Gas chromatographic method for ethylene dibromide in grains and grain-based products: collaborative study

Nine laboratories analyzed samples of whole grain, intermediate, and ready-to-eat products for ethylene dibromide (EDB) residues. Supplied samples of wheat, rice, and flour contained both fortified and incurred EDB; corn bread mix, baby cereal, and bread contained only fortified EDB. The whole grains and intermediates were analyzed by the same basic procedural steps as in the official method for multifumigants: They were extracted by soaking in acetone-water (5 + 1). The baby cereal and bread were analyzed by a modification of the Rains and Holder hexane co-distillation procedure. EDB was determined by electron capture gas chromatography operated with an SP-1000 column. All products contained 3 different levels of EDB and were analyzed as blind duplicates. Overall mean recoveries ranged from 85.2% for 69.6 ppb to 105.0% for 4.35 ppb, both in baby cereal. Interlaboratory relative standard deviations ranged from 5.7% for 869 ppb in wheat to 20.2% for 69.6 ppb in baby cereal, both fortified. Mean levels of incurred EDB in wheat, rice, and flour were 926.7, 982.0, and 49.9 ppb, respectively; corresponding relative standard deviations were 9.9, 7.7, and 13.1%. The method was adopted official first action.     

Limited survey of deoxynivalenol in wheat and corn in the United States

A limited survey was conducted over a 2-year period to determine the incidence and levels of deoxynivalenol (DON) in corn and wheat grown in selected areas of the United States. Samples of corn (198) and wheat (247) were collected and analyzed by a gas chromatographic method. Sixty-six percent of the corn samples collected in 1984 and 30% of the corn samples collected in 1985 contained DON. The average concentration of DON in corn, by state, ranged from 0.11 to 1.20 micrograms/g; the maximum concentration was 2.47 micrograms/g. Only 2 of the 247 samples of wheat contained DON at a concentration greater than 2 micrograms/g, which is the level of concern suggested by the Food and Drug Administration for wheat entering the milling process for human consumption.     

Comparative study of methods for the extraction of eleven organophosphorus pesticide residues in rice.

Several extraction methods are compared for the simultaneous analysis of organophosphorus pesticides in unpolished rice. Four stationary phases were used for the subsequent gas-liquid chromatographic (GLC) determination of the selected pesticides. Using 3 different GLC columns, 11 pesticides were completely separated and identified. The efficiency of the cleanup and the sensitivity of the analytical method were evaluated by using powdered unpolished rice samples fortified with the pesticides and also wheat and dried bean samples. Average recoveries ranged from 74.7% for disulfoton to 97.4% for malathion in unpolished rice and from 68.1% for disulfoton to 108.3% for malathion in other crops. The method described is applicable to the analysis of selected organophosphorus pesticide residues in unpolished rice, wheat, buckwheat, and dried beans.     

Phytate, calcium, iron, and zinc contents and their molar ratios in foods commonly consumed in China

A total of 60 food samples commonly consumed in China were analyzed for phytate using the anion-exchange method and for calcium, iron, and zinc using atomic absorption spectrophotometry. The foods analyzed included those based on cereal grains and soybean. Phytate contents expressed on a wet weight basis ranged from 0 for foods made from starches to 1878 mg/100 g for dried stick-shaped soybean milk film. The calcium contents were between 2.08 mg/100 g for ground corn and 760.67 mg/100 g for diced fried soybean curd. The lowest values of iron and zinc were 0.04 mg/100 g for Panjin pearl rice cooked with discarding extra water and 0.08 mg/100 g for potato and bean starches, while the highest values of iron and zinc were observed in dried stick-shaped soybean milk film. Although many foods were relatively rich in calcium, zinc, and iron, many also contained a higher level of phytate. Of the 60 food samples, 34 foods had a phytate/calcium molar ratio >0.24, 53 foods had a phytate/iron molar ratio >1, 31 foods had a phytate/zinc molar ratio >15, and only 7 foods had a phytate x calcium/zinc >200. Phytate in foods impair the bioavailability of calcium, iron, and zinc, which to some extent depends upon food processing and cooking methods.     

