Antioxidant capacity of potato chips and snapshot trends in acrylamide content in potato chips and cereals on the Canadian market

The concentration of acrylamide was measured in selected varieties of five brands of potato chips and breakfast cereals over a 5-year period. Most of the products were purchased in one locality in Canada. Samples were analysed by an isotope dilution (¹³C₃) acrylamide method. They were extracted with water, partitioned with dichloromethane, filtered through a 5 kDa centrifuge filter, cleaned-up on HLB Oasis polymeric and Accucat mixed mode anion and cation exchange SPE columns, and analysed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The acrylamide concentration in potato chips varied from 106 to 4630 ng g^?1, while values in cereals varied from 50 to 347 ng g^?1. Wide variations were observed between brands, within brands over time, and between lots of the same brand. A subset of potato chip samples was analysed for in vitro antioxidant activity. No relationship was found between antioxidative capacity of potato chips and their acrylamide content.     

Risk assessment for furan contamination through the food chain in Belgian children

Young, old, pregnant and immuno-compromised persons are of great concern for risk assessors as they represent the sub-populations most at risk. The present paper focuses on risk assessment linked to furan exposure in children. Only the Belgian population was considered because individual contamination and consumption data that are required for accurate risk assessment were available for Belgian children only. Two risk assessment approaches, the so-called deterministic and probabilistic, were applied and the results were compared for the estimation of daily intake. A significant difference between the average Estimated Daily Intake (EDI) was underlined between the deterministic (419?ng?kg^?1 body weight (bw) day^?1) and the probabilistic (583?ng?kg^?1 bw day^?1) approaches, which results from the mathematical treatment of the null consumption and contamination data. The risk was characterised by two ways: (1) the classical approach by comparison of the EDI to a reference dose (RfD_chronic-oral) and (2) the most recent approach, namely the Margin of Exposure (MoE) approach. Both reached similar conclusions: the risk level is not of a major concern, but is neither negligible. In the first approach, only 2.7 or 6.6% (respectively in the deterministic and in the probabilistic way) of the studied population presented an EDI above the RfD_chronic-oral. In the second approach, the percentage of children displaying a MoE above 10,000 and below 100 is 3–0% and 20–0.01% in the deterministic and probabilistic modes, respectively. In addition, children were compared to adults and significant differences between the contamination patterns were highlighted. While major contamination was linked to coffee consumption in adults (55%), no item predominantly contributed to the contamination in children. The most important were soups (19%), dairy products (17%), pasta and rice (11%), fruit and potatoes (9% each).     

Australian survey of acrylamide in carbohydrate-based foods

A method was developed and validated for the determination of acrylamide in carbohydrate-based foods. Solid-phase extraction employing a mixed-bed anion and cation exchange cartridge in series with a C18 extraction disk was used to clean-up water extracts of food samples before analysis by liquid chromatography coupled with tandem mass spectrometry detection. The limit of detection was calculated as approximately 25 μg kg^-1 and the limit of reporting was 50 μg kg^-1. The average method recovery for 84 samples from a range of matrices reporting was 99% with a relative standard deviation of 11.2%. A survey was conducted of 112 samples of carbohydrate-based foods composited from 547 products available in the Australian market. The analytical results were used in conjunction with Australian food consumption data derived from the 1995 National Nutrition Survey (NNS) to prepare preliminary dietary exposure estimates of Australians to acrylamide through only the food groups examined. Mean dietary exposure to acrylamide resulting from consumption of the foods tested, for Australians aged 2 years and above, was estimated as 22-29 µg day^-1 (equivalent to 0.4-0.5 µg kg^-1 bodyweight day^-1) and between 73 and 80 µg day^-1 (1.4 and 1.5 µg kg^-1 bodyweight day^-1) for 95th percentile consumers. Young children (2-6 years) consuming acrylamide-containing foods had a higher acrylamide exposure on a per kilogram bodyweight basis (mean 1.0-1.3 µg kg^-1 bodyweight day^-1). The estimated exposure of Australians to acrylamide is similar to that estimated for other countries.     

