Aspergillus section Flavi and aflatoxins in dried figs and nuts in Algeria

The presence of Aspergillus section Flavi and aflatoxin (AF) contamination was investigated in 112 samples of peanuts, almonds and dried figs collected in Algeria. The occurrence of aflatoxin B1 (AFB1), B2 (AFB2), G1 (AFG1) and G2 (AFG2) in different commodities has been determined with a sensitive method based on high performance liquid chromatography (HPLC) coupled with fluorescence detection with post-column photochemical derivatisation. Analytical results indicated that 28 samples of peanuts, 16 samples of almonds and 26 samples of dried figs contained detectable levels of AFs. A total of 69 samples (61.6%) were contaminated with AFB1 ranging from the limit of quantification to 174 µg kg^?1. AFB2 was found in 12 samples (10.7%) and varied from 0.18 to 193 µg kg^?1. Seven samples revealed AF concentrations lower than the limit of quantification. Eleven peanut and fourteen dried fig samples exceeded the European maximum limits for AFB1.     

Determination of 2,6-diisopropylnaphthalene (DIPN) and n-dibutylphthalate (DBP) in food and paper packaging materials from US marketplaces

A gas chromatography-ion-trap tandem mass spectrometry procedure was developed for the determination of 2,6-diisopropylnaphthalene (DIPN) and n-dibutylphthalate (DBP) in domestic and imported paper packages and food sold in US marketplaces. The procedure involved ultrasonic extraction with dichloromethane, followed by analysis with the gas chromatography-ion-trap tandem mass spectrometry. Calibration curves for DIPN and DBP were achieved with concentrations ranging from 0.01 to 10 µg ml^?1 and the corresponding r ² values were 0.9976 and 0.9956, respectively. In most of the fortified samples the recoveries were higher than 80% with a relative standard deviation (RSD) <10%. Using this procedure, it was found that less than 20% of the tested domestic packages and more than 60% of the tested imported food packages contained both DIPN and DBP. The concentrations of DIPN and DBP ranged from 0.09 to 20 mg kg^?1 and 0.14 to 55 mg kg^?1, respectively, with most of the DINP and DBP levels lower than 20 mg kg^?1. DIPN was not detected (<0.01 mg kg^?1) in 41 food samples and DBP was only detected in two domestic and four imported food samples with concentrations ranging from <0.01 to 0.81 mg kg^?1.     

Nutritional quality of uncultivated cereal grains utilised as famine foods in western sudan as measured by chemical analysis

The chemical composition of the grain of a number of wild grasses used as food in times of famine in western Sudan are reported and compared with those of the local staple cereals. Species investigated were Cenchrus hiflorus (haskaneet). Dactyloctenium aegyptium (korecb). Echinochloa colona (difra) and Oryza punctata (ruz el wadi). The grain of all species except C biflorus had protein contents in the range 110-140 g kg^?1, broadly similar to those of the local staple cereals. The grain of C biflours possessed a particularly high protein content of 210 g kg^?1. The nutritional quality of the proteins as measured by their chemical scores ranged from 20 for C biflorus to 65 for O punctata compared with scores of 36 and 58 for the local sorghum var Kurgi and millet respectively. Lysine was the limiting amino acid in all cases. Whilst the composition of the protein of sorghum var Kurgi was typical of sorghum generally, that of var Karamaka was unusual in possessing lysine levels of 3.4 g (16 g N)¹, double that normally present in sorghum and raising the chemical score of the protein to 62. The ash content of the famine food cereals was high, attaining 113 g kg¹ in C biflorus. All grains were good sources of zinc and D aegyptium. O punctata and E colonum additionally contained comparatively high levels of iron. Remarkably for a cereal D aegyptium was also very rich in calcium, containing over 10 g kg¹, and possessed high concentrations of manganese. Carbohydrate content of the famine food grains varied from 617 g kg¹ for C biflorus to 748 g kg¹ for O punctata, values comparable to the local staple cereals. The proportion of starch in this fraction ranged from 650 g kg¹ in D aegyptium to 940 g kg¹ in O punctata. Seed oils of all species were predominantly unsaturated containing mainly oleic and linoleic acids.     

