Chemometric evaluation of trace metal concentrations in some nuts and seeds

Seventeen trace metals in acid digests of nuts and seeds were determined using inductively coupled plasma-atomic emission spectrometry and inductively coupled plasma-mass spectrometry. The data were subjected to chemometric evaluation using principal component analysis (PCA), linear discriminant analysis (LDA) and cluster analysis (CA) in an attempt to classify the samples. Hazelnuts (raw and dry roasted), almonds (raw and dry roasted), sunflower seeds (black and white), peanuts (raw and dry roasted), cashew nuts, Brazil nuts, walnuts, chickpeas (raw and dry roasted), pumpkin seeds (raw and dry roasted), and pistachio nuts were used as samples. The samples were classified into seven groups by PCA and CA. All group members determined using PCA and CA were found by LDA to be correctly classified in the predicted groups. Interestingly, the chemometric evaluation indicated that the raw and roasted nuts are very close to each other even though some originated from different countries.     

A novel real-time polymerase chain reaction (PCR) method for the detection of hazelnuts in food

A real-time PCR (polymerase chain reaction)-based method for the detection of hazelnuts (nuts of Corylus avellana or C. maxima) in confectionery and bakery products is described. The method consists of DNA isolation by chaotropic solid phase extraction and the subsequent PCR with hazelnut-specific primers and a TaqMan fluorescent probe. The primers and the probe are targeted to the hsp1 gene encoding for a low molecular weight heat-shock protein. The method was positive for five hazelnut varieties approved in Slovakia and negative for all other tested plant materials used in food industry including peanuts, walnuts, almonds, pistachio nuts, cashews and chestnuts. The intrinsic detection limit of the method was 13 pg hazelnut DNA, which corresponds to approximately 27 genome equivalents (1C). Using a series of model pastry samples with defined hazelnut contents, a practical detection limit of 0.01% (w/w) hazelnut was determined. Practical applicability of the PCR method was tested by the analysis of 20 food samples (confectionery and bakery products) along with ELISA. For all of the food samples, identical results were obtained by both methods, which conformed to the labelling. The presented PCR method is useful for sensitive and selective detection of hazelnuts in food samples and can be performed in one working day.     

Influence of different non-grain commodities on the population growth of Trogoderma granarium Everts (Coleoptera: Dermestidae)

We investigated the population growth of the khapra beetle, Trogoderma granarium Everts (Coleoptera: Dermestidae) on seven groups of commercially available non-grain commodities. Six powdered spices were used in the first group of experiments: black pepper, clove, nutmeg, allspice, cinnamon and turmeric. The second group of the tested commodities included seven animal products: powdered cow milk, powdered pork zelatin, sheeps’ wool, goat’s skin, ducks’ feathers, dry dog food and dry cat food. The third group of commodities was consisted of six herbs: oregano, spearmint, basil, coriander, laurel and marjoram. The fourth group of commodities contained ten pulses: chickpeas, lentil, split peas, black-eyed peas, beans, soybean flour and whole kernels, lima beans, mung bean, and broad beans. The fifth group comprised six dried fruits: Corinthian currants, sultanas, banana chips, melons, apricots and figs. The sixth group was consisted of five non-grain commodities: cottonseed cake, tobacco, black tea, Turk kahvesi and potato flour. The seventh group included seven nuts: sunflower seeds, pumpkin seeds, pistachios, roasted chickpeas (yellow), almonds, hazelnuts and walnuts. Finally, six cracked containment categories (0% cracked kernels, 5% cracked kernels and 95% intact kernels, 10% cracked kernels and 90% intact kernels, 25% cracked kernels and 75% intact kernels, 50% cracked kernels and 50% intact kernels and 100% cracked kernels) from five pulses were tested: chickpeas, black-eyed peas, mung bean, soybean and split peas. The highest progeny production (3.01 individuals per vial) was recorded on powdered cow milk. On pistachios, split peas, sunflower seeds, soybean flour, pumpkin seeds, walnuts, almonds and coriander, T. granarium built high population densities rapidly, while on roasted chickpeas, cottonseed cake, hazelnuts, chickpeas, dog food and lentils, its population growth was much less. Broad beans, melons, figs, lima beans, beans, Corinthian currants, pork zelatin and potato flour were less suitable diets for the development of this species. On black pepper, clove, nutmeg, allspice, cinnamon, turmeric, sheeps’ wool, goat’s skin, ducks’ feathers, cat food, sultanas, banana chips, apricots, tobacco, black tea, Turk kahvesi, oregano, spearmint, basil, laurel and marjoram, no progeny production was recorded. The proportion of 100% cracked black-eyed peas or mung beans was more suitable for the population growth of T. granarium. Also, the percentages of 50% cracked chickpeas or 10% cracked soybeans enhanced the development of the species, in comparison with the whole kernels of each pulse. Our study indicated that several non-grain commodities are beneficial for the population growth of T. granarium, a fact that should be seriously taken into account in international trade, as this pest may utilize them as “vehicles” of expansion.     