Study of Viscoelastic Properties of Wheat Kernels Using Compression Load Method

The method to measure hardness and other viscoelastic properties of intact wheat kernels is presented. Wheat with 9.3% moisture showed high elastic behavior compared with wheat tempered at 22.5% moisture that showed a plastic behavior. Load‐deformation curves showed that bread wheat behaves as a more plastic material than durum wheat, which is a more elastic material. Yield point of all the wheat samples was ≈18.5 N, independent of wheat type and moisture content. The height of the wheat kernel increased linearly, and the compression area increased exponentially, with increasing moisture content. The modulus of elasticity of wheat ranged from 99.2 MPa for 22.5% moisture content to 394.8 MPa for 9.3% moisture content. Young's modulus range for soft wheat such as Salamanca, Saturno, and Cortazar cultivars was 232.2–308.5 MPa, as compared with Rayón bread wheat at 321.5 MPa and the Altar, Sofía, and Rafi cultivars of durum wheat that had elastic moduli of 438.7–485.8 MPa. The compression force and final stress decreased from 69.9 N and 40.1 MPa in soft wheat to 90 N and 78.9 MPa in durum, respectively. Total work range was 14.7 MPa/sec in soft wheat to 19.7 MPa/sec for durum wheat and, as expected, was higher in the durum and bread wheat than in soft wheat. The plastic part ranged from 2.4 MPa/sec in soft wheat to 0.6 MPa/sec in durum wheat.     

Determination of total dietary fiber in foods and food products: collaborative study

A collaborative study was conducted to determine the total dietary fiber (TDF) content of food and food products, using a combination of enzymatic and gravimetric procedures. The method was basically the same as published earlier (J. Assoc. Off. Anal. Chem. (1984) 67, 1044-1052), with changes in the concentration of alcohol and buffers, time of incubation, sample preparation, and some explanatory notes, all with the intent of decreasing the coefficient of variation (CV) of the method. Duplicate blind samples of soy isolate, white wheat flour, rye bread, potatoes, rice, wheat bran, oats, corn bran, and whole wheat flour were analyzed by 9 collaborators. TDF was calculated as the weight of the residue minus the weight of protein and ash. CV values of the data from all laboratories for 7 of the samples ranged from 1.56 to 9.80%. The rice and soy isolate samples had CV values of 53.71% and 66.25%, respectively; however, each sample contained only about 1% TDF. The enzymatic-gravimetric method for determining TDF has been adopted official first action.     

Analysis of bisphenol A and alkylphenols in cereals by automated on-line solid-phase extraction and liquid chromatography tandem mass spectrometry

An on-line solid-phase extraction (SPE) following a liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) method was established for the simultaneous analysis of bisphenol A (BPA), nonylphenol (NP), and octylphenol (OP) in cereals (including rice, maize, and wheat). The target compounds were extracted by acetonitrile, purified by an automated on-line SPE cartridge, and analyzed by LC-MS/MS under the negative-ion mode. Mean recoveries fortified at three concentration levels ranged from 81.6 to 115.7%, and the coefficient of variation ranged from 4.6 to 19.9% (n = 6). The limits of quantification (LOQs) of the method were 0.5, 0.5, and 0.25 μg/kg for BPA, NP, and OP, respectively, in both rice and maize, while the LOQs in wheat were 0.5, 1.25, and 0.5 μg/kg for BPA, NP, and OP, respectively. This method was applied in the analysis of rice, maize, and wheat from a local market. As a result, NP occurred in all cereal samples at the concentration range of 9.4-1683.6 μg/kg and BPA was detected in a few samples.     

Use of electrochemical biosensor and gas chromatography for determination of dichlorvos in wheat

Two methods for the determination of dichlorvos in durum wheat by electrochemical assay and gas chromatography, respectively, have been developed. Dichlorvos, an organophosphorus anticholinesterase pesticide, was extracted from wheat with hexane, and the filtered extract was directly analyzed by gas chromatography with nitrogen-phosphorus flame detection (NPD). Recoveries of dichlorvos from milled wheat spiked at 0.25-1.5 microg/g ranged from 96.5 to 100.9%, and the limit of detection was 0.02 microg/g. The electrochemical assay was based on the detection of choline, the acetylcholinesterase product, via a choline oxidase biosensor. An aliquot of the filtered hexane extract was partitioned with phosphate buffer solution, and the organic layer was evaporated prior to electrochemical analysis. A limit of detection of 0.05 microg/g of dichlorvos was obtained with mean recoveries of 97-103% at spiking levels of 0.25-1.5 microg/g. A good correlation (r = 0.9919) was found between the results obtained with the electrochemical and those obtained with the gas chromatographic methods. The electrochemical method was peer-validated in two laboratories that analyzed 10 blind samples (5 duplicates), including a blank and 4 spiked samples with dichlorvos at levels of 0.25, 0.60, 1.00, and 1.50 microg/g. Within-laboratory repeatability (RSDr) and between-laboratory reproducibility (RSDR) ranged from 5.5 to 7.8% and from 9.9 to 17.6%, respectively.     

