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Rapid and sensitive determination of zearalenone in cereals by high-performance liquid chromatography with fluorescence detection.

Zearalenone, an estrogenic mycotoxin of Fusarium species, in cereals can be extracted with acetonitrile-water (3:1), purified on a Florisil column, resolved by high-performance liquid chromatography (HPLC) with a Nucleosil 50-10 column using 90% water-saturated chloroform-cyclohexane-acetonitrile-ethanol (50:15:2:1) and quantitated by fluorescence measurement. This method is rapid, simple and reproducible, and detects zearalenone in wheat, barley, corn and other cereals with picogram sensitivity. A combination of this HPLC method with a gas-liquid chromatographic method for trichothecenes may be applied to the simultaneous detection of Fusarium mycotoxins (zearalenone, nivalenol and deoxynivalenol) in cereals.

Chromatography, High Pressure Liquid↗

Selective identification by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of different types of gluten in foods made with cereal mixtures.

The gluten toxic fractions responsible for the mucosal damage in coeliac disease (CD), so-called gliadins, hordeins, secalins and avenins from a large number (30-40) of wheat, barley, rye and oats cultivars respectively, have been mass analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Gliadin, secalin and avenin characteristic mass profiles are nearly identical amongst distinct cultivars from the corresponding cereal, while hordeins profiles show more variability depending on the particular barley cultivar. On the basis of these four distinguishable characteristic mass patterns spreading within the 20,000-40,000 Da range, MALDI-TOF-MS has permitted the direct and simultaneous visualization of gliadins, hordeins, secalins and avenins in foods elaborated with cereal mixtures of wheat, barley, rye and oats. This capacity has been demonstrated by mass analyzing foods made with these four cereals in varying ratios. Thus MALDI-TOF-MS can be preliminarily established as a unique system with the ability to discriminate the specific type of gluten toxic fractions present in food samples.

Avena↗

Rapid determination of glyphosate in cereal samples by means of pre-column derivatisation with 9-fluorenylmethyl chloroformate and coupled-column liquid chromatography with fluorescence detection.

A rapid procedure for the determination of glyphosate in cereals has been developed. Convenient sample pretreatment is carried out by (i) a overnight standing extraction of 1.0 g homogenized sample with 20 ml of water, (ii) centrifugation of the samples, (iii) a passing of 2.5 ml of the clear layer through a 100 mg C18 solid-phase extraction cartridge and (iv) collection of the last 1.5 ml of the eluent into a calibrated tube. For the instrumental analysis, the efficient approach developed earlier for environmental water samples [J.V. Sancho, F. Hernández, F.J. LUpez, E.A. Hogendoorn, E. Dijkman, P. van Zoonen, J. Chromatogr. A, 737 (1996) 75] was successfully adopted for the determination of glyphosate in the obtained cereal extracts. The procedure includes a 15 min derivatisation step of the analyte with 9-fluorenylmethyl chloroformate and a 16 times dilution step prior to instrumental analysis employing coupled-column LC with fluorescence detection. The developed procedure has a sample throughput of more than 25 samples per day and a limit of quantification of 0.5 mg/kg. The method was validated by analyzing freshly spiked cereal samples and samples with aged residues at levels between 1.0 and 10 mg/kg. The overall recovery of the freshly spiked samples was 86% (n = 10) with a repeatability of 6.5% and a reproducibility of 9.5%. For samples with aged residues recoveries performed at different time intervals (range 80-150 days) did not differ significantly; the overall recovery (n = 10) was 74% with a repeatability and reproducibility of 14 and 20%, respectively.

Chromatography, High Pressure Liquid↗

Cereal-based oral rehydration therapy. I. Clinical studies.

Studies of "improved" oral rehydration solutions, in which glucose polymers (starch) derived from rice or other cereals were added to improve cotransport of sodium and to promote sodium and water absorption, have been reported. These solutions were administered to decrease diarrhea volume and duration, reduce vomiting, and replace volume loss in stools. In clinical trials of children and adults with high-output diarrhea, such as in cases of cholera, the use of cereal-based oral rehydration solutions (ORS) compared with glucose-based ORS produced significant (20% to 53%) reductions in stool volumes. In one study the duration of diarrhea was shortened by 30%. In noncholera diarrhea in children, cereal-based ORS was as effective as glucose-based ORS. Although the amino acid transport systems were intact in patients with cholera, the addition of glycine to glucose-based or rice-based ORS did not reduce stool volume or duration of diarrhea. The exception was alanine, which reduced stool output and ORS requirements. More research is needed to determine the optimal mix of starch, amino acids, oligopeptides, and proteins that would utilize the absorptive active transport systems maximally to reduce fluid losses and duration of diarrhea.

