Occurrence of fumonisins B1 and B2 in Portuguese maize and maize-based foods intended for human consumption

Fumonisin B₁ (FB₁) and fumonisin B₂ (FB₂) are mycotoxins mainly produced by Fusarium verticillioides and Fusarium proliferatum, which are field pathogens of maize. A survey was conducted on the incidences of FB₁ and FB₂ in both maize and derived products purchased in Portugal. The analytical method involved extraction with methanol-water, clean-up by immunoaffinity column and derivatization with naphthalene-2,3-dicarboxaldehyde. Determination was carried out by high-performance liquid chromatography (HPLC) with spectrofluorimetric detection, with liquid chromatography/mass spectrometry (LC/MS) confirmation. The presence of FB₁ and FB₂ was determined in 67 samples of maize and maize-based foods, such as flour, semolina, starch, sweet maize, cornflakes and other breakfast cereals, and snacks collected in 2005. FBs were found in 15 samples at concentrations ranging from 113 to 2026 µg kg^-1. Two of the samples showed higher contamination levels than the limits established by the European Commission Regulation. None of the samples contained levels of fumonisins that would lead to an exposure exceeding the tolerable daily intake (TDI).     

Occurrence of mycotoxins and ergosterol in maize harvested over 5 years in Northern Italy

Maize samples collected from storage bins and feed mills in Northern Italy between 1995 and 1999 were surveyed for the occurrence of aflatoxin B₁ (AFB₁), zearalenone (ZEA), deoxynivalenol (DON) and fumonisin (FB₁); further, ergosterol was analysed as a fungal growth marker. The incidence and mean content of AFB₁ were generally low; nevertheless, a remarkable contamination was found in two samples (109 and 158 μg kg^-1), while five others exceeded 20 μg kg^-1. DON and ZEA mean levels were significantly higher in 1996 (2716 and 453 μg kg^-1) with respect to the other years, when mean contents ranged from 7 to 30% and from 3 to 17%, respectively, expressed in per cent of 1996 contents. FB₁ was present in all samples and was by far the most remarkable mycotoxin in Northern Italian maize, with the exception of samples from 1996. The average level was 3064 μg kg^-1, 69.6% of samples resulted over 1000 μg kg^-1 and 16.9% over 5000 μg kg^-1. Significant correlations were found between ergosterol and the major mycotoxin(s) in each year (FB₁ in 1995 and 1997-99; ZEA + DON in 1996). Consequently, ergosterol seems to be a good index of the toxicological quality of maize. Climatic conditions influenced the growth of different fungal species. In 1996, the first 20 days of October were extremely rainy; these weather conditions delayed the harvest until the first week of November and favoured the growth of DON and ZEA producing fungi and the synthesis of mycotoxins. On the contrary, the temperate and dry climate of the other years supported the growth of FB₁-producing fungi.     

Aflatoxin and ochratoxin A content of spices in Hungary

In October and November 2004, 91 spice samples (70 ground red pepper, six black pepper, five white pepper, five spice mix and five chilli samples), the majority of which originated from commercial outlets, were analysed for aflatoxins B₁, B₂, G₁ and G₂ (AFB1, AFB2, AFG1, AFG2) and ochratoxin A (OTA) content by high-performance liquid chromatography (HPLC) after immunoaffinity column clean-up. Eighteen of the 70 ground red pepper samples contained AFB1, seven of them in a concentration exceeding the 'maximum level' of 5 µg kg^-1 (range 6.1-15.7 µg kg^-1). Of the other spices assayed, the AFB1 contamination of one chilli sample exceeded 5 µg kg^-1 (8.1 µg kg^-1). Thirty-two of the 70 ground red pepper samples contained OTA, eight of them in a concentration exceeding the 10 µg kg^-1 'maximum level' (range 10.6-66.2 µg kg^-1). One chilli sample was contaminated with OTA at 2.1 µg kg^-1. The AFB1 and OTA contamination of ground red pepper exceeding the 'maximum level' (5 and 10 µg kg^-1, respectively) was obviously the consequence of mixing imported ground red pepper batches heavily contaminated with AFB1 and OTA with red pepper produced in Hungary. This case calls attention to the importance of consistently screening imported batches of ground red pepper for aflatoxin and ochratoxin A content and strictly prohibiting the use of batches containing mycotoxin concentrations exceeding the maximum permitted level.     

