Estimation of daily exposure to 4-hydroxy-2-alkenals in Korean foods containing n-3 and n-6 polyunsaturated fatty acids

4-Hydroxy-2-alkenals are cytotoxic aldehydes generated from the oxidation of n-3 and n-6 polyunsaturated fatty acids. The compounds have shown various biological effects via Schiff base adducts or Michael addition adducts at levels higher than physiological ones. To assess human exposure to 4-hydroxy-2-alkenals in the diet, 4-hydroxy-2-alkenals in vegetable oils, fish and shellfish were monitored using GC/MS/SIM. 2001 National Health and Nutrition Survey (2002) data were employed for the dietary intake pattern. The Korean daily exposure to 4-hydroxy-2-alkenals, excluding a possible one from fried food, was 4.3 µg day^-1, constituted of 1.6 µg 4-hydroxy-2-hexenal (HHE) and 2.7 µg 4-hydroxy-2-nonenal (HNE). It was calculated that Koreans could be additionally exposed to more than 11.8 µg day^-1 4-hydroxy-2-alkenal from fried foods. Thus, the combined exposure would be 16.1 µg day^-1, which corresponds to 0.3 µg kg^-1 body weight day^-1 for a 60 kg Korean adult. In spite of the biological toxicity of 4-hydroxy-2-alkenals, the risk for human could not be quantified due to the lack of a virtually safe dose of the compounds. However, considering the basal level of 4-hydroxy-2-alkenals in many tissues, the present value from the diet may not pose a significant risk for human health.     

Development of a Sensitive and Accurate Stable Isotope Dilution Assay for the Simultaneous Determination of Free 4-Hydroxy-2-(E)-Nonenal and 4-Hydroxy-2-(E)-Hexenal in Various Food Matrices by Gas Chromatography–Mass Spectrometry

An analytical method suitable for the determination of 4-hydroxy-2-(E)-nonenal (HNE) and 4-hydroxy-2-(E)-hexenal (HHE) in various food matrices was developed and validated. The method involves the use of deuterated HNE and HHE as internal standards, extraction of the analytes from the matrices followed by derivatization and detection with gas chromatography–mass spectrometry. Four different food matrices were chosen as model systems including vegetable oils, unprocessed meat, fried potato crisps, and infant formula and three different extraction techniques suitable for the different matrices were applied including the Quick Easy Cheap Effective Rugged Safe method. The simplicity of the extraction techniques allows the method to be applied for routine analysis of a large amount of samples. The results verify the accuracy and reproducibility of the analytical technique and its ability to provide reliable quantification of both analytes at concentrations as low as 12.8 ng g^?1 in meat samples. Furthermore, a short overview of the levels of HNE and HHE in several products available in the Belgian market is presented.     

Formation of malondialdehyde, 4-hydroxy-hexenal and 4-hydroxy-nonenal during deep-frying of potato sticks and chicken breast meat in vegetable oils

The present work investigated the formation of three toxic aldehydes, malondialdehyde (MDA), 4-hydroxy-hexenal (HHE) and 4-hydroxy-nonenal (HNE), during deep-frying of different foods in corn oil (CO), rapeseed oil (RO) and linseed oil (LO). Besides, their contents in French fries (FF) and fried chicken breast meat (FCBM) were also explored. Results showed that the MDA/HHE/HNE levels in frying groups were all lower than those in control, indicating the incorporation into the fried foods. Apart from the different levels, MDA/HHE/HNE in FF and FCBM possessed similar variation tendencies, revealing the matrix-mediated distribution. The combined exposure assessment based on the threshold of toxicological concern (TTC) levels was conducted, and risks of exceeding the limitation values (1.5 μg kg bw⁻¹ day⁻¹) for HNE in CO and HHE in LO were exhibited. The consumption of both FF and FCBM should be regulated, considering the levels of MDA, HHE and HNE these fried foods may contain.     

Fumonisins in Brazilian corn-based foods for infant consumption.

