 2005, Felton, Knize et al. 2007). As well as the amounts of heterocyclic amine produce in meats cooking depend upon at higher temperatures and time periods of cooking (Zheng and Lee 2009). Although, another factor related to heterocyclic amine produce, is type of meats with cooking method was used (Cross and Sinha 2004). In some studies described and detected most available heterocyclic amines in cooking meat such as, 2amino-1-metthyl-6-phennylimidazo (4, 5 -b) pyridine (PhIP) and 2-amino-3,8- dimmethylimidazo(4, 5 f)quinnoxaline (MeIQx). Moreover, described and detected other carcinogens in cooking meats, like 2-amino-3,4,8-trimmethylimidazo(4,5-f)quinnoxaline (DiMeIQx) (Cross and Sinha 2004, Knize and Felton 2005, Felton, Knize et al. 2007). Heterocyclic amines are generally very harmful chemicals which produce in cooked meat by the Maillard reaction, which Maillard reaction occurs between amino acids and reducing sugars that gives browned food almost foods undergo this reaction. When meat or fish broiled, smoked, or barbecued the amounts of amino acids dispersed in air which causes to forming of heterocyclic amines, which depend on the chemical structures of amino acids, for example creatine grouped into amino carboline group, or amino-imidazo-azaarene group. The pyrolytic heterocyclic amines is amino carboline group formed by pyrolysis of amino acids reactions at higher temperature > 300 degree centigrade (Felton, Knize et al. 2007). There are also, aminoimmidazo-azaarrene are referred to heterocyclic amines, the imidazo is functional group which attached to quinolone, or pyridine, or quinoxaline. Pyridine ( IQ ) and pyrazine ( IQx ) are produced by Maillard reaction which occurs between hexose sugar and free amino acid (Hollman and Katan 1999). On the other hand, transformation of aldehyde and creatine, degradation of Imidazoquuinoline and imidazoquinnoxaline which causes to free amino acid will be form. Heterocyclic amines formation in food, depend on the some points like types of amino acids, types of sugars, creatine with temperature degree, and moisture, and cooking type process, and time of cooking, and presence of antioxidant (Ohgaki, Takayama et al. 1991). Heterocyclic amines are organic chemical compounds which forming at cooking meats from high temperatures with longer time, in hotplate and when surfaces of meat brown occurs(Humans 1993). Polycyclic Aromatic Hydrocarbons are generally products by pyrolysis of fat at coking meat and fish when smoked or grilled. When directly meat and fish added on over an open fire, then fat drip onto the fire, Polycyclic Aromatic Hydrocarbons are flames and break the fat into intermediate products, after it is back to the surface of the meat and fish. Polycyclic Aromatic Hydrocarbons qualitative and quantitative detect in foods, smoke of cigarette, silencers in car exhaust fumes , crude oil industrial ,and petrochemicals industrial (Cross and Sinha 2004). Polycyclic Aromatic Hydrocarbons and Heterocyclic amines formed are affected by causes food type, and cooking method, and time of cooking (Jaegerstad and Skog 2005). HCAs and PAHs have been reported that about bioactivation, which DNA damage capacity and Cancer risks occurred, then specific enzymes worked to metabolism process in the body (Sinha, Rothman et al. 1995, Moonen, Engels et al. 2005). Polycyclic aromatic hydrocarbons are one types of organic chemical, which consist of two fused aromatic rings or more containing carbon atoms and hydrogen atoms. Polycyclic aromatic hydrocarbons are detection in cooking foods process, as well as major source of Polycyclic aromatic hydrocarbon is smoking of fish in foods (Guillen, Sopelana et al. 1997). Fats in foods occurs when fat pyrolysis or incomplete thermal decomposition, which causes to PAHs are production, when meat or fish are grilled, roasted, smoked or barbecued (Hydrocarbons 2006). When fats in the meat or fats in the fish combustion