Determination of the mycotoxin fumonisin B1 in maize by reversed-phase thin-layer chromatography: a collaborative study

A simple and cost-effective method using thin-layer chromatography for the determination of the mycotoxin fumonisin B₁ in maize is described. The analytical method consisted of the extraction of ground maize by shaking with methanol/water (75:25) for 60 min and clean-up of the resultant extract by means of strong anion exchange solid-phase extraction. The purified residue, formed by evaporation of the elution solvent, was reacted with fluorescamine and the fumonisin B₁-derivative was separated by reversed-phase thin-layer chromatography using a developing solution of methanol/aqueous 4% potassium chloride (70:30). The derivatized FB₁ was readily visualized as a greenish-yellow spot under long wavelength ultraviolet light and quantified by visual comparison with a set of similarly derivatized standards in the range 20-300 ng FB₁ spotted on plate. Based on visual comparison, levels down to 0.5 mg kg^-1 were successfully estimated. The method was collaboratively studied in 14 laboratories using four duplicate maize meal samples (including a blank) and a spiked sample for determination of recovery. No significant difference was observed between mean FB₁ levels by high-performance liquid chromatography or thin-layer chromatography. Based on within-laboratory relative standard deviations of 27.1-41.7% and between-laboratory relative standard deviations of 35.0-63.3%, the method can be considered semiquantitative. The mean recovery achieved by participants at a spiking level of 2.00 mg kg^-1 was 74.5%.     

Consumers' ability to discriminate aflatoxin-contaminated Brazil nuts

The objectives of the study were to investigate the extent to which consumers can separate nuts with a high content of aflatoxin from sound nuts, and whether sorting results can be improved by information or whether they are affected by certain factors. A test panel consisting of 100 subjects was asked to crack 300 g Brazil nuts and to sort the nuts into those they considered edible and inedible. The test showed that consumers can, on current behaviour, discriminate aflatoxin-contaminated Brazil nuts to a significant extent. The median and the 95th percentile of the total concentrations of aflatoxins (B₁, B₂, G₁, G₂) in the samples before sorting were 1.4 and 557 µg kg^-1, respectively, and in the edible fractions after sorting 0.4 and 56 µg kg^-1, respectively. Given that levels of aflatoxins before sorting exceed either 2 µg aflatoxin B₁ kg^-1 or totally 4 µg aflatoxins kg^-1, there was no effect of aflatoxin concentrations before sorting on the probability of exceeding these thresholds in the edible fraction. This means that similar sorting results were obtained for samples with aflatoxin levels exceeding either of the two thresholds, irrespective of if the thresholds were exceeded with a few µg kg^-1 or up to more than 1000 µg kg^-1. None of the tested factors (such as sex, age, level of education, ethnic background or knowledge of mycotoxins) had any effects on the probability of exceeding either of the two aflatoxin thresholds.     

Natural co-occurrence of aflatoxin B1, fumonisin B1 and ochratoxin A in barley and corn foods from Korea

A survey for aflatoxin B₁ (AFB₁), fumonisin B₁ and ochratoxin A (OTA) was conducted on 127 samples that included 30 food-grade barley, 32 barley foods, 18 food-grade corn and 47 corn foods, randomly collected during 1998-99 in Seoul, Korea. The presence of mycotoxins was analysed by direct competitive enzyme-linked immunosorbent assay (ELISA), and most of the positive samples from ELISA were confirmed using high-performance liquid chromatography (HPLC). Recoveries of AFB₁ and OTA spiked at 10 ng g -1 and FB₁ spiked at 50 ng g^-1 were 106, 87 and 105% by ELISA, whereas those by HPLC were 80, 79 and 84%, respectively. Detection limits by ELISA for AFB₁, FB₁ and OTA were 1, 5 and 5 ng g^-1, and those by HPLC were 0.6, 35 and 1 ng g^-1. Naturally occurring AFB₁, FB₁ and OTA were found in 4/32 (12%), 2/32(6%) and 4/32 (12%) samples of barley foods with an average of 26, 16 and 9 ng g^-1, respectively. AFB₁ and FB₁ in corn foods were detected in 4/47 (8%) and 9/47 (19%) samples with the average being 20 and 74 ng g^-1, while no OTA was found in any corn foods samples. No AFB₁, FB₁ or OTA was detected in any of food-grade barley and corn samples. This is the first report on the natural co-occurrence of AFB₁ and FB₁ in barley and corn foods as well as on surveillance of OTA in Korea.     

