Characterization and differentiation of diverse transgenic and nontransgenic soybean varieties from CE protein profiles

Nowadays, soybeans are commercialized in a wide variety of colors and tones. Moreover, some pigmented seeds are being commercialized as soybeans while, on other occasions, these seeds are labeled as mung beans, azuki beans or soybean frijoles generating confusion on their identity. In this work, CE has been applied for the first time for the characterization and differentiation of different pigmented beans commercialized as soybeans. Other seeds commercialized as azuki, mung green soybeans or soybean frijoles were also analyzed. Borate buffer (at pH 8.5) containing 20% v/v ACN was used as the separation media and solution containing ACN/water (75:25 v/v) with 0.3% v/v acetic acid was used to solubilize the proteins from the samples. A 50 cm bare fused-silica capillary was employed for obtaining adequate separations in about 12 min. The CE protein pattern observed for yellow soybeans was different from that corresponding to green and red soybeans. The seeds commercialized as black soybean presented electropherograms identical or similar to those yielded by the yellow seeds with the exception of the sample labeled as black soybeans frijoles that presented a totally different pattern. In addition, CE protein profiles obtained for azuki and mung green soybeans were very similar to those corresponding to red soybeans and green soybeans, respectively. Finally, the CE method was also applied to differentiate transgenic and nontransgenic soybean varieties. Discriminant analysis, using several protein peak areas as variable, was used to successfully classify these samples.     

Capillary electrophoresis for the analysis of food proteins of animal origin

In recent years, capillary electrophoresis (CE) has become an analytical technique with many applications in the study of food proteins and peptides. This review describes the existing CE methods of analysis of milk, egg, meat and fish proteins and peptides. The major developments in the application of CE to solve different problems in food technology, such as the assessment of technological processes, quality, and authenticity control of animal foods, are considered. A section dealing with future directions on the analysis of food proteins by CE is also included.     

Detection and quantitation of additions of soybean proteins in cured-meat products by perfusion reversed-phase high-performance liquid chromatography

Perfusion liquid chromatography has been applied in this work to the determination of soybean proteins in commercially available cured meat products, enabling the detection of additions of soybean proteins in cured meat products to which the addition of these vegetable proteins is forbidden and the quantitation of soybean proteins in cured meat products to which the addition of these proteins is allowed up to a certain limit. The analytical methodology is based on a sample treatment (fat extraction and soybean protein solubilization) prior to chromatographic analysis. Fat extraction with acetone and soybean protein solubilization with a buffer solution at basic pH (pH 10 or 9) were necessary to obtain selective and sensitive conditions. Use of water-acetonitrile-trifluoroacetic acid or water-tetrahydrofuran-trifluoroacetic acid linear binary gradients at a flow rate of 3 mL/min, a temperature of 50 degrees C, and UV detection at 280 nm enabled chromatographic analysis of soybean proteins in cured meat products in less than 3 min.     

Measurement of Antioxidant Capacity of Meat and Meat Products: Methods and Applications

At present, a wide variety of analytical methods is available to measure antioxidant capacity. However, this great diversity is not reflected in the analysis of meat and meat products, as there are a limited number of studies on determining this parameter in this complex food matrix. Despite this, and due to the interest in antioxidants that prevent oxidation reactions, the identification of antioxidants in meat and meat products is of special importance to the meat industry. For this reason, this review compiled the main antioxidant capacity assays employed in meat and meat products, to date, describing their foundations, and showing both their advantages and limitations. This review also looked at the different applications of antioxidant properties in meat and meat products. In this sense, the suitability of using these methodologies has been demonstrated in different investigations related to these foods.     

