Distribution of aflatoxin in pistachios. 7. Sequential sampling

Sequential sampling for aflatoxin testing in pistachios is evaluated using the aflatoxin distribution and Monte Carlo results previously obtained (J. Agric. Food Chem. 1999, 47, 3771-3775). The sequential protocol is modeled on the current EU test protocol by applying a three-step sampling, using 10, 20, and 30 kg sample averages. An acceptance level of 15 ng/g of total aflatoxin, under consideration for U.S. standards, is applied. Optimization leads to indifference regions of 2-30 ng/g for the first two steps. The resulting OC curve approximates that for a single 50 kg sample. The sequential protocol is applied to the results for a set of 1293 lots of the 1998 crop year, each tested with a single 10 kg sample. Ninety-five percent of the lots would have been accepted on the basis of the single test and 1.5% would have been rejected, whereas 3.5% of the lots would have required retesting.     

Distribution of aflatoxin in almonds. 2. Distribution in almonds with heavy insect damage

The aflatoxin distribution function in individual insect-damaged NePlus Ultra almonds was determined and found to be the sum of two distributions. Substantially all almonds exhibited a positive aflatoxin level between 0.02 ng/g (the detection level) and 0.3 ng/g, the precise form of this distribution depending on the lot studied. In addition, 1/1000 of the nuts showed contamination between 60 and 600 000 ng/g, independent of the lot. The latter distribution showed a smooth decrease with log concentration in this range, with no evidence of a minimum, as had been found previously for pistachios. No distribution data between 0.3 and 60 ng/g could be obtained. The distribution below 0.3 ng/g was assigned to contamination during post-harvest storage. The distribution above 60 ng/g was tentatively assigned to navel orange worm damage occurring when insects enter the kernel during split hulls late in the growing season. Considerable additional work will be required to verify these assignments.     

Sample preparation and presampling of pistachios

A theory has been developed to quantify the reduction of subsample variance of aflatoxin contamination, which is observed when granular materials are wet slurried, rather than dry ground, during subsample homogenization. A coefficient of variation, based on particle size distribution, subsample size, and probability of contamination, is predicted. The theory is tested with dry ground and with wet slurried pistachios, and excellent quantitative agreement is obtained. A 32% increase in the mean aflatoxin level is observed as well when wet slurrying is applied. Although no statistical explanation for this effect can be found, it is suggested that it is related to physiochemical binding between the nut matrix, which is (partly) broken by wet slurrying, and aflatoxin, making the extraction of more toxin possible. Other parameters that may affect slurrying results have been investigated as well.     

Survey of U.S. wheat for ochratoxin and aflatoxin.

A total of 291 hard red winter wheat samples, 286 hard red spring wheat samples, and 271 soft red winter wheat samples were analyzed for the presecne of ochratoxin and aflatoxin. Samples in all grades came from those collected during crop years 1970-1973 for grade determinations by the Agricultural Marketing Service, U.S. Department of Agriculture. Sensitivity limits of the analytical method as carried out were 1-3 ppb aflatoxin B1 and 15-30 ppb ochratoxin A. No aflatoxin was detected in any sample. Three samples of hard red winter wheat (Grades U.S. No. 4 and 5 and Sample Grade) contained ochratoxin A (trace, 35, and 25 ppb, respectively). Eight of the hard red spring wheats contained ochratoxin A (15-115 PPB); these were in Grades U.S. No. 4 and 5 and Sample Grade.     

Laser fluorometric determination of aflatoxin B1 in corn.

A 2-step chromatographic separation, using both thin layer chromatography (TLC) and high pressure liquid chromatography (HPLC), in conjunction with the high sensitivity of laser fluorometry permits extension of the detection limits of aflatoxin contamination in corn to 0.1 ppb (microgram/kg) with a 26% root mean square variation. Aflatoxin B1 is extracted from corn with water-methanol and cleaned up by TLC. The recovery of aflatoxin from the TLC plates was linear from 10 to 1000 pg. Aflatoxin B1 is converted to the more highly fluorescent B2A derivative by treatment with 1N HCl. Experiments with aflatoxin B1 standard establish a constant conversion to B2A over approximately 3 orders of magnitude in B1 concentration. An extract of the B2A aflatoxin derivative is injected onto a reverse phase HPLC column. A flowing droplet of eluant is irradiated by an amplitude-modulated 325 nm He-Cd ion laser beam, and fluorescence from the droplet is detected by a lock-in amplifier in phase with the laser modulation. Several chromatograms are presented that demonstrate the capability of this procedure for removing interfering components in the corn extract.     

