Synthesis of stable isotope labeled 3-acetyldeoxynivalenol

Stable isotope labeled 3-acetyldeoxynivalenol (3-AcDON) was synthesized in excellent yield from deoxynivalenol as starting material. This is the first synthesis of a stable isotope labeled type-B trichothecene suitable as internal standard for HPLC-MS/MS or GC-MS analysis of trichothecene mycotoxins. The isotopic purity of the 3-d(3)-AcDON was determined to be 94.9%.     

Occurrence of Fusarium species and trichothecenes in Nigerian maize

A total of 180 maize samples meant for human consumption from four maize-producing states of southwestern Nigeria were screened for twelve major Fusarium mycotoxins (trichothecenes). Mycological examination of the samples showed that Fusarium verticillioides was the most commonly isolated fungi (71%), followed by F. sporotrichioides (64%), F. graminearum (32%), F. pallidoroseum (15%), F. compactum (12%), F. equiseti (9%), F. acuminatum (8%), F. subglutinans (4%) and F. oxysporum (1%). The trichothecenes include deoxynivalenol (DON), 3, mono-acetyldeoxynivalenol (3-AcDON), 15, mono-acetyldeoxynivalenol (15-AcDON), nivalenol (NIV), HT-2 toxin (HT-2), neosolaniol (NEO), T-2 toxin (T-2), T-2 tetraol and T-2 triol, diacetoxyscirpenol (DAS), MAS-monoacetoxyscirpenol (MAS) and fusarenone-X. Quantification was by high performance liquid chromatography coupled with mass spectroscopy (HPLC/MS); the detection limits for each of the mycotoxins varied between 20 and 200 microg kg(-1). Sixty six samples (36.3%) were contaminated with trichothecenes, DON (mean: 226.2 microg kg(-1); range: 9.6-745.1 microg kg(-1)), 3-AcDON (mean: 17.3 microg kg(-1); range: 0.7-72.4 microg kg(-1)) and DAS (mean: 16.0 microg kg(-1); range: 1.0-51.0 microg kg(-1)) were detected in 22%, 17% and 9% of total samples respectively. There were no 15-AcDON, NIV, HT-2, NEO, T-2, T-2 tetraol, T-2 triol, MAS and fusarenone-X detected. This is the first comprehensive report about the natural occurrence of DON, AcDON and DAS in maize for direct human consumption in Nigeria.     

Influence of environmental factors on the biosynthesis of type B trichothecenes by isolates of Fusarium spp. from Spanish crops

Various species of Fusarium can produce trichothecene mycotoxins that contaminate food commodities and can represent a risk for human and animal health. In this paper, a full factorial design was applied to study the influence of incubation temperature, water activity (a(w)) and type of isolate on the production of deoxynivalenol (DON), nivalenol (NIV) and 3-acetyldeoxynivalenol (3-AcDON) in corn kernel cultures by three isolates of Fusarium graminearum and three isolates of Fusarium culmorum from crops grown in Spain. The tested temperatures were 15, 20, 28 and 32 degrees C. The a(w)-values were 0.960, 0.970 and 0.980. Moisture of cultures (within the studied range) did not affect significantly production of trichothecenes; however, the temperature affected significantly mycotoxin production and the optimal values were 28, 20 and 15 degrees C for DON, NIV and 3-AcDON, respectively. Four additional isolates of F. graminearum and two additional isolates of F. culmorum were examined for production of these mycotoxins at the optimal temperatures. Of the seven isolates of F. graminearum, four produced DON (0.88-3.97 microg/g), seven produced NIV (1.53-124 microg/g), and three produced 3-AcDON (0.65-10.6 microg/g). Of the five isolates of F. culmorum, four produced DON (1.20-4.93 microg/g), four produced NIV (6.94-701 microg/g), and four produced 3-AcDON (0.83-7.70 microg/g). Practically all isolates seem to belong to the NIV-chemotype. This is the first study done with regard to interaction between strain and ecological variables on type B trichothecene production by isolates of these two species from crops grown in Spain.     