A survey of 196 samples of corn-based infant foods from 13 cities of Sao Paulo State, Brazil, was carried out to investigate the fumonisin contamination in the products. Based on their ingredients, the products were divided into seven groups: infant cereal designated as types A-D, corn meal, corn starch and instant cereal baby food. Although certain infant food samples were free of fumonisin contamination (<20 microg kg(-1); corn starch and infant cereals of type A, B and D), contamination levels in the other products (corn meal, instant corn-based baby food and cereal type C) were of concern, particularly those in corn meal. All samples in these categories contained fumonisins. The mean level for total fumonisins (FB1 + FB2 + FB3) in corn meal was 2242 microg kg(-1) (maximum 8039 microg kg(-1)), in instant corn-based baby food was 437 (maximum 1096) microg kg(-1) and in infant cereal type C was 664 (maximum 1753) microg kg(-1).     

Oxidation products of polyunsaturated fatty acids in infant formulas compared to human milk--a preliminary study

Information about lipid oxidation in fresh and stored human milk compared with infant formulas is scarce. We aimed to assess n-6 and n-3 PUFA oxidation in these milks by measuring the 4-hydroxynonenal (4-HNE) and 4-hydroxyhexenal (4-HHE) content. Human milk samples (n = 4), obtained from volunteer mothers, were analyzed fresh and after 1 wk at 4 degrees C or 24 h at 18 degrees C. Vitamin E and malondialdehyde (MDA) were measured by HPLC and fatty acid profile by GC. The 4-HHE and 4-HNE contents were measured by GC-MS. Infant formulas (n = 10) were tested; their fat droplet size was measured by laser light scattering and observed by confocal laser scanning microscopy. Human milk samples contained 31.0 +/- 6.3 g/L of lipids and 1.14 +/- 0.26 mg/L of vitamin E. Fat droplets were smaller in infant formulas than reported in human milk. The (4-HHE/n-3 PUFA) ratio was 0.19 +/- 0.01 microg/g in fresh human milk (unchanged after storage) versus 3.6 +/- 3.1 microg/g in dissolved powder formulas and 4.3 +/- 3.8 microg/g in liquid formula. (4-HNE/n-6 PUFA) was 0.004 +/- 0.000 microg/g in fresh milk (0.03 +/- 0.01 microg/g after storage) versus 1.1 +/- 1.0 microg/g in dissolved powder formulas and 0.2 +/- 0.3 microg/g in liquid formula. Infant formulas also contained more MDA than human milk. n-3 PUFA were more prone to oxidation than n-6 PUFA. Whether threshold levels of 4-HHE and 4-HNE would be of health concern should be elucidated.     

Influence of fatty acids compositions and manufacturing type on the formation of 4-hydroxy-2-alkenals in food lipids

Correlations between 4-hydroxy-2-hexenal (HHE) or 4-hydroxy-2-nonenal (HNE) and fatty acids were statistically investigated in 73 kinds of commercially available food sources of n-3 or n-6 polyunsaturated fatty acids (PUFA). The analysis revealed that the level of HNE was obviously correlated with the contents of PUFA (r=0.462), n-6 PUFA (r=0.512), and saturated fatty acids (r=0.535). Significant correlation was also observed between HHE and HNE (r=0.486) although whose source fatty acids were different. Regarding the sesame oils and perilla oils currently available on the Korean markets, no appreciable difference in the levels of HHE or HNE was found depending on their manufacturing types.     

Potential of selected infant food formulas for production of Bacillus cereus emetic toxin, cereulide

Cereulide producing Bacillus cereus was isolated from randomly chosen commercial infant foods. The cereulide production in infant food formulas was investigated. When the reconstituted foods were inoculated with >10(5) cfu ml(-1) of cereulide producing B. cereus, 2 to 200 microg of cereulide per 100 ml of food accumulated during 24 h of non-refrigerated storage. The amount of cereulide measured in the foods by the accurate chemical assay (LC-MS) matched with that found by sperm micro assay, proving the cereulide was the sole heat stable toxin in the foods and present in its toxic form. The infant formulas containing both cereal and dairy ingredients were the most supportive for cereulide production. Cereulide accumulation was affected by the infant food composition as well as by the handling of the food. Diluting the reconstituted food with water resulted in increased toxin production expressed as mug per volume. More cereulide was accumulated when the food was incubated stationary compared with moderate shaking. The amount of cereulide accumulated within 24 h at room temperature per 100 ml of cereal and dairy or in rice-nondairy reconstituted infant formulas, inoculated with >or=10(5) cfu ml(-1) of B. cereus strain F4810/72, was higher or similar to the amounts reported for foods implicated in emetic type of food poisonings. Thus mishandling and temperature abuse of infant foods may cause food poisoning when emetic B. cereus is present.     