Validation by collaborative trial of an isotope dilution liquid chromatographic tandem mass spectrometric method to determine the content of acrylamide in roasted coffee

A method for the determination of acrylamide in roasted coffee was subjected to method validation by collaborative trial. The aim was to extend the scope of a method already standardized for the determination of acrylamide in bakery and potato products to include roasted coffee. Modifications of the already standardized method were therefore kept to a minimum. The method was based on aqueous extraction of the roasted coffee matrix and solid-phase extraction (SPE) clean-up followed by isotope dilution high-performance liquid chromatography with tandem mass spectrometric detection (LC-MS/MS). The test portion of the sample was spiked with stable isotope-labelled acrylamide and extracted on a mechanical shaker with n-hexane and water for 1 h. The sample extract was centrifuged, the organic phase discarded, and a portion of the aqueous extract further cleaned-up by SPE on an Isolute Multimode cartridge followed by a second clean-up step on an Isolute ENV + cartridge. The volume of the acrylamide-containing fraction eluted from the second SPE column was reduced by evaporation and analysed by LC-MS/MS. Three coffee samples and one aqueous acrylamide standard solution were sent to eleven laboratories from eight European Union Member States. All samples were sent as blind duplicates. Based on the reported results, the relative standard deviations for reproducibility (RSD_R) were 11.5% at a level of 160 µg kg^?1, 10.1% at a level of 263 µg kg^?1, and 9.6% at a level of 585 µg kg^?1. The values for repeatability (RSD_r) in those materials ranged from 1.0% to 3.5%. The method performance parameters satisfied internationally accepted criteria. Hence, the method would be suitable for the enforcement of regulatory limits.     

The chemistry and analysis of annatto food colouring: a review

Annatto food colouring (E160b) has a long history of use in the food industry for the colouring of a wide range of food commodities. The principal colouring component of annatto is the oil-soluble diapo carotenoid bixin, which is the methyl ester of the dicarboxylic acid norbixin and soluble in aqueous alkali. Bixin and norbixin, therefore, exhibit not only physicochemical properties normally associated with carotenoids, but also certain anomalous properties that have an impact on the stability, food colouring applications, and importantly the analysis of annatto. This review summarizes the key aspects of the structural determination of bixin (and norbixin) with special attention to cis-trans isomerization and how this links with its chemical structure, spectroscopic properties, and stability. The oxidative, thermal, and photo-stability of annatto and the subsequent implications for its use in the colouring of foods, food processing, and the analysis of foods and beverages are discussed along with important mechanistic, thermodynamic and kinetic aspects. The main analytical techniques used for the chemical characterization of annatto, i.e. spectrophotometry, nuclear magnetic resonance (NMR), chromatography (particularly high-performance liquid chromatography (HPLC)) and mass spectrometry are reviewed in detail and other methods are discussed. This links in with a review of the methods available for the detection and measurement of annatto in colour formulations and foods and beverages, which highlights the importance of the need for a good understanding and knowledge of the chemistry of bixin and norbixin in order to meet new analytical challenges.     