Characterization and application of isotope-substituted (13C15)-deoxynivalenol (DON) as an internal standard for the determination of DON

The powerful combination of liquid chromatography and mass spectrometry (MS) is often limited by matrix effects during ionization in the MS ion source. The use of fully isotope-substituted (¹³C₁₅)-deoxynivalenol ((¹³C₁₅)-DON) as an internal standard (IS) corrects matrix effects and improves the accuracy of analytical methods using mass spectrometry for the quantitative determination of the Fusarium mycotoxin deoxynivalenol (DON). The IS was characterized with respect to its chromatographic purity by liquid chromatography-ultraviolet light and its isotope distribution by time-of-flight mass spectrometry. Its low-energy collision-induced dissociation behaviour was compared with DON. Moreover, this work describes the successful application of (¹³C₁₅)-DON as IS for the determination of DON in maize using high-performance liquid chromatography (HPLC) electrospray (ESI) with tandem mass spectrometry. The results demonstrate that the IS can successfully correct for fluctuations during extraction and clean-up of the sample as well as the ionization of DON in the MS ion source. Random variations in ionization affect the IS in the same way as the analyte. Recoveries for DON in maize of 76% ± 1.9% (external calibration) or 101% ± 2.4% (internal calibration) were reached, respectively, after sample clean-up.     

Analysis of deoxynivalenol, masked deoxynivalenol, and Fusarium graminearum pigment in wheat samples, using liquid chromatography-UV-mass spectrometry

Tolerable limits set for deoxynivalenol (DON) do not consider DON conjugates such as DON-3-glucoside. Conjugates may be metabolized in vivo to DON. Such masked mycotoxins and the potentially toxic Fusarium pigment are not routinely analyzed in cereals. We quantified DON, DON-3-glucoside, and a red Fusarium pigment in hard red spring wheat, using a new liquid chromatography-mass spectrometry method. Extraction protocols using centrifugation and shaking, and methanol-methylene chloride (50:50 [vol/vol]) or acetonitrile-water (84:16 [vol/vol]) were assessed. Purposively and randomly selected hard spring wheat samples were extracted with solvent filtered through a C18 column and analyzed using liquid chromatography-UV-mass spectrometry. Isocratic mobile phase (70% methanol) was used. Recoveries were 96.4% (DON) and 70.0% (DON-3-glucoside), while limits of detection were 1 microg/kg (MS) and 10 microg/kg (UV), and limits of quantification were 1 microg/kg (UV) and 0.5 microg/kg (MS), respectively. The pigment limits of quantification and limits of detection on the MS were 4.3 and 0.0005 microg/kg, respectively. The purposively selected samples had DON, DON-3-glucoside, and pigment averages of 3.4 +/- 4.0 microg/g, 3.8 +/- 8.3 microg/g, and 0.31 +/- 3.71 g/kg, respectively. The randomly selected spring wheat had lower mean levels of DON (1.4 +/- 2.3 microg/g), DON-3-glucoside (0.2 +/- 1.0 microg/g), and pigment (147.93 +/- 247.84 microg/g). Analytical tools such as this new liquid chromatography-UV-mass spectrometry method can be used to quantify masked and parent mycotoxins, plus a potentially toxic pigment for risk assessment.     

Deoxynivalenol and its acetyl derivatives in bread and biscuits in Shandong province of China

In the present study, 100 commercial breads and biscuits were investigated for the occurrence of deoxynivalenol (DON) and its acetylated derivatives 3-acetyldeoxynivalenol (3-Ac-DON) and 15-acetyldeoxynivalenol (15-Ac-DON). The target mycotoxins were determined by isotope dilution ultrahigh performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS). DON was determined in 95% of the test samples with a mean value of 153.3 µg/kg. Compared with DON, 3-Ac-DON and 15-Ac-DON were far less frequently detected, with mean values of 1.14 and 0.37 µg/kg, respectively. The estimated daily intakes of the sum of DON and its derivatives in breads and biscuits were 0.0059 and 0.0313 µg/kg bw/day, respectively, which was within the group provisional tolerable daily intake of 1.0 µg/kg bw/day set by the Joint FAO/WHO Expert Committee on Food Additives. In the future, systematic monitoring programmes of DON and its derivatives in relevant foodstuffs are still necessary for food safety and human health.     