Study of fumonisins B1 and B2 contamination of baby food

Fumonisins content were studied in the baby foods, grits and flour by HPLC. FB1 +FB2 were found in 95% of 129 samples of grits and flour in quantities from 30 to 4350 microg/kg (mean: 580 microg/kg). Fumonisins were found in baby food in 52% of snacks in the range from 10 to 720 microg/kg (mean: 35 microg/kg). Processed cereal-based and cereal and dairy-based food contained FB1+FB2 in 39% of cases in the amount from 10 to 9200 microg/kg (mean: 244 microg/kg). Fumonisins were not found in canned baby food on the cereal-based with meat, fish or fruit products.     

The distribution of 4-hydroxy-hexenal and 4-hydroxy-nonenal in different vegetable oils and their formation from fatty acid methyl esters

The formation of 4-hydroxy-hexenal (HHE) and 4-hydroxy-nonenal (HNE) in eight vegetable oils was investigated at 180 °C. HHE was only detectable in soybean (SBO), rapeseed (RO) and linseed oils (LO). HNE was measured in all tested oils, but was found mainly in corn (CO), sunflower (SO) and soybean oil (SBO). Oil-dependent formation of HHE/HNE was remarkably observed. Furthermore, different fatty acid methyl esters in tricaprylin, as model oil systems, were constructed to demonstrate their characteristic contribution to HHE/HNE formation. As expected, HHE and HNE originated from the oxidative degradation of methyl linolenate (MLN) and methyl linoleate (ML) respectively. Whereas low concentrations of MLN (<5.0%) and ML (<1.0%) produced no detectable HHE/HNE. The results suggested MLN/ML could induce both HHE/HNE formation and pro-oxidation at higher concentrations. Unexpectedly, methyl stearate and methyl oleate slightly promoted HHE/HNE formation, which might be attributed to free radical transfer mechanisms during thermal oxidation.     

Rapid and improved determination of furan in baby foods and infant formulas by headspace GC/MS

Furan is a 5-membered ring compound with high volatility. The U.S. Food and Drug Administration (FDA) has recently published a report on the occurrence of furan in a large number of thermally processed foods. However, the FDA's analytical method, using standard curve addition, is not suitable for high-throughput routine laboratory operations. We developed a rapid and improved method for determination of furan in foods by headspace GC/MS. Quantification was achieved by using an internal standard of d4-furan and an external calibration curve of furan normalized against the internal standard. The incubation temperature for equilibration was set at 60 degrees C to avoid the formation of furan during analysis. The levels of furan in baby foods and infant formulas were determined with this method. Validation data showed good precision and accuracy. The LOD and LOQ were 0.2-0.5 ng/g and 0.5-2 ng/g for various food matrixes, respectively. The level of furan detected was in the range of 1.4 to 90 ng/g in baby foods and in the range of non-detectable to 36 ng/g in infant formulas.     

Ochratoxin A in conventional and organic cereal derivatives: a survey of the Italian market, 2001-02.

