Effect of brewing process and sugar content on 5-hydroxymethylfurfural and related substances from Turkish coffee

The effect of brewing process on 5-hydroxymethylfurfural, 2-furylmethylketone, and 2-furoic acid levels of traditionally prepared and instant Turkish coffee samples containing different amounts of table sugar (0, 2, 4, or 8 g in 20 mL of coffee) were analyzed by high pressure liquid chromatography with diode array detector. The highest change at the levels of furfurals was observed in sample of traditional and instant Turkish coffee named T4 and S4 both of containing 8 g of sugar, respectively. The results showed that 5-hydroxymethylfurfural, 2-furylmethylketone, and 2-furoic acid concentrations in both traditionally prepared and instant Turkish coffee samples increased with increasing sugar concentration. The brewing method and sugar concentration had a significant effect on furfural contents of Turkish coffee (p < 0.05). Daily intakes of furfurals for Turkish population were calculated as 8.14–13.54 and 9.36–10.25 µg kg^?1 body weight for traditionally prepared and instant Turkish coffee samples, respectively, and daily intakes of furfurals were lower than the acceptable daily intake value of 0.5 mg kg^?1 body weight.     

Survey on acrylamide in roasted coffee and barley and in potato crisps sold in Italy by a LC–MS/MS method

ABSTRACT

A survey on the occurrence of acrylamide (AA) in roasted coffee, barley, and potato crisps was carried out using an intra-lab validated liquid chromatography (LC)–MS (mass spectrometry)/MS method. Over the years 2015–2016, 66 samples of coffee, 22 of roasted barley, and 22 of potato crisps were collected from retail outlets in Italy. AA was detected in almost all samples. In roasted coffee, the level exceeded 450 µg kg^?1, the limit recommended by the European Commission (EC), in 36.4% of the samples. In roasted barley, mean contamination was slightly lower than in coffee and no sample exceeded the EC limit of 2000 µg kg^?1. The AA contamination in potato crisps was remarkable. A percentage of 36.4 (n = 8) showed a value higher than the EC limit of 1000 µg kg^?1. Considering the average consumption of coffee and potato crisps by Italian people, AA exposure is significant and should be decreased.     

Furanic compounds and furfural in different coffee products by headspace liquid-phase micro-extraction followed by gas chromatography–mass spectrometry: survey and effect of brewing procedures

In this study, the levels of furan, 2-methylfuran, 2,5-dimethylfuran, vinyl furan, 2-methoxymethyl-furan and furfural in different coffee products were evaluated. Simultaneous determination of these six furanic compounds was performed by a head space liquid-phase micro-extraction (HS-LPME) method. A total of 67 coffee powder samples were analysed. The effects of boiling and espresso-making procedures on the levels of furanic compounds were investigated. The results showed that different types of coffee samples contained different concentrations of furanic compounds, due to the various processing conditions such as temperature, degree of roasting and fineness of grind. Among the different coffee samples, the highest level of furan (6320 µg kg^?1) was detected in ground coffee, while coffee-mix samples showed the lowest furan concentration (10 µg kg^?1). Levels in brewed coffees indicated that, except for furfural, brewing by an espresso machine caused significant loss of furanic compounds.     

A deep eutectic solvent based liquid phase microextraction for the determination of caffeine in Turkish coffee samples by HPLC-UV

ABSTRACT

A novel liquid-phase microextraction procedure for the determination and extraction of caffeine using a deep eutectic solvent based liquid-phase microextraction method (DES-LPME) was developed. A deep eutectic solvent consisting of choline chloride-phenol (1:3) and tetrahydrofuran as a dilution solvent were used for the extraction of caffeine from Turkish coffee samples. The quantitative recoveries were obtained when the DES volume was 400 µL, and the volume of tetrahydrofuran that was used as an emulsifier solvent was 800 µL. The limit of detection and the limit of quantification were found to be 0.12 and 0.40 µg mL^?1, respectively. Linear dynamic range was 0.5–100 µg mL^?1, and coefficient of determination (R²) was 0.9998. The relative standard deviation (RSD) was 2.20% when the standard solution concentration was 1.0 µg mL^?1. The fact that the determination coefficient obtained from the calibration curve was greater than 0.9998 gives information about the accuracy of the method. Also, in order to determine the accuracy of the developed method, selected coffee samples were spiked with 25 and 50 µg mL^?1 caffeine.     