Duplex real-time PCR method for the detection of sesame (Sesamum indicum) and flaxseed (Linum usitatissimum) DNA in processed food products

The development of a duplex real-time polymerase chain reaction (PCR) method allowing the simultaneous detection of sesame and flaxseed DNA in commercial food products is described. This duplex real-time PCR technique is based in the design of sesame- and flaxseed-specific primers based on the ITS1 region and two TaqMan fluorescent probes. The method was positive for sesame and flaxseed, and showed no cross-reactivity for all other heterologous plant and animal species tested. Sesame and flaxseed could be detected in a series of model samples with defined raw and heat-treated sesame in flaxseed, and flaxseed in sesame, respectively, with detection limits of 1.3 mg kg^?1 for sesame and 1.4 mg kg^?1 for flaxseed. The applicability of the assay for determining sesame and flaxseed in different food matrices was investigated by analysing a total of 238 commercial foodstuffs. This PCR method is useful for highly selective and sensitive detection of traces of sesame and flaxseed in commercial food products.     

Aflatoxin B1, ochratoxin A and zearalenone in sorghum grains marketed in Tunisia

A total of 64 samples of sorghum (37 Tunisian sorghum samples and 27 Egyptian sorghum samples) were collected during 2011–2012 from markets in Tunisia. Samples were analysed for contamination with aflatoxin B1, ochratoxin A and zearalenone by High-Performance Liquid Chromatography Coupled with Fluorescence Detection (HPLC-FLD). Aflatoxin B1 was found in 38 samples in the range 0.03–31.7 µg kg^?1. Ochratoxin A was detected in 24 samples with concentrations ranging from 1.04 to 27.8 µg kg^?1. Zearalenone was detected in 21 samples and the concentration varied between 3.7 and 64.5 µg kg^?1. ANOVA analysis of the influence of the country of origin on the incidence and concentration of mycotoxins in the samples studied showed no significant difference (P > 0.05) between the two batches of samples for each of the three mycotoxins studied. The studied mycotoxins contaminate sorghum and may also co-exist because of the diversity of the mycobiota in this cereal.     

Influence of PROP taster status on the consumer acceptability of food made from tannin sorghums

BACKGROUND: Condensed tannins in sorghum are powerful antioxidants, beneficial for health. However, tannin sorghums are believed to be unpalatable. The objective was to determine the influence of 6‐n‐propylthiouracil (PROP) taster status on consumer acceptability of food from tannin sorghums. Consumers (n = 194) classified by PROP taster status (super, medium and non) evaluated the appearance, flavour, overall liking and texture of sorghum rice from two tannin‐free sorghums, PAN 8564 and Phofu, and two tannin sorghums, PAN 3860 (82 g catechin equivalents (CE) kg^?1) and NS 5511 (18 g CE kg^?1), with high antioxidant activity. RESULTS: The PROP tasters could distinguish differences among the sorghum cultivars varying in tannin content levels, finding PAN 3860 less acceptable than the other sorghums. The non tasters preferred all the cultivars equally, presumably because they could not detect taste (bitter and astringent) differences between the sorghums. With the exception of appearance, tannin sorghum NS 5511 was generally equally preferred by PROP tasters to tannin‐free sorghums. CONCLUSION: There appear to some tannin sorghums that are palatable, even to PROP tasters, because the level of tannins is below a sensitivity threshold (ca 20 g CE kg^?1). Hence such tannin sorghums could be used to produce acceptable high‐antioxidant activity foods. Copyright © 2009 Society of Chemical Industry     

Characterisation and functional properties of watermelon (Citrullus lanatus) seed proteins

BACKGROUND: People in developing countries depend largely on non‐conventional protein sources to augment the availability of proteins in their diets. Watermelon seed meal is reported to contain an adequate amount of nutritional proteins that could be extracted for use as nutritional ingredients in food products. RESULTS: Osborne classification showed that globulin was the major protein (≥500 g kg ^?1) present in watermelon seed meal, followed by albumin and glutelin. Sodium dodecyl sulfate polyacrylamide gel electrophoresis indicated that the polypeptides had low molecular weights ranging from 35 to 47 kDa. Isoelectric focusing revealed that the isoelectric point of most proteins was in the acidic range 4–6. These proteins are rich in aspartic acid, glutamic acid and serine. An increase in pH (5–9) significantly (P < 0.05) decreased the denaturation enthalpy of these proteins. Among functional properties, albumin exhibited a much higher dispersibility index (810.3–869.6 g kg^?1) than globulin (227.8–245.4 g kg^?1), glutelin (182.1–187.7 g kg^?1) and prolamin (162.3–177.7 g kg^?1). Digestibility was in the ranges 760.6–910.0 and 765.5–888.5 g kg^?1 for Mateera and Sugar Baby watermelon protein fractions respectively, while surface hydrophobicity ranged from 126.4 to 173.2 and from 125.8 to 169.3 respectively. The foaming and emulsifying properties of albumin were better than those of the other proteins studied. CONCLUSION: The good nutritional and functional properties of watermelon seed meal proteins suggest their potential use in food formulations. Copyright © 2010 Society of Chemical Industry     

