Natural occurrence of fumonisins B(1) and B(2) in domestic maize of Taiwan.

Samples of maize grown in various districts of Taiwan were collected and analyzed for the presence of fumonisin B(1) (FB(1)) and fumonisin B(2) (FB(2)) using high-performance liquid chromatography. Forty-nine (44.5%) and 2 (1.8%) of 110 samples were found to contain FB(1) (109-1148 ng/g) and FB(2) (222-255 ng/g), respectively. The frequency of detection and also the maximum FB(1) concentration were found in samples from Penton (2/2, 262 ng/g), followed by Chiayi (18/26, 264 ng/g), Tainan (8/16, 160 ng/g), Hualinen (5/14, 1148 ng/g), Taitung (7/20, 109 ng/g), and Yunlin (9/26, 361 ng/g). Of the 110 samples examined, only 2 samples from Hualinen had been detected containing FB(2). During an analysis of the distribution pattern of FB(1), it became apparent that >79% of tested samples had FB(1) concentrations <100 ng/g, whereas 2.7% (or 3 samples) contained FB(1) >300 ng/g. These results clearly illustrated that domestically produced maize for human consumption is frequently contaminated with FB(1).     

Determination of N-(carboxymethyl)fumonisin B(1) in corn products by liquid chromatography/electrospray ionization--mass spectrometry

It is well-known that fumonisin B(1) (FB(1)) in corn meal decreases during baking, frying, and cooking, but it is still not exactly clear how heating affects the formation of N-(carboxymethyl)fumonisin B(1) (NCM-FB(1)), the reaction product of FB(1) and reducing sugars. In model experiments corn grits were spiked with FB(1) (2 mg/kg) and D-glucose (50 g/kg) or sucrose (50 g/kg) and manufactured into extrusion products at various temperatures (160--180 degrees C) and moisture levels (16--20%). A liquid chromatography/electrospray ionization-mass spectrometry method using isotopically labeled fumonisin FB(1)-d(6) as an internal standard was developed for the determination of NCM-FB(1). For sample cleanup solid-phase C18 cartridges were used. The detection limit achieved with this method was 10 ng/g (signal-noise ratio = 3:1) using the protonated molecule [M + H](+) signal of NCM-FB(1) (m/z 780) in the selected ion monitoring mode. Low concentrations of NCM-FB(1) (29-97 ng/g) were detected in all samples spiked with D-glucose and FB(1), whereas those spiked with FB(1) and sucrose showed only NCM-FB(1) in samples produced at 180 degrees C (NCM-FB(1) = 27 ng/g). Various corn-containing food samples from the German market were analyzed for the presence of NCM-FB(1), FB(1), and hydrolyzed fumonisin B(1) (HFB(1)). All samples were contaminated with FB(1) (22--194 ng/g) and HFB(1) (5--247 ng/g). Six of nine samples contained NCM-FB(1) in low concentrations ranging from 10 to 76 ng/g. From these data and the low toxicity of NCM-FB(1) it can be concluded that the significance of NCM-FB(1) in food seems to be a minor one.     

Analysis of fumonisin B(1) in Fusarium proliferatum-infected asparagus spears and garlic bulbs from Germany by liquid chromatography-electrospray ionization mass spectrometry

Fusarium proliferatum is one of a group of fungal species that produce fumonisins and is considered to be a pathogen of many economically important plants. The occurrence of fumonisin B(1) (FB(1)) in F. proliferatum-infected asparagus spears from Germany was investigated using a liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS) method with isotopically labeled fumonisin FB(1)-d(6) as internal standard. FB(1) was detected in 9 of the 10 samples in amounts ranging from 36.4 to 4513.7 ng/g (based on dry weight). Furthermore, the capability of producing FB(1) by the fungus in garlic bulbs was investigated. Therefore, garlic was cultured in F. proliferatum-contaminated soil, and the bulbs were screened for infection with F. proliferatum and for the occurrence of fumonisins by LC-MS. F. proliferatum was detectable in the garlic tissue, and all samples contained FB(1) (26.0-94.6 ng/g). This is the first report of the natural occurrence of FB(1) in German asparagus spears, and these findings suggest a potential for natural contamination of garlic bulbs with fumonisins.     