Acrylamide, 5-hydroxymethylfurfural and Nε-carboxymethyl-lysine in coffee substitutes and instant coffees

Sensitive analytical methods were developed and validated for the quantification of acrylamide, N^ε-carboxymethyl-lysine (CML) and 5-hydroxymethylfurfural (HMF) in 24 commercial coffee substitutes (CSs) and 12 instant coffees (ICs). Acrylamide levels varied widely from 200 to 4940 µg kg^–1 with higher levels in CSs. Only two out of 24 CSs had a level of acrylamide above the indicative value set for this food category by the European Commission (4000 µg kg^–1). None of the ICs tested in this study exceeded the indicative value set for this foodstuff (900 µg kg^–1). CML ranged from 0.17 to 47 mg kg^–1 and it increased in proportion to the protein content of the samples. The highest concentrations were found in IC partly due to the relatively high protein content of this food group. HMF was the most abundant neoformed compound (NFC) found in the tested commercial samples. It was found between 0.59 and 13 g kg^–1. Among other food categories IC and CS could appear to be major contributors to the exposure to NFCs if consumed on a daily basis. Further investigations are needed to elucidate the acrylamide formation during processing and to determine the daily intake level of frequent consumers of these products.     

Occurrence of patulin and 5-hydroxymethylfurfural in apple sour,which is a traditional product of Kastamonu,Turkey

Apple sour is a traditional product of Kastamonu, Turkey. It is consumed by spreading on bread or drinking after diluting with water. The aim of this study was to determine patulin (PAT) and 5-hydroxymethylfurfural (HMF) in apple sour. This study is the first to evaluate the occurrence of PAT and HMF in apple sour. The samples were extracted with ethyl acetate using liquid-liquid extraction technique. PAT and HMF were determined by HPLC with UV detection. PAT was detected in all samples, and the PAT level in 94.9% of samples was found to be equal or greater than the legal limit for juice concentrates. The mean value for PAT was found to be 284 ± 307 μg kg^?1. PAT levels in 13 of 39 samples were in the range of 100 ≤ x < 200 μg kg^?1, two samples were in the range of 0 ≤ x < 50 μg kg ?1 and two samples were in the range of 1000 ≤ x < 1500 μg kg¹. HMF levels of all samples were above the legal limit for solid molasses. The mean value for HMF was found to be 16215 ± 13317 mg kg^?1. HMF levels of 10 of 39 samples were determined to be in the range of 10000 ≤ x < 20000 mg kg^?1, eight samples were in the range of 20000 ≤ x < 30000 mg kg^?1 and only three samples were in the range of 100 ≤ x < 1000 mg kg^?1. There was a significant and inverse relationship between HMF and pH of the samples. These results indicate that consumption of apple sour is a considerable risk in terms of HMF and PAT toxicity.     

Improved sensitivity of ochratoxin A analysis in coffee using high-performance liquid chromatography with hybrid triple quadrupole-linear ion trap mass spectrometry (LC-QqQLIT-MS/MS)

A novel and sensitive method utilising high-performance liquid chromatography coupled to triple quadrupole-linear ion trap mass spectrometry (LC-QqQLIT-MS/MS) was developed in order to analyse the content of ochratoxin A (OTA) in coffee samples. The introduction of the triple-stage MS scanning mode (MS³) has been shown to increase greatly sensitivity and selectivity by eliminating the high chromatographic baseline caused by interference of complex coffee matrices. The analysis included the sample preparation procedure involving extraction of OTA using a methanol–water mixture and clean-up by immunoaffinity columns and detection using the MS³ scanning mode of LC-QqQLIT-MS/MS. The proposed method offered a good linear correlation (r² > 0.998), excellent precision (RSD < 2.9%) and recovery (94%). The limit of quantification (LOQ) for coffee beans and espresso beverages was 0.010 and 0.003 µg kg^?1, respectively. The developed procedure was compared with traditional methods employing liquid chromatography coupled to fluorescent and tandem quadrupole detectors in conjunction with QuEChERS and solid-phase extraction. The proposed method was successfully applied to the determination of OTA in 15 samples of coffee beans and in 15 samples of espresso coffee beverages obtained from the Latvian market. OTA was found in 10 samples of coffee beans and in two samples of espresso in the ranges of 0.018–1.80 µg kg^?1 and 0.020–0.440 µg l^?1, respectively. No samples exceeded the maximum permitted level of OTA in the European Union (5.0 µg kg^?1).     