incompletely, which causes to produce chemical compounds intermediate as known Polycyclic aromatic hydrocarbons that deposited on the food surfaces. Polycyclic aromatic hydrocarbon Produce in meat and fish surfaces, when smoking, barbecued, grilled meat over charcoal is based on the fat content in meat or fish as well as depend on distance to heat source (Kazerouni, Sinha et al. 2001). Several PAHs in fish roasted, grilled, smoked or barbecued are formed such as benzo[a]pyrene, anthracene, chrysene, benzo[a]anthracene, indeno[1,2,3- c,d]pyrene (Guillén and Sopelana 2005, Akpambang, Purcaro et al. 2009, Palm, Carboo et al. 2011)(Alonge 1988, Lijinsky 1991). In the three area of smoked fish shop and smoked meat shop from Nigeria detect large quantities of benzo[a]anthrracene and benzo[a]pyrene as a Polycyclic aromatic hydrocarbons (Borokovcova, Dofkova et al. 2005). Research works showed by period exposure to Polycyclic Aromatic Hydrocarbons and Heterocyclic amines causes to increases cancer risks from animal laboratory (Emerole 1980). HCAs which causes to increasing level of tumors in the body like breast cancer, colon cancer, liver cancer, skin cancer, lung cancer, and prostate cancer(Ohgaki, Hasegawa et al. 1986, Kato, Ohgaki et al. 1988, Sugimura, Wakabayashi et al. 2004). As well as Polycyclic Aromatic Hydrocarbons causes to developed leukemia, and lung tumors (Dubuisson, Murph et al. 2001).
Fig (1): The Principal heterocyclic amines
Fig (2): The Principal Polycyclic aromatic hydrocarbons 2. Carcinogenic effect of PAHs with HCAs Carcinogenic of some heterocyclic amines were studied in laboratory by uses some animal tests. So as a result determine many heterocyclic amines are causing carcinogenic effect, such as colon cancer, and breast cancer, and prostate cancer. Heterocyclic amines was detected that, when pyridine uses as a dosed to monkeys, then showed developed liver cancer in monkeys body, and heterocyclic amines are carcinogenic effects to other mammals (Diet and Cancer 1982). The World Health Organization (WHO) and International Agency for Research on Cancer (IARC) established seven heterocyclic amines as possible carcinogens to humans. These seven heterocyclic amines include: and methylPyridine (MeIQ) , and methyl-pyrazine (MeIQx), and Glu- P-1, and Glu-P-2, and PhIP, and AaC, and MeAaC and Pyridine (IQ) (Ohgaki, Takayama et al. 1991). Carcinogenicity of polycyclic aromatic hydrocarbons has been several studied on the period exposure polycyclic aromatic hydrocarbons of workers, and in those studies showed high relationship between long time exposure polycyclic aromatic hydrocarbons and risks, like risks on the skin, lung, and gastrointestinal cancers. However till now don’t have any studies which directly showed cancer with exposure to polycyclic aromatic hydrocarbons and worked simultaneous with to other cancer direct exposure like heterocyclic amines (Humans 1993). Other chemical carcinogenic polycyclic aromatic hydrocarbons in foods has been discovered like, Benzo[a]pyrene which used as biomarker. Then several study have classification of some polycyclic aromatic hydrocarbons and shown as carcinogenic compounds in animal laboratory. The EPA are shown and classifications of seven polycyclic aromatic hydrocarbons related to human carcinogens, like benz[a]anthrracene, benzo[a]pyrrene, benzo[b]fluorannthene, benzo[k]fluorannthene, chryseene, dibenz[a,h]anthrracene, and indenol[1,2,3cd]pyrene (Mumtaz, George et al. 1996). As well as, According to EPA polycyclic aromatic hydrocarbons are grouped together so considered of greatest concern of their carcinogenicity (Adeyeye 2020). PAHs generally discovered that do not effect to cancer only but have effects to cancer when different polycyclic aromatic hydrocarbons mixture linked together. Moreover, PAHs makes the reasonable impact of each polycyclic aromatic hydrocarbons is very difficult on the human health (Adeyeye 2020).
