Toxicity of Selenium in Brazil Nuts to Rats

Nuts from Bertholletia excelsa, commonly known as Brazil nuts, were analyzed for selenium. Of 529 nuts from one commercial source, 6% were found to contain 100 ppm selenium or more. The mean value for all nuts was 29.6 ppm and the median value was 13.4 ppm. Hexane extracted high-selenium Brazil nut meal in a corn-based diet fed to rats produced toxicity similar to that obtained from seleniferous corn, selenomethionine or sodium selenite as assessed by weight gain, visually scored liver damage, and liver, kidney, and spleen weights. Selenium contents of liver, kidney, spleen, and blood were also determined. It is suggested that the selenium in Brazil nuts is as biologically potent as that from other sources.     

Determination of aflatoxin risk components for in-shell Brazil nuts

A study was conducted on the risk from aflatoxins associated with the kernels and shells of Brazil nuts. Samples were collected from processing plants in Amazonia, Brazil. A total of 54 test samples (40?kg) were taken from 13 in-shell Brazil nut lots ready for market. Each in-shell sample was shelled and the kernels and shells were sorted in five fractions: good kernels, rotten kernels, good shells with kernel residue, good shells without kernel residue, and rotten shells, and analysed for aflatoxins. The kernel?:?shell ratio mass (w/w) was 50.2/49.8%. The Brazil nut shell was found to be contaminated with aflatoxin. Rotten nuts were found to be a high-risk fraction for aflatoxin in in-shell Brazil nut lots. Rotten nuts contributed only 4.2% of the sample mass (kg), but contributed 76.6% of the total aflatoxin mass (µg) in the in-shell test sample. The highest correlations were found between the aflatoxin concentration in in-shell Brazil nuts samples and the aflatoxin concentration in all defective fractions (R ^2?=?0.97). The aflatoxin mass of all defective fractions (R ^2?=?0.90) as well as that of the rotten nut (R ^2?=?0.88) were also strongly correlated with the aflatoxin concentration of the in-shell test samples. Process factors of 0.17, 0.16 and 0.24 were respectively calculated to estimate the aflatoxin concentration in the good kernels (edible) and good nuts by measuring the aflatoxin concentration in the in-shell test sample and in all kernels, respectively.     

Determination of aflatoxin risk components for in-shell Brazil nuts

A study was conducted on the risk from aflatoxins associated with the kernels and shells of Brazil nuts. Samples were collected from processing plants in Amazonia, Brazil. A total of 54 test samples (40 kg) were taken from 13 in-shell Brazil nut lots ready for market. Each in-shell sample was shelled and the kernels and shells were sorted in five fractions: good kernels, rotten kernels, good shells with kernel residue, good shells without kernel residue, and rotten shells, and analysed for aflatoxins. The kernel:shell ratio mass (w/w) was 50.2/49.8%. The Brazil nut shell was found to be contaminated with aflatoxin. Rotten nuts were found to be a high-risk fraction for aflatoxin in in-shell Brazil nut lots. Rotten nuts contributed only 4.2% of the sample mass (kg), but contributed 76.6% of the total aflatoxin mass (μg) in the in-shell test sample. The highest correlations were found between the aflatoxin concentration in in-shell Brazil nuts samples and the aflatoxin concentration in all defective fractions (R(2)=0.97). The aflatoxin mass of all defective fractions (R(2)=0.90) as well as that of the rotten nut (R(2)=0.88) were also strongly correlated with the aflatoxin concentration of the in-shell test samples. Process factors of 0.17, 0.16 and 0.24 were respectively calculated to estimate the aflatoxin concentration in the good kernels (edible) and good nuts by measuring the aflatoxin concentration in the in-shell test sample and in all kernels, respectively.     

One-step multiplex PCR method for the determination of pecan and Brazil nut allergens in food products

BACKGROUND: A one‐step polymerase chain reaction (PCR) method for the simultaneous detection of the major allergens of pecan and Brazil nuts was developed. Primer pairs for the amplification of partial sequences of genes encoding the allergens were designed and tested for their specificity on a range of food components. RESULTS: The targeted amplicon size was 173 bp of Ber e 1 gene of Brazil nuts and 72 bp of vicilin‐like seed storage protein gene in pecan nuts. The primer pair detecting the noncoding region of the chloroplast DNA was used as the internal control of amplification. The intrinsic detection limit of the PCR method was 100 pg mL^?1 pecan or Brazil nuts DNA. The practical detection limit was 0.1% w/w (1 g kg^?1). The method was applied for the investigation of 63 samples with the declaration of pecans, Brazil nuts, other different nut species or nuts generally. In 15 food samples pecans and Brazil nuts allergens were identified in the conformity with the food declaration. CONCLUSION: The presented multiplex PCR method is specific enough and can be used as a fast approach for the detection of major allergens of pecan or Brazil nuts in food. Copyright © 2011 Society of Chemical Industry     