Survey of Fusarium toxins in foodstuffs of plant origin marketed in Germany

A total of 219 samples of foodstuffs of plant origin, consisting of grain-based food, pseudocereals and gluten-free food as well as vegetables, fruits, oilseeds and nuts, were randomly collected during 2000 and 2001 in food and health food stores. A spectra of 13 trichothecene toxins including diacetoxyscirpenol (DAS), 15-monoacetoxyscirpenol (MAS), scirpentriol (SCIRP), T-2 and HT-2 toxins (T-2, HT-2), T-2 triol, T-2 tetraol, neosolaniol (NEO) of the A-type as well as deoxynivalenol (DON), 3- and 15-acetyl-DON (3-, 15-ADON), nivalenol (NIV), and fusarenon-X (FUS-X) of the B-type were determined by gas chromatography/mass spectrometry. Analysis of zearalenone (ZEA), alpha- and beta-zearalenol (alpha- and beta-ZOL) was made by high-performance liquid chromatography with fluorescence and UV-detection. Detection limits ranged between 1 and 19 microg/kg. Out of 84 samples of cereal-based including gluten-free foods, 60 samples were positive for at least one of the toxins DON, 15-ADON, 3-ADON, NIV, T-2, HT-2, T-2 tetraol and ZEA, with incidences at 57%, 13%, 1%, 10%, 12%, 37%, 4% and 38%, respectively, whereas SCIRP and its derivatives MAS and DAS, T-2 triol, Fus-X as well as alpha- and beta-ZOL were not detected in any sample of this subgroup. Contents of DON ranged between 8 and 389 microg/kg, for all other toxins determined concentrations were below 100 microg/kg. The pseudocereals amaranth, quinoa and buckwheat were free of the toxins investigated. Ten of 85 samples of vegetables and fruits were toxin positive. ZEA and the type A trichothecenes MAS, SCIRP, DAS, HT-2 were detected in 7, 3, 2, 1 and 1 samples, respectively. Out of 35 samples of oilseeds and nuts, 7 samples were toxin positive. HT-2, T-2 and ZEA were detected in 4, 3 and 4 samples, respectively. In vegetables and fruits as well as in oilseeds and nuts, toxin levels were below 50 microg/kg. None of the B-type trichothecenes analysed was found for both subgroups.     

Contents of Fusarium toxins in Finnish and imported grains and feeds

The aim of the present study was to investigate the levels of trichothecenes in representative samples of Finnish and imported cereals and feeds from the 1987 and 1988 crops. Trichothecene concentrations were determined in samples as trimethylsilylether derivatives by a gas chromatograph equipped with a mass selective detector (GC-MSD) using selected ion monitoring (SIM) which proved to be a reliable and sufficiently sensitive technique. Representative samples of Finnish and imported oats, wheat, barley, rye, maize gluten, soy granules, rapeseed, turnip rapeseed, fish meal, poultry feed and pig feed were analysed employing 19-nortestosterone as an internal standard. Almost all grain and feed samples contained deoxynivalenol (DON) from 7 to 300 micrograms/kg and smaller amounts (13-120 micrograms/kg) of 3-acetyldeoxynivalenol (3-AcDON). The most toxic trichothecenes, T-2, HT-2 toxins and nivalenol (NIV) and also zeralenone (ZEN) were found at low concentrations in some samples. Six lots of oats containing 1.3-2.6 mg/kg of DON and 0.2-0.6 mg/kg of 3-AcDON were found in the Finnish grain samples. Silo samples of almost all imported grains contained DON but at levels below 50 micrograms/kg. Imported grains contained no other trichothecenes. DON, 3-AcDON and ZEN were found in imported feedstuffs at low concentrations.     