Detection of 5-hydroxymethyl-2-furfural Levels in Selected Chinese Foods by Ultra-High-Performance Liquid Chromatograph Analytical Method

In this study, an ultra-high-performance liquid chromatography method was developed to detection the levels of 5-hydroxymethyl-2-furfural (HMF) in 227 selected food products obtained from the Chinese markets. The performance of the analysis method was evaluated by some quality parameters such as limit of detection (LOD), limit of quantification (LOQ), linearity, recovery, and run-to-run (n?=?6) and day-to-day (n?=?18) precisions. The LOD and the LOQ of the method in different food matrices ranged from 0.15 to 0.50 and from 0.35 to 1.20 mg kg^?1, respectively. The results from this study showed that HMF was mostly detected in all samples selected. HMF contents in different samples varied greatly according to the raw materials and processing conditions. The highest level of HMF was found in preserved fruits and ground coffee, with average levels of 409.6 and 409.9 mg kg^?1, respectively. Preliminary estimates of HMF exposure from foods in the Chinese population was estimated to be 0.12 mg kg^?1 body weight day^?1, which is relatively low compared with the result reported by JECFA and European Food Safety Authority.     

Simultaneous Determination of Alkaloids in Green Coffee Beans from Ethiopia: Chemometric Evaluation of Geographical Origin

The alkaloid compositions of 99 green coffee (Coffea arabica L.) bean samples comprising eight varieties (Harar, Jimma, Kaffa, Wollega, Sidama, Yirgachefe, Benishangul and Finoteselam) from the major production regions of Ethiopia were investigated. High performance liquid chromatography was applied for the simultaneous determination of four coffee alkaloids in the aqueous extracts of the beans. The limits of detection for the method were established as 13 mg kg^?1 for trigonelline, 7 mg kg^?1 for theobromine, 8.5 mg kg^?1 for caffeine and 4 mg kg^?1 for theophylline in the dry coffee beans. Theophylline was not detected in any of the samples. The determined concentrations (% w/w dry coffee beans) ranged from 0.98 to 1.32 % for trigonelline, 0.0048 to 0.0094 % for theobromine and 0.87 to 1.38 % for caffeine. The concentrations of the alkaloids varied significantly, depending on the geographical origin of the beans. Theobromine was not detected in coffee beans from the East (Harar coffees), and its absence in samples can be used to ascertain whether the coffee originates from this region. Coffee beans from the Northwest were characterized by higher concentrations of caffeine. Application of linear discriminant analysis provided 75 % correct classification of samples into the respective production regions, with a 74 % prediction success rate. The moderate classification efficiency obtained when using alkaloid data demonstrates the potential of using this class of compounds in discriminant models for determination of the geographical origin of green coffee beans from Ethiopia.     

Post-harvest practices linked with ochratoxin A contamination of coffee in three provinces of Cordillera Administrative Region,Philippines

One of the emerging concerns in the Cordillera Administrative Region, Philippines is ochratoxin A (OTA) contamination in coffee. During 2015 to 2016, a total of 51 Arabica (Coffea arabica) coffee samples from Benguet province and 71 Robusta (Coffea canephora var. Robusta) coffee samples from the provinces of Ifugao and Kalinga were analysed for OTA contamination. The OTA-producing fungal contaminants during drying and storage of Arabica and Robusta coffee were Aspergillus niger and Aspergillus ochraceus. Ochratoxin A was more commonly detected in Robusta coffee (36.6%) than in Arabica coffee (21.6%). Among the contaminated samples, Robusta coffee cherries in the drying yard had the highest mean OTA level (120.2 μg kg^?1, n = 10) while roasted Robusta coffee beans had the lowest mean level (4.8 μg kg^?1, n = 9). The onset of contamination of Arabica coffee occurred during storage, with a mean OTA level of 46.7 μg kg^?1 (n = 9). Roasted coffee had lower OTA content although five samples had levels >5.0 μg kg^?1. Pearson Chi-square analysis (χ²) and Fisher’s exact test revealed that several post-harvest practices involving non-removal of the husk or hull and mixing of defective coffee were significantly associated with the occurrence of OTA during drying and storage (p < 0.05). No significant associations, however, were identified during roasting. This study suggests that the post-harvest practices in Cordillera Administrative Region should focus on the removal of defective coffee in all stages of post-harvest and rapid reduction of moisture content particularly during drying.     