Determination of furan levels in commercial samples of baby food from Brazil and preliminary risk assessment

Commercial baby food samples available on the Brazilian market (n = 31) were analysed for furan content using a gas chromatography-mass spectrometry method preceded by solid-phase microextraction. A limit of detection of 0.7 µg kg^?1, a limit of quantitation of 2.4 µg kg^?1, mean recoveries varying from 80% to 107%, and coefficients of variation ranging from 5.6% to 9.4% for repeatability and from 7.4% to 12.4% for within-laboratory reproducibility were obtained during an in-house validation. The levels of furan found in the samples were from not detected to 95.5 µg kg^?1. Samples containing vegetables and meat showed higher furan levels as compared with those containing only fruits. An exposure assessment showed furan intakes up to 2.4 µg kg^?1 body weight day^?1 (99th percentile) for babies fed exclusively with commercial baby foods. Margins of exposure obtained from intakes estimated in this work indicated a potential public health concern.     

Aflatoxin B1 and total fumonisin contamination and their producing fungi in fresh and stored sorghum grain in East Hararghe,Ethiopia

Natural contamination of sorghum grains by aflatoxin B₁ and total fumonisin and their producing toxigenic fungi has been studied. A total of 90 sorghum grain samples were collected from small-scale farmers’ threshing floors and 5–6 months later from underground pits during 2013 harvest from three districts of East Hararghe, Ethiopia. Mycotoxin analysis was done using enzyme-linked immunosorbent assay (ELISA). The limits of detection were in the range 0.01–0.03 μg kg^–1. The results revealed that all sorghum grain samples were contaminated with both Aspergillus and Fusarium species. Aflatoxin B₁ was detected at levels ranging from ?1 grain. There were marked variations in aflatoxin B₁ concentrations between fresh and stored samples, with much higher levels in the latter. Total fumonisin levels varied between 907 and 2041 µg kg^?1 grain across the samples. Lowest total fumonisin was recorded in freshly harvested sorghum grain samples. Sorghum is a main staple cereal in the studied districts and its consumption per day per person is high. Daily intake of low doses of mycotoxin-contaminated food stuff over a period of time could lead to chronic mycotoxicosis.     

Occurrence of Aspergillus spp. and aflatoxin B1 in Malaysian foods used for human consumption

Malaysian population widely consumes the cereal-based foods, oilseeds, nuts, and spices in their daily diet. Mycotoxigenic fungi are well known to invade food products under storage conditions and produce mycotoxins that have threat to human and animal health. Therefore, determining toxigenic fungi and aflatoxin B(1) (AFB1) in foods used for human consumption is of prime importance to develop suitable management strategies and to minimize risk. Ninety-five food products marketed in Penang, Malaysia were randomly collected from different supermarkets and were analyzed for presence of Aspergillus spp. by agar plate assay and AFB1 by enzyme-linked immunosorbent assay (ELISA). A. flavus was the dominant fungi in all foods followed by A. niger. Fifty-five A. flavus strains were tested for their ability to produce aflatoxins on rice grain substrate. Thirty-six (65.4%) strains out of 55 produced AFB1 ranging from 1700 to 4400 μg/kg and 17 strains (31%) produced AFB2 ranging from 620 to 1670 μg/kg. Natural occurrence of AFB1 could be detected in 72.6% food products ranging from 0.54 to 15.33 μg/kg with a mean of 1.95 μg/kg. Maximum AFB1 levels were detected in peanut products ranging from 1.47 to 15.33 μg/kg. AFB1 levels detected in all food products were below the Malaysian permissible limits (<35 μg/kg). Aspergillus spp. and AFB1 was not detected in any cookies tested. Although this survey was not comprehensive, it provides valuable information on aflatoxin levels in foods marketed in Malaysia.     