Surveillance of fumonisins in maize-based feeds and cereals from spain.

A survey has been carried out to determine the levels of fumonisins in 171 samples of maize-based feeds and cereals available in Spain. Also, the samples were examined for mold count and fungal species. Aspergillus, Penicillium, and Fusarium were the most frequent genera, and Fusarium and Aphanocladium had the highest individual percentage counts. Regarding Fusarium species, F. moniliforme (47. 4%) was the predominant species; F. proliferatum (5.3%) and F. subglutinans (7.0%) were isolated at low frequency. The high-performance liquid chromatography-o-phthaldialdehyde fluorescence method was used for the analysis of fumonisins. The highest levels of fumonisins were detected in maize. Overall, fumonisin B(1) (FB(1)) and fumonisin B(2) (FB(2)) were detected in 79.5 and 14.6%, of samples respectively, with average FB(1) levels of 3.3 microg/g and average FB(2) levels of 1.7 microg/g. Low levels of fumonisins in wheat, barley, and soybean were detected. This would appear to be the first report of concentrations of fumonisins in these commodities.     

Natural occurrence of fumonisins in corn from Iran

Corn collected in the Mazandaran and Isfahan Provinces of Iran was analyzed for fumonisin B(1) (FB(1)), fumonisin B(2) (FB(2)), and fumonisin B(3) (FB(3)). The samples from Mazandaran Province, situated on the Caspian littoral of Iran, were random samples from farmers' corn lots collected in September 1998, whereas those from Isfahan Province, situated further south in the center of Iran, were bought as corn cobs in the local retail market during October 1998. All 11 samples from Mazandaran showed high levels of fumonisin contamination with FB(1) levels between 1.270 and 3.980 microg/g, FB(2) levels between 0.190 and 1.175 microg/g, and FB(3) levels between 0.155 and 0.960 microg/g. Samples from Isfahan showed lower levels of contamination with eight of eight samples having detectable FB(1) (0.010-0.590 microg/g), two of eight samples having detectable FB(2) (0.050-0.075 microg/g), and two of eight samples having detectable FB(3) (0.050-0.075 microg/g). This is the first report of fumonisin contamination of corn from Iran, in which samples from the area of high esophageal cancer on the Caspian littoral have been shown to contain high levels of fumonisins.     

Simultaneous determination of fumonisin B(1) and hydrolyzed fumonisin B(1) in corn products by liquid chromatography/electrospray ionization mass spectrometry

A method for the simultaneous determination of fumonisin B(1) (FB(1)) and its major hydrolysis product (HFB(1)), which is known to be formed during alkaline treatment of fumonisin-containing corn meal, was devised to analyze the levels of these mycotoxins in corn products available on the German market. Liquid chromatography/electrospray mass spectrometry in combination with selected ion monitoring (SIM) was used for unambiguous detection of FB(1) and HFB(1) after extraction of samples with acetonitrile/methanol/water (25:25:50) and solid-phase C18 cleanup. Quantitation was carried out using labeled fumonisin FB(1)-D(6) as an internal standard. The detection limits achieved with this method were 8 ng/g for HFB(1) (signal-noise ratio = 5:1) and 5 ng/g for FB(1) (s/n = 5:1) using the protonated molecule signals m/z 406 and 722 in the SIM mode. A screening of several corn-containing foodstuffs, among them extrusion products and alkali-processed corn food such as tortilla chips, showed HFB(1) and FB(1) contamination with levels of 8-80 and 5-450 ng/g, respectively.     