Five-year survey of ochratoxin A in processed sultanas from Turkey

The results of surveillance for ochratoxin A (OTA) in 1885 samples of sultanas taken during five crop years between 1999 and 2003 are reported. The analytical method was based on extraction with methanol + sodium bicarbonate and clean-up by immunoaffinity column chromatography followed by high-performance liquid chromatography with fluorescence detection. The limit of detection for OTA was 0.3 µg kg^-1. The results show that 9.3% of the samples contained no detectable levels of OTA, whereas 0.6% had concentrations exceeding 10 µg kg^-1; the remaining 90.3% had levels within the range 0.3-10 µg kg^-1. The overall mean OTA concentration in the total number of 1885 samples taken was 1.36 ± 2.91 µg kg^-1; the overall median was calculated as 0.90 µg kg^-1.     

Occurrence of fumonisin B1 and B2 in beer

A total of 29 nationally distributed brands of beer, representing 25 domestic US and four imported brands, were purchased in retail outlets in Lincoln, Nebraska and analysed for concentrations of fumonisin B₁ (FB₁) and B₂(FB₂). Immunoaffinity column extraction and cleanup of fumonisins from the beer samples, coupled with detection and analysis by gradient high performance liquid chromatography (HPLC), provided a limit of quantitation for each toxin of 0.3ng/ml. Of the brands of beer sampled, 86% were positive for FB and 41% were positive for FB2. No beer contained a detectable quantity of FB without a detectable quantity of FB1. The total fumonisin (FB₁ + FB₂) content of positive samples ranged from 0.3 to 12.7ng/ml, with a mean concentration for all positive samples of 4.0 ± 3.4ng/ml (n = 25). Considering that the level of fumonisin contamination of corn in recent harvest years has been minimal, the results of this limited survey could represent levels associated with current agricultural and brewing practices.     

In vitro microbial metabolism of fumonisin B1

There is a lack of information on the effect of swine caecal microbiota on fumonisin metabolism. In this in vitro study, the biotransformation of fumonisin B₁ (FB₁) by the gut microbiota of adult, healthy pigs was examined. Suspensions of caecal contents and McDougall buffer solution were incubated anaerobically with pure FB₁ for 0, 12, 24, 48 and 72 h. After 48 h, the conversion of FB₁ to partially hydrolysed FB₁ (46%) was nearly equal to the percentage ratio of FB₁, while by 72 h it was 49%. In vitro, the conversion of fumonisin B₁ to aminopentol was less than 1%. The results show that the caecal microbiota are capable of transforming fumonisin B₁ to the above metabolites. Further studies on FB₁ metabolism in the small intestine are clearly justified.     

Determination of aflatoxin B1 in tiger nut-based soft drinks

An analytical method for the determination of aflatoxin B₁ in a tiger nut-based soft drinks named 'horchata' is described. The method is based on an immunoaffinity clean-up, followed by HPLC separation and fluorescence detection after electrochemical post-column derivatization. The detection limit (S/N = 3) and the quantification limit (S/N = 10) were 0.02 and 0.06 µg kg^-1, respectively. The mean recovery at a level of 2 µg l^-1 was 88% (n = 6) and the coefficient of variation was 9%. The method was applied to conduct a small market survey for a beverage named 'horchata' that is frequently consumed by parts of the population in Southern Europe. Twenty-two samples from Spanish and Belgian supermarkets were analysed. As a result, only one sample was found to contain aflatoxin B₁ at the limit of quantitation of the method.     

Wheat kernel black point and fumonisin contamination by Fusarium proliferatum

Fusarium proliferatum is a major cause of maize ear rot and fumonisin contamination and also can cause wheat kernel black point disease. The primary objective of this study was to determine whether nine F. proliferatum strains from wheat from Nepal can cause black point and fumonisin contamination in wheat kernels. For comparison, the study included three Fusarium strains from US maize. In test 1, all the strains but one produced significant symptoms of kernel black point; two strains decreased kernel yield; and four strains contaminated kernels with fumonisins B₁, B₂ and B₃ as determined by liquid chromatography-mass spectroscopy. Strain Ggm202 from Nepal, which produced the highest levels of fumonisins (mean = 49 µg g^-1) on five wheat cultivars in test 1, was confirmed to produce fumonisins (mean = 38 µg g^-1) on two cultivars in test 2. The data indicate a potential for fumonisin contamination of wheat infected with F. proliferatum.     