Estimate of intake of benzoic acid in the Belgian adult population

An exposure assessment was performed to estimate average daily benzoic acid intake for Belgian adults. Food consumption data were retrieved from the national food-consumption survey. As a first step, individual food-consumption data were multiplied with the maximum permitted use levels for benzoic acid per food group (Tier 2). As a second step, a label survey to identify the foods where benzoic acid is effectively used as an additive and a literature review of the possible occurrence of benzoic acid as a natural substance were performed. With this information, a refined list of foods was drafted for the quantification of benzoic acid, which was performed by a high-performance liquid chromatography (HPLC) method, optimized and validated for this purpose. Individual food-consumption data were then multiplied with the actual average concentrations of benzoic acid per food group (Tier 3). Usual intakes were calculated using the Nusser method. The mean benzoic acid intake was 1.58 mg kg^?1 body weight day^?1 (Tier 2) and 1.25 mg kg^?1 body weight day^?1 (Tier 3). In Tier 2, men exceeded the acceptable daily intake (ADI) of 5 mg kg^?1 body weight day^?1 at the 99th percentile. The greatest contributors to the benzoic acid intake were soft drinks. Benzoic acid as a natural substance represents only a small percentage of the total intake. The results show that actual benzoic acid intake is very likely to be below the ADI. However, there is a need to collect national food-consumption data for children as they might be more vulnerable to an excessive intake.     

Pyrrolizidine alkaloids in honey: comparison of analytical methods

Pyrrolizidine alkaloids (PAs) are a structurally diverse group of toxicologically relevant secondary plant metabolites. Currently, two analytical methods are used to determine PA content in honey. To achieve reasonably high sensitivity and selectivity, mass spectrometry detection is demanded. One method is an HPLC-ESI-MS-MS approach, the other a sum parameter method utilising HRGC-EI-MS operated in the selected ion monitoring mode (SIM). To date, no fully validated or standardised method exists to measure the PA content in honey. To establish an LC-MS method, several hundred standard pollen analysis results of raw honey were analysed. Possible PA plants were identified and typical commercially available marker PA-N-oxides (PANOs). Three distinct honey sets were analysed with both methods. Set A consisted of pure Echium honey (61–80% Echium pollen). Echium is an attractive bee plant. It is quite common in all temperate zones worldwide and is one of the major reasons for PA contamination in honey. Although only echimidine/echimidine-N-oxide were available as reference for the LC-MS target approach, the results for both analytical techniques matched very well (n?=?8; PA content ranging from 311 to 520?µg?kg^?1). The second batch (B) consisted of a set of randomly picked raw honeys, mostly originating from Eupatorium spp. (0–15%), another common PA plant, usually characterised by the occurrence of lycopsamine-type PA. Again, the results showed good consistency in terms of PA-positive samples and quantification results (n?=?8; ranging from 0 to 625?µg?kg^?1 retronecine equivalents). The last set (C) was obtained by consciously placing beehives in areas with a high abundance of Jacobaea vulgaris (ragwort) from the Veluwe region (the Netherlands). J. vulgaris increasingly invades countrysides in Central Europe, especially areas with reduced farming or sites with natural restorations. Honey from two seasons (2007 and 2008) was sampled. While only trace amounts of ragwort pollen were detected (0–6.3%), in some cases extremely high PA values were detected (n?=?31; ranging from 0 to 13019?µg?kg^?1, average?=?1261 or 76?µg?kg^?1 for GC-MS and LC-MS, respectively). Here the results showed significantly different quantification results. The GC-MS sum parameter showed in average higher values (on average differing by a factor 17). The main reason for the discrepancy is most likely the incomplete coverage of the J. vulgaris PA pattern. Major J. vulgaris PAs like jacobine-type PAs or erucifoline/acetylerucifoline were not available as reference compounds for the LC-MS target approach. Based on the direct comparison, both methods are considered from various perspectives and the respective individual strengths and weaknesses for each method are presented in detail.     

Determination of ethylenethiourea (ETU) and propylenethiourea (PTU) in foods by high performance liquid chromatography-atmospheric pressure chemical ionisation-medium-resolution mass spectrometry

A robust and sensitive method for the determination of ethylenethiourea (ETU) and iso-propylenethiourea (i-PTU) in foods is reported. ETU and i-PTU were extracted by blending with dichloromethane (DCM) in the presence of sodium sulphate, sodium carbonate, thiourea and ascorbic acid. ²H₄-ETU and n-PTU were used as internal standards. After filtration the DCM was removed by rotary evaporation and the extract re-dissolved in water before analysis by reversed-phase liquid chromatography with detection by atmospheric pressure chemical ionization-mass spectrometry using a double focusing mass spectrometer at a resolution of 5000. Mean recoveries of ETU and i-PTU from fruit-based, cereal-based and meat-based infant foods, potato chips and tinned potatos at 0.01 mg kg^-1 and from pizza and yoghurt at 0.02-0.1 mg kg^-1 were 95% and 97% respectively. Precision, including both repeatability and internal reproducibility, was in the range of 3.1-13.1%.     