Incidence of trichothecenes and zearalenone in poultry feed mixtures from Slovakia

A total of 50 samples of poultry feed mixtures of Slovakian origin were analyzed for eight toxicologically significant Fusarium mycotoxins, namely zearalenone (ZON), A-trichothecenes: diacetoxyscirpenol (DAS), T-2 toxin (T-2) and HT-2 toxin (HT-2) and B-trichothecenes: deoxynivalenol (DON), 3-acetyl-deoxynivalenol (3-ADON), 15-acetyl-deoxynivalenol (15-ADON) and nivalenol (NIV). The A-trichothecenes and the B-trichothecenes were detected by means of high pressure liquid chromatography with tandem mass spectrometry detection (HPLC-MS/MS) and gas chromatography electron capture detection (GC-ECD), respectively. Reversed phase-high performance liquid chromatography with a fluorescence detector (RP-HPLC-FLD) was used for ZON detection. The most frequent mycotoxin detected was T-2, which was found in 45 samples (90%) in relatively low concentrations ranging from 1 to 130 microg kg(-1) (average 13 microg kg(-1)), followed by ZON that was found in 44 samples (88%) in concentrations ranging from 3 to 86 microg kg(-1) (average 21 microg kg(-1)). HT-2 and DON were detected in 38 (76%) and 28 (56%) samples, respectively, in concentrations of 2 to 173 (average 18 microg kg(-1)) for HT-2 and 64 to 1230 microg kg(-1) sample (average 303 microg kg(-1)) for DON. The acetyl-derivatives of DON were in just four samples, while NIV was not detected in any of the samples investigated. In as many as 22 samples (44%), a combination of four simultaneously co-occurring mycotoxins, i.e. T-2, HT-2, ZON and DON, was revealed. Despite the limited number of samples investigated during this study poultry feed mixtures may represent a risk from a toxicological point of view and should be regarded as a potential source of the Fusarium mycotoxins in Central Europe. This is the first reported study dealing with zearalenone and trichothecene contamination of poultry mixed feeds from Slovakia.     

Quantitation of type B-trichothecene mycotoxins in foods and feeds by a multiple stable isotope dilution assay

For quantitation of the trichothecene mycotoxins deoxynivalenol (DON), 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol and 4-acetylnivalenol (Fusarenon X), stable isotope dilution assays (SIDAs) were developed by using analogues labelled with carbon-13 as the internal standards. The SIDAs presented here are the first ones reported for type B-trichothecenes. The synthesis of labelled acetyl derivatives was accomplished by [¹³C₂]-acetylation of unlabelled trichothecenes and subsequent hydrolysis of the products. The mycotoxins were quantified in foods by LC tandem MS after cleanup on multifunctional columns. The method revealed an excellent sensitivity with low detection and quantification limits for all mycotoxins in the low μg/kg range, and good precision in inter-assay stu- dies. The analysis of a certified reference material (maize flour) resulted in a low bias of 2.1% from the certified value for DON and verified excellent trueness of the new method. To prove the suitability of SIDA, a number of cereals and cereal products were analysed and revealed DON contents ranging from 2 to 300 μg/kg. Acetylated derivatives occurred in lower contents and mainly in corn products.     

Distribution of deoxynivalenol and 3-acetyldeoxynivalenol in naturally contaminated and Fusarium culmorum infected triticale samples

Eight triticale genotypes, naturally contaminated (C) and collected from heads inoculated with two separate isolates Fusarium culmorum (I1 and I2) kernels were analysed for accumulation of Fusarium toxins. Triticale kernels were contaminated with deoxynivalenol (DON) and 3-acetyldeoxynivalenol (3-AcDON). Statistical analysis revealed significant differences between naturally contaminated samples (C) and those inoculated with isolate I1 and I2. The following accumulation of toxins in kernels (mean mg/kg) was found: C, I1, I2--DON 0.06, 14.45, 8.09; 3-AcDON--0.01, 0.20, 0.49. Distribution of DON and 3-AcDON was studied in four size fractions of kernels (> 2.8 mm; < or = 2.8-2.5 mm, < 2.5-2.2 mm and < 2.2 mm). The highest concentration of DON and 3-AcDON was found in the smallest fraction of damaged kernels. The percentage distribution of DON and 3-AcDON in fraction < 2.2 mm was (C) 66%, 77%, (I1) 38%, 27%, (I2) 46%, 40%; and in fraction < 2.5 mm: (C) 85%, 94%, (I1) 57%, 51%, (I2) 80%, 76%. The naturally contaminated kernel fraction < 2.2 mm (although only 2% of all) contained 33% DON and 59% 3-AcDON, for more toxigenic isolate I1 10%, 15%, 14%, less toxigenic isolate I2 4%, 13%, 11%, respectively, calculated on dry matter basis. No significant correlation between examined characteristics was found for either I1 or I2 isolate. The biosynthesis mechanism of toxic metabolites was to approximate (significantly correlated) to pair objects C/I2.     