Aspergillus section Flavi and aflatoxins in dried figs and nuts in Algeria

The presence of Aspergillus section Flavi and aflatoxin (AF) contamination was investigated in 112 samples of peanuts, almonds and dried figs collected in Algeria. The occurrence of aflatoxin B1 (AFB1), B2 (AFB2), G1 (AFG1) and G2 (AFG2) in different commodities has been determined with a sensitive method based on high performance liquid chromatography (HPLC) coupled with fluorescence detection with post-column photochemical derivatisation. Analytical results indicated that 28 samples of peanuts, 16 samples of almonds and 26 samples of dried figs contained detectable levels of AFs. A total of 69 samples (61.6%) were contaminated with AFB1 ranging from the limit of quantification to 174 µg kg^?1. AFB2 was found in 12 samples (10.7%) and varied from 0.18 to 193 µg kg^?1. Seven samples revealed AF concentrations lower than the limit of quantification. Eleven peanut and fourteen dried fig samples exceeded the European maximum limits for AFB1.     

Tocopherols and total phenolics in 10 different nut types

The study was conducted to assess the content of tocopherols (α-, β-, γ- and δ-) and carotenoids (α- and β-carotene, zeaxanthin, lutein, cryptoxanthin and lycopene) in the unsaponifiable matter as well as the amount of total phenols of 10 different types of nuts. Tocopherols and carotenoids were analysed with HPLC, total phenols photometrically. The mean value of α-tocopherol equivalents ranged from non-detectable (macadamias) to 33.1 mg/100 g extracted oil (hazelnuts). Among all nuts, almonds and hazelnuts had the highest mean α-tocopherol content (24.2 and 31.4 mg/100 g extracted oil, respectively). β- and γ-tocopherols were prevalent in Brazil nuts, cashews, peanuts, pecans, pines, pistachios and walnuts. Mean values oscillated between 5.1 (cashews) and 29.3 (pistachios). Traces of δ-tocopherol (<4 mg/100 g extracted oil) were analysed in cashews, hazelnuts, peanuts, pecans, pines, pistachios and walnuts. There were no carotenoids detected in the tested nuts with the exception of pistachios. The mean content of total phenolics varied between 32 mg gallic acid equivalents/100 g (pines) and 1625 mg (walnuts). The results show the heterogenic amounts of antioxidants in nuts, which emphasises the recommendation of a mixed nuts intake.     

A novel real-time polymerase chain reaction method for the detection of macadamia nuts in food

A real-time PCR-based method for the detection of macadamia nuts (fruits of Macadamia integrifolia or M. tetraphylla or their hybrids) in food products is described. The method consists of DNA isolation by chaotropic solid phase extraction and subsequent PCR with macadamia-specific primers and a TaqMan fluorescent probe. The primers and the probe were targeted to the gene encoding for vicilin precursor. The method was positive for M. integrifolia and M. tetraphylla and negative for 16 other plant species used in food industry, including peanuts, walnuts, hazelnuts, almonds, pistachio nuts, cashew nuts, Brazil nuts, and chestnuts. The DNA-based detection limit of the method was 1.45 pg. Using a series of model samples with defined macadamia nut contents, a practical detection limit of 0.02% (w/w) macadamia nuts was determined. Practical applicability of the PCR method was tested by the analysis of 14 confectionery samples. For all of the samples, results conforming to the labeling were obtained. The presented PCR method is useful for relatively fast, highly selective, and moderately sensitive detection of macadamia nuts in food samples.     