Rapid multi-class multi-residue method for the confirmation of chloramphenicol and eleven nitroimidazoles in milk and honey by liquid chromatography-tandem mass spectrometry (LC-MS)

A confirmatory method was developed to allow for the analysis of eleven nitroimidazoles and also chloramphenicol in milk and honey samples. These compounds are classified as A6 compounds in Annex IV of Council Regulation 2377/90 (European Commission 1990) and therefore prohibited for use in animal husbandry. Milk samples were extracted by acetonitrile with the addition of NaCl; honey samples were first dissolved in water before a similar extraction. Honey extracts underwent a hexane wash to remove impurities. Both milk and honey extracts were evaporated to dryness and reconstituted in initial mobile phase. These were then injected onto a liquid chromatography-tandem mass spectrometry (LC-MS/MS) system and analysed in less than 9 min. The MS/MS was operated in multiple reaction monitoring (MRM) mode with positive and negative electrospray ionization. The method was validated in accordance with Commission Decision 2002/657/EC and is capable of analysing metronidazole, dimetridazole, ronidazole, ipronidazole and there hydroxy metabolites hydroxymetronidazole, 2-hydroxymethyl-1-methyl-5-nitroimidazole, and hydroxyipronidazole. The method can also analyse for carnidazole, ornidazole, ternidazole, tinidazole, and chloramphenicol. A recommended level of 3 µg l^?1/µg kg^?1 for methods for metronidazole, dimetridazole, and ronidazole has been recommended by the Community Reference Laboratory (CRL) responsible for this substance group, and this method can easily detect all nitroimidazoles at this level. A minimum required performance level of 0.3 µg l^?1/µg kg^?1 is in place for chloramphenicol which the method can also easily detect. For nitroimidazoles, the decision limits (CCα) and detection capabilities (CCβ) ranged from 0.41 to 1.55 µg l^?1 and from 0.70 to 2.64 µg l^?1, respectively, in milk; and from 0.38 to 1.16 µg kg^?1 and from 0.66 to 1.98 µg kg^?1, respectively, in honey. For chloramphenicol, the values are 0.07 and 0.11 µg l^?1 in milk and 0.08 and 0.13 µg kg^?1 in honey. Validation criteria of accuracy, precision, repeatability, and reproducibility along with measurement uncertainty were calculated for all analytes in both matrices.     

Fate of trichothecene mycotoxins during the processing: Milling and baking

Toxic secondary metabolites produced by fungi representing Fusarium genus are common contaminants in cereals worldwide. To estimate the dietary intake of these trichothecene mycotoxins, information on their fate during cereal processing is needed. Up-to-date techniques such as high-performance liquid chromatography (HPLC) coupled with tandem mass spectrometry (LC-MS/MS) was used for the analysis of seven trichothecenes (deoxynivalenol, nivalenol, HT-2 toxin, T-2 toxin, 15- and 3-acetyldeoxynivalenol, and fusarenon-X) in bread production chain (wheat grains, intermediate products collected during milling and baking process, breads). Regardless of whether the grains were naturally infected or artificially inoculated by Fusarium spp. in the field, the fractions obtained from the grain-cleaning procedure contained the highest mycotoxin levels. During milling the highest concentrations of deoxynivalenol were found in the bran, the lowest in the reduction flours. Baking at 210°C for 14 min had no significant effect on deoxynivalenol levels. The rheological properties of dough measured by fermentograph, maturograph, oven rise recorder, and laboratory baking test were carried out, and based on the obtained results the influence of mycotoxin content on rheological behaviour was investigated.     