Absorption, distribution and elimination of fumonisin B1 metabolites in weaned piglets

The absorption, distribution and elimination of fumonisin B₁ (and B₂) after oral administration of Fusarium verticillioides (MRC 826) fungal culture, mixed into the experimental feed for 10 days, was studied in weaned barrows. In order to determine the absorption of FB₁ from the feed marked by chromium oxide, a special T-cannula was implanted into the distal part of pigs' ileum. During the feeding of toxin-containing diet (45 mg FB₁ kg^-1) and until the tenth day after the end of treatment, the total quantity of urine and faeces was collected and their toxin content analysed. At the end of the trial, samples of lung, liver, kidney, brain, muscle, and fat were also collected and their fumonisin content analysed by LC-MS. The fumonisins appeared to decrease the reduced glutathione content in blood plasma and red blood cell haemolysate, possibly associated with in vivo lipid peroxidation. From a data set of 80 individual data and the concentration and rate of C_r and fumonisins (FB₁, partially hydrolysed FB₁ and aminopentol) in the chymus, it could be established that the accumulative absorption of fumonisin B₁ was 3.9% ± 0.7%. In the chymus, the FB₁ conversions into aminopentol and partially hydrolysed FB₁ were 1.0 and 3.9%, respectively. The degree of metabolism in faeces was variable, although the main product was the partially hydrolysed form, with very small amounts of the aminopentol moiety being recovered. In the investigated tissues the FB₁ conversion to aminopentol and partially hydrolysed FB₁ was 30 and 20%, respectively.     

Determination of ochratoxin A in virgin olive oils of Greek origin by immunoaffinity column clean-up and high-performance liquid chromatography

An improved specific analytical method for ochratoxin A (OTA) determination in olive oil is described, using a methanolic-aqueous extraction, an immunoaffinity column clean up step and high-pressure liquid chromatography with fluorescence detection. The mean recovery was found at 108% (relative standard deviation, RSD = 4.7%) and the detection limit (DL) was estimated at 4.6 ng kg^-1. Along with OTA, aflatoxin B₁ (AFB₁) was determined using the same extract. The recovery factor was 84.8% (RSD = 17.8%) and the DL was 56 ng kg^-1 olive oil. Both determinations were applied in 50 samples of olive oil originated from representative regions of Greece. Results revealed the presence of OTA in 88% of samples tested (n = 44, mean 267 ng kg^-1). Among them, 10 were contaminated with more than 500 ng kg^-1 (median 568 ng kg^-1), 10 with 200-500 ng kg^-1 (median 260 ng kg^-1), 15 with 100-200 ng kg^-1 (median 140 ng kg^-1), nine with DL-100 (median 60 ng kg^-1) and in six samples, OTA was not detectable. Interestingly, most contaminated samples were from Southern Greece. Results of AFB₁ determination showed the presence of aflatoxin B₁ (60 ng kg^-1) in only one olive oil sample also from Southern Greece. The levels of OTA found in Greek olive oil were relatively low as compared with other commodities such as cereals or wine reported in the literature.     

Quantitative analysis of Fusarium mycotoxins in maize using accelerated solvent extraction before liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometry

A method for the simultaneous quantitative determination of deoxynivalenol (DON), fumonisin B1 (FB1) and zearalenone (ZEN) in maize by liquid chromatography-atmospheric pressure chemical ionization tandem mass spectrometry (LC-APCIMS/MS), using stable isotopically labelled and structural analogues internal standards, is described. The procedure involves accelerated solvent extraction followed by two solid-phase clean-up steps on strong anion exchange resin and a Mycosep® column. Typical recoveries were calculated by spiking blank maize at three different concentrations for deoxynivalenol (200, 400 and 1000 μg kg^-1) at 70%, for fumonisin B1 (100, 200 and 1000 μg kg^-1) at 90%, and for zearalenone (50, 100 and 200 μg kg^-1) at 40%. LC-APCIMS/MS analyses were realized in collision-induced dissociation on an ion-trap instrument to provide a high degree of selectivity and sensitivity. Extraction of ions from two transition reactions, monitored by LC-APCIMS/MS for each analyte, enabled a limit of detection for DON, FB1 and ZEN at, respectively, 10, 20 and 3 μg kg^-1, and a limit of quantification at, respectively, 50, 50 and 10 μg kg^-1. The robustness of the method was also evaluated with the analysis of wheat samples.     