VARIATION IN THE SELENIUM CONTENT OF INDIVIDUAL BRAZIL NUTS

Brazil nuts are high in selenium content relative to other human foods. Since the limits between human selenium essentiality and toxicity are narrow, it is important to know the variation in selenium content of individual nuts. Analysis was performed on 72 individual nuts obtained in stores as shelled nuts in bulk and shelled and unshelled nuts in packages. Their average selenium content was 14.66 ppm with a range of 0.2 to 253.0. The agronomic and health aspects of these findings are discussed.     

Speciation of Se in Bertholletia excelsa (Brazil nut): A hard nut to crack?

A separation method based on ion-pairing liquid chromatography was combined with both elemental (inductively coupled plasma mass spectrometry (ICP-MS)) and molecular (electrospray tandem mass spectrometry (ES-MS-MS)) mass spectrometry in order to unravel the identity of the Se-species present in the complex matrix of Brazil nuts rich in Se. Via enzymatic digestion, Se-species were released from the matrix. Subsequently the species were separated and the Se was monitored on-line by ICP-MS. By HPLC–ES-MS-MS, the species were identified based on their molecular mass and their specific product ions. The main compound was identified as Se-Methionine. Another compound was identified as Se-Cystine, partly on the basis of the isotopic pattern of Se. This research was further extended to the analyses of in vitro gastrointestinal digests of the Brazil nuts. These digests were analyzed for their Se-content and screened for the presence of the different Se-species by HPLC–ICP-MS. In both the gastric and the intestinal digests, we were able to identify the Se-species as Se-Methionine and Se-Cystine by HPLC–ES-MS-MS. By coupling HPLC to both elemental and molecular mass spectrometry, the species present in Brazil nuts and supposedly extractable by our body were fully characterized.     

Predominant mycobiota and aflatoxin content in Brazil nuts

The Brazil nut (Bertholletia excelsa) is an economically important product to the Brazilian Amazon. Currently, its marketing is being affected by the high incidence of aflatoxins (AF) produced by potentially aflatoxigenic fungi associated with its seeds. In this context, the purpose of this study was to determine which part of the nut contributes to contamination by aflatoxins and to identify the mycobiota in Brazil nut samples. Unshelled and shelled nuts were analyzed by measuring the total count of filamentous fungi (Aspergillus sections Flavi, Nigri and Circumdati) in sanitised and non-sanitised treatments. The isolates identified as Aspergillus section Flavi, the major producers of AF, were plated for determination of their aflatoxigenic potential. To perform the AF analysis, samples of Brazil nuts were treated separately. The AF from the shell and kernel were extracted by chloroform and analysed by the HPLC-FD system in isocratic mode. The Aspergillus section Flavi count was 21.67% lower. The production of AF by the isolated fungi was 30% for sanitised and 23.8% for non-sanitised samples. The concentrations obtained of AFB₁ and AFG₁ were higher than those of AFB₂ and AFG₂. The AFB₁ concentrations of shelled nuts and shell samples were 35.0 and 1.78 μg/kg, respectively. AFB₂ and AFG₂ were detected only in shelled nut samples. The HPLC-FD analysis presented limits of detection (LOD) and quantification (LOQ) of 0.2 and 0.4 μg/kg, respectively.     

Predominant mycobiota and aflatoxin content in Brazil nuts

The Brazil nut (Bertholletia excelsa) is an economically important product to the Brazilian Amazon. Currently, its marketing is being affected by the high incidence of aflatoxins (AF) produced by potentially aflatoxigenic fungi associated with its seeds. In this context, the purpose of this study was to determine which part of the nut contributes to contamination by aflatoxins and to identify the mycobiota in Brazil nut samples. Unshelled and shelled nuts were analyzed by measuring the total count of filamentous fungi (Aspergillus sections Flavi, Nigri and Circumdati) in sanitised and non-sanitised treatments. The isolates identified as Aspergillus section Flavi, the major producers of AF, were plated for determination of their aflatoxigenic potential. To perform the AF analysis, samples of Brazil nuts were treated separately. The AF from the shell and kernel were extracted by chloroform and analysed by the HPLC-FD system in isocratic mode. The Aspergillus section Flavi count was 21.67% lower. The production of AF by the isolated fungi was 30% for sanitised and 23.8% for non-sanitised samples. The concentrations obtained of AFB₁ and AFG₁ were higher than those of AFB₂ and AFG₂. The AFB₁ concentrations of shelled nuts and shell samples were 35.0 and 1.78???g/kg, respectively. AFB₂ and AFG₂ were detected only in shelled nut samples. The HPLC-FD analysis presented limits of detection (LOD) and quantification (LOQ) of 0.2 and 0.4???g/kg, respectively.     