A survey of styrene monomer levels in foods and plastic packaging by coupled mass spectrometry—automatic headspace gas chromatography

An extensive UK survey of styrene monomer levels in styrene based plastic packaging materials and their contained foods (133 samples) has been carried out, examining a wide range of retail foods of different brand names and including yogurts, creams, salads, coleslaws, soft cheeses, margarines, hot and cold beverages from dispensing machines, spreads, fresh and cooked meats, candied fruits, fresh strawberries, and take-away ‘fast’ foods. Analysis by headspace gas chromatography of styrene levels in the plastic containers showed levels of monomer ranging from 16 to 1300 mg kg^?1 although the majority of containers (73%) had styrene levels in the range 100–500 mg kg^?1 and only five plastic tubs had levels exceeding 1000 mg kg^?1. Analysis of the food contents of the plastic containers by automated headspace gas chromatography with single ion monitoring mass spectrometric detection showed levels of the monomer ranging from < 1 μg kg^?1 to 200 μg kg^?1, although the majority of foods (77%) had styrene levels below 10 μg kg^?1, and 26% of the total number analysed had levels below 1 μg kg^?1.     

Evolution of fungal population and mycotoxins in sorghum silage

Silage, one of the most important feed sources for cattle, is vulnerable to contamination by spoilage moulds and mycotoxins because ensilage materials are excellent substrates for fungal growth. The aim of this study was to identify the mycobiota of sorghum silages, to determine the presence of aflatoxins and fumonisins, and to correlate these results with physical parameters of the silage. A total of 275 samples of sorghum were collected from dairy farms in the south-west region of Uruguay were silage practices are developed. The presence of fungi was observed in all of the sorghum samples with values varying from 0.2 × 10⁴ to 4085 × 10⁴ UFC g^?1. Significant difference were detected in the total number of fungi during the storage period; at six months there is a high risk of fungal spoilage. The most frequent genera isolated from sorghum samples were Penicillium (70%), Aspergillus (65%), Absidia (40%), Fusarium (35%), Paecilomyces (35%) and Alternaria, Cladosporium, Gliocadium and Mucor (30%). The toxigenic species most frequently found were Penicillium citrinum, Aspergillus flavus and Fusarium nygamai. Only two samples were contaminated by AFB1 with levels of 1 and 14 µg kg^–1. Fumonisin was detected in 40% of freshly harvest samples with levels ranged from 533 µg kg^–1 to 933 µg kg^–1. The use of silo bags seems to be an effective tool to store sorghum. However, the presence of toxigenic fungi show that regular screening for mycotoxins levels in silages must be performed to avoid the exposure of animals to contaminated feed and the introduction of these compounds into the food chain.     

Inexpensive,rapid screening method for aflatoxins in peanuts and peanut products

Surveys of major Brazilian foodstuffs demonstrated that peanuts and peanut products continue to be very susceptible to aflatoxin contamination. To prevent, or at least minimise, the problem the aflatoxins need to be monitored by a rapid and inexpensive screening method. The AOAC Romer method has been used and found highly reliable. However, the clean-up step utilises anhydrous FeCl₃ and basic CuCO₃ which are expensive and not readily available in Brazil. Thus, the extraction (with a mixture of 270 ml methanol plus 30ml 40 g litre^?1 aqueous KCl) and clean-up (150 ml 100 g litre^?1 aqueous CuSO₄) steps of the method of Soares and Rodriguez-Amaya (1985) were combined with the AOAC minicolumn to provide a rapid, inexpensive screening technique. Fifty-two sample lots of peanuts and peanut products were screened by this and Romer's method, and the results were in complete agreement: 28 samples were negative, four < 20 μg kg^?1, 12 in the range 20–50 μg kg^?1, three in the range 50–100 μg kg^?1 and 5 >100 μg kg^?1. The results also agreed well when the extracts obtained by the two methods were submitted to quantitative TLC.     

Simultaneous detection of DNA from 10 food allergens by ligation-dependent probe amplification

The simultaneous detection of DNA from different allergenic food ingredients by a ligation-dependent probe amplification (LPA) system is described. The approach allows detection of several targets in a one-tube assay. Synthetic oligonucleotides were designed to detect DNA from peanuts, cashews, pecans, pistachios, hazelnuts, sesame seeds, macadamia nuts, almonds, walnuts and brazil nuts. The specificity of the system was tested with DNA from more than 50 plant and animal species. The sensitivity of the method was suitable to detect allergenic ingredients in the low mg kg^?1 range. The limit of detection (LOD) for single allergens in different food matrices was 5 mg kg^?1. The novel analytical strategy represents a useful tool for the surveillance of established legislation on food allergens within the European Union.     