Fusarium toxins in wheat from an area in Henan Province, PR China, with a previous human red mould intoxication episode.

Wheat samples of the 1998 and 1999 crops from Puyang, an area in Henan Province, PR China with a previous human red mould intoxication episode, were analysed for trichothecenes and zearalenone (ZEA). For the 1998 Puyang crop, deoxynivalenol (DON) was the predominant toxin detected abundantly and frequently at a level of up to 14,000 microg kg(-1) (mean 2850 microgkg(-1)) in 30 of 31 (97%) wheat samples. Among these were 21 (70%) with a DON level that exceeded the Chinese regulation of 1,000 microg kg(-1). Nivalenol (NIV) and 15-acetyl-DON (15-ADON) were also found at 578 microg kg(-1) (one sample) and 59-1,800 microg kg(-1) (mean 365 microg kg(-1), 20 samples), respectively. ZEA co-occurred in 21 samples at 9-1,400 microg kg(-1) (mean 209 microg kg(-1)). Twenty-five (89%) wheat samples from Zhumadian, a region without a history of human red mould intoxication in the same province, contained low levels of DON (53-1240, mean 223 microg kg(-1)) with seven (25%) co-contaminated with ZEA (10-217, mean 108 microg kg(-1)). All were free from 15-ADON and NIV. Significant differences in DON, 15-ADON and ZEA concentrations between both areas were found. DON (<1000 microg kg(-1)) and ZEA (5-111 microg kg(-1)) were also detected in the 1999 Puyang wheat. Proper environmental conditions for Fusarium species surviving winter combined with unusual high precipitation during wheat flowering were responsible for a high concentration of Fusarium mycotoxins in the 1998 Puyang wheat.     

An epimer of deoxynivalenol: purification and structure identification of 3-epi-deoxynivalenol

In an investigation of deoxynivalenol (DON)-transformation products by Devosia mutans 17-2-E-8, the major product was identified as 3-epi-DON. This DON-transformation product was analysed by liquid chromatography and identified by congruent retention time and UV/Vis spectrum, as well as mass spectrometric data. Nuclear magnetic resonance (NMR) experiments including correlation spectroscopy (COSY), heteronuclear single quantum coherence (HSQC) and nuclear overhauser effect (NOE) were conducted for structural characterisation of 3-epi-DON. High-speed counter-current chromatography (HSCCC) was applied to scale up the separation of 3-epi-DON from DON in a D. mutans 17-2-E-8 culture. From the culture where 100 mg DON was applied, 56 mg of 3-epi-DON (purity of 96.8%) was obtained from the HSCCC. The purified 3-epi-DON will be used for toxicological characterisation studies of this chemical.     

Cross-reactivity features of deoxynivalenol (DON)-targeted immunoaffinity columns aiming to achieve simultaneous analysis of DON and major conjugates in cereal samples

Immunoaf?nity columns (IACs) are a well-established tool in the determination of regulated mycotoxins in food and feed commodities. However, they also have the potential to become attractive pre-concentration and clean-up materials for the determination of masked (also called modified) mycotoxins, which have been recognised as important contributors to the toxicological hazard deriving from fungal spoilage of goods. However, the information available in the literature concerning the cross-reactivity of DON-IACs against the major conjugates (DON-3-G, 15-AcDON and 3-AcDON) is incomplete and often contradictory. We have carried out a detailed characterisation of the cross-reactivity of the four main IACs brands against DON and its conjugates as well as an assessment of the competition among the analytes. Only one IAC enabled the simultaneous analysis of all relevant DON forms while two missed 15-AcDON and the fourth one missed DON-3-G and 3-AcDON. In the case of the multivalent IAC, the analytes modified at the C-3 position compete for the antibody binding with preference for 3-AcDON (less spatially hindered) while DON-3-G has the more-hindered access to the active sites. Taking into consideration the levels of DON conjugates existing in real samples, the cross-reactivity of one DON-IAC allows a quantitative analysis of all of these analytes. Important but rather neglected aspects such as the continuous supply of IACs with identical characteristics, and of columns which are strictly blank, are also addressed in this paper.     