Occurrence of furan in commercial processed foods in Brazil

Selected commercial processed foods available in the Brazilian market (306 samples) were analysed for furan content using a validated gas chromatography-mass spectrometry method preceded by headspace solid phase micro-extraction (HS-SPME-GC/MS). Canned and jarred foods, including vegetable, meat, fruit and sweet products, showed levels up to 32.8?µg?kg^?1, with the highest concentrations observed in vegetables and meats. For coffee, furan content ranged from 253.0 to 5021.4?µg?kg^?1 in the roasted ground coffee and from not detected to 156.6?µg?kg^?1 in the beverage. For sauces, levels up to 138.1?µg?kg^?1 were found. In cereal-based products, the highest concentrations (up to 191.3?µg?kg^?1) were observed in breakfast cereal (corn flakes), cracker (cream crackers) and biscuit (wafer). In general, these results are comparable with those reported in other countries and will be useful for a preliminary estimate of the furan dietary intake in Brazil.     

Oxytetracycline,tetracycline, chlortetracycline and doxycycline in pasteurised cow’s milk commercialised in Brazil

The aim of this study was to investigate the presence of tetracycline residues in pasteurised cow’s milk using high-performance liquid chromatography coupled with UV/VIS detection to determine the exposure of Brazilian’s population to antibiotic residues. One hundred samples collected from the State of Paraná, Brazil, were analysed. Three of these samples were contaminated at the following concentrations: 121.8 µg·kg^?1 for oxytetracycline, 93.5 µg·kg^?1 for tetracycline and 134.6 µg·kg^?1 for chlortetracycline (61.6 µg·kg^?1) and doxycycline (73.0 µg·kg^?1). The median tetracycline residue concentration found in the samples was 42.3 µg·kg^?1, and the estimated daily intake (EDI) was 0.05 µg Kg^?1 bw day^?1 in Brazil. These results demonstrate that the occurrence of tetracycline in Brazilian milk was low (3%) and only for 2% above the maximum residue limit, so the risk to the population from the presence of these residues in milk was low (<1% of the acceptable daily intake).     

Characterization of Chlorogenic Acids in Coffee by Flow-Through Chronopotentiometry

In this study, flow-through chronopotentiometry (FTCP) has been developed as an electroanalytical method for characterization (identification and quantification) of chlorogenic acids (CGAs) in coffees. The characterization of CGAs in coffee was based on the electrochemical behavior of the main chlorogenic acid (CGAs) isomers presented in coffee (caffeoylquinic acids (CQAs), dicaffeoylquinic acids (diCQAs), and feruloylquinic acids (FQAs)) and the spiking of CGAs standards in coffee samples. The FTCP study has shown that electrochemical properties of CGAs strongly depend on their chemical structure and electronic properties, particularly on the presence of electron-donating ?OH, ?CH═CH? and ?OCH₃ groups and strong electron-withdrawing ester (?COOR) group presented in their structure. The FTCP measurements of coffee samples show that their electrochemical behavior is very similar to that of CGAs. Therefore, FTCP can be used for characterization of CGAs and determination of their content in coffees. 5-O-Caffeoylquinic acid (5-CQA), prevailed CGAs in coffees, was used as a standard for quantification of total CGA content in coffee. The linear calibration curve of 5-CQA was observed within the concentration range of 5 to 100 μmol L^?1 with the limit of detection 5.7·10^?7 mol L^?1. The total CGA content of coffees has been expressed in 5-CGA equivalents per 100 g of coffee. It was shown that FTCP is a very sensitive, precise, and acurate method for determination of total CGA content in coffee. It should be noted that in presented investigation, FTCP was for the first time used for the study of electrochemical properties of polyphenolic antioxidants (including CGAs) and their characterization in some of the food samples.     

Survey of acrylamide levels in Chinese foods

A survey of levels of acrylamide (AA) in 349 food products obtained from the Chinese market was conducted. AA was determined by an liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. The limit of detection (LOD) and the limit of quantification (LOQ) of the method in four different matrices ranged from 0.8 to 10 µg kg^?1 and from 4.0 to 25 µg kg^?1, respectively. The results from this survey indicated that AA was present in all samples except drinking water and tea. AA contents in different samples varied greatly according to the raw materials and processing conditions. The highest level of AA was found in potato products, with an average level of 1467 µg kg^?1. Preliminary estimates of AA exposure and risk assessment of AA from foods in the Chinese population were performed using a combination of data obtained in the present survey and 2002 dietary consumption survey carried out in 2002 for the Chinese population. The average dietary exposure of AA was estimated to be 0.38 µg kg^?1 body weight day^?1, which is relatively low compared with the result reported by the Food and Agricultural Organization/World Health Organization (FAO/WHO). Furthermore, the margin of exposure for neurotoxicity, reproductive toxicity, and carcinogenicity of AA was calculated to be 1318, 5250, and 787, respectively.     