Identification,mycotoxin risk and pathogenicity of Fusarium species associated with fig endosepsis in Apulia,Italy

In a survey carried out on 87 rotted fig fruits samples collected in the Apulia region of Italy, the authors isolated 126 Fusarium strains identified as F. ramigenum (69 strains), F. solani (49), F. proliferatum (five) and three not identified. Investigation on the fertility of the strains belonging to F. proliferatum and F. ramigenum revealed that only strains of F. proliferatum were fertile. The identity of F. ramigenum strains was confirmed by sequencing a portion of the translation elongation factor-1α gene. When Fusarium species were analysed for their toxigenicity, 37/69 strains of F. ramigenum produced fusaric acid (FA) up to 525 mg kg^?1; 30 strains produced beauvericin (BEA) up to 190 mg kg^?1; 60 strains produced fumonisin B₁ (FB₁) and fumonisin B₂ (FB₂) up to 1575 mg kg^?1 of total FBs; and two strains produced fusaproliferin (FUP) up to 345 mg kg^?1; all five strains of F. proliferatum produced FA at low levels; two strains produced BEA up to 205 mg kg^?1; one strain produced FB₁ and FB₂, 1100 and 470 mg kg^?1, respectively; and one strain produced FUP, 820 mg kg^?1; F. solani (30 strains) produced FA, 13 strains up to 215 mg kg^?1. Few fungal extracts showed high toxicity toward brine shrimp larvae and in some cases in relation to BEA and FA content. A pathogenic assay on fig fruits showed that all three species were pathogenic, with higher virulence of F. ramigenum. These data report for the first time the production of BEA and FB₁/FB₂ by F. ramigenum and show that it is a main agent of fig endosepsis in Apulia and can contribute to fumonisin contamination of fresh and dried figs.     

Evaluation of some essential oils as botanical fungitoxicants for the protection of stored food commodities from fungal infestation

Essential oils from different parts of 18 plants belonging to 11 families were extracted and tested against two toxigenic strains of Aspergillus flavus Link through the poisoned food technique. The oil of Mentha arvensis was found to be effective against both strains of A. flavus and completely stopped the radial mycelial growth of A. flavus at 0.10 mg mL^?1. It was found to be superior over the synthetic fungicides tested and showed a broad fungitoxic spectrum against A. niger, A. fumigatus, Botryodiplodia theobromae, Cladosporium cladosporioides, Fusarium oxysporum, Helminthosporium oryzae, Macrophomina phaseolina and Sclerotium rolfsii at 0.10 mg mL^?1. The oil completely inhibited the aflatoxin B1 production by the toxigenic strain of A. flavus at 0.05 mg mL^?1. Moreover, the Mentha oil also exhibited potent antioxidant activity in 2,2′‐azino‐bis‐3‐ethylbenzothiazoline‐6‐sulphuric acid (ABTS) bioassay. Keeping in mind the side effects of synthetic pesticides and the global interest in botanical pesticides for plant protection due to their biodegradable nature, M. arvensis oil may be used as a botanical fungitoxicant against fungal attack to stored food commodities. The antiaflatoxigenic and antioxidant nature of the oil suggest the possibility of its exploitation for enhancing the shelf life of stored food commodities. Copyright © 2007 Society of Chemical Industry     

Aflatoxins,ochratoxins and zearalenone in sorghum and sorghum products in Sudan

This survey examined 60 samples of sorghum and 30 samples of sorghum products from three states (Khartoum, Kordofan and Gadarif) of Sudan for aflatoxin B1, B2, G1 and G2 (AFB1, AFB2, AFG1, AFG2), ochratoxin A and B (OTA, OTB) and zearalenone (ZEN), using high performance liquid chromatography with fluorescence detection. The limits of detection and limits of quantification were in the range 0.01–0.6 µg kg^–1 and 0.03–2.0 µg kg^?1, respectively. The frequency of contaminated samples with AFB1 from Khartoum, Gadarif and Kordofan state was 38.1%, 22.2% and 23.8%, respectively. Only two samples of sorghum from Khartoum state were contaminated with OTA (3.3%). Concentrations of OTA and OTB were low and may not cause problems. No sample of sorghum or sorghum products was contaminated with ZEN. Some sorghum samples contained AFB1 concentrations above the European Union regulatory limits. The highest contaminated samples were found in Khartoum state.     