Determination of selected mycotoxins in mould cheeses with liquid chromatography coupled to tandem with mass spectrometry

A simple and feasible method is described for analysing nine mycotoxins in cheese matrix. The method involves liquid extraction followed by high performance liquid chromatographic separation and mass spectrometric detection of the analytes, and allows the determination of aflatoxins B1, B2, G1, G2 and M1, ochratoxin A, mycophenolic acid, penicillic acid and roquefortine C simultaneously. Average recoveries of the mycotoxins from spiked samples at concentration levels of 5-200 µg kg^-1 ranged from 96-143%. Within-day relative standard deviations at these concentration levels varied from 2.3-12.1%. The limit of quantification for aflatoxin M1 was 0.6 µg kg^-1 and for the other compounds 5 µg kg^-1. The method developed was applied for analysing these mycotoxins in blue and white mould cheeses purchased from Finnish supermarkets. Roquefortine C was detected in all of the blue mould cheese samples in concentrations of 0.8-12 mg kg^-1. One blue cheese contained also 0.3 mg kg^-1 mycophenolic acid. The other investigated mycotoxins were absent in the samples.     

Occurrence of fumonisins B1 and B2 in asparagus from Shandong province, P.R. China

Thirty samples of asparagus spears were collected from the fields in Shandong province, China, in July 2004, and were analysed for the occurrence of fumonisins B₁ and B₂ (FB₁ and FB₂) by HPLC coupled with electrospray ionization tandem mass spectrometry. Twenty-four samples (80%) contained fumonisins, ranging from 24 to 670 ng g^-1 (average 123 ng g^-1) and 17 to 138 ng g^-1(average 35 ng g^-1) for FB₁ and FB₂, respectively. The total amount of fumonisins (FB₁ and FB₂) in all samples ranged from 47 to 714 ng g^-1 (average 158 ng g^-1) (based on dry weight). This is the first report on the natural occurrence of FB₁ and FB₂ in asparagus spears in China.     

Residue formation of fumonisin B1 in porcine tissues

The residues derived from the uptake of fumonisin B1, a toxic metabolite of Fusarium verticillioides frequently occurring in corn and corn products, were determined in growing pigs. After oral administration of 100 mg FB₁/animal/day for 5-11 days, serum, bile, lung, liver, kidney, brain, spleen, pancreas, heart, muscle, eye, and fat samples were collected immediately and analysed by LC-MS. The highest values were measured in kidney (833 ±1329 μg kg^-1, mean ±SD), liver (231 ±163 μg kg^-1), lung (170 ±311 μg kg^-1) and spleen (854 ±2212 μg kg^-1). Muscle contained 26 ±41 μg kg^-1, while in fat only 2 ±3 μg kg^-1 were traceable. Despite the potential accumulation over extended feeding periods as well as the large variations in the residue formation of FB₁, a carry-over in edible tissues from swine was considered not to be of toxicological relevance.     

Contaminants in organically and conventionally produced winter wheat (Triticum aestivum) in Belgium

A database has been compiled with the levels of important contaminants (mycotoxins, heavy metals and pesticides) measured from 2002 to 2005 in winter wheat (Triticum aestivum) grown in Belgium according to the organic and conventional farming systems. Assuming no further change in contaminant levels during cereal processing and during the preparation of foodstuffs, conservative intakes are estimated for the consumers of cereal-based products such as flour, bread, breakfast cereals, dough and pastry. The results show that for the consumer of organic foodstuffs, estimated daily intakes are 0.56 µg deoxynivalenol (DON), 0.03 µg zearalenone (ZEA), 0.19 µg Cd, 0.28 µg Pb and 0.0006 µg Hg kg^-1 body weight, taking into account the average contaminant levels in unprocessed grains and the average cereal products consumptions in Belgium. For the consumers of conventional foodstuffs, the corresponding estimated daily intakes are 0.99 µg DON, 0.06 µg ZEA, 0.17 µg Cd, 0.12 µg Pb and 0.0007 µg Hg kg^-1 body weight. In addition, it appears that for the consumers of conventional products, intakes of some post-harvest insecticides have to be taken into account (0.11 µg chlorpyriphos-methyl, 0.2 µg dichlorvos and 0.24 µg pirimiphos-methyl kg^-1 bw). When expressed as a percentage of the tolerable/acceptable daily intake (TDI/ADI), it seems that the corresponding estimated (conservative) intakes are the highest for DON (56% for organic and 99% for conventional cereal products), ZEA (16% for organic and 32% for conventional cereal products), and Cd (19% for organic and 17% for conventional cereal products), all other estimated intakes of contaminants (including pesticides) being lower than 10% of the TDI/ADI.     