Administration, Oral↗

Cereal-based oral rehydration therapy. II. Strategic issues for its implementation in national diarrheal disease control programs.

Field studies in Bangladesh demonstrated that after proper training, village mothers were able to prepare and use rice-based, salt-enriched oral rehydration solutions containing safe concentrations of sodium, and were capable of achieving significantly fewer treatment failures and a reduction in the duration of diarrhea than with glucose-based oral rehydration solutions (ORS). An additional longitudinal study showed that improved growth and weight gain occurred with the consistent use of ORS; the effect was greater when rice-based ORS were used. In addition, the following possible limitations and benefits of cereal-based oral rehydration therapy, which are relevant to the strategies for its implementation in national diarrheal disease control programs, are discussed: safety, osmolarity, hypernatremia, spoiling, effectiveness, rehydration ability, reduction in diarrhea volume and duration, nutritional effects, effect on food intake, acceptance and usage by care givers, training of health workers, self-reliance of families, effect on other child survival activities, costs, potential problems in changing to cereal-based ORS, and the role of industrial production in packaged cereal-based ORS.

Bangladesh↗

Comparison of genes among cereals.

Comparison of partially sequenced cereal genomes suggests a mosaic structure consisting of recombinationally active gene-rich islands that are separated by blocks of high-copy DNA. Annotation of the whole rice genome suggests that most, but not all, cereal genes are present within the rice genome and that the high number of reported genes in this genome is probably due to duplications. Within the cereals, macrocolinearity is conserved but, at the level of individual genes, microcolinearity is frequently disrupted. Preliminary evidence from limited comparative analysis of sequenced orthologous genomic segments suggests that local gene amplification and translocation within a plant genome may be linked in some cases.

Gene Amplification↗

Sterol oxidation in infant milk formulas and milk cereals.

7-Ketocholesterol and 7-ketositosterol were chosen as reliable markers of the oxidation of cholesterol and phytosterols in infant milk formulas and infant milk cereals. A reversed-phase HPLC method was developed to measure them simultaneously in infant formulas. This method was then tested on a wide range of infant milk formulas and milk cereals on sale in Italy whose lipid composition is representative of the most common commercial formulas. The analytical results revealed no significant differences in the extent of oxidation of cholesterol and sitosterol. As the level of 7-ketocholesterol often followed the cholesterol level, a cholesterol content similar to that of human milk produced amounts of cholesterol oxides with possible negative effects on infant health. In contrast, the low cholesterol content of milk cereals never produced amounts of cholesterol oxides high enough to cause concern. The contents of phytosterols and hence their oxides were always low.

Animals↗

Enteropathogenic bacteria in faecal swabs of young children fed on lactic acid-fermented cereal gruels.

The influence of consumption of a lactic acid-fermented cereal gruel togwa with pH < or = 4 on the presence of faecal enteric bacteria such as campylobacter, enterohaemorrhagic Escherichia coli (EHEC:O157), enterotoxigenic Escherichia coli (ETEC), salmonella and shigella was evaluated. Under 5 years old healthy children listed in an ascending order of age were alternatively assigned and given either a lactic-acid fermented cereal gruel togwa (test diet) or an unfermented cereal gruel uji (control diet) once a day for 13 consecutive days. The presence of the enteropathogens was examined in rectal swabs collected from the children at baseline (before feeding session started), on days 7 and 13, and additionally 14 days (follow-up day) after the feeding session had stopped. The swabs were cultured on to different optimal media for respective enteropathogen and confirmed by standard microbiological and serological methods. Campylobacter spp. dominated among the enteropathogens (62% out of total) followed by Salmonella spp., ETEC and Shigella spp. Children with isolated enteropathogens in the togwa group was significantly reduced (P < 0.001) from 27.6% at baseline to 7.8, 8.2 and 12.7% on days 7, 13 and follow-up day, respectively. The effect was more pronounced in those children taking togwa > 6 times during the study period. In the control group, there was a slight decrease from 16.7% at baseline to 11.4% on day 7 and 8.1% on day 13. On the follow-up day, enteropathogens were found in 22.6% of the children, which was significantly higher than in those children taking togwa > 6 times. We conclude, that regular consumption of togwa with pH < or = 4, once a day, three times a week may help to control intestinal colonization with potential diarrhoea-causing pathogens in young children.