Bioactive peptides from nuts: A review

Oxidative stress occurs because of an imbalance in the production of reactive oxygen species. Nuts are rich in bioactive compounds, including phenolic compounds, tannins, and phytosterols. Although nuts are widely consumed because of their beneficial effects on nutrition and health, there is limited information about bioactive peptides from nuts. Pine nut and walnut-derived peptides are the most studied because these nuts contain a higher amount of protein. Different biological activities have been demonstrated for nut peptides; many of them exhibit antioxidant, anticancer, antihypertensive, antidiabetic, immunomodulatory, antiseizure, or neuroprotective activity. Recent studies have focused on increasing the bioactivity of identified bioactive peptides by applying new technologies and chemosynthetic strategies. Research tendency points to the generation of peptides with specific sequences for application in specific diseases. Nut bioactive peptides can become key functional ingredients for food, pharmaceuticals, or cosmetics.     

Microbiological and aflatoxin evaluation of Brazil nut pods and the effects of unit processing operations

Harvesting of Brazil nuts not only helps to preserve the Amazon rainforest but also provides income to individuals who would otherwise have little means of making a livelihood. Recently, the European Community has tightened the quality requirements for Brazil nuts, particularly with regard to aflatoxin levels and microbiological contamination. The objectives of this research were to gain a better understanding of the origin of aflatoxins on Brazil nuts and to microbiologically evaluate some of the operations involved in processing. In this regard, five Brazil nut pods were aseptically picked from trees located in each of three concessions of the Peruvian Amazon rainforest (Madre de Dios province). The exteriors of the pods and the nuts were examined for yeast and molds, including Aspergillus flavus and Aspergillus parasiticus, and for bacteria, including Salmonella and Escherichia coli. Brazil nuts obtained from various commercial process operations located in Peru were similarly evaluated. Exteriors of all Brazil nut pods did not contain A. parasiticus, and only pods from one concession yielded A. flavus isolates. All isolates tested were aflatoxigenic (630 to 915 ppb total aflatoxin). Coliforms, E. coli, and salmonellae were not recovered from any of the pods. Whole, in-shell nuts obtained after opening the pods yielded no A. flavus or A. parasiticus. Aflatoxins were not detected (detection limit 1.75 ppb) in any of the nuts. Whole, in-shell and shelled nuts from various process operations were all positive for A. flavus but negative for E. coli and salmonellae. Soaking of whole, in-shell nuts before cracking or shelling increased coliform numbers, whereas levels of A. flavus decreased. In order to gain a better understanding of the sanitary performance of the unit process operations, additional evaluations should be conducted on product lots processed on different days. Also, the microbiology of product processed from common lots should be followed through the various unit operations and compared.     

Processing effects on tree nut allergens: A review

“Tree nut” is a broad term for classification of nuts that include cashews, almonds, hazelnuts, etc. Reports of mild to adverse immune reactions following the consumption of these nuts has been on a rise in recent years. Currently, about 1.2–2% of the world's population suffer from sensitivity to tree nuts. The only solution is complete abstinence from the allergy causing tree nut which is not feasible in most cases due to issues like cross contamination or their presence in the form of hidden ingredients in processed foods. Various studies have shown that food processing can effectively vary the secondary structures of the allergenic protein which in turn influences their functional properties. But, the impact of these processing methods on tree nuts allergens is mixed. This review gives an update on the recent findings on how conventional and novel processing methods influence the tree nut allergens.     