Determination and surveillance of hydrocortisone and progesterone in livestock products by liquid chromatography-tandem mass spectrometry

A simple analytical method for the determination of hydrocortisone and progesterone in bovine, swine, and chicken muscle and eggs was developed. Hydrocortisone and progesterone were extracted with acetonitrile and subsequently cleaned-up using an Oasis^® HLB mini-cartridge. The method was validated in accordance with Japanese guidelines and exhibited trueness from 86.6% to 104.3% and precision (relative standard deviations (RSDs) of repeatability and within reproducibility were under 8.7% and 11.7%, respectively). The method was applied to 103 bovine muscle, 137 swine muscle, 69 chicken muscle and 52 egg samples that were commercially available in Tokyo, Japan. The hydrocortisone concentration was 0.9–41.2 µg kg^?1 in all bovine muscle samples, with an average of 7.7 µg kg^?1 and a median of 6.2 µg kg^?1. The progesterone concentration in 50 samples exceeded the limit of quantification (LOQ) and reached a maximum of 95.4 µg kg^?1. Hydrocortisone was also detected in all swine muscle samples at concentrations of 2.0–56.0 µg kg^?1. Its average and median concentrations amounted to 13.1 and 11.3 µg kg^?1, respectively. Twenty-three samples contained progesterone levels surpassing the LOQ, with a maximum concentration of 107.0 µg kg^?1. No chicken muscle samples contained any of the analytes. The progesterone concentration was 15.5–200.0 µg kg^?1 in all egg samples, with an average of 95.4 µg kg^?1 and a median of 90.5 µg kg^?1.     

Evaluation of bioactive amine and mineral levels in Turkish coffee

Coffee is one of the most popular drinks across the world and its enormous commercial and social importance is obvious. The levels of nine bioactive amines and five minerals, as well as pH, color, total ash and dry matter characteristics, were determined in different brands of ground and brewed Turkish coffees for the first time. Turkish coffee is a method of preparation, not a kind of coffee. Furthermore, this paper reported the estimated daily mineral intake of five elements. The amines were extracted with trichloroacetic acid (TCA) and quantified by reversed-phase high performance liquid chromatography (HPLC), pre-column derivatization with benzoyl chloride. Of the nine bioactive amines under study putrescine, cadaverine, tyramine and serotonin were detected in all coffee samples. Serotonin was the prevailing bioactive amine in both ground and brewed coffees. The total amine levels in the ground coffee varied from 126.0 mg/kg to 352.2 mg/kg. The total amine levels in the brewed coffee varied from 5.679 mg/L to 48.88 mg/L. The concentrations of five minerals (Mg, Mn, Zn, Na, K) were determined after dry mineralization of both ground and brewed coffee samples. The total mineral contents were analyzed by high-resolution continuum source flame atomic absorption spectrometry (HR-CS-FAAS). Potassium was the prevailing mineral in both ground and brewed coffees. The mean total mineral contents of ground and brewed coffees were 12673.2 mg/kg and 1014.8 mg/L, respectively. Significant differences were obtained between bioactive amines, minerals (except Na), color values (L*, a*, b*), total ash and total dry matter contents of ground and brewed Turkish coffee samples (P < 0.05).     

Aflatoxins and ochratoxin A in export quality raisins collected from different areas of Pakistan

During 2012–2014, 170 samples of export quality raisins were collected from different vendors in Pakistan. The collected samples were analysed for the presence of aflatoxins (AFs) and Ochratoxin A (OTA) contamination using high-performance liquid chromatography technique. The limit of detection and limit of quantification of AFs/OTA were 0.12/0.10 and 0.36/0.30 µg kg^?1, respectively. Only 5% of the samples were contaminated with AFs, ranging 0.15–2.58 µg kg^?1 with a mean of 0.05 ± 0.26 µg kg^?1. None of the raisin samples exhibited AFs contamination above the maximum limit (ML = 4 µg kg^?1) as set by the European Union (EU). About 72% of the samples were contaminated with OTA, ranging 0.14–12.75 µg kg^?1 with a mean of 2.10 ± 1.9 µg kg^?1. However, in 95.3% of the tested samples, OTA level was lower than the ML of 10 µg kg^?1 as regulated by the EU. Apparently, a strict and continuous monitoring plan, including regulatory limits, improves food safety and quality for all types of commodities.     