Assessment of Pelargonium graveolens oil as plant‐based antimicrobial and aflatoxin suppressor in food preservation

BACKGROUND: Contamination of stored food commodities by moulds and mycotoxins results in qualitative as well as quantitative losses. Most of the synthetic antimicrobials used for preservation of stored food items produce side effects in the form of residual and mammalian toxicity. Recently some higher plant products have been recommended as safe alternatives of such synthetic antimicrobials. In the present investigation antifungal efficacy of some essential oils was evaluated against two toxigenic strains of Aspergillus flavus with special reference to the oil of Pelargonium graveolens to investigate its potential to inhibit aflatoxin B₁ secretion. RESULTS: Essential oil of P. graveolens exhibited absolute fungitoxicity against both the toxigenic strains of A. flavus. The minimum inhibitory concentration of the oil was found to be 0.75 g L^?1 and exhibited a fungistatic nature. It was found superior over the synthetic fungicides tested and exhibited a broad fungitoxic spectrum. The oil showed excellent anti‐aflatoxigenic efficacy as it completely inhibited aflatoxin B₁ production even at 0.50 g L^?1. CONCLUSION: This is the first report on the aflatoxin B₁ inhibitory nature of P. graveolens oil. It may be recommended as a novel plant‐based antimicrobial as well as aflatoxin B₁ suppressor over synthetic preservatives in food protection. Copyright © 2008 Society of Chemical Industry     

Sensitive determination of bisphenol A and bisphenol F in canned food using a solid-phase microextraction fibre coated with single-walled carbon nanotubes before GC/MS

A reliable and sensitive method for simultaneous determination of bisphenol A (BPA) and bisphenol F (BPF) in canned food by gas chromatography-mass spectrometry (GC/MS) is described after extraction and pre-concentration by a new solid-phase microextraction (SPME) adsorbent. The potential of single-walled carbon nanotubes (SWCNTs) as SPME adsorbent for the pre-concentration of environmental contaminants has been investigated in recent years. This work was carried out to investigate the feasibility of SWCNTs as a headspace SPME adsorbent for the determination of bisphenol derivatives in canned food. Potential factors affecting the extraction efficiency, including extraction time, extraction temperature, desorption time, desorption temperature, and salinity were optimized. Calibration curves were linear (r ²≥ 0.994) over the concentration range from 0.30 to 60 µg kg^?1. For both target analytes, the limit of detection (LOD) at signal-to-noise (S/N) ratio of 3 was 0.10 µg kg^?1. In addition, a comparative study between the SWCNT and a commercial polydimethylsiloxane (PDMS) SPME fibre for the determination of bisphenol derivatives in canned food was conducted. SWCNT fibre showed higher extraction capacity, better thermal stability (over 350°C) and longer life span (over 150 times) than the commercial PDMS fibre. The method was successfully applied to determine BPA in canned food samples which were purchased from local markets. BPA was found in some of the samples within the concentration range from 0.5 to 5.2 µg kg^?1.     

Occurrence of 3-MCPD fatty acid esters in human breast milk

A series of twelve breast milk samples were analysed by gas chromatography-mass spectrometry (GC/MS) operated in selected ion monitoring mode for 3-chloropropane-1,2-diol (3-MCPD). Whilst none of the samples contained 3-MCPD above the limit of detection of 3 μg kg^?1 milk, all contained high amounts of 3-MCPD esterified with higher fatty acids. The levels of 3-MCPD released by hydrolysis of these esters (bound 3-MCPD) ranged from the limit of detection (300 μg kg^?1, expressed on a fat basis) to 2195 μg kg^?1; with a mean level of bound 3-MCPD of 1014 μg kg^?1, which corresponded to 35.5 μg kg^?1 milk. The presence of bound 3-MCPD was confirmed using orthogonal gas chromatography coupled with high-speed time-of-flight mass spectrometry analysis for four randomly selected breast milk samples. Six breast milks collected from one of the nursing mothers 14–76 days after childbirth contained bound 3-MCPD within the range of 328–2078 μg kg^?1 fat (mean 930 μg kg^?1 fat). The calculated bound 3-MCPD content of these samples was within the range of 6 and 19 μg kg^?1 milk (mean of 12 μg kg^?1 milk). The major types of 3-MCPD esters were the symmetric diesters with lauric, palmitic, and oleic acids, and asymmetric diesters with palmitic acid/oleic acid among which 3-chloro-1,2-propanediol 1,2-dioleate prevailed.