坚果的主要生物活性成分及其保健作用

坚果作为高档零食,其营养价值和保健功能也越来越受到人们的关注?本论文综述了常见坚果(如澳洲坚果、核桃、榛子、杏仁、松子、板栗、山核桃、开心果,日本核桃、巴西坚果、花生等)中的酚类(原花青素、酚酸、黄酮、单宁、烷基酚、维生素E等)?酸类?甾醇类、色素(类胡萝卜素、叶绿素等)、硒、糖苷、功能性脂肪酸等营养物质和生物活性成分,并阐述了食用坚果与预防心血管疾病?延年益寿?预防糖尿病?控制体重等保健作用的关系?这对于进一步研究坚果与健康的关系、正确挑选与消费坚果均具有较好的指导意义。     

Investigation of major and trace elements and their distributions between lipid and non-lipid fractions in Brazil nuts by inductively coupled plasma atomic optical spectrometry

Major (Ca, Mg, P) and trace (As, Al, Ba, Cd, Cr, Cu, Fe, Hg, Mn, Ni, Se, Zn) elements were determined in Brazil nuts by means of inductively coupled plasma atomic optical spectrometry (ICP-OES) and five of them were measured for the first time. For measurement of As, Hg and Se levels, hydride generation was used as the sample treatment method. The element concentrations were compared with recommended dietary allowances and upper tolerable levels. The distributions of the elements between lipid and lipid-free fractions were investigated with the use of solvent extraction. Two extractants (petroleum ether and chloroform:methanol 2:1) were applied. Most of the Cr, Fe and Ni contents were found in the lipid fraction, while Ba, Ca, Cu, Mn, P and Zn were mainly bonded with defatted nut residue. Al, Mg, Se, Sr were only present in the defatted fraction.     

Phenolic compounds and antioxidant activity of Brazil nut (Bertholletia excelsa)

Brazil nuts were shelled and separated as kernel and brown skin; whole nuts were also used. Soluble phenolics from each portion as well as the whole nut were extracted using 70% acetone under reflux conditions. Insoluble-bound phenolics were subsequently extracted into diethyl ether–ethyl acetate mixture (1:1, v/v) after alkaline hydrolysis. Both soluble and insoluble-bound phenolic extracts were separately examined for their total phenolics content; antioxidant activities were evaluated by trolox equivalent antioxidant capacity (TEAC), 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and hydroxyl radical scavenging activities using electron paramagnetic resonance (EPR), reducing power, and oxygen radical scavenging capacity (ORAC). Soluble phenolics in brown skin was 1236.07 as compared to 406.83 in kernel and 519.11 mg/100 g in whole nut. Bound phenolics content of brown skin was also 86- and 19-folds higher than kernel and whole nut, respectively. Similarly extracts from the brown skin exhibited the highest antioxidant activity. Free- and bound phenolics were identified and quantified; these included nine phenolic acids and flavonoids and their derivatives (gallic acid, gallocatechin. protocatechuic acid, catechin, vanillic acid, taxifolin, myricetin, ellagic acid, and quercetin). However, some phenolics were present only in the bound form. Furthermore, the phenolics were dominant in the brown skin.     

Improvement of a sample preparation procedure for multi-elemental determination in Brazil nuts by ICP-OES

Various sample preparation procedures, such as common wet digestions and alternatives based on solubilisation in aqua regia or tetramethyl ammonium hydroxide, were compared for the determination of the total Ba, Ca, Cr, Cd, Cu, Fe, Mg, Mn, Ni, P, Pb, Se, Sr and Zn contents in Brazil nuts using inductively coupled plasma optical emission spectrometry (ICP-OES). For measurement of Se, a hydride generation technique was used. The performance of these procedures was measured in terms of precision, accuracy and limits of detection of the elements. It was found that solubilisation in aqua regia gave the best results, i.e. limits of detection from 0.60 to 41.9 ng ml^?1, precision of 1.0–3.9% and accuracy better than 5%. External calibration with simple standard solutions could be applied for the analysis. The proposed procedure is simple, reduces sample handling, and minimises the time and reagent consumption. Thus, this can be a vital alternative to traditional sample treatment approaches based on the total digestion with concentrated reagents. A phenomenon resulting from levels of Ba, Se and Sr in Brazil nuts was also discussed.     

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.     

电感耦合等离子体质谱法测定食品中硒元素的条件优化

目的优化电感耦合等离子体质谱法(inductively coupled plasma mass spectrometer, ICP-MS)测定食品中硒元素含量的条件,并对河北省食品中硒元素含量情况进行调查分析。方法选取8种不同基质的国家标准物质作为检测样品对实验条件进行优化,同时选择80Se作为检测目标,采用碰撞模式并加入内标和5%异丙醇的优化条件下对样品中硒元素含量进行检测。通过对比结果,分析河北省膳食样品中硒元素含量,对居民通过膳食补充硒元素给予指导。结果通过采用碰撞模式及内标添加法可以有效降低基质干扰,同时加入异丙醇可极大提高硒元素的信号强度;其在5.00~100.00μg/L范围内呈现良好的线性关系(r2=0.999),检出限为0.002mg/kg;坚果及籽类、豆类及豆制品和肉及肉制品含硒较高,坚果及籽类样品中鲍鱼果含量最高为1.472mg/kg,豆类及豆制品中黄豆含量最高为0.123mg/kg,肉及肉制品中鸭肉含量最高为0.169 mg/kg。结论在检测的6类食品中,坚果及籽类、豆类及豆制品和肉及肉制品中硒含量相对较高,缺硒人群可适量增加食用。     