Co-occurrence of mycotoxins in maize and maize-derived food in China and estimation of dietary intake

ABSTRACT

A total of 576 samples marketed in China, including maize, maize flour, maize grits and maize meal, was determined for the simultaneous presence of 12 mycotoxins (FB₁, FB₂, FB₃, DON, 3-DON, 15-DON, ZEN, AFB₁, AFB₂, AFG₁, AFG₂ and OTA) using a validated UPLC-MS/MS for multi-mycotoxin method. DON were the most widespread mycotoxins (63%), followed by FB₁ (57%) and ZEN (46%). 78% of the samples was contaminated with at least one of these mycotoxins. Risk assessment indicated that maize and maize-derived food intake does not pose a potential risk for general adult population with respect to individual mycotoxin. However, two or more mycotoxins were detected in 60% of all samples, and a combination of up to seven different mycotoxins was found. A particular attention should be paid to the combined exposure of mycotoxins, in this cases the estimated daily intake might increase greatly due to the high frequency of co-occurrence.     

Investigation of ochratoxin a, B and citrinin contamination in various commercial foods

Ochratoxin A (OTA), ochratoxin B (OTB) and citrinin (CIT) in commercial foods were simultaneously determined and confirmed with high-performance liquid chromatography (HPLC) and liquid chromatography coupled with tandem mass spectrometry (LC/MS/MS). The samples examined were made up of cereal, fruit, coffee, and cacao products. The limits of quantification (S/N> or =10) of OTA, OTB and CIT were 0.1 microg/kg or less. Aflatoxins (AF), deoxynivalenol (DON) and fumonisins were also surveyed. Of 157 samples examined, 44 were contaminated with OTA at levels of 0.11 to 4.0 microg/kg. At least 2 positive samples were labeled as domestics. In most positive samples, the OTA level was low, less than 1 microg/kg. The highest incidence of OTA was observed in cacao powder (10/12), followed by instant coffee (5/7), cocoa (5/8) and raisin (6/13). OTB was found in fruit and cacao products containing relatively high levels of OTA. Co-occurrence of OTA, CIT and DON was found in cereal products, and co-occurrence of OTA and AF was found in cacao products. Approximately 30% of naturally contaminated OTA in roasted coffee bean moved into the extract solution when brewed with paper filter.     

超高效液相色谱-串联质谱法测定牛奶中24 种真菌毒素

运用基质分散固相萃取净化,建立牛奶中24 种真菌毒素（黄曲霉毒素B1、黄曲霉毒素B2、黄曲霉毒素G1、黄曲霉毒素G2、黄曲霉毒素M1、赭曲霉毒素A、玉米赤霉烯酮、玉米赤霉酮、α-玉米赤霉烯醇、β-玉米赤霉烯醇、α-玉米赤霉醇、β-玉米赤霉醇、T-2毒素、HT-2霉素、伏马毒素B1、伏马毒素B2、伏马毒素B3、脱氧雪腐镰刀菌烯醇、3-乙酰脱氧雪腐镰刀菌烯醇、15-乙酰脱氧雪腐镰刀菌烯醇、交链孢霉单甲基醚、交链孢酚、腾毒素、细交链孢菌酮酸）多残留检测的超高效液相色谱-串联质谱检测方法。样品经80%乙腈溶液（体积分数）提取,通过基质分散固相萃取净化,氮吹至近干,1 mL 50%乙腈溶液（体积分数）复溶,采用超高效液相色谱-串联质谱进行测定。经ACQUITY UPLC HSS T3反相柱（2.1 mm×100 mm,1.8 μm）分离,梯度洗脱,采用电喷雾离子源-多反应监测模式采集。24 种目标物的相关系数（R2）均大于0.985,加标回收率为71.0%～123.0%,相对标准偏差均小于10%。该方法具有操作简单、重复性好、灵敏度高、杂质干扰小等特点,可以用于牛奶中24 种真菌毒素的检测。     