Aflatoxin levels in raw and processed hazelnuts in Turkey

Aflatoxin levels in hazelnut samples obtained from exporter companies were monitored over a 3-year period. A total of 3188 samples of raw and processed hazelnuts were analysed using an HPLC method. The total aflatoxin content of the contaminated samples was in the range of 0.02–78.98?µg?kg^?1 for hazelnut kernels, 0.07–43.59?µg?kg^?1 for roasted hazelnut kernels, 0.02–39.17?µg?kg^?1 for roasted sliced hazelnut kernels, and 0.02–11.20?µg?kg^?1 for hazelnut purees, respectively, showing that the variations of aflatoxin contamination were very high. The results of aflatoxin analysis revealed that the aflatoxin contamination in the hazelnut samples was at a tolerable level. A total of 3147 samples were contaminated with aflatoxins, although below the legal limits. However, the aflatoxin contents of 41 samples exceeded the legal limits. Therefore, aflatoxin contents of hazelnuts should be monitored regularly to minimise the risk of aflatoxin hazard, and pre- and post-harvest strategies should be developed to prevent aflatoxin formation.     

Simultaneous detection of DNA from 10 food allergens by ligation-dependent probe amplification

The simultaneous detection of DNA from different allergenic food ingredients by a ligation-dependent probe amplification (LPA) system is described. The approach allows detection of several targets in a one-tube assay. Synthetic oligonucleotides were designed to detect DNA from peanuts, cashews, pecans, pistachios, hazelnuts, sesame seeds, macadamia nuts, almonds, walnuts and brazil nuts. The specificity of the system was tested with DNA from more than 50 plant and animal species. The sensitivity of the method was suitable to detect allergenic ingredients in the low mg kg^?1 range. The limit of detection (LOD) for single allergens in different food matrices was 5 mg kg^?1. The novel analytical strategy represents a useful tool for the surveillance of established legislation on food allergens within the European Union.     

Peanut and Walnut Rancidity: Effects of Oxygen Concentration and Relative Humidity

Accelerated tests for oxidative rancidity of blanched peanuts, blanched dry-roasted peanuts, blanched oil-roasted peanuts and shelled Persian walnuts were performed at high and low oxygen content at controlled intermediate and low relative humidities. The results confirmed and quantified the importance of oxygen content, relative humidity and roasting process in the oxidative rancidity of peanuts and walnuts. There is a potential to extend shelf-life of roasted peanuts and walnuts by edible coatings with low oxygen permeability or nitrogen-flushing with oxygen barrier packaging. Static headspace chromatography was useful to monitor oxidative rancidity in walnuts and roasted peanuts.     

Effect of application of nontoxigenic strains of Aspergillus flavus and A. parasiticus on subsequent aflatoxin contamination of peanuts in storage