Quantitative determination of mycotoxins in urine by LC-MS/MS

Naturally occurring mycotoxins are responsible for a wide array of adverse health effects. The measurement of urinary mycotoxin levels is a useful means of assessing an individual's exposure, but the development of sensitive and accurate analytical methods for detecting mycotoxins and their metabolites in urine samples is challenging. Urinary mycotoxins are present in low pg ml^–1 concentrations, and the chromatographic identification of their metabolites can be obscured by other endogenous metabolites. We developed an analytical method focused on the selection of two appropriate multiple-reaction monitoring transition for unambiguous identification and quantification of carcinogenic aflatoxin M₁ (AFM₁), ochratoxin A (OTA) and fumonisin B₁, B₂ (FB₁, FB₂) in urine samples from a small volunteer group in a pilot study. AFM₁, OTA, FB₁ and FB₂ were concentrated selectively, interfering substances were removed using an immunoaffinity column (IAC), and mycotoxins were measured using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in combination with a stable-isotope standard-dilution assay (SIDA). The method was sensitive enough to measure mycotoxins and their metabolites at pg ml^–1 levels in urine. The combination of LC-MS/MS and SIDA was critical to distinguishing pseudo-OTα interference from genuine OTα. Twelve urine samples contained OTA ranging from 0.013 to 0.093 ng ml^?1 (mean = 0.031 ng ml^?1). AFM₁ were detected in one sample at a 0.002 ng ml^?1 level, while FB₁ and FB₂ were undetectable in all 12 samples. None of the samples in this pilot study contained a detectable level of OTα, despite the presence of OTA, and this may suggest the need for further epidemiological investigation of OTA exposure in the Korean population.     

Pattern of intake of food additives associated with hyperactivity in Irish children and teenagers

A double-blind randomized intervention study has previously shown that a significant relationship exists between the consumption of various mixes of seven target additives by children and the onset of hyperactive behaviour. The present study set out to ascertain the pattern of intake of two mixes (A and B) of these seven target additives in Irish children and teenagers using the Irish national food consumption databases for children (n = 594) and teenagers (n = 441) and the National Food Ingredient Database. The majority of additive-containing foods consumed by both the children and teenagers contained one of the target additives. No food consumed by either the children or teenagers contained all seven of the target food additives. For each additive intake, estimates for every individual were made assuming that the additive was present at the maximum legal permitted level in those foods identified as containing it. For both groups, mean intakes of the food additives among consumers only were far below the doses used in the previous study on hyperactivity. Intakes at the 97.5th percentile of all food colours fell below the doses used in Mix B, while intakes for four of the six food colours were also below the doses used in Mix A. However, in the case of the preservative sodium benzoate, it exceeded the previously used dose in both children and teenagers. No child or teenager achieved the overall intakes used in the study linking food additives with hyperactivity.     

Analysis of isothiazolinone biocides in paper for food packaging by ultra-high-performance liquid chromatography–tandem mass spectrometry

A novel and simple method to detect isothiazolinone-type biocides (2-methyl-3-isothiazolinone (MI), 5-chloro-2-methyl-3-isothiazolinone (CMI), 1,2-benzisothiazolinone (BIT) and 2-octyl-3-isothiazolinone (OIT)) in paper used for food packaging by ultrasonic extraction coupled with UPLC-MS/MS was developed. Parameters affecting process efficiency such as extraction solvents, UPLC mobile phase, gradient elution procedure and MS/MS conditions were studied to optimise the operating conditions. Using the optimised gradient elution procedure, the retention time was less than 6?min. The limits of detection (LODs) were found to be between 0.001 and 0.010?mg?kg^?1, which was validated using actual concentrations. After diluting the standard solution with a blank matrix, the linear calibration curve ranges were 0.002–1.000?mg?kg^?1 for BIT and OIT, 0.005–1.000?mg?kg^?1 for MI, and 0.020–1.000?mg?kg^?1 for CMI, with correlation coefficients higher than 0.9985 (n?=?6). A good level of precision with a mean recovery greater than 81.3% and a relative standard deviation (RSD) less than 6.2% were also obtained. A methodology has been proposed for the analysis of isothiazolinones in paper.     