3. Formation of HCAs and polycyclic aromatic hydrocarbons in cooking foods Several studies has been shown which HCAs and PAHs in foods occurs. Some points directly related to Heterocyclic amines formation in meat and fish such as, types of meat, methods of cooking, temperature, and distance meat on the heating sours as well as cooking time. The meat contains lower level in fat and water causes to forming higher concentrations of Heterocyclic amines at cooking time which compere to meat that has higher fat and water content, causes to forming lower concentrations of Heterocyclic amines at cooking time. Heterocyclic amines are more produce that at temperatures > 220 C, when meat most frying or grilling. As well as Heterocyclic amines at lower temperatures produce especially at roasting of meat for long time. When meat browned or burnt crusts, the Heterocyclic amines concentrations was found are higher (Skog, Johansson et al. 1998). The pan drippings and meat bits after frying left more Heterocyclic amines at high concentrations. Sausages meat was contained high level fat and water, so concentration of Heterocyclic amines is low (Augustsson, Lindblad et al. 1999). The meat of beef patties contained smaller amounts of Heterocyclic amines, when regularly storage meat until uses (Knize and Felton 2005). Frozen of beef patties no difference significant recorded in Heterocyclic amines levels compared to room-temperature while, meat grilled or barbecued (Knize and Felton 2005). Several Heterocyclic amines have been identified which great effects to human healthsuch as, pyridine, MeIQ, MeIQx,GluP-1, Glu-P-2, AaC, Trp-P1, TrpP2, PhIP, and MeAaC. After that investigation which can controlling formation of Heterocyclic amines in cooking chicken by controlling temperature pan-frying. Then Heterocyclic amines quantities determination in chicken cooked by controlling of temperature for DMIP, MeIQx, 4,8-DiMeIQx, PhIP, determined in ranged from 0.93 to 27.52 ng / g, when some Heterocyclic amines not detected in the control sample, such as pyridine (IQ), MeIQ, AaC, MeAaC, Trp-P1 and Trp-P2. Also in chicken cooked Heterocyclic amines was found in the ranged from 0.14 and 19.57 ng / g1 to DMIP, MeIQx, 4,8- DiMeIQx, and PhIP, as well as IQ and MeIQ cannot detect because below the limit of quantification, and also AaC, and MeAaC, and Trp-P1, and Trp-P2 were not detected in chicken meat samples. Furthermore, Heterocyclic amines formation was discovered which increase by reduced harman and norharman after adding to food. Although Heterocyclic amines increasing by adding edible oil, which due to using of pepper, garlic, tomato, and paprika. Some Natural spices causes to increasing formation Heterocyclic amines like, tomato, onion, garlic, ginger, and pepper when camel meat was cooked (Khan 2015, Khan, Naushad et al. 2017). The samples of meat control without natural produced Heterocyclic amines at higher quantities such as MeIQx, DiMeIQx, and PhIP while, some Heterocyclic amines not detected because lower than quantification limit like, MeIQ and IQ. As well as in cooked camel meat found different Heterocyclic amines like, MeIQx, DiMeIQx, and PhIP, as well as some Heterocyclic amines not detected because lower than quantification limit like MeIQ and IQ. The camel meat cooking with different natural spices, when Heterocyclic amines low production and could be attributed to high amounts of antioxidants, which antioxidants in such natural spices worked to scavenge free radicals from the meat, when during meat cooking process. camel meats Cooking methods have different types like frying, griddling, stewing, and barbequing which causes to Heterocyclic amines formation, analyzing under controlled temperature by solid-phase extraction (SPE) extract and quantitative by high performance liquid chromatography with mass spectrometry detector (Khan, Naushad et al. 2015). Pyridine (IQ) and MeIQ not found in all samples of meats analyzed. They identified most noteworthy amounts of MeIQx, 4,8-DiMeIQx, and PhIP in seared examples, while the griddled and grilled examples created moderate measures of MeIQx, 4,8-DiMeIQx, and PhIP. The stewing strategy created most minimal measures of PhIP. While MeIQx and 4,8-DiMeIQx were not distinguished in the examples. They were of the assessment that the low degrees of Heterocyclic amines in stewing strategy could be because of the way that the examples don't have direct contact with the cooking pot or blast influences the development of Heterocyclic amines (Khan, Bertus et al. 2009). Additionally some studies the working on the warm preparing on Heterocyclic amines development in meat offal, for example, hamburger liver, and sheep kidney and meat tongue. They revealed norharman and harman as the most plentiful amines in the items, while PhIP was found at lower amount. DMIP, MeIQx, and 4,8-DiMeIQx were recognized in cooked kidney and tongue among the remainder of Heterocyclic amines broke down. They likewise examined the jobs of added substances on Heterocyclic amines development, and recognized norharman and harman in liver cooked with added substances. They were of assessment that offal items exposed to delayed warm medicines of over 200C delivered the most minimal measures of Heterocyclic amines (Adeyeye 2020).