Simple method for simultaneous detection of aflatoxin and zearalenone in corn.

A method is described for the simultaneous detection of alfatoxin and zearalenone in corn at 5 and 200 ppb, respectively. No evaporation of solvent is required and the procedure is simple enough to be considered for use at marketing locations. The presence of absence of these myocotoxins can be determined in 10-20 min/sample. The procedure involves an initial blender extraction with methanol, partitioning of fat and pigments into 1-1,2-trichlorotrifluoroethane (Freon-113) from an aqueous ammonium sulfate layer, followed by extraction of aflatoxin from the aqueous layer with chlorobenzene. The chlorobenzene extract can be spotted directly onto a thin layer chromatographic plate which requires only 4 min development. Concentrations of aflatoxin and zearalenone can be estimated by visual comparison of sample spots with standards.     

Fluorometric measurement of aflatoxin adsorbed on florisil in minicolumns.

Filter fluorometers have been adapted to measure the fluorescence intensity of aflatoxin absorbed on a Florisil layer in minicolumns. The relationship between concentration and intensity is near linear in the aflatoxin range from 10 to 100 ng. Although individual aflatoxin fractions cannot be resolved, since the measure is one of total intensity, fluorometric measurements advance the minicolumn screening procedure to a semiquantitative level. The detection of 1 ng aflatoxin B1 is well within the limits of a filter fluorometer with a photomultiplier detector. A precision, expressed as per cent coefficient of variation, ranging from 1.2 to 4.2%, was obtained for standard B1 columns.     

Formation of aflatoxin derivatives on thin layer chromatographic plates.

Rapid confirmation of the presence of aflatoxins B-1 and G-1 in foods is provided by reaction with trifluoroacetic acid at the origin of a thin layer chromatographic plate. The procedure has been used successfully with various nuts, grains, coffee and cocoa beans, and other foods.     

A quantitative method for determination of aflatoxin B in roasted corn.

Roasting aflatoxin-contaminated corn will reduce toxin levels. A quantitative analysis for aflatoxin in roasted corn has been developed by modifying a cleanup technique for green coffee extracts approved as official first action by the AOAC. A chloroform extract is partially purified on a Florisil column, and thin layer chromatographic (TLC) plates are developed with methylene chloride-chloroform-isoamyl alcohol-formic acid (81+15+3+1). Recoveries average 101% and the sensitivity limit is 5 ppb aflatoxin B1. A 2-dimensional TLC procedure can also be used to separate the aflatoxins from background interferences.     

西藏尼洋河水体重金属分布特征及风险评价

重金属含量是影响水生态环境安全的重要因素,该文意在探索西藏河流水体重金属变化规律。研究了雅鲁藏布江第二大支流尼洋河的Zn、Cu、Mn、Cd、Fe 5种重金属含量时空变化特征和相关性,以及迁移变化机理,对重金属富集程度和污染风险进行了评价。结果表明：尼洋河重金属含量源头较低,中上游位置有突变。Fe、Mn、Cd是影响水质变化的敏感因子。Zn、Cu、Mn含量平水期＞枯水期＞丰水期。丰水期重金属含量主要受人类活动或水文气象活动影响,平水期及枯水期的Zn、Cu、Mn、Fe含量主要受自然过程影响,枯水期Cd主要受人类活动影响。水库对重金属富集有一定的影响。河源和中下游河段污染较轻,上游河段重金属综合污染指数最高。     