Experiments were conducted to determine the potential for biological control of aflatoxin contamination of peanuts during storage. Florunner peanuts were treated in field plots by applying competitive, nontoxigenic strains of Aspergillus flavus and A. parasiticus, at 76 and 67 days after planting in 1998 and 1999, respectively. After harvest, half the peanuts from both treated and control plots were sprayed with an aqueous conidial suspension containing the nontoxigenic strains; the other half of the peanuts from each group were not sprayed. The peanuts were then placed in separate compartments of a miniature warehouse. Therefore, storage treatments consisted of peanuts that were (1) not treated at all; (2) treated prior to storage only; (3) field-treated only; (4) treated both in the field and prior to storage. Peanuts were stored for 3-5 months under high temperature and relative humidity conditions designed to promote aflatoxin contamination. In 1998, peanuts were not contaminated with aflatoxins prior to storage. After storage, peanuts that were never treated with the competitive fungi contained an average of 78.0 ppb of aflatoxins. Peanuts not treated in the field but receiving the spray treatment before storage contained 48.8 ppb. Peanuts treated in the field only averaged 1.4 ppb, and peanuts treated both in the field and prior to storage contained 0.8 ppb. In 1999, peanuts suffered from late-season drought and were contaminated with aflatoxins at harvest, with controls averaging 516.8 ppb compared with 54.1 ppb in treated peanuts. After storage, non-field-treated peanuts averaged 9145.1 ppb compared with 374.2 ppb for peanuts that had been field-treated, a 95.9% reduction. Spraying of pods with the nontoxigenic strains postharvest but prior to storage provided no additional protection against aflatoxin contamination. Results demonstrated that field application of the nontoxigenic strains had a carry-over effect and reduced aflatoxin contamination that occurred in storage.     

基于植物DNA条形码技术对杏仁露中花生源性成分的鉴别研究

杏仁露含有丰富的植物蛋白,深受广大消费者的喜爱,拥有广阔的消费市场,同时其成分与配料表是否相符也备受消费者关心。本研究基于植物DNA条形码技术与PCR技术,设计、筛选同时能对杏仁、花生、核桃、大豆、芝麻和榛子六个物种进行扩增的通用引物,并将其应用于杏仁露中花生源性成分的检测。试验表明引物ITS2-2和trn H-psb A-1分别对六个物种的扩增成功率和测序成功率均较高;通过计算杏仁、花生的基因组DNA提取率,设计掺假模型在提取的杏仁基因组DNA中掺入花生基因组DNA,引物ITS2-2对于掺入85.80%的花生检测结果为杏仁,trn H-psb A-1对于掺入6.94%的花生检测结果为花生。引物ITS2-2和trn H-psb A-1可作为鉴别杏仁露中花生源性成分的植物DNA条形码组合。本研究为该类食品检测提供了新的思路,可作为相关研究的参考。     

The occurrence of aflatoxins in peanuts imported into Czechoslovakia for human consumption

Results of 492 analyses for aflatoxin in raw shelled peanuts imported into Czechoslovakia during 1982-1984 are presented. Most samples (55.3%) had aflatoxin content less than the detection limit of the radioimmunochemical screening method (0.8 micrograms/kg). Further analyses showed that 239 out of 410 samples of roasted peanuts contained aflatoxin below the detection limit. Only 1.9% of all peanut samples were found to have contamination level more than 5 micrograms/kg aflatoxin. The highest levels of aflatoxin observed were in a raw peanut sample containing 202.1 micrograms/kg and in a roasted peanut sample containing 32.6 micrograms/kg.     

Aflatoxins in nuts assayed by immunological methods

 Different kinds of raw and processed nuts available in the local retail market were investigated for aflatoxin content. Total aflatoxins and aflatoxin B₁ content were determined by immunoaffinity chromatography with fluorescence detection after reaction with bromine solution and by immunoenzymatic test kits. Of 29 investigated samples, 38% were contaminated. The total aflatoxin content in contaminated samples was between 1.20 μg/kg for peanuts and 5.50 μg/kg for walnuts. The concentration of aflatoxin B₁ found in contaminated samples was between 0.35 μg/kg for cashew nuts and 4.04 μg/kg for walnuts. The mean recovery of total aflatoxins was 95% for the Ridascreen test and 92% for immunoaffinity chromatography with fluorescence detection. For aflatoxin B₁ the mean recovery was 84%. Received: 4 March 1999 / Revised version: 17 May 1999     