Barcode High Resolution Melting analysis for forensic uses in nuts: A case study on allergenic hazelnuts (Corylus avellana)

Tree nuts are nutritionally important foods, being rich sources of vitamins, minerals, healthy fats and proteins and having cardio protective effects. Despite their healthy attributes, some tree nuts are also known for triggering adverse immune responses to individuals. Thus, an ongoing challenge in molecular biology is to develop rapid, accurate and reliable techniques to identify organisms and their processed foods, particularly those which could be contaminated with allergenic compounds. Lately, in addition to different chemical and immunological methods used, DNA based methods were also applied for this purpose. We describe here a novel, specific and highly sensitive High Resolution Melting analysis based method, using chloroplast barcoding regions (Bar-HRM) in order to a) obtain barcoding information for the major tree nut species (we have also included peanuts and sesame in our study) and b) to quantitatively identify one such allergenic tree nut species, like hazelnut in processed foods. The results show that the proposed method i) is capable of distinguishing among different tree nut species, ii) can be applied to processed tree nut products such as biscuits and iii) can detect a ratio of 0.01% of hazelnut in biscuits. In conclusion, it is demonstrated that BAR-HRM is suitable for the rapid, closed-tube and accurate screening of large numbers of tree nuts and/or of their processed foods and for the identification of allergenic tree nut species contained in processed foods.     

Development of a Competitive ELISA for the Detection of Pecan (Carya illinoinensis (Wangenh.) K. Koch) Traces in Food

Pecan nuts are included among the so-called big-eight food allergens, which are responsible for 90% of food allergies. However, unlike other tree nuts such as Brazil nut and hazelnut, studies on pecan from the point of view of allergies are scarce, and only a few analytical methods to detect its presence in food have been published. The objective of the present study was to develop an enzyme-linked immunosorbent assay (ELISA) method to detect traces of pecan in foods. Therefore, pecan-specific polyclonal antibodies were developed in rabbits and characterized for their specificity to pecan proteins and for their selectivity in challenging studies with other food ingredients and allergens. In addition, the research was complemented with more specific analyses like sodium dodecyl sulfate polyacrylamide gel electrophoresis to characterize the protein standard and western blot to identify the reactive proteins. By applying the selected conditions, the data were fitted to a four parameter logistic curve, which gave a R square of 0.999, and IC50 and IC80 values of 5.0 and 2.2 ng/mL, respectively. This highly sensitive method allows the detection of trace amounts of pecan protein within complex food matrices, which can provide the food industry with a useful tool to comply with the requirements enforced by regulatory agencies.     

Nuts, especially walnuts, have both antioxidant quantity and efficacy and exhibit significant potential health benefits

Free and total (after basic hydrolysis) polyphenols in nine types of raw and roasted nuts and two types of peanut butter (54 commercial samples) were analyzed after methanol extraction by a single step Folin-Ciocalteu reagent using catechin as standard. Walnuts had the highest free and total polyphenols in both the combined raw and roasted samples. Total polyphenols in the nuts were significantly higher than free polyphenols. Roasting had little effect on either free or total polyphenols in nuts. Raw and roasted walnuts had the highest total polyphenols. The efficacy of raw and roasted nut antioxidants was assessed by measuring the ability of the free polyphenol nut extracts to inhibit the oxidation of lower density lipoproteins (LDL + VLDL). A nut polyphenol, catechin, was measured after binding of three nut extracts to lower density lipoproteins. Walnut polyphenols had the best efficacy among the nuts and also the highest lipoprotein-bound antioxidant activity. Based on USDA availability data, the per capita total polyphenols was 162 mg from nuts per day in 2008. This corresponds to 19% of the total polyphenols from fruits and vegetables, nuts, grains, oils and spices in the US diet. Nuts provided 158 mg of polyphenols per day to the European Union diet. Nuts are high in polyphenol antioxidants which by binding to lipoproteins would inhibit oxidative processes that lead to atherosclerosis in vivo. In human supplementation studies nuts have been shown to improve the lipid profile, increase endothelial function and reduce inflammation, all without causing weight gain. These qualities make nuts a nutritious healthy snack and food additive.