4. Reducing HCA and PAH Several techniques was used to reducing polycyclic aromatic hydrocarbons and Heterocyclic amines production in cooking meats method. Although reducing polycyclic aromatic hydrocarbons and Heterocyclic amines causes to good production, and minimum consumption of cooked meats.
Presence distance between direct meat contact with an open flame or hot metal surface, which causes to reduce the effect on pyrolysis of fat and reducing polycyclic aromatic hydrocarbons, as well as avoiding high temperatures cooking and avoiding high time which causes to reducing formation of polycyclic aromatic hydrocarbons and Heterocyclic amines. Using microwave oven to reduce cooking time of meat and decrease meat exposure to high temperatures which causes to reducing formation of Heterocyclic amines. Continuously turning of meat over heating at high temperature source causes to reducing formation of Heterocyclic amines. Because continuously turning of meat causes to reduce the reaction rate creatine with, amino acids and sugars in the meat cooking. Prevent using or eating of meats which contained large concentration of polycyclic aromatic hydrocarbons and Heterocyclic amines (Adeyeye 2020)
5. Analysis of PAHs with HCAs in foods The methods to determination of PAHs with HCAs in meat foods, used in research laboratories and for improvements routine detecting of PAHs with HCAs in foods at last years. Nowadays, no one procedure accepted to detections, because the difficulties solve different associated with quantitative isolation of heterocyclic amines and polycyclic aromatic hydrocarbons in foods material. As well as purification of the extraction analyte without significant loss (Stołyhwo and Sikorski 2005). Several method has used For analysing of heterocyclic amines in meat or foods such as, high-performance liquid chromatography combined with mass spectrophotometer detectors (Stavric, Lau et al. 1997)(Richling, Decker et al. 1997), or combined with diode array detection (Janoszka, Błaszczyk et al. 2009), are commonly used. As well as gas chromatography- mass spectrophotometer (GC-MS) (Richling, Kleinschnitz et al. 1999), GC-electrospray tandem MS (Hoenicke, Gatermann et al. 2004), and HPLC-electrospray tandem MS (Andrzejewski, Roach et al. 2004) are used for detection of polycyclic aromatic hydrocarbons in food. The IUPAC through shared examinations delivered what could be known as the principal official logical system for benzo[a]pyrene for prepared food items like fish, meat, cheddar, refreshments, and smoked food items. The insightful methodology was extensive and use to benzo[a]pyrene as well as the whole polycyclic sweet-smelling hydrocarbons in food items could be resolved. Individual polycyclic aromatic hydrocarbons in handled nourishments could be acquired using bright spectrophotometry, affirmed by the spectrrophotofluorommetry (ISO 2005). reverse phase HPLC fluorescence detection (FLD) after tidy up on alumina and this method has been applied by distinguish sixteen polycyclic aromatic hydrocarbons in creature and vegetable fats and oils, notwithstanding, this procedure doesn't give quantitative outcomes unpredictable mixes like naphthalene, acenaphthene, and fluorene. Moreover the solid phase extraction (SPE) method includes extraction and decontamination of polycyclic aromatic hydrocarbons, at that point recognizable proof by reverse-phase HPLC-FLD (Zelinkova and Wenzl 2015).