Detection of microbial food contaminants and their products by capillary electromigration techniques

This work reviews the different analytical strategies based on capillary electromigration techniques developed for detecting microbial contaminants and their products in food matrices. Namely, this work presents an exhaustive and critical review, including works published till March 2007, on CE methods developed to detect and identify contaminants of microbial origin that represent a hazard to humans in foods. First, an overview on the strategies adopted for the analysis of intact microorganisms is presented. Next, CE methodologies based on the analysis of microbial constituents, including those based on genomics and proteomics approaches, are described. Finally, CE methods developed for detecting microbial toxins are discussed.     

CE: a useful analytical tool for the characterization of Maillard reaction products in foods

The Maillard reaction (MR) is a complex series of nonenzymatic reactions between reducing compounds and amines, amino acids, peptides, or proteins that play an important role in the formation of flavors and colors in foods during processing and storage. Also, the antioxidant properties of some Maillard reaction products (MRP) was an additional benefit reported. On the other hand, these reactions decrease the nutritional quality of foods and may result in the formation of toxic MRP. Although, research to assess the risks and benefits associated with the consumption of MRP in the diet is still awaiting for new analytical methodologies to be developed. Structural characterization of MRP has been very challenging due to the chemical diversity of these compounds which present a wide range of polarities and molecular weights, making analyses difficult. CE is a technique that has gained popularity for the separation of complex mixtures that have otherwise proved difficult to analyze. Thus, the purpose of this overview is to give the reader an appreciation of some of the CE analytical developments on the analysis of MRP in model systems and foods, and to address the potential of CE on the characterization of this complex group of compounds.     

The application of capillary electrophoresis to the analysis of vitamins in food and beverages

Capillary electrophoretic (CE) methods have been used to separate and determine a wide range of water-soluble vitamins in pharmaceutical preparations, but has found limited application in determining vitamins at naturally occurring levels in food and beverages. CE has been used to determine vitamin C in fruits and beverages, niacin in a range of foods and thiamine in samples of meat and milk. The CE methodologies used to determine vitamins in pharmaceutical preparations and biological fluids are also included as examples of the effectiveness of CE in vitamin analysis.     

Review: Authentication and traceability of foods from animal origin by polymerase chain reaction-based capillary electrophoresis

This work presents an overview of the applicability of PCR-based capillary electrophoresis (CE) in food authentication and traceability of foods from animal origin. Analytical approaches for authenticating and tracing meat and meat products and fish and seafood products are discussed. Particular emphasis will be given to the usefulness of genotyping in food tracing by using CE-based genetic analyzers.     

Characterization of nutraceuticals and functional foods by innovative HPLC methods

In recent years there is a growing interest in food and food ingredient which may provide health benefits. Food as well as food ingredients containing health-preserving components, are not considered conventional food, but can be defined as functional food. To characterise such foods, as well as nutraceuticals specific, high sensitive and reproducible analytical methodologies are needed. In light of this importance we set out to develop innovative HPLC methods employing reversed phase narrow bore column and high-performance anion-exchange chromatographic methods coupled with pulsed amperometric detection (HPAEC-PAD), which are specific for carbohydrate analysis. The developed methods were applied for the separation and quantification of citrus flavonoids and to characterize fructooligosaccharide (FOS) and fructans added to functional foods and nutraceuticals.     

High Performance Liquid Chromatography versus Stacking-Micellar Electrokinetic Chromatography for the Determination of Potentially Toxic Alkenylbenzenes in Food Flavouring Ingredients

Alkenylbenzenes, including eugenol, methyleugenol, myristicin, safrole, and estragole, are potentially toxic phytochemicals, which are commonly found in foods. Occurrence data in foods depends on the quality of the analytical methodologies available. Here, we developed and compared modern reversed-phase high performance liquid chromatography (HPLC) and stacking-micellar electrokinetic chromatography (MEKC) methods for the determination of the above alkenylbenzenes in food flavouring ingredients. The analytical performance of HPLC was found better than the stacking-MEKC method. Compared to other HPLC methods found in the literature, our method was faster (total run time with conditioning of 15 min) and able to separate more alkenylbenzenes. In addition, the analytical methodology combining an optimized methanol extraction and proposed HPLC was then applied to actual food flavouring ingredients. This methodology should be applicable to actual food samples, and thus will be vital to future studies in the determination of alkenylbenzenes in food.     