Ochratoxin A is a mycotoxin produced mainly by Penicillium verrucosum and Aspergillus ochraceus. Although typically considered a cereal contaminant, it has also been detected in dried fruit, nuts, meat and derivatives. To estimate the quantity of ochratoxin A that might be ingested by Italian consumers from these foods, 211 cereal derivatives (flours and bakery products) were analysed by high-performance liquid chromatography. Products were from conventional and organic agriculture and from integrated pest management agriculture. All commercial flours and derivatives examined contained ochratoxin A at concentrations very much below the legal limit (3 microg kg(-1)): the highest value, 0.816 microg kg(-1), was detected in a sample of spelt whole flour from organic agriculture. In many samples, the ochratoxin content was below the limit of detection; only rarely did values exceed 0.5 microg kg(-1). In baby foods, four samples were above the particularly restrictive Italian legal limit of 0.5 microg kg(-1). Although some significant differences were found between samples from conventional and organic agriculture when some product categories were examined (namely, baby foods as semolina and rice creams), no important difference was found between the two types of agricultural practice when all types of cereal derivatives were considered together.     

Understanding the difference of 4-hydroxyhexenal and 4-hydroxynonenal formation in various vegetable oils during thermal processes by kinetic and thermodynamic study

The formation of two toxic aldehydes, 4-hydroxyhexenal (HHE) and 4-hydroxynonenal (HNE) were monitored in five vegetable oils during thermal processing (100–200 °C) with the results modelled. HHE was only determined in rapeseed and linseed oil, while HNE was determined in all tested oils and the contents varied significantly depending on the oil type. For HHE/HNE-detected oils, the evolution during heating was well described by the pseudo-first-order kinetic model. The equilibrium contents of HHE/HNE increased and the duration until equilibrium decreased as a function of temperature. The calculated reaction rate constants followed the Arrhenius law. The formation rate of HHE/HNE was remarkably oil-dependent, which was also in accordance with the calculated kinetic and thermodynamic parameters. Accordingly, the above results can contribute to a prediction of HHE/HNE formation in different vegetable oils during thermal processing, especially the temperature/time required during frying, which is generally higher than 100 °C and 6 h., respectively.     

Effect of Antioxidants and Cooking on Stability of n-3 Fatty Acids in Fortified Meat Products

ABSTRACT: This study was carried out to determine the effect of processing and cooking on n-3 polyunsaturated fatty acid (PUFA) stability and to determine the efficacy of antioxidants (ANTI) to minimize lipid oxidation in cooked n-3 PUFA-fortified meat products. An emulsion of n-3 PUFAs (25% algal oil) was incorporated into ground turkey, pork sausages, or restructured hams (500 mg n-3 PUFA/110 g meat) with or without an "antioxidant cocktail" containing citrate (0.5% w/w), erythorbate (1 g/kg product), and rosemary (0.2% w/w). Ground turkey and pork sausages were frozen 2 d, then cooked to 71°C, and stored at 4°C for 2 d. Cooked, restructured hams were sliced, vacuum packaged, and stored at 4°C, or frozen and thawed with subsequent storage at 4°C. Treatments were CON (control), n-3 (n-3 PUFAs), CON + ANTI and n-3 + ANTI. Products were analyzed for color, lipid oxidation (TBARS and peroxide values), and n-3 PUFA profile. TBARS of n-3 PUFA-fortified treatments in ground turkey and pork sausages increased with storage ( P < 0.05); there were no changes in TBARS for CON + ANTI and n-3 + ANTI groups ( P > 0.05). For restructured hams nitrite curing appeared to delay lipid oxidation such that antioxidant treatment effects were unobservable ( P > 0.05). Overall recovery of n-3 PUFAs after heat processing was 69% to 85%, and there was no effect of storage on n-3 PUFA concentration in raw or cooked products ( P > 0.05). Sensory scores for n-3 treated restructured hams were lower than controls ( P < 0.05). Results suggested that cooking resulted in some losses of n-3 PUFAs in fortified meat products and that an "antioxidant cocktail" protected against lipid oxidation during subsequent storage in non-cured meat products.     