Transfer of aflatoxin B1 and fumonisin B1 from naturally contaminated raw materials to beer during an industrial brewing process

The aim of this research was to determine the fate of aflatoxins (AFs) and fumonisins (FBs) naturally occurring in raw materials (maize grit and malted barley) during four industrial brewing processes. The aflatoxin B₁ (AFB₁) level in raw materials varied from 0.31 to 14.85 µg kg^?1, while the fumonisin B₁ (FB₁) level (only in maize grit) varied from 1146 to 3194 µg kg^?1. The concentration in finished beer ranged from 0.0015 to 0.022 µg l^?1 for AFB₁ and from 37 to 89 µg l^?1 for FB₁; the other aflatoxins and fumonisin B₂ were not found in beer samples. The average percentage of toxins recovered in finished beer, referring to the amounts contained in raw materials, were 1.5% ± 0.8% for AFB₁ and 50.7% ± 4.7% for FB₁. These results were mainly due to the different solubility of the two mycotoxins during the mashing process. If raw materials comply with the limits fixed by European Commission Regulations, the contribution of a moderate daily consumption of beer to AFB₁ and FB₁ intake does not contribute significantly to the exposure of the consumer.     

Evaluation of fumonisin biomarkers in a cross-sectional study with two high-risk populations in China

Levels of serum and urinary sphinganine (Sa) and sphingosine (So), the Sa/So ratio, and urinary-free fumonisin B₁ (FB₁) were determined in a cross-sectional study consisting of 43 adults in Huaian and 34 adults in Fusui, China. Home-produced corn had 100% contamination with FB₁. There were 93.0% (40/43) of Huaian subjects and 52.9% (18/34) of Fusui subjects with daily FB₁ intakes greater than 2 µg kg^?1 body weight, which showed a significant difference (p < 0.01). Levels of sphinganine and sphingosine and the Sa/So ratio were not correlated with levels of dietary exposure. The median level of the serum Sa/So ratio in Huaian subjects (0.41, range = 0.14–0.85) was significantly lower than that in Fusui subjects (0.78, range = 0.57–1.08) (p < 0.01). The median level of the urinary Sa/So ratio was also significantly lower in Huaian subjects (0.31, range = 0.08–1.33) than in Fusui subjects (0.57, range = 0.03–2.52) (p < 0.01). Urinary-free FB₁ was detected in 83.7% (36/43) of Huaian samples and in 82.4% (28/34) of Fusui urine samples (p > 0.05). However, the median level of urinary-free FB₁ in Huaian subjects, 3.91 (range = 0.06–253.61) ng mg^?1 creatinine, was significantly higher than 0.39 (range = 0.01–3.72) ng mg^?1 creatinine found in Fusui subjects (p < 0.01). These results suggest that urinary-free FB₁ may be a potential biomarker for human fumonisin exposure, while further validation is needed in human epidemiological and intervention studies.     