湖州市土壤重金属元素分布及潜在生态风险评价

在对湖州市土壤重金属污染状况进行调查后,按照有关标准和方法对该市土壤重金属潜在生态风险进行了评价。结果表明:单因子污染指数最小的是Pb,最大的是Cd。土壤中Pb含量均达标,其余各重金属元素有部分土壤超标。68.9%的土壤处于安全等级,21.6%的土壤处于警戒限,轻污染土壤占8.1%,重污染土壤占1.4%。68.9%的土壤处于低风险等级,27.0%的土壤处于中等风险等级,强风险等级土壤占2.7%,很强风险等级土壤占1.4%。林地的重金属含量变化幅度普遍大于耕地。耕地土壤污染程度轻于林地。土壤中各重金属对污染的贡献大小为:CdNiCrHgZnCuAsPb。     

Effect of season and variety on the differentiation of geographic growing origin of pistachios by stable isotope profiling

The objectives of this study were to demonstrate if seasonal or variety differences affected the feasibility of stable isotope profiling methods to differentiate the geographical growing regions of pistachios (Pistachia vera). Bulk carbon and nitrogen isotope analyses of approximately 150 pistachios samples were performed. Isotope ratios were determined using a stable isotope mass spectrometer. The pistachio samples analyzed were from the three major pistachio-growing regions: Turkey, Iran, and the United States (California). Geographic regions were well separated on the basis of isotope ratios. Seasonal effects were found to affect some isotopes for some regions. Pistachio varieties within specified geographic regions were not found to affect the discriminating power of stable isotopes, for the varieties tested. This paper reports the development of a simple chemical profiling method using bulk stable isotope ratios that may be widely applied to the determination of the geographic origin of foods.     

Inhibition of aflatoxin B(1) biosynthesis in Aspergillus flavus by anthocyanidins and related flavonoids.

Anthocyanidins and precursors or related flavonoids were tested at concentrations from 0.3 to 9.7 mM ( approximately 0.1-3.0 mg/mL) for activity against growth and aflatoxin B(1) biosynthesis by Aspergillus flavus Link:Fr. NRRL 3357. Aflatoxin B(1) production was inhibited by all anthocyanidins tested, and 3-hydroxy compounds were more active than 3-deoxy forms. Monoglycosides of cyanidin were 40% less inhibitory than the aglycon, whereas a monoglucoside and a diglucoside of pelargonidin were 80 and 5%, respectively, as active as the aglycon. Of eight flavonoids tested, only kaempferol was moderately active, whereas luteolin and catechin were weakly inhibitory. Binary combinations of delphinidin and three other aflatoxin inhibitors acted independently of each other. Results with an aflatoxin pathway mutant indicated that anthocyanidin inhibition occurred before norsolorinic acid synthesis.     

Sources of resistance to aflatoxin production in maize

Drought-tolerant maize genotypes (Huffman, Z08-004, Tuxpan, PH 9, NRC 5348, Chunco, Saint Croix, and Arizona) were compared in the field and laboratory to toxin-resistant GT-MAS:gk and Yellow Creole. SDS-PAGE, scanning electron microscopy of kernel cuticle, amount of kernel wax, Aspergillus flavus kernel colonization, Aspergillus ear rot, insect damage, aflatoxin production, and their relationships were examined. SDS-PAGE showed the presence of a 14 kDa trypsin inhibitor in the kernels of all genotypes except Chunco, which contains a protein of a larger molecular weight. The 14 kDa trypsin inhibitor protein content in these genotypes was higher than in GT-MAS:gk and Yellow Creole. Scanning electron microscopy revealed that Arizona, Huffman, and Chunco genotypes had abundant wax deposits on kernel surfaces and the amount of pericarp wax was equal to or above that from GT-MAS:gk and Yellow Creole. Differences in Aspergillus ear rot ratings, fungal colonization, and insect damage by corn earworm were observed in all drought-tolerant maize genotypes as well as in the controls. Kernel screening assays showed that aflatoxin B(1) levels in inoculated drought-tolerant genotypes differed significantly from those in GT-MAS:gk and Yellow Creole (LSD = 576). Aflatoxin B(1) levels in the inoculated genotypes differed significantly from those of GT-MAS:gk or Yellow Creole (LSD = 1389) when grown under drought stress conditions. Pearson correlation coefficients were significant between ear rot ratings and insect damage (r = 0.75; P = 0.01) and between Aspergillus ear rot and aflatoxin levels (r = 0.54; P = 0.05). On the basis of the parameters studied, there are indications that these genotypes were potential sources of A. flavus resistance.     