Beverages↗

Problems associated with measuring phytate in infant cereals.

The inositol hexaphosphate (IP6) content of commercially available dried infant cereals was measured by ion pair high-pressure liquid chromatography (ion pair HPLC) and ion exchange high-pressure liquid chromatography (ion exchange HPLC). Large differences between methods were apparent: ion pair HPLC gave values 14 to 190-fold lower than the values from ion exchange HPLC. Poor recoveries of added IP6 (25 to 60%) by ion pair HPLC suggested that some component of the infant cereal was responsible for the difference. Further experimentation suggested that an excess of minerals (approximately 11 mg/g calcium and 0.3 mg/g iron) in these samples sequestered the endogenously low phytate content. This problem may be unique to samples with low IP6 and high mineral content as wheat bran was not problematic. These results suggest that ion exchange HPLC is the method of choice for measuring inositol phosphates in infant cereals.

Artifacts↗

Rapid prediction of gross energy and utilizable energy in cereal food products using near-infrared reflectance spectroscopy.

Near-infrared (NIR) spectroscopy has been used in foods for the rapid assessment of several macronutrients; however, little is known about its potential for the evaluation of the utilizable energy of foods. Using NIR reflectance spectra (1104-2494 nm) of ground cereal products (n = 127) and values for energy measured by bomb calorimetry, chemometric models were developed for the prediction of gross energy and available energy of diverse cereal food products. Standard errors of cross-validation for NIR prediction of gross energy (range = 4.05-5.49 kcal/g), energy of samples after adjustment for unutilized protein (range = 3.99-5.38 kcal/g), and energy of samples after adjustment for unutilized protein and insoluble dietary fiber (range = 2.42-5.35 kcal/g) were 0.053, 0.053, and 0.088 kcal/g, respectively, with multiple coefficients of determination of 0.96. Use of the models on independent validation samples (n = 58) gave energy values within the accuracy required for U.S. nutrition labeling legislation. NIR spectroscopy, thus, provides a rapid and accurate method for predicting the energy of diverse cereal foods.

Calibration↗

Normal phase high-performance liquid chromatography method for the determination of tocopherols and tocotrienols in cereals.

The eight vitamers of vitamin E (alpha-, beta-, gamma-, and delta-tocopherols and -tocotrienols) have different antioxidant and biological activities and have different distributions in foods. Some cereals, especially oat, rye, and barley, are good sources of tocotrienols. A fast procedure for the determination of tocopherols and tocotrienols (tocols) in cereal foods was developed. It involves sample saponification and extraction followed by normal phase high-performance liquid chromatography (HPLC). The results have been compared with those found by direct extraction without saponification. The method is sensitive and selective enough to be tested on a wide variety of cereal samples. The highest tocol levels were found in soft wheat and barley ( approximately 75 mg/kg of dry weight). beta-Tocotrienol is the main vitamer found in hulled and dehulled wheats (from 33 to 43 mg/kg of dry weight), gamma-tocopherol predominates in maize (45 mg/kg of dry weight) ), and alpha-tocotrienol predominates in oat and barley (56 and 40 mg/kg of dry weight, respectively).

Chromatography, High Pressure Liquid↗

Near-infrared reflectance model for the rapid prediction of total fat in cereal foods.

AOAC method 996.01, used in cereal foods to determine total fat as defined by the U.S. Nutrition Labeling and Education Act (NLEA), is laborious and time-consuming and utilizes hazardous chemicals. Near-infrared (NIR) reflectance spectroscopy, a rapid and environmentally benign technique, was investigated as a potential method for the prediction of total fat using AOAC method 996.01 as the reference method. Near-infrared reflectance spectra (1104-2494 nm) of ground cereal products (n = 72) were obtained using a dispersive spectrometer, and total fat was determined according to AOAC method 996.01. Using multivariate analysis, a modified partial least-squares model was developed for total fat prediction. The model had a SECV of 1.12% (range = 0.5-43.2%) and a multiple coefficient of determination of 0.99. The model was tested with independent validation samples (n = 36); all samples were predicted within NLEA accuracy guidelines. The results indicate that NIR reflectance spectroscopy is an accurate means of determining the total fat content of diverse cereal products for nutrition labeling.