Occurrence of zearalenone-4-beta-D-glucopyranoside in wheat

An LC-MS method was developed for the analysis of zearalenone-4-beta-D-glucopyranoside and zearalenone in wheat (Triticum aestivum). The limit of determination for zearalenone-4-beta-D-glucopyranoside and zearalenone was 10 microg/kg. The recovery rates were calculated to be 69% and 89% at a concentration of 100 microg/kg for zearalenone-4-beta-D-glucopyranoside and zearalenone, respectively. Twenty-four Bavarian wheat samples from a 1999 harvest were analyzed. Zearalenone was present in 22 of 24 field samples, the levels ranged from 11 to 860 microg/kg. Zearalenone-4-beta-D-glucopyranoside was found in 10 of the zearalenone positive samples (42%) at levels ranging from 17 to 104 microg/kg. The amounts of zearalenone-4-beta-D-glucopyranoside were correlated to those of zearalenone (r2 = 0.86, b = 0.10). After gastrointestinal hydrolyzation, zearalenone-4-beta-glucopyranoside might be implicated in the development of a zearalenone-syndrome. Therefore, more attention should be focused on conjugated mycotoxins in food and feed.     

小麦籽粒铅污染来源的同位素解析研究

以中国西北地区某工业区为研究区域,采集大气降尘、耕层土壤样品(0～20 cm),及对应农田位置的小麦籽粒样品,用电感耦合等离子质谱(ICP-MS)测定铅元素质量比及铅同位素比率值。运用普通块克里金法(Ordinary Kriging)预测大气铅沉降通量分布、耕层土壤和小麦籽粒铅质量比空间分布;结合二元混合模型计算各污染源对小麦籽粒铅质量比的贡献率。结果表明:1)耕层土壤铅质量比范围为21.8～40.0 mg/kg,平均值为27.1 mg/kg,高于当地土壤背景值(21.4 mg/kg),说明耕层土壤受到一定程度的污染。2)研究区域小麦籽粒铅质量比范围为0.269～0.768 mg/kg,平均值为0.430 mg/kg,超标率达100%。3)大气降尘和耕层土壤对小麦籽粒铅质量比的贡献率分别为90%～99%和1%～10%。通过研究耕层土壤和小麦铅污染的范围及程度,探讨小麦籽粒铅污染的来源,解析各污染源的相对贡献率,可为制定控制食品中重金属污染,保障食品安全措施提供依据。     

An analytical method for the simultaneous determination of butachlor and benoxacor in wheat and soil

Butachlor is a chloroacetanilide herbicide successfully employed in weeding some important crops, and benoxacor is a safening compound able to induce the enzymatic mechanism of chloroacetanilide detoxification in plants. A practical method for a simultaneous detection of butachlor and benoxacor residues in wheat and in soil is described. The procedure can be performed by GC and HPLC. They were extracted with methanol and cleaned up by solid phase extraction (SPE). The analytes were satisfactorily separated via both GC and HPLC techniques, and no interferences were observed coming from plant or soil matrixes or reagents. The limit of quantitation was found to be 5.0 ng by GC and 20.0 ng by HPLC for butachlor and 2.5 ng by GC and 15.0 ng by HPLC for benoxacor. Butachlor recovery tests ranged from 85.4% to 91.7% in wheat shoots and 84.0% to 93.2% in soil; benoxacor recovery tests ranged from 86.5% to 90.8% in wheat shoots and 85.7% to 90.7% in soil. The reproducibility and the accuracy make this method a selective and sensitive tool for routine analyses.     

In vitro assay for protein digestibility: interlaboratory study

True protein digestibilities of 17 protein sources were estimated by 6 laboratories using an in vitro, 3-enzyme digestion system in a pH stat. Samples from animal, vegetable, and mixed food sources were freeze-dried (if not already dried), ground, mixed, and shipped to each collaborator along with a sodium caseinate standard and trypsin, chymotrypsin, and peptidase. The uptake of titrant during enzymatic digestion was used to calculate estimates of digestibility. Digestibilities ranged from 100% for casein to 89.9% for whole wheat cereal. Mean relative standard deviations for repeatability were 1.4% for rolled oats and less than 1% for the remaining 16 samples. Mean relative standard deviations for reproducibility ranged from 5.0 to 0.8%; values were less than 2.5% for 13 of the 17 samples.     