5.1. HPLC analysis of PAHs with HCAs in foods High-performance liquid chromatography (HPLC) is a most common type’s in liquid chromatography which has applied to determine organic compounds, depend on polarity of compounds. The more types of High-performance liquid chromatography columns available to determination of air pollutants. Determination of organic compounds in HPLC based on stationary phase which have normal phases and reversed phase, which as reversed phase is most used in HPLC. The C18 columns are reversed phase column is most uses in HPLC. The separation of heterocyclic amines and polycyclic aromatic hydrocarbons by HPLC depend on designed columns which available commercially. For example the column Vydac 5 ml C-18, and 150 * 4.6 mm at 30 C used to separation of 16 polycyclic aromatic hydrocarbons from a pure, or standard mixture, when programmed to eluent from 1v :1v water to acetonitrile at 30 min, and 1 ml/min flow rate. The FLD most frequently detectors in high-performance liquid chromatography was used to controlled wavelength of emission and excitation, and fluorescence spectra. The fluorescence detector in high-performance liquid chromatography used for low limit of detection of each of 15 polycyclic aromatic hydrocarbons in the ranges of picogramme, when working with FLD must be oxygen-free solvent uses to avoid quenching of some polycyclic aromatic hydrocarbons at fluorescence, for example pyrene (de Boer and Law 2003). At possible fluorescence detectors in high-performance liquid chromatography measurement of most polycyclic aromatic hydrocarbons at 100 times lower concentrations than diode array detector. Some polycyclic aromatic hydrocarbons not detected by fluorescence like acenaphthylene, because acenaphthylene does not emit any fluorescence. As well as in some cases UV–visible uses to detection individual polycyclic aromatic hydrocarbons. The UV–visible spectra of polycyclic aromatic hydrocarbons are more precise in HPLC than the fluorescence spectra. The UV detector higher reliability to identification of the polycyclic aromatic hydrocarbons. Several improvement of the analysis of polycyclic aromatic hydrocarbons in real sample matrices can be achieved by using high-performance liquid chromatography- mass spectroscopy. Although high-performance liquid chromatography- mass spectroscopy instrumentation is high cost, so cannot used to routine work (Stołyhwo and Sikorski 2005).
5.1.1. Determination of PAHs The polycyclic aromatic hydrocarbons determine in meat or food by HPLC. For example, weighted 5g meat sample and add to 15ml 1.0 M NaOH solution for 60 min until homogenised. The sample after homogenized was mixed with 17g Extrelut compound and loaded in an Extrelut-20 column. Polycyclic aromatic hydrocarbons Elution started from columns, by 60 ml of dichloromethane containing 5% toluene which absorbed on the propyl sulphonic acid (PRS) cartridges. Then dichloromethane solution was preconcentrate by vacuum evaporator to 0.5-1ml. Then residue repeat dissolved in 1ml n-hexane and transferred to the silica-gel glass packed column, and polycyclic aromatic hydrocarbons eluted by (mixture of 25 ml of n-hexane with 60 ml of 60 v:40 v (n hexane–dichloromethane) and loaded to HPLC analysis. The recovery rate of method was measuring before extraction to 2 standard mixtures spiked into samples which each sample added 50 ng. The determination concentration of each polycyclic aromatic hydrocarbons depend on using standard calibration curves. This method HPLC combined with a fluorescence detector, which mobile phase consist of 84% acetonitrile with 16% water, as well as the temperature in this system applied in 40 C used to separations with 1.0 ml/min flow rate (Janoszka 2011).