Analysis of Variance↗

Methodological aspects of measuring phytase activity and phytate phosphorus content in selected cereal grains and digesta and feces of pigs.

This study was to examine the time course of sample-specific linearity of intrinsic phytase hydrolysis in major cereal grains and in ileal digesta and fecal samples and to determine the time course of the microbial phytase-catalyzed hydrolysis of various sources of phytate for estimating phytate phosphorus (P) content. The intrinsic phytase activity in barley, corn, oat, and wheat samples was measured over multiple time points from 0 to 120 min at 1.5 mmol.L(-1) of sodium phytate at pH 5.5 and 37 degrees C. Time courses of hydrolysis of purified phytate and phytate associated with the cereal grain samples and the pig digesta and fecal samples were examined with the Natuphos microbial phytase over multiple time points from 0 to 48 h of incubation. The intrinsic phytase hydrolysis was linear (P < 0.05) for up to 120 min for the barley, corn, and wheat samples, whereas in the oat sample the hydrolysis was linear (P < 0.05) for only up to 30 min of incubation. The intrinsic phytase activities (phytase unit: mumol.kg(-1) of dry matter.min(-1)) for the barley, corn, and wheat samples were estimated to be 693, 86, and 1189 by linear regression analysis. Intrinsic phytase activity (412 phytase units) for the oat sample based on a 30-min incubation was considerably higher than the value (103 phytase units) determined from the 120-min incubation for the same oat sample. There were quadratic with plateau relationships (P < 0.05) between the hydrolytic release of inorganic P from various sources of phytate and the incubation time. The minimal incubation times required for the complete hydrolysis of phytate were estimated to be 4, 3, and 11 h for the purified phytate, the cereal grain samples, and the pig digesta and feces, respectively. It was concluded that multiple time point experiments need to be conducted to determine valid intrinsic phytase activity and phytate P content in samples through intrinsic and microbial phytase hydrolysis incubations.

6-Phytase↗

Literature data may underestimate the actual antioxidant capacity of cereals.

Several recent articles have reported a significant antioxidant capacity of cereal products, determined in methanolic and ethanolic extracts. The aim of this work was to conduct an assessment of the antioxidant capacity of cereals using both chemical and in vitro digestive enzymatic extraction of antioxidants. Ferric reducing power (FRAP) and free radical scavenging capacity (DPPH) methods were used to determine the antioxidant capacity in wheat flour, bread, raw and boiled rice, wheat bran, and oat bran. The most efficient antioxidant extraction was achieved by using successively acidic methanol/water (50:50 v/v, pH 2) and acetone/water (70:30 v/v). The antioxidant capacity in these extracts ranged from 1.1 to 4.4 micromol Trolox/g dw. A significant amount of hydrolyzable phenolics with a high antioxidant capacity (from 5 to 108 micromol Trolox/g dw) was found in the residues of this aqueous-organic extraction. The antioxidant capacities of these nonextractable polyphenols are usually ignored in the literature, although they may have an antioxidant role in the gastrointestinal tract, especially after colonic fermentation, and may be fermentated to active metabolites. On the other hand, in vitro digestive enzymatic extracts obtained by enzymatic treatments that mimic conditions in the gastrointestinal tract showed that the amount of antioxidants released by the cereal matrix into the human intestine may be higher than the one that can be expected from measurements in the usual aqueous-organic extracts.

Antioxidants↗

Browning indicators in model systems and baby cereals.

In many countries, baby cereals are the first solid food given to 3-4-month-old babies after weaning and to infants aged 6-12 months. Various simple technologies are traditionally used in the processing of cereals, including toasting, hydrolysis, and drying. In this study color and hydroxymethylfurfural (HMF) assays have been used to evaluate heat effects induced during the manufacture of these foods. The baby cereals analyzed were wheat, rice, oat, and four mixtures of flours. No HMF was detected in the raw flours. Toasting the flours increased HMF values by between 1.1 and 4.53 mg/kg and color (DeltaE) values by 2.51-9.34. The drying step increased HMF values by between 1.14 and 19.60 mg/kg. High values of HMF coincided with the addition of ingredients containing HMF. Color and HMF contents in sugar-amino acid model systems were much higher than in sugar systems at temperatures >100 degrees C and low moisture content.