Gastric Digestion of Raw and Roasted Almonds In Vivo

Almonds are an important dietary source of lipids, protein, and α‐tocopherol. It has been demonstrated that the physical form of almond kernels influences their digestion and absorption, but the role of thermal processes on the digestion of almonds has received little attention. The objectives of this study were to examine the gastric emptying and nutrient composition of gastric chyme from pigs (used as a model for the adult human) fed a single meal of either raw or roasted almonds over a 12‐h postprandial period (72 pigs total, 6 pigs at each diet–time combination). Concentrations of glucose, triacylglycerols, and α‐tocopherol in peripheral plasma during the 12‐h postprandial period were determined. For dry matter and lipid, the gastric emptying profile was not different between raw and roasted almonds. Roasting almonds also did not influence gastric pH, or plasma glucose or triacylglycerols levels. In contrast, the gastric emptying of protein was more rapid for raw almonds compared to roasted almonds (P < 0.01) and intragastric protein content exhibited segregation (P < 0.001) throughout the stomach, with raw almonds having a higher level of segregation compared to roasted almonds. Postprandial plasma α‐tocopherol levels were, on average 33% greater (P < 0.001) after consumption of raw almonds, most likely as a result of the higher concentration of α‐tocopherol in raw almonds compared to roasted almonds. Roasting of almonds did not influence the overall gastric emptying process, but did lead to differences in the distribution of protein in the stomach and to the gastric emptying of protein.     

Effect of different roasting methods on the proximate by composition,flow properties,amino acid compositions,colour, texture,and sensory profile of the chickpeas

Roasting is a common process for chickpeas to improve their texture, palatability, appearance, shelf-life, physical, and functional properties. This study aimed to determine the effect of different roasting methods (conventional, microwave, and microwave + conventional) on the proximate and amino acid compositions, powder properties, texture, and sensorial properties of the chickpeas. For this purpose three different roasting times (3, 5, and 7 min), microwave powers (100, 300, and 600 W), and microwave roasting + conventional roasting treatment (100 W + 250 °C, 300 W + 250 °C, and 600 W + 250 °C) were applied to raw chickpea samples. The moisture content and water activity values of roasted chickpeas were found to be lower than 7% (w.b.) and 0.50, respectively. The lower ash and protein contents, hygroscopicity value, wettability time and higher fat content and L* value were observed for control compared to roasted samples. The flowability behaviour of the samples was found at a fair level. Roasting methods significantly affected the amount of amino acids in chickpeas but do not reduce the nutritional quality of their proteins. The hardness value of chickpea samples from the suture and cheek angle was decreased parallel to the increase in the roasting temperature and time. The highest sensory scores in terms of general appeal were obtained from the combined group (300 W–250 °C) for 3 min.     

Manual sorting to eliminate aflatoxin from peanuts

A manual sorting procedure was developed to eliminate aflatoxin contamination from peanuts. The efficiency of the sorting process in eliminating aflatoxin-contaminated kernels from lots of raw peanuts was verified. The blanching of 20 kg of peanuts at 140 degrees C for 25 min in preheated roasters facilitated the manual sorting of aflatoxin-contaminated kernels after deskinning. The manual sorting of raw materials with initially high aflatoxin contents (300 ppb) resulted in aflatoxin-free peanuts (i.e., peanuts in which no aflatoxin was detected). Verification procedures showed that the sorted sound peanuts contained no aflatoxin or contained low levels (<15 ppb) of aflatoxin. The results obtained confirmed that the sorting process was effective in separating contaminated peanuts whether or nor contamination was extensive. At the commercial level, when roasters were not preheated, the dry blanching of 50 kg of peanuts for 45 to 55 min facilitated the proper deskinning and subsequent manual sorting of aflatoxin-contaminated peanut kernels from sound kernels.     