Occurrence of pyrrolizidine alkaloids in animal- and plant-derived food: results of a survey across Europe

Pyrrolizidine alkaloids (PAs) are secondary metabolites of plant families such as Asteraceae or Boraginaceae and are suspected to be genotoxic carcinogens. Recent investigations revealed their frequent occurrence in honey and particularly in tea. To obtain a comprehensive overview of the PA content in animal- and plant-derived food from the European market, and to provide a basis for future risk analysis, a total of 1105 samples were collected in 2014 and 2015. These comprised milk and milk products, eggs, meat and meat products, (herbal) teas, and (herbal) food supplements collected in supermarkets, retail shops, and via the internet. PAs were detected in a large proportion of plant-derived foods: 91% of the (herbal) teas and 60% of the food supplements contained at least one individual PA. All types of (herbal) teas investigated were found to contain PAs, with a mean concentration of 460 µg kg^?1 dry tea (corresponding to 6.13 µg L^?1 in [herbal] tea infusion). The highest mean concentrations were found in rooibos tea (599 µg kg^?1 dry tea, 7.99 µg L^?1 tea infusion) and the lowest in camomile tea (274 µg kg^?1 dry tea, 3.65 µg L^?1 tea infusion). Occurrence of PAs in food supplements was found to be highly variable, but in comparable ranges as for (herbal) tea. The highest concentrations were present in supplements containing plant material from known PA-producing plants. In contrast, only 2% of the animal-derived products, in particular 6% of milk samples and 1% of egg samples, contained PAs. Determined levels in milk were relatively low, ranged between 0.05 and 0.17 µg L^?1 and only trace amounts of 0.10–0.12 µg kg^?1 were found in eggs. No PAs were detected in the other animal-derived products.     

Metal levels in Trachurus trachurus and Cyprinus carpio in Turkey

Concentrations of five toxic metals were determined in two fish species from Turkish cities during 2010–2011. The obtained lead concentrations for all of the studied Trachurus trachurus (mean 777 μg kg^?1) and Cyprinus carpio (mean 439 μg kg^?1) samples were found to be higher than the maximum level (ML) of 300 μg kg^?1, while Cd concentrations in the same samples were lower than the ML. Mean chromium (501 μg kg^?1), Ni (272 μg kg^?1) and Cu (785 μg kg^?1) concentrations in T. trachurus were significantly higher than in C. carpio (336 μg Cr kg^?1, 229 μg Ni kg^?1 and 394 μg Cu kg^?1), similar to those of Pb and Cd. Measured Pb concentrations in T. trachurus tissues are significantly higher than the ML, while those of Cd in both T. trachurus and C. carpio species were lower than the ML values.     

Surveys of aflatoxin B1 contamination of retail Turkish foods and of products intended for export between 2007 and 2009

Surveys were carried out between 2007 and 2009 to determine the aflatoxin B₁ content of 3345 commercial Turkish foodstuffs supplied by producers for testing for their own purposes or for export certification. To simplify the reporting of data, foods were categorized as: 1, high sugar products with nuts; 2, nuts and seeds; 3, spices; 4, grain; 5, cocoa products; 6, dried fruit and vegetables; 7, processed cereal products; 8, tea; and 9, baby food and infant formula. Aflatoxin analysis was carried out by high-performance liquid chromatography with fluorescence detection after immunoaffinity column clean-up, with a recoveries ranging from 91% to 99%, depending on the matrix. Of the 3345 samples analysed, 94% contained aflatoxin B₁ below the European Union limit of 2 µg kg^?1, which applies to nuts, dried fruit, and cereals products. The 6% of the 206 contaminated samples were mainly nuts and spices. For pistachios, 24%, 38%, and 42% of the totals of 207, 182, and 24 samples tested for 2007, 2008 and 2009, respectively, were above 2 µg kg^?1, with 50 samples containing aflatoxin B₁ at levels ranging from 10 to 477 µg kg^?1.