Incidence of Fusarium verticillioides and levels of fumonisins in corn from main production areas in Iran

A total of 52 corn samples collected in 2000 from four main corn production provinces of Iran (Fars, Kermanshah, Khuzestan, and Mazandaran) were analyzed for contamination with Fusarium verticillioides and fumonisins (FB(1), FB(2), FB(3), and 3-epi-FB(3)). The mean incidence of F. verticillioides (percent of kernels infected) for these four areas was 26.7, 21.4, 24.9, and 59.0%, respectively. The incidence in Mazandaran was significantly (p < 0.05) above that of the other areas. All samples from Mazandaran were contaminated with fumonisins with a mean level of total fumonisins of 10674 microg/kg. In contrast, the incidence of fumonisin contamination above 10 microg/kg was 53 (8/15), 42 (5/12), and 57% (8/14) in the samples from Fars, Kermanshah, and Khuzestan, respectively, and the corresponding mean total fumonisin levels were 215, 71, and 174 microg/kg, respectively. No statistical differences (p > 0.05) were observed in the fumonisin levels of the corn samples from these three provinces, which were significantly (p < 0.05) lower than the fumonisin contamination in samples from Mazandaran.     

Evaluation of fumonisin contamination in cornflakes on the Belgian market by "flow-through" assay screening and LC-MS/MS analyses

A total of 205 cornflake samples collected in Belgian retail stores during 2003-2004 were surveyed for the natural occurrence of fumonisin B1 (FB1), B2 (FB2), and B3 (FB3). These cornflake samples, originating from conventional as well as from organic production, were analyzed using an intralaboratory-validated LC-MS/MS method. Additionally, 90 cornflake samples were subjected to rapid screening using a flow-through enzyme immunoassay method to demonstrate the practicability of a screening test coupled to a validated confirmatory LC-MS/MS method for the management of food safety risks. FB(1) concentrations ranged from not detected (nd) [LOD (FB1) = 20 microg/kg] to 464 microg/kg with mean and median concentrations of respectively 104 +/- 113 and 54 microg/kg. For FB2 and FB3, the concentration ranges varied respectively from nd [LOD (FB2) = 7.5 microg/kg] to 43 microg/kg and from nd [LOD (FB3) = 12.5 microg/kg] to 90 microg/kg. Mean concentrations for FB2 and FB3 were respectively 12 +/- 8 and 21 +/- 15 microg/kg, while the median concentration was 11 microg/kg for FB2 and 19 microg/kg for FB3. From the statistical tests (chi2 and ANOVA model III), it could be concluded that the agricultural practice did not have any significant effect on the fumonisin concentrations but that the variation between different batches was significant (p < 0.0001).     

Fluorescence polarization as a means for determination of fumonisins in maize

Fumonisins, mycotoxins produced by certain species of Fusaria, are commonly found worldwide as contaminants in maize. This paper reports the development of a rapid, portable fluorescence polarization-based assay for fumonisins in maize. The assay was based on the competition of unlabeled fumonisin, from a sample, with a fluorescently tagged fumonisin (FB(1)-FL) for a fumonisin-specific monoclonal antibody in solution. The fluorescence polarization (FP) of the tagged fumonisin was increased upon binding with the antibody. In the presence of free toxin, less of the FB(1)-FL was bound and the polarization signal was decreased. The assays were very simple to perform, requiring only mixing of an aqueous extract of maize with the tagged fumonisin and antibody, and required <2 min per sample, excluding extraction time. Two permutations of the assay were tested, one with each sample matrix serving as its own blank, and the other with all of the samples compared relative to a PBS blank with normalization of the data similar to an ELISA. The limit of detection, defined as the toxin content associated with a fluorescence polarization signal 5 standard deviations from that of a fumonisin-free control, was 0.5 microg of FB(1)/g in spiked maize. Recoveries from spiked maize over the range of 0.5-20 ppm averaged 94.3 +/- 13.8%. Forty-eight samples of field-contaminated maize were tested by the FP and an established HPLC method, with a good correlation between the two (r(2) = 0.85-0.88). For these samples, the two variations of the FP assay also compared well to one another (r(2) = 0.97), suggesting the assay principle is very robust. The results, combined with the speed and ease of use for the assay, suggest that this technology has substantial potential as a screening tool for mycotoxins in foods.     