Lead and cadmium in meat and meat products consumed by the population in Tenerife Island, Spain

The aim of this study was to determine the levels of lead and cadmium in chicken, pork, beef, lamb and turkey samples (both meat and meat products), collected in the island of Tenerife (Spain). Lead and cadmium were measured by graphite furnace atomic absorption spectrometry (GFAAS). Mean concentrations of lead and cadmium were 6.94 and 1.68 µg kg^-1 in chicken meat, 5.00 and 5.49 µg kg^-1 in pork meat, 1.91 and 1.90 µg kg^-1 in beef meat and 1.35 and 1.22 µg kg^-1 in lamb meat samples, respectively. Lead was below the detection limit in turkey samples and mean cadmium concentration was 5.49 µg kg^-1. Mean concentrations of lead and cadmium in chicken meat product samples were 3.16 and 4.15 µg kg^-1, 4.89 and 6.50 µg kg^-1 in pork meat product, 6.72 and 4.76 µg kg^-1 in beef meat product and 9.12 and 5.98 µg kg^-1 in turkey meat product samples, respectively. The percentage contribution of the two considered metals to provisional tolerable weekly intake (PTWI) was calculated for meat and meat products. Statistically significant differences were found for lead content in meats between the chicken and pork groups and the turkey and beef groups, whereas for cadmium concentrations in meats, significant differences were observed between the turkey and chicken, beef and lamb groups. In meat products, no clear differences were observed for lead and cadmium between the various groups.     

Analysis of heat-processed corn foods for fumonisins and bound fumonisins

Thirty retail samples of heat-processed corn foods, i.e. corn flakes, corn-based breakfast cereals, tortilla chips and corn chips, were analysed for fumonisins — fumonisin B₁ (FB₁), fumonisin B₂ (FB₂) and hydrolysed FB₁ (HFB₁) — as well as for protein- and total-bound FB₁. Bound (hidden) fumonisins cannot be detected by conventional analysis. Improved methods for the determination of bound FB₁ were developed. The protein-bound FB₁ was extracted with 1% sodium dodecylsulfate (SDS) solution. The SDS, which interfered with high-performance liquid chromatography (HPLC) analysis, was then separated from protein-bound FB₁ by complexing with methylene blue followed by solvent extraction and hydrolysis with 2 N KOH. To measure total-bound FB₁, the sample itself was hydrolysed with KOH. In both cases, clean-up was accomplished on an OASIS polymeric solid-phase extraction column and the bound fumonisins were determined by HPLC measurement of HFB₁. Fourteen of 15 samples of corn flakes and other corn-based breakfast cereals analysed contained detectable levels of FB₁ with a mean in positive samples of 67 ng g^-1 (13-237 ng g^-1). Two samples also had detectable levels of FB₂ (21-23 ng g^-1). Bound FB₁ was found in all samples; the mean protein-bound FB₁ measured was 58 ng g^-1 (22-176 ng g^-1) and the mean total-bound FB₁ measured was 106 ng g^-1 (28-418 ng g^-1), reported as FB₁ equivalents after correction for recoveries of HFB₁. There was an average of about 1.3 times more FB₁ in the bound form compared with extractable FB₁, and this was about twice as much as protein-bound FB_1. Seven of the 15 samples of alkali-processed corn-based foods, such as tortilla chips and corn chips, contained FB₁ and three contained HFB₁ with means in measurable positive samples of 78 (48-134) and 29 (13-47) ng g^-1, respectively. Five of these alkali-processed corn foods contained bound FB₁; the mean measurable protein-bound FB₁ was 42 ng g^-1 (39-46 ng g^-1) and the mean measurable total-bound FB₁ was 100 ng g^-1(54-209 ng g^-1). HFB₁ derived from bound FB₁ in selected samples was confirmed by HPLC with mass spectrometry (MS).