Chromatography, High Pressure Liquid↗

Development of a sensitive ELISA for the determination of fumonisin B(1) in cereals.

Monoclonal fumonisin B(1) antibodies with high titer were raised by using FB(1)-glutaraldehyde-keyhole limpet hemocyanin immunogen prepared by a short cross-linker reagent (glutaraldehyde). Mean cross-reactivities of the selected monoclonal antibody for FB(1), FB(2), and FB(3) were 100, 91.8, and 209%, respectively; no reactivity was found with hydrolyzed fumonisin. A direct, competitive enzyme-linked immunosorbent assay for the quantitative determination of FB(1) in cereals has been developed with this antibody. Fifty percent acetonitrile-based solvent with some additives was used for extraction of cereals, and the diluted extracts were used without cleanup in the test. The mean within-assay and interassay coefficients of variation for the standard curve were <10%. The measuring range of this test is 10-500 ng/g, with a detection limit of 7.6 ng/g FB(1). The toxin recovery from cereals infected with 50-200 ng/g of FB(1) varied between 61 and 84%. According to the comparable results of naturally infected maize samples, this test proved to be suitable for the rapid screening of food and feed samples for the presence of FBs.

Carboxylic Acids↗

A survey of Fusarium toxins in cereal-based foods marketed in an area of southwest Germany.

A total of 237 commercially available samples of cereal-based foods including bread and related products, noodles, breakfast cereals, baby and infant foods, rice and other foods were randomly collected in southwest Germany during the first six months of 1998. The trichothecenes deoxynivalenol (DON), 3- and 15-acetyl-deoxynivalenol (3-,15-ADON), nivalenol (NIV), fusarenon-X (FUS-X), T-2 toxin (T-2) and HT-2 toxin (HT-2) were determined by gas chromatography/mass spectrometry following clean-up by a two stage solid-phase extraction. Detection limits ranged between 2 and 12 micrograms/kg. Based on all samples, the incidence of DON, HT-2, T-2, 3-ADON, 15-ADON, and NIV was at 71, 18, 4, 4, 4 and 2%, respectively; the average contents in positive samples were at 103, 16, 14, 17, 24 and 109 micrograms/kg, respectively. Fus-X was not detected in any sample. A lower (P < 0.05) DON content was found in baby and infant foods as well as in cookies and cakes compared to bread. Overall, based on the incidence and level of all six toxins, the degree of contamination was lowest in baby and infant foods. Foods produced from either white or whole grain flour did not differ (P > 0.05) with regard to the incidence and level of DON. In foods produced from cereals of organic production both the incidence and median content of DON was lower compared to conventional production. Zearalenone, alpha- and beta-zearalenol were determined by high performance liquid chromatography in 20 selected samples, mostly baby and infant foods. These toxins were not present in excess of the detection limit in any sample.

Bread↗

Acceptability of supplementary foods based on popped cereals and legumes suitable for rural mothers and children.

Eight types of supplementary foods based on popped cereals (wheat, ragi, bajra and sorghum) blended with legumes (soy and bengal gram) and fortified with essential vitamins and minerals were developed on a pilot plant scale. Four of the supplements were prepared with cereals, soy flour (SF) and bengal gram (BG) dhal and the other four were prepared with combinations of cereals and SF. These blends were mixed with jaggery (obtained by boiling juice out of sugarcane) syrup and pressed into compact form. One hundred gram portions of these foods provided 370+/-20 kilocalories and 11+/-1 g protein. Moisture, crude protein, total carbohydrates, total lipids, ash, dietary fiber and energy contents, of all the developed supplements were within the ranges prescribed by the Indian Standards Institute for processed weaning foods and could satisfactorily meet one-third of the Recommended Dietary Allowance (RDA) of these nutrients per day for preschool children. Organoleptic evaluation and feeding trials revealed that the foods were well accepted by rural mothers and children.

Child, Preschool↗