Analysis of food proteins and peptides by chromatography and mass spectrometry

The research topics and the analytical strategies dealing with food proteins and peptides are summarized. Methods for the separation and purification of macromolecules of food concern by both high-performance liquid chromatography (HPLC) on conventional packings and perfusion HPLC are examined. Special attention is paid to novel methodologies such those based on multi-dimensional systems that comprise liquid-phase based protein separation, protein digestion and mass spectrometry (MS) analysis of food peptide and proteins. Recent applications of chromatography and MS-based techniques for the analysis of proteins and peptides in food are discussed.     

CE methods applied to the analysis of micronutrients in foods

This article reviews the applications of CE that are relevant to the analysis of small molecules in foods. CE has been applied to a wide range of important areas of food analysis and is rapidly being established as an alternative technique to chromatographic methods including HPLC and GC within analytical food and research laboratories. In recent years the analysis of food by CE has become more frequent and important and as such a variety of compounds have been separated and quantified. Although many other analytes have been detected by CE, this review will highlight areas relating primarily to the rather broad chemical classes of free amino acids, carbohydrates, organic acids, vitamins and a variety of antioxidants. In addition, information relating to the analyte, sample matrix, mode of CE employed, scope of the methodology and the detection and derivatization of the small molecules are considered and discussed.     

Recent Discoveries in the Field of Lipid Bio-Based Ingredients for Meat Processing

Current culture and pace of lifestyle, together with consumer demand for ready-to-eat foods, has influenced the food industry, particularly the meat sector. However, due to the important role that diet plays in human health, consumers demand safe and healthy food products. As a consequence, even foods that meet expectations for convenience and organoleptic properties must also meet expectations from a nutritional standpoint. One of the main nutritionally negative aspects of meat products is the content and composition of fat. In this sense, the meat industry has spent decades researching the best strategies for the reformulation of traditional products, without having a negative impact in technological processes or in the sensory acceptance of the final product. However, the enormous variety of meat products as well as industrial and culinary processes means that a single strategy cannot be established, despite the large volume of work carried out in this regard. Therefore, taking all the components of this complex situation into account and utilizing the large amount of scientific information that is available, this review aims to comprehensively analyze recent advances in the use of lipid bio-based materials to reformulate meat products, as well as their nutritional, technological, and sensorial implications.     

A metallomics approach discovers selenium-containing proteins in selenium-enriched soybean

Our previous study found that high-molecular-weight selenium (Se) species make up 82% of the total Se in the bean of Se-enriched soybean plants (Chan et al. 2010, Metallomics, 2(2): p. 147–153). The Se species have been commonly seen in other plants in addition to soybean, but their identities remain unresolved. The present study employs a multi-technique metallomics approach to characterize the proteins containing Se in the beans of Se-enriched soybean plants. Two main categories of proteins, maturation proteins and protease inhibitors, were found in Se-containing high-performance liquid chromatography (HPLC) fractions. The proteins were screened by two-dimensional HPLC-inductively coupled plasma mass spectrometry, size-exclusion chromatography, and anion-exchange chromatography, and the Se-containing fractions were then identified by peptide mapping using HPLC-Chip-electrospray ion trap mass spectrometry. Based on the belief that Se goes into proteins through non-specific incorporation, a new method was designed and applied for the Se-containing peptide identification. The Se-containing peptide KSDQSSSYDDDEYSKPCCDLCMCTRS, part of the sequence of protein Bowman–Birk proteinase isoinhibitor (Glycine max), was found in one of the Se-containing fractions. The nutritional value of the Se-containing proteins in Se-enriched soybeans will be an interesting topic for the future studies.     