Development and Application of a Method for the Analysis of 9 Mycotoxins in Maize by HPLC‐MS/MS

A reliable and sensitive liquid chromatography/tandem mass spectrometry (LC‐MS/MS) method was developed for the simultaneous determination of aflatoxins (AFB₁, AFB₂, AFG₁, and AFG₂), ochratoxin A (OTA), deoxynivalenol (DON), zearalenone (ZEA), fumonisin B₁ (FB₁), and T2‐toxin in maize. The samples were first extracted using acetonitrile: water: acetic acid (79 : 20 : 1), and then further cleaned‐up using OASIS HLB cartridge. Optimum conditions for the extraction and chromatographic separation were investigated. The mean recoveries of mycotoxins in spiked maize ranged from 68.3% to 94.3%. Limits of detection and quantification ranged from 0.01 to 0.64 μg/kg and from 0.03 to 2.12 μg/kg, respectively. The LC‐MS/MS method has also been successfully applied to 60 maize samples, which were collected from Shaanxi Province of China. Twenty‐four of the total 60 samples (40%) were contaminated with at least 1 of these 9 mycotoxins. Occurrence of mycotoxins were 6.7%, 1.7%, 3.3%, 6.7%, 1.7%, 23.3%, and 3.3% for AFB₁, AFB₂, OTA, ZEA, DON, FB₁, and T2‐toxin, respectively. The results demonstrated that the procedure was suitable for the simultaneous determination of these mycotoxins in maize matrix.     

Exposure of consumers to deoxynivalenol from consumption of white bread in Hungary

In view of the frequent occurrence of mycotoxins in cereals, a study was initiated to assess the exposure of the Hungarian adult population. Consumption data for 1360 individuals, based on a 3-day questionnaire, indicated that white bread accounted for the major intake of cereal-based products. Various cereal products were analysed for 16 mycotoxins by a LC/MS/MS multi-toxin method with LOD of 16 μg kg?1 and LOQ of 50 μg kg?1. Deoxynivalenol (DON) was most frequently detected, but no acetyl-deoxynivalenol was present in detectable concentrations. Consumer exposure was calculated with standard Monte Carlo probabilistic modelling and point estimates, taking into account bread consumption and DON contamination in independently taken wheat flour and wheat grain samples. Over 55% of cases the DON intake were below 15% of the provisional maximum tolerable daily intake (PMTDI) of 1 μg/(kg bw)/day. However, in 5-15% of cases, the intake from bread consumption alone exceeded the PMTDI. Wheat grain data led to the higher percentage. Intakes estimated from both data sets were at or below the acute reference dose (ARfD) of 8 μg/(kg bw)/day in 99.94-99.97% of cases.     

Acrylamide levels in Finnish foodstuffs analysed with liquid chromatography tandem mass spectrometry

Sample clean-up and HPLC with tandem mass spectrometric detection (LC-MS/MS) was validated for the routine analysis of acrylamide in various foodstuffs. The method used proved to be reliable and the detection limit for routine monitoring was sensitive enough for foods and drinks (38 microg/kg for foods and 5 microg/L for drinks). The RSDs for repeatability and day-to-day variation were below 15% in all food matrices. Two hundred and one samples which included more than 30 different types of food and foods manufactured and prepared in various ways were analysed. The main types of food analysed were potato and cereal-based foods, processed foods (pizza, minced beef meat, meat balls, chicken nuggets, potato-ham casserole and fried bacon) and coffee. Acrylamide was detected at levels, ranging from nondetectable to 1480 microg/kg level in solid food, with crisp bread exhibiting the highest levels. In drinks, the highest value (29 microg/L) was found in regular coffee drinks.