5.1.2. Determination of HCAs Separation, and extraction of heterocyclic amines were applied according to the procedures. For example for determination of heterocyclic amines in surface of meatballs, about 2mm depth of surface from meatballs and weighted 3g meat sample mixed well with 12ml 1.0 M NaOH. Then transferred into an Extrelut-20 column. The heterocyclic amines were eluted in column with 60ml ethyl acetate , and transferred into solid phase extraction which uses to purification. After that the adsorbed sample washed with 6ml 0.1M hydrochloric acid, as well as elute 15ml methanol with 18 ml of 0.1M HCl and 2ml pure water by mixture to remove interferences effect on the absorbed heterocyclic amines in solid phase extraction. The heterocyclic amines were eluted by 20 ml 0.5M ammonium acetate (pH 8.5). Then, heterocyclic amines eluted with 1ml methanol/ammonium hydroxide (9/1, v/v) and submitted for HPLC analysis. In this system standard compounds like spiked was used, such as mixing 1ml standard compounds which containing 50ng/ml IQ, MeIQ, MeIQx, 4, 8-DiMeIQx and PhIP mixed to samples before extraction to recovery rate measurement. The HPLC conjugated with a diode array detector used for this determination. As well as, reversed-phase Luna 5u C18 column (250 × 4.60 mm, 5 μm, 100A) was used with 0.01 M trimethylamine like Mobil phase (Puangsombat, Gadgil et al. 2011)(Lu, Kuhnle et al. 2018). 5.2. Gas chromatography Gas chromatography (GC) is a chromatography techniques depend on partition and absorption of diseased compound present in air samples. Determination in GC based on the volatility of organic compounds. Capillary columns in most applied in GC. The Gas chromatography have most used in analyzing of organic compounds like polycyclic aromatic hydrocarbons and Heterocyclic amines. Capillary columns in Gas chromatography have a great efficiency of the order of 60,000 plates in 30 meter. The separation and extraction of components by Gas chromatography is much better than high-performance liquid chromatography. The sample of smoked fish contains polycyclic aromatic hydrocarbons injection to Gas chromatography can separated over 100 peaks. The many number of polycyclic aromatic hydrocarbons extracts from smoked meat or fish products and quantification by flame ionization detector by measurement of retention times without multistage purification procedures to remove interference. As well as, for quantification and detection of polycyclic aromatic hydrocarbons in smoke by mass spectroscopy combined with Gas chromatography (Lawrence and Weber 1984). Selected ion mode technique used to detection of polycyclic aromatic hydrocarbons at 100 times lower concentrations compare to high- performance liquid chromatography- fluoresces detector. Nowadays, mass spectroscopy combined with Gas chromatography most used to determine ten polycyclic aromatic hydrocarbons in smoked meats. The gel-permeation chromatography method used to extraction and determination of polycyclic aromatic hydrocarbons (Jira 2004). Determined 15 polycyclic aromatic hydrocarbons in coking wastewater, which in coking wastewater by DLME coupled with GC- mas spectroscopy. Studied general parameters in DLME such as type and concentration solvent of extraction, types of dispersive solvent, pH of solution, and time of extraction. The optimum conditions of extraction were selected to be: 15 microliter extraction solvent consist of mixture of 2:1 v/v carbon tetrachloride and chlorobenzene, dispersive solvent is 0.75 mL of acetonitrile, the pH of solution is 8, and time of extraction is 2 min. The enrichment factors for 15 polycyclic aromatic hydrocarbons ranged from 452 to 685, and the linear ranges were obtained from 0.1 to 100mg/L, , and the relative recoveries of this method to determine 15 polycyclic aromatic hydrocarbons ranged from 67.5 to 103.5% with RSDs of 4.0–9.1% (Song, Zhu et al. 2013).
6. Conclusions In conclude, Heterocyclic amines (HCA) and polycyclic aromatic hydrocarbons (PAH) are two well-known organic compounds, which contain in food samples such as meat, fish, when at high temperatures products will be form. Heterocyclic amine compounds are produce in the resulting high temperatures of cooking of amino acids reacts with creatine. Formation of heterocyclic amines and polycyclic aromatic hydrocarbons in meat, when resulting from cooking methods contamination at high temperatures like grilling cooking method, or smoking cooking method or barbecuing cooking method of chicken, meat and fish. The polycyclic aromatic hydrocarbons was formed at open fire when meat and fish smoked, grilled and barbecued cooking method. Heterocyclic amines related to forming several carcinogenic in the human body like, colon cancer, breast cancer, and prostate cancer, although polycyclic aromatic hydrocarbons has do not effect to the cancer in the human body only. Several techniques was used to reducing polycyclic aromatic hydrocarbons and Heterocyclic amines production in cooking meats method. Although reducing polycyclic aromatic hydrocarbons and Heterocyclic amines causes to good production, and minimum consumption of cooked meats, like Avoidance direct contact of meats with an open flame or a hot metal surface, and using microwave to heating, and continuous turning meat over fire. Nowadays, the several methods used to determination of PAHs with HCAs in meat, and foods, high-performance liquid chromatography combined with mass spectrophotometer detectors, or combined with diode array detection are commonly used. As well as gas chromatography- mass spectrophotometer (GC-MS), GC-electrospray tandem MS, and HPLC-electrospray tandem MS.
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