Assessment of PCDD/F, PCB, OCP and BPA dietary exposure of non-breast-fed European infants

The aim of this study is to obtain data on the exposure of non-breast-fed infants to polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/F), polychlorinated biphenyls (PCB), organochlorine pesticides (OCP), and bisphenol A (BPA) and its chlorinated derivatives through consumption of commercial infant foods with largest shares of the market in 22 European Union countries. The E-Screen bioassay was employed to assess the oestrogenicity of the baby foods and the ethoxyresorufin-O-deethylase (EROD) induction was measured to determine the levels of PCDD/F and PCB. Consequently, the highest total effective xenoestrogen burden (TEXB) of 73.60?pM?Eeq?g(-1) was found in the soy-based formula and the EROD bioassay was always below the limit of quantification (LOQ) (3.5?pg?g(-1)). Overall, the estimated dietary exposure to BPA via commercial baby foods was lower than the tolerable daily intake (TDI) of 50?μg?kg(-1) body weight (bw). Furthermore, the findings indicated that the dietary exposure of 0-9-month-old infants through the products investigated here does not exceed the maximum TDI of 4?pg WHO-TEQ (toxic equivalents)?kg(-1) bw. However, exposure to more than 2?pg WHO-TEQ?kg(-1)?bw?day(-1) might occur for 0-4-month-old infants consuming 'starting' hypoallergenic formula. Moreover, analysis of OCP indicated that the dietary exposure of non-breast-fed infants was not harmful. Considering the importance of early development and the vulnerability of infants and children, it is essential to determine their dietary exposure to contaminants in order to decide which efforts of risk reduction should receive highest priority.     

Estimated US infant exposures to 3-MCPD esters and glycidyl esters from consumption of infant formula

A dietary exposure assessment was conducted for 3-monochloropropane-1,2-diol (3-MCPD) esters (3-MCPDE) and glycidyl esters (GE) in infant formulas available for consumption in the United States. 3-MCPDE and GE are food contaminants generated during the deodorisation of refined edible oils, which are used in infant formulas and other foods. 3-MCPDE and GE are of potential toxicological concern because these compounds are metabolised to free 3-MCPD and free glycidol in rodents and may have the same metabolic fate in humans. Free 3-MCPD and free glycidol have been found to cause adverse effects in rodents. Dietary exposures to 3-MCPDE and GE from consumption of infant formulas are of particular interest because formulas are the sole or primary food source for some infants. In this analysis, US Food and Drug Administration data on 3-MCPDE and GE concentrations (as 3-MCPD and glycidol equivalents, respectively) in a small convenience sample of infant formulas were used to estimate exposures from consumption of formula by infants 0–6 months of age. 3-MCPDE and GE exposures based on mean concentrations in all formulas were estimated at 7–10 and 2 µg/kg bw/day, respectively. Estimated mean exposures from consumption of formulas produced by individual manufacturers ranged from 1 to 14 µg/kg bw/day for 3-MCPDE and from 1 to 3 µg/kg for GE.     

Natural co-occurrence of deoxynivalenol and fumonisins B1 and B2 in Italian marketed foodstuffs

A survey was performed to obtain the frequency and levels of contamination by deoxynivalenol (DON) and fumonisins B₁ and B₂ (FB1, FB2) mycotoxins in Italian marketed foods. Of 202 samples investigated, including raw materials and processed cereal foods (bread, pasta, breakfast cereals, biscuits, baby and infant foods), 84% were contaminated with DON at levels from 0.007 to 0.930 μg g^-1 (median 0.065 μg g^-1); 26% contained FB1 ranging from 0.010 to 2.870 μg g^-1 (0.070 μg g^-1); 35% contained FB2 at 0.010-0.790 μg g^-1 (0.080 μg g^-1). The highest levels of DON and FB1 were detected in raw cereals and wholemeal flours. The highest levels of FB2 were detected in durum wheat pasta. A widespread DON contamination was found in baby and infant foods at levels varying from 0.007 to 0.166 μg g^-1.     

Novel approach to fast determination of multiple pesticide residues using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS)

A rapid, high-throughput method employing ultra-performance liquid chromatography with tandem quadrupole mass spectrometry (UPLC-MS/MS) was developed and optimized for simultaneous quantification and confirmation of 64 pesticide residues and their toxic metabolites in fruit extracts prepared by a buffered QuEChERS procedure. The total time required for UPLC-MS/MS analysis was 8 min plus 2 min for re-equilibration to the initial UPLC conditions. Performance characteristics were determined for apple extracts spiked at 10 microg kg(-1). The repeatability of measurements expressed as relative standard deviations was in the range 1.5-13% at this level for most analytes. Thanks to very low limits of quantification (<10 microg kg(-1)for the majority of pesticides), an optimized method allows for the reliable control of not only common maximum residue limits (MRLs) set by European Union regulation for various pesticides/fruit combinations, but also of a uniform MRL of 10 microg kg(-1)endorsed for baby food.     