Application of a modified Haug and Lantzsch method for the rapid and accurate photometrical phytate determination in soybean, wheat, and maize meals

A modified version of the Haug and Lantzsch method for rapid photometrical phytate determination was applied for the analysis of phytate in soybean, wheat, and maize meals. In comparison to the original protocol, the amount of the toxic reagent thioglycolic acid is reduced substantially to minimize potential health risks for laboratory personnel. Different extraction conditions for soybean meal were tested, and boiling for at least 30 min was found to be necessary to remove an interfering compound in soybean meal extracts. Phytate contents determined according to the modified Haug and Lantzsch method did not differ from those obtained by measuring total precipitated phosphorus or by sensitive and specific high-performance ion chromatography. Applicability and accuracy of the modified method for phytate analysis in major feed substrates, including soy-based textured vegetable protein, were demonstrated.     

Residue analysis of glyphosate and its principal metabolite in certain cereals, oilseeds, and pulses by liquid chromatography and postcolumn fluorescence detection.

A postcolumn liquid chromatographic method to determine the extractable residues of glyphosate (GLYPH) and its principal metabolite, (aminomethyl)phosphonic acid (AMPA), in various cereals and beans is described. The finely ground sample is extracted with a mixture of chloroform and water, and the resulting aqueous layer is passed through a cation exchange column. The eluate is adjusted to pH 7-10 and passed through an anion exchange column. The second column is eluted with 0.3M HCl solution and the resulting acidic eluate is analyzed with liquid chromatography coupled with postcolumn fluorescence detection. The mean recoveries for GLYPH in barley, canola, dry pea, flax, soybean, wheat, and white bean ranged from 90.0 to 98.1%, with coefficients of variation (CV) from 2.9 to 10.0% and limits of detection (LOD) from 0.07 to 0.14 ppm. Similarly, mean recoveries for AMPA in the same crops ranged from 87.4 to 98.9%, with CV from 4.6 to 7.7 and LOD from 0.05 to 0.12 ppm. Using this method, an analyst can routinely analyze 6 samples per 1.5 days. The advantages of this procedure are discussed.     

Effects of Sampling and Wheat Grade on Precision and Accuracy of Kernel Features Determined by Digital Image Analysis

The effect of sampling on the precision and accuracy of digital image analysis of different commercial sample grades of Canada Western Red Spring (CWRS) wheat was investigated. Kernel perimeter, length, width, and area measurements were used to determine mean and dispersion statistics for composite railcar CWRS samples of No. 1, 2, and 3 grades; the numbers of railcars sampled were 27, 40, and 36, respectively. Sample sizes ranged from 10 to 2,000 kernels. Instrumental measurement precision was routinely better than 0.1 mm for macroview images, with a resolution of 0.0054 cm² per pixel. Computed mean kernel feature measurements and dispersion statistics were highly dependent on sample size and grade. Comparative analysis of wheat samples by digital imaging of individual kernels required a sample of no less than 300–500 kernels, depending on sample grade, for accurate representation of the parent sample. This level of sampling resulted in detection of significant differences (P < 0.05) in mean kernel features that, on average, differed by <1%. Except for some samples containing low numbers of kernels, lower grade wheat had more variable kernel features compared with higher grade samples. In relative terms, for comparably sized samples (≥133 kernels), variance in No. 2 grade wheat was 6–11% higher that for No. 1 grade wheat, depending on kernel feature. Similarly, variance in No. 3 grade wheat was 13–23% higher than for No. 2 grade wheat and 20–37% higher than for No. 1 grade wheat, indicating that wheat grading has a predictable effect on and is influenced by the uniformity of kernel characteristics in a sample. The ability of digital image analysis to detect these effects reflects the potential of this technology for use in objective classification of wheat according to grade.     