A survey of the bacteriological quality of preroasted peanut, almond, cashew, hazelnut, and brazil nut kernels received into three Australian nut-processing facilities over a period of 3 years

There is little information about bacteriological quality of preroasted kernels available in the public domain. An investigation of the bacteriological quality of preroasted peanut, almond, cashew, hazelnut, and Brazil nut kernels received into three Australian nut-processing facilities was performed over a period of 3 years. A total of 836 samples were analyzed for aerobic plate count, and 921 samples for Salmonella and Escherichia coli. The 921 samples included 653 peanut, 100 cashew, 60 almond, 60 Brazil nut, and 48 hazelnut kernels. There was no E. coli detected in any sample. Salmonella subsp. II (Fremantle) was detected in one raw almond sample. The aerobic plate count percentages of positive samples with counts above the detection level of the plating method used (100 CFU/g) for peanuts, almonds, cashews, hazelnuts, and Brazil nuts were 84, 78, 74, 50, and 45%, respectively. Of the samples containing more than this detection limit, the means were 4.5, 4.4, 3.1, 2.5, and 3.8 log CFU/g respectively. Although roasted kernel quality was not within the scope of this survey, raw microbial bioload would be expected to reduce on roasting. The bacteriological quality of preroasted peanut, almond, cashew, hazelnut, and Brazil nut kernels received into nut-processing facilities in Australia does not appear to suggest a public health concern.     

A sandwich enzyme-linked immunosorbent assay for the detection of almonds in foods

An enzyme-linked immunosorbent assay was developed to detect almonds as potential allergenic contaminants in food. Polyclonal antibodies directed against roasted almonds were partially purified from immunized sheep and rabbits and used as capture and secondary antibodies, respectively, in a sandwich-type, 96-well plate format. Food samples and almond-spiked samples were extracted 1:10 in phosphate-buffered saline at 60 degrees C for 2 h, centrifuged, and applied to wells coated with sheep anti-almond antibody. After incubation, washing, and the addition of rabbit anti-almond antibody, the amount of almond present was detected with the subsequent addition of goat anti-rabbit immunoglobulin G-alkaline phosphatase conjugate and p-nitrophenyl phosphate substrate. Plate absorbances were read at 410 nm, and standard curves were developed in all matrices to quantify unknowns. Antibodies developed were specific for almond; however, some cross-reactivity was observed with extracts of some tree nuts and sesame seeds. Sodium dodecylsulfate-polyacrylamide gel electrophoresis and Western immunoblotting indicated that sheep anti-almond antibody recognized proteins extracted from black walnuts, Brazil nuts, cashews, hazelnuts, macadamia nuts, pistachios, and sesame seeds in addition to those from almond. The assay was optimized to detect less than 1 ppm of almond and was used successfully to determine almond residues in cereal and chocolate without cross-reacting interferences. A retail survey of 20 brands of cereal demonstrated that the assay produced statistically consistent results. This assay provides a useful quality control tool for the food industry for the protection of consumers allergic to almonds.     

High-resolution Orbitrap™-based mass spectrometry for rapid detection of peanuts in nuts

Peanut represents one of the most harmful allergenic foods capable of triggering severe and sometimes lethal reactions in allergic consumers upon ingestion of even small amounts. Several proteins capable of inducing allergic reactions that have been recognised by patients’ IgE antibodies have been identified from this nut source. Methods mainly based on ELISA assays have been developed in order to detect peanuts in several food commodities. In addition LC-MS/MS methods based on different mass analysers have also been devised for tracing peanut contamination in different foods achieving low limits of detection. The applicability of a benchtop high-resolution Exactive? mass spectrometer has never been investigated for the rapid screening of peanut contamination in complex food matrices like mixtures of nuts. We report in this paper the design of suitable peanut markers and the development of an high-resolution Orbitrap? mass spectrometer-based method for peanut detection in a mixture of nuts species. With this aim, different types of samples were prepared: (1) nuts-based powder made up of a mixture of hazelnuts, pistachios, almonds and walnuts; and (2) nuts powder fortified with peanuts. Different levels of fortifications were produced and the applicability of the method was tested. Finally, a subset of six peptides fulfilling specific analytical requirements was chosen to check the suitability of the method tailored to the detection of peanuts in nuts-based products, and two of them, peptides VYD and WLG, were selected as quantitative markers. The method proved to be a suitable screening tool to assess the presence of traces of peanuts in other tree nuts with a limit of detection as low as 4 µg of peanuts proteins or 26 µg of peanuts in 1 g of matrix.