Novel approach to fast determination of multiple pesticide residues using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS).

A rapid, high-throughput method employing ultra-performance liquid chromatography with tandem quadrupole mass spectrometry (UPLC-MS/MS) was developed and optimized for simultaneous quantification and confirmation of 64 pesticide residues and their toxic metabolites in fruit extracts prepared by a buffered QuEChERS procedure. The total time required for UPLC-MS/MS analysis was 8 min plus 2 min for re-equilibration to the initial UPLC conditions. Performance characteristics were determined for apple extracts spiked at 10 microg kg(-1). The repeatability of measurements expressed as relative standard deviations was in the range 1.5-13% at this level for most analytes. Thanks to very low limits of quantification (<10 microg kg(-1)for the majority of pesticides), an optimized method allows for the reliable control of not only common maximum residue limits (MRLs) set by European Union regulation for various pesticides/fruit combinations, but also of a uniform MRL of 10 microg kg(-1)endorsed for baby food.     

Bisphenol A in canned foods in New Zealand: an exposure assessment.

Exposure to bisphenol A (BPA) from the consumption of canned and bottled food has been determined for New Zealand adults. Eighty different canned foods purchased from retail outlets in Christchurch, New Zealand, between November 2003 and February 2004 were analysed for BPA concentration by gas chromatography/mass spectrometry. BPA was detected in all foods analysed except for soft drinks. Concentrations ranged from < 10 to 29 microg kg(-1), except for individual samples of tuna, corned beef and coconut cream, which were 109, 98 and 191 microg kg(-1) , respectively. The limit of quantitation was <10 microg kg(-1) for foods of low fat content (< 1%) and <20 microg kg(-1) for foods containing >1% fat. Mean concentration data were combined with 24-h dietary recall information for 4399 individual consumers. Mean and maximum exposures were 0.008 and 0.29 microg kg(-1) bw day(-1), respectively, well below the temporary tolerable daily intake of 10 microg kg(-1) bw day(-1) given by the European Commission in 2002. The results of the present survey suggest that the levels of BPA identified in canned foods are unlikely to be of concern to adult health, and there is no reason for consumers to change their consumption patterns as a result of these findings. When the concentration data found in the current survey are applied to an oestrogenicity model for an adult male, the contribution of BPA to the total oestrogenicity from 16 food components is 7%. The impact of this level of oestrogenicity remains unclear.     

In-house direct cELISA for determining aflatoxin B1 in Thai corn and peanuts.

To overcome the problem of aflatoxins (AF) in Thai foods, a sensitive in-house direct cELISA using monoclonal antibody (mAb) was established and compared with a commercial ELISA kit and thin-layer chromatography for the determination of AFB(1) levels in corn and peanuts. Among eight in-house mAbs (AF1-8), AF5 was used in the direct cELISA owing to its excellent specificity and sensitivity with the detection limit of 4 microg kg(-1). The recovery of AFB(1) spiked at 5, 10, 20, 40 and 80 microg kg(-1) ranged from 88.1 to 99.5%. Correlation coefficients of the ELISA with the commercial ELISA kit and thin-layer chromatography were 0.912 and 0.802 for corn, and 0.941 and 0.832 for peanuts, respectively (p<0.05). The cost per sample was estimated to be about 16 times lower than the commercial ELISA kit. Subsequently, the in-house direct cELISA was successfully applied to screen the contamination of AFB(1) in Thai corn and peanuts. Mean levels of AFB(1) (per cent positive) were 73 microg kg(-1) (85.7%) in corn and 102 microg kg(-1) (67.9%) in peanuts, for which 46.4% of both foods were above the Thailand regulation limit (20 microg kg(-1)).