UNE-EN ISO/IEC 17025:2005-accredited method for the determination of pesticide residues in fruit and vegetable samples by LC-MS/MS

A rapid, simple and sensitive multi-residue method was developed and validated for the simultaneous quantification and confirmation of 69 pesticides in fruit and vegetables using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The samples were extracted following the quick, easy, cheap, effective, rugged and safe method known as QuEChERS. Mass spectrometric conditions were individually optimised for each analyte in order to achieve maximum sensitivity in multiple reaction monitoring (MRM) mode. Using the developed chromatographic conditions, 69 pesticides can be separated in less than 17 min. Two selected reaction monitoring (SRM) assays were used for each pesticide to obtain simultaneous quantification and identification in one run. With this method in SRM mode, more than 150 pesticides can be analysed and quantified, but their confirmation is not possible in all cases according to the European regulations on pesticide residues. Nine common representative matrices (zucchini, melon, cucumber, watermelon, tomato, garlic, eggplant, lettuce and pepper) were selected to investigate the effect of different matrices on recovery and precision. Mean recoveries ranged from 70% to 120%, with relative standard deviations (RSDs) lower than 20% for all the pesticides. The proposed method was applied to the analysis of more than 2000 vegetable samples from the extensive greenhouse cultivation in the province of Almería, Spain, during one year. The methodology combines the advantages of both QuEChERS and LC-MS/MS producing a very rapid, sensitive, accurate and reliable procedure that can be applied in routine analytical laboratories. The method was validated and accredited according to UNE-EN-ISO/IEC 17025:2005 international standard (accreditation number 278/LE1027).     

Exposure assessment of the Belgian population to pesticide residues through fruit and vegetable consumption

Exposure of the Belgian consumer to pesticide residues from the consumption of fruit and vegetables was determined based on data collected in the Belgian food consumption survey performed by the Scientific Institute for Public Health and data from the Belgian Federal Agency for the Safety of the Food Chain 2005 monitoring programme. A first screening of pesticide residue exposure was performed by a deterministic approach. For most pesticide residues studied, the exposure was 100 times lower than the acceptable daily intake (ADI). However, for a high consumer (97.5th percentile of consumption) the intake could reach 23% of the ADI for imazalil, 15% for chlorpropham, 14% for the dithiocarbamates, 10% for dimethoate and lambda-cyhalothrin, and 9% for chlorpyriphos. Nevertheless, probabilistic exposure assessment performed on these pesticides in a second phase of the study indicated that, except for chlorpropham, the probability to exceed the ADI is much lower than 0.1%.     

Gynaecomastia linked to the intake of a herbal supplement fortified with diethylstilbestrol

This study reports the findings of a supplement marketed on the Internet for prostate problems. The supplement was orally taken by a 60-year-old man with divergent hormonal levels and who was surgically treated for gynaecomastia: development of abnormally large mammary glands in males. The supplement showed a strong effect in a yeast oestrogen bioassay, expressing a yeast-enhanced green fluorescent protein (yEGFP) upon exposure to oestrogens. Using both nuclear magnetic resonance (NMR) and a gradient liquid chromatographic time-of-flight mass spectrometric (LC/TOF-MS) method, the response was shown to be caused by very high levels of diethylstilbestrol, known for causing gynaecomastia. The gynaecomastia was most probably caused by this orally taken ‘natural’ herbal supplement, as the patient's hormonal levels also returned to normal again when stopping the use of it. This case demonstrates that physicians need to be aware of the use of supplements with illegal components that may be responsible for unwanted side-effects.     

Occurrence of deoxynivalenol and its 3-β-D-glucoside in wheat and maize

Deoxynivalenol-3-β-D-glucoside (D3G), a phase II plant metabolite of the mycotoxin deoxynivalenol (DON), occurs in naturally Fusarium-contaminated cereals. In order to investigate the frequency of occurrence as well as the relative and absolute concentrations of D3G in naturally infected cereals, 23 wheat samples originating from fields in Austria, Germany and Slovakia as well as 54 maize samples from Austrian fields were analysed for DON and D3G by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Both analytes were detected in all the 77 field samples. DON was found at levels from 42 to 4130 ng g^?1 (977 ± 1000 ng g^?1 on average). The D3G concentrations in all cereal samples were in the range 10–1070 ng g^?1 (216 ± 253 ng g^?1 on average), corresponding to about 5–46 mol% of their DON concentrations (15 ± 8 mol% on average).