洛川苹果林地重金属分布特征和污染评价

通过取样调查和试验分析,选用As、Cr、Cu、Ni、Pb,Zn、Mn、Co.V等9种重金属元素研究了洛川苹果林地典型剖面(LC剖面)的重金属分布特征,并利用地累积指数法、重金属富集指数法、重金属潜在生态危害指数法,对洛川苹果林地重金属的污染状况和金属元素富集规律进行了初步研究.结果表明:As、Cr,Ni、Mn、V含量变化走势基本相同,总体上由表层向下波动递增,Cu、Pb、Zn、Co4元素含量垂向变化特征基本相似,由表层向下总体呈先减少后增多趋势;除Pb和Zn外,As、Cr、Cu、Ni、Mn,Co和V均有一定程度的富集.As的生态危害指数(Eri)较高,在9种重金属元素中最大,潜在生态危害指数(RJ)为39.9,但小于轻微生态危害的阈值150,表明该区苹果林地重金属尚未构成污染危害.     

Fluorimetric determination of aflatoxin M1 in cheese

A simple and sensitive method is proposed for the determination of aflatoxin M1 in cheese. The ground cheese sample is extracted with acetone-water (3 + 1). Acetone is evaporated under vacuum, and the aqueous phase is passed through a C18 disposable cartridge. After the cartridge is washed with acetonitrile-water (1 + 9), the toxin is eluted with acetonitrile. The extract is then cleaned up on a silica cartridge. Final analysis is performed by 2-dimensional thin layer chromatography (TLC) combined with fluorodensitometry or by liquid chromatography on a reverse phase C18 column with fluorescence detection. Recovery is greater than 90%, and the coefficient of variation is 6% or less. The detection limit is in the range of 10 ng/kg. The identity of aflatoxin M1 is confirmed by formation of the M2a or acetyl-M1 derivative and rechromatography.     

Decontamination of aflatoxin-forming fungus and elimination of aflatoxin mutagenicity with electrolyzed NaCl anode solution.

Electrolysis of a 0.1% (17.1 mM) solution of NaCl using separate anode and cathode compartments gives rise to solutions containing active chemical species. The strongly acidic "anode solution" (EW+) has high levels of dissolved oxygen and available chlorine in a form of hypochlorous acid (HOCl) with a strong potential for sterilization, which we have investigated here. Exposing Aspergillus parasiticus at an initial density of 10(3)spores in 10 microL to a 50-fold volume (500 microL) of EW+ containing ca. 390 micromol HOCl for 15 min at room temperature resulted in a complete inhibition of fungal growth, whereas the cathode solution (EW-) had negligible inhibitory effects. Moreover, the mutagenicity of aflatoxin B(1) (AFB(1)) for Salmonella typhimurium TA-98 and TA-100 strains was strongly reduced after AFB(1) exposure to the EW+ but not with the EW-. In high-performance liquid chromatography analysis, the peak corresponding to AFB(1) disappeared after treatment with the EW+, indicating decomposition of the aflatoxin. In contrast, the routinely used disinfectant sodium hypochlorite, NaOCl, of the same available chlorine content as that of EW+ but in a different chemical form, hypochlorite (OCl-) ion, did not decompose AFB(1) at pH 11. However, NaOCl did decompose AFB(1) at pH 3, which indicated that the principle chemical formula to participate in the decomposition of AFB(1) is not the OCl- ion but HOCl. Furthermore, because the decomposition of AFB(1) was suppressed by pretreating the EW+ with the OH radical scavenger thiourea, the chemical species responsible for the AFB(1)-decomposing property of the EW+ should be at least due to the OH radical originated from HOCl. The OH in EW+ was proved by electron spin resonance analysis.