Survey of 1975 wheat and soybeans for aflatoxin, zearalenone, and ochratoxin

Wheat samples (102 lots) were collected from Virginia, North Carolina, southeastern Missouri, southern Illinois, and Kentucky. Soybean samples (180 lots) were collected from Virginia, Illinois, Iowa, Minnesota, Nebraska, Alabama, Arkansas, and Texas. Samples of both commodities were analyzed for zearalenone, aflatoxin, and ochratoxin by the Eppley method. None of the 3 mycotoxins was detected in soybeans. Aflatoxins and ochratoxin A were not detected in wheat, but zearalenone was detected in 19 of 42 samples collected in Virginia. Half of the Virginia samples were collected because they were mold-damaged. Zearalenone levels ranged from 0.36 to 11.05 ppm; the identity of the zearalenone was confirmed by gas-liquid chromatography and mass spectroscopy. Gibberella zea infection (6-60%) was detected in all of the zearalenone-positive samples; 6-60% of the kernels in the samples tested contained G. zea.     

Quantitative analysis of phytate globoids isolated from wheat bran and characterization of their sequential dephosphorylation by wheat phytase

Wheat phytase was purified to investigate the action of the enzyme toward its pure substrate (phytic acid - myo-inositol hexakisphosphate) and its naturally occurring substrate (phytate globoids). Phytate globoids were purified to homogeneity from wheat bran, and their nutritionally relevant parameters were quantified by ICP-MS. The main components of the globoids were phytic acid (40% w/w), protein (46% w/w), and several minerals, in particular, K > Mg > Ca > Fe (in concentration order). Investigation of enzyme kinetics revealed that K(m) and V(max) decreased by 29 and 37%, respectively, when pure phytic acid was replaced with phytate globoids as substrate. Time course degradation of phytic acid or phytate globoids using purified wheat phytase was followed by HPIC identification of inositol phosphates appearing and disappearing as products. In both cases, enzymatic degradation initiated at both the 3- and 6-positions of phytic acid and end products were inositol and phosphate.     

Gas chromatographic determination of deoxynivalenol in wheat

Modifications to a published method are described for the determination of deoxynivalenol (DON) in wheat by gas chromatography with electron capture quantitation of the heptafluorobutyrate derivative. In the modified method, DON is extracted by shaking the sample with methanol-water on a wrist-action shaker, followed by filtration through rapid flow paper. One concentration step is eliminated, and a hexane wash is incorporated to remove toluene from the silica gel column. Recoveries of DON from wheat samples spiked at 0.1, 0.5, and 1.0 ppm ranged from 77.3 to 86.3% and averaged 81.5%.     

LC/ESI/MS method for the quantitative detection of guazatine residues in cereals

Guazatine is a fungicide used in agriculture to control a wide range of seed-borne diseases of cereals and other vegetable foods. In this work, a LC-ESI-MS method was developed for the quantitative detection of guazatine residues in maize and hard wheat. Quantitative data were determined for the residues of the main diamines, triamines, and tetramines that cover more than 87% of the total contents of the mixture. The mean recoveries from the fortified cereals at 0.050 mg/kg ranged from 81 to 86%, with the coefficients of variation (CVs) ranging from 0.9 to 5.5% (n = 5). At 0.025 mg/kg, the recoveries ranged from 78 to 87%, with the CVs ranging from 0.8 to 6.3% (n = 5). The limits of quantification have been estimated to be 0.010, 0.004, 0.002, 0.002, 0.005, and 0.002 mg/kg, respectively, for GN, GG, GNG, GGN, GGG, and GGGG in maize and hard wheat (S/N ratio >10).     

The importance of lactic acid bacteria for phytate degradation during cereal dough fermentation

Lactic acid fermentation of cereal flours resulted in a 100 (rye), 95-100 (wheat), and 39-47% (oat) reduction in phytate content within 24 h. The extent of phytate degradation was shown to be independent from the lactic acid bacteria strain used for fermentation. However, phytate degradation during cereal dough fermentation was positively correlated with endogenous plant phytase activity (rye, 6750 mU g(-1); wheat, 2930 mU g(-1); and oat, 23 mU g(-1)), and heat inactivation of the endogenous cereal phytases prior to lactic acid fermentation resulted in a complete loss of phytate degradation. Phytate degradation was restored after addition of a purified phytase to the liquid dough. Incubation of the cereal flours in buffered solutions resulted in a pH-dependent phytate degradation. The optimum of phytate degradation was shown to be around pH 5.5. Studies on phytase production of 50 lactic acid bacteria strains, previously isolated from sourdoughs, did not result in a significant production of intra- as well as extracellular phytase activity. Therefore, lactic acid bacteria do not participate directly in phytate degradation but provide favorable conditions for the endogenous cereal phytase activity by lowering the pH value.