Liquid chromatographic determination of fumonisins B1, B2, and B3 in corn silage

Corn silage was dried, ground, and then extracted with 0.1 M ethylenediaminetetraacetic acid. The filtrate was applied to a FumoniTest immunoaffinity column. Fumonisins were derivatized with naphthalene-2,3-dicarboxaldehyde, separated on a C(18) liquid chromatographic column, and detected by fluorescence. The detection limits for fumonisin B(1), fumonisin B(2), and fumonisin B(3) were 50, 25, and 25 ng/g of dried silage, respectively. Recoveries of fumonisin B(1), fumonisin B(2), and fumonisin B(3) from wet and dried corn silage spiked over the range of 100-5000 ng/g averaged 91-106%. The method was applied to corn silage samples collected from the midwestern area of the United States during 2001-2002. Of 89 corn silage samples, fumonisin B(1), fumonisin B(2), and fumonisin B(3) were found in 86 (97%), 64 (72%), and 51 (57%) of the samples. The mean positive levels of fumonisin B(1), fumonisin B(2), and fumonisin B(3) were 615, 93, and 51 ng/g, respectively, in dried silage. This suggests that fumonisins may be frequent low level contaminants in corn silage.     

Occurrence of Fusarium spp. and fumonisin in durum wheat grains

A survey was carried out to determine Fusarium species and fumonisin contamination in 55 durum wheat (Triticum turgidum L. var. durum) samples collected during two harvest seasons (2007 and 2008) using HPLC and further LC-MS/MS confirmation. All samples showed Fusarium contamination with infection levels ranging from 8 to 66%, F. proliferatum being the species most frequently isolated during 2007 and the second most frequently isolated one during the 2008 harvest season, respectively. Natural contamination with fumonisins was found in both harvest seasons. In 2007, 97% of the samples showed total fumonisin (FB(1) + FB(2)) levels ranging from 10.5 to 1245.7 ng/g, while very low levels of fumonisins were detected in samples collected during 2008. These results could be explained by differences in the amount of rainfall during both periods evaluated. A selected number (n = 48) of F. proliferatum isolates showed fumonisin production capability on autoclaved rice. This is the first report of the presence of natural fumonisins in durum wheat grains.     

Mycoflora and fumonisin contamination in Brazilian corn from sowing to harvest

The present study aimed to analyze the mycoflora and potential mycotoxin contamination of soil and corn samples collected at different plant maturity stages in Cap?o Bonito and Ribeir?o Preto, two regions of the State of S?o Paulo, Brazil. In addition, the data obtained were correlated with the occurrence of wind-dispersed fungi and the predominant climatic conditions of the two regions studied. Corn mycoflora profiles showed that Fusarium verticillioides prevailed in 35% of the samples from Cap?o Bonito and in 49% of the samples from Ribeir?o Preto. Examination of wind-dispersed fungi also revealed a high incidence of F. verticillioides. Soil mycoflora analyses showed that Penicilliumwas the most prevalent genus, although F. verticillioides was present in 55.5% of Cap?o Bonito's samples and in 26.7% of Ribeir?o Preto's samples. With respect to water activity, the corn kernels most contaminated with F. verticillioides had water activity levels of 0.70-0.80. HPLC analysis of fumonisins revealed that 88.5% of Cap?o Bonito's kernels were contaminated with fumonisin B(1) (FB(1)) (0.09-10.87 microg/g) and 53.8% with fumonisin B(2) (FB(2)) (0.05-0.52 microg/g); Ribeir?o Preto's kernels presented contamination levels of 93.5% for FB(1) (0.11-17.69 microg/g) and 61.3% for FB(2) (0.05-5.24 microg/g). No aflatoxins were detected by thin-layer chromatography in corn grains of either region. The concomitant occurrence of F. verticillioides and fumonisins in most of the field corn assayed demonstrates the importance of an effective control of cultivation throughout the plant maturity stages.     