Application of Pomegranate by-Products in Muscle Foods: Oxidative Indices,Colour Stability,Shelf Life and Health Benefits

In recent years, considerable importance is given to the use of agrifood wastes as they contain several groups of substances that are useful for development of functional foods. As muscle foods are prone to lipid and protein oxidation and perishable in nature, the industry is in constant search of synthetic free additives that help in retarding the oxidation process, leading to the development of healthier and shelf stable products. The by-products or residues of pomegranate fruit (seeds, pomace, and peel) are reported to contain bioactive compounds, including phenolic and polyphenolic compounds, dietary fibre, complex polysaccharides, minerals, vitamins, etc. Such compounds extracted from the by-products of pomegranate can be used as functional ingredients or food additives to harness the antioxidant, antimicrobial potential, or as substitutes for fat, and protein in various muscle food products. Besides, these natural additives are reported to improve the quality, safety, and extend the shelf life of different types of food products, including meat and fish. Although studies on application of pomegranate by-products on various foods are available, their effect on the physicochemical, oxidative changes, microbial, colour stabilizing, sensory acceptability, and shelf life of muscle foods are not comprehensively discussed previously. In this review, we vividly discuss these issues, and highlight the benefits of pomegranate by-products and their phenolic composition on human health.     

The development and application of capillary electrophoresis methods for food analysis

Capillary electrophoresis (CE) offers the analyst a number of key advantages for the analysis of the components of foods. CE offers better resolution than, say, high-performance liquid chromatography (HPLC), and is more adept at the simultaneous separation of a number of components of different chemistries within a single matrix. In addition, CE requires less rigorous sample cleanup procedures than HPLC, while offering the same degree of automation. However, despite these advantages, CE remains under-utilized by food analysts. Therefore, this review consolidates and discusses the currently reported applications of CE that are relevant to the analysis of foods. Some discussion is also devoted to the development of these reported methods and to the advantages/disadvantages compared with the more usual methods for each particular analysis. It is the aim of this review to give practicing food analysts an overview of the current scope of CE.     

Multispecies Identification of Oilseed- and Meat-Specific Proteins and Heat-Stable Peptide Markers in Food Products

Consumer demand for both plant products and meat products enriched with plant raw materials is constantly increasing. Therefore, new versatile and reliable methods are needed to find and combat fraudulent practices in processed foods. The objective of this study was to identify oilseed species-specific peptide markers and meat-specific markers that were resistant to processing, for multispecies authentication of different meat and vegan food products using the proteomic LC-MS/MS method. To assess the limit of detection (LOD) for hemp proteins, cooked meatballs consisting of three meat species and hemp cake at a final concentration of up to 7.4% were examined. Hemp addition at a low concentration of below 1% was detected. The LOD for edestin subunits and albumin was 0.9% (w/w), whereas for 7S vicilin-like protein it was 4.2% (w/w). Specific heat-stable peptides unique to hemp seeds, flaxseed, nigella, pumpkin, sesame, and sunflower seeds, as well as guinea fowl, rabbit, pork, and chicken meat, were detected in different meat and vegan foods. Most of the oilseed-specific peptides were identified as processing-resistant markers belonging to 11S globulin subunits, namely conlinin, edestin, helianthinin, pumpkin vicilin-like or late embryogenesis proteins, and sesame legumin-like as well as 2S albumins and oleosin isoforms or selected enzymic proteins.     

Recent advances in the application of capillary electromigration methods for food analysis

This article reviews the latest developments in the application of capillary electromigration methods for the analysis of foods and food components. Nowadays, methods based on CE techniques are becoming widely used in food analytical and research laboratories. This review covers the application of CE to analyze amino acids, biogenic amines, peptides, proteins, DNAs, carbohydrates, phenols, polyphenols, pigments, toxins, pesticides, vitamins, additives, small organic and inorganic ions, chiral compounds, and other compounds in foods, as well as to investigate food interactions and food processing. The use of