Fumonisin mycotoxins in traditional Xhosa maize beer in South Africa

The production and consumption of home-brewed Xhosa maize beer is a widespread traditional practice in the former Transkei region of South Africa. HPLC determination of fumonisins B1 (FB1), B2 (FB2), and B3 (FB3) in maize beer samples collected in two magisterial areas, Centane and Bizana, showed a wide range of levels. All samples were positive for FB(1), with a mean level of 281 +/- 262 ng/mL and a range from 38 to 1066 ng/mL. Total fumonisins (FB1 + FB2 + FB3) ranged from 43 to 1329 ng/mL, with a mean of 369 +/- 345 ng/mL. Data on the consumption of home-brewed beer are not available. On the basis of published data for the consumption of commercial beer in South Africa, the fumonisin exposure in these districts among the consumers of maize beer was found to be well above the provisional maximum tolerable daily intake of 2 mug/kg of body weight/day set by the Joint FAO/WHO Expert Committee on Food Additives.     

Rapid determination of fumonisin B1 in food samples by enzyme-linked immunosorbent assay and colloidal gold immunoassay

A rapid enzyme-linked immunosorbent assay (ELISA) test (microwell plate) and a membrane-based colloidal gold immunoassay in flow-through and lateral-flow formats for the rapid detection of fumonisin B1 (FB1) were developed. The rapid microwell assay can be completed within 20 min with the detection limit of 0.5 +/- 0.2 microg/L. Membrane-based colloidal gold immunoassays had a visual detection limit of 1.0 microg/L for FB1 with the detection time of <10 min. Matrix interference was eliminated by 15-fold dilutions of methanol extracts with buffer. These immunoassays can be used as quantitative or qualitative tools for the rapid detection of FB1 residues in 10-20 min on-site.     

Mycoflora and fumonisin mycotoxins associated with cowpea [Vigna unguiculata (L.) Walp] seeds

Cowpea seed samples from South Africa and Benin were analyzed for seed mycoflora. Fusariumspecies detected were F. equiseti, F. chlamydosporum, F. graminearum, F. proliferatum, F. sambucinum, F. semitectum, and F. subglutinans. Cowpea seed from South Africa and Benin and F. proliferatum isolates from Benin, inoculated onto maize patty medium, were analyzed for fumonisin production. Samples were extracted with methanol/water and cleaned up on strong anion exchange solid phase extraction cartridges. HPLC with precolumn derivatization using o-phthaldialdehyde was used for the detection and quantification of fumonisins. Cowpea cultivars from South Africa showed the presence of fumonisin B(1) at concentrations ranging between 0.12 and 0.61 microg/g, whereas those from Benin showed no fumonisins. This is believed to be the first report of the natural occurrence of FB(1) on cowpea seed. Fumonisin B(1), B(2), and B(3) were produced by all F. proliferatum isolates. Total fumonisin concentrations were between 0.8 and 25.30 microg/g, and the highest level of FB(1) detected was 16.86 microg/g.     

Comparison of extraction buffers for the detection of fumonisin B(1) in corn by immunoassay and high-performance liquid chromatography

The Associatian of Official Analytical Chemists approved method for quantification of fumonisin B(1) (FB(1)) in corn meal or corn-based food products includes extraction into methanol (MeOH)/water (3:1, v/v). Disposal of the extraction medium can pose safety and environmental problems. To secure a rapid and inexpensive screen for FB(1) contamination, a sensitive competitive ELISA using a rabbit polyclonal antibody was developed. This assay was used in a comparative study measuring the extraction efficiency of FB(1) in aqueous or organic solvent buffers using 16 field corn samples. An aqueous phosphate buffer was found to be suitable for extracting FB(1), thus eliminating the need for organic solvents. HPLC and ELISA determinations compared well in fortified samples at known concentrations between 1 and 50 microg/mL of extract. Overestimation at levels >50 microg/mL were common. The characteristics and application of the ELISA for screening purposes are discussed.     

Reduction of fumonisin B1 in corn grits by single-screw extrusion

This study was designed to determine the efficacy of extrusion in reducing fumonisin B1 in corn flaking grits in the presence and absence of glucose. In addition, degradation products of fumonisin B1 during extrusion were identified and quantitated with a mass balance approach. Uncontaminated clean corn grits, grits spiked with 30 microg/g fumonisin B1, and grits fermented with Fusarium verticillioides M-2552 (40-50 microg/g fumonisin B1) were extruded in the presence and absence of glucose (10%, w/w) using a single-screw extruder. Extrusion decreased fumonisin B1 by 21-37%, whereas the same process with added glucose further decreased fumonisin B1 by 77-87%. LC-fluorescence and LC-MS showed that most fumonisin in the extruded samples without added glucose was the fumonisin B1 form, whereas the main degradation product in grits extruded with glucose was N-(deoxy- d-fructos-1-yl)fumonisin B1. The formation of hydrolyzed fumonisin B1 was not significant during extrusion. Results suggest that extrusion in the presence of glucose may reduce fumonisin B1 in corn grits significantly.     

Extraction of fumonisins B1 and B2 from white rice flour and their stability in white rice flour,cornstarch, cornmeal,and glucose

To extract fumonisin B1 (FB1) and fumonisin B2 (FB2) from Thai white rice flour, different solvent mixtures, temperatures, pH values, and addition of enzymes or ethylenediaminetetraacetic acid disodium salt (Na2EDTA) were examined. Three extractions with 0.1 M Na2EDTA achieved the highest recoveries. Initial recoveries of fumonisins added to white rice flour, cornstarch, cornmeal, and glucose varied with commodity. Fumonisins disappeared in Thai white rice flour after 12 h, but 55% remained in another white rice flour. With cornstarch 20-30% fumonisins remained after 24 h; only 43% of 14C-labeled FB1 materials extracted from cornstarch was eluted with methanol from an immunoaffinity column. Fumonisins were stable in cornmeal for 24 h but only approximately 50% remained after 30 days. With glucose, 25% of FB1 and FB2 remained 24 h after addition; N-(1-deoxy-D-fructos-1-yl)FB(1) andN-(carboxymethyl)FB(1) were detected in lower amounts than residual FB(1) after 3 months.     

Evaluation of conventional and organic italian foodstuffs for deoxynivalenol and fumonisins B(1) and B(2)

Two lots of human foodstuffs from conventional and organic brand foods were purchased from supermarkets and analyzed for three Fusarium toxins, deoxynivalenol, by GC-ECD, and fumonisins B(1) and B(2) (FB(1)-FB(2)), by LC-MS. The occurrence of deoxynivalenol contamination was higher than 80% in both organic and conventional foods; fumonisin B(1) was found in 20% of organic foods and in 31% of conventional ones and fumonisin B(2) in more than the 32% of the food samples from both the agricultural practices. The highest median concentration of deoxynivalenol occurred in conventional rice-based foodstuffs (207 microg/kg): that of fumonisin B(1) in conventional maize-based foods (345 microg/kg) and that of fumonisin B(2) in organic wheat-based foods (210 microg/kg).     

A validated multianalyte LC-MS/MS method for quantification of 25 mycotoxins in cassava flour, peanut cake and maize samples

This study was designed to develop a sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the simultaneous detection and quantification of 25 mycotoxins in cassava flour, peanut cake and maize samples with particular focus on the optimization of the sample preparation protocol and method validation. All 25 mycotoxins were extracted in a single step with a mixture of methanol/ethyl acetate/water (70:20:10, v/v/v). The method limits of quantification (LOQ) varied from 0.3 μg/kg to 106 μg/kg. Good precision and linearity were observed for most of the mycotoxins. The method was applied for the analysis of naturally contaminated peanut cake, cassava flour and maize samples from the Republic of Benin. All samples analyzed (fifteen peanut cakes, four maize flour and four cassava flour samples) tested positive for one or more mycotoxins. Aflatoxins (total aflatoxins; 10-346 μg/kg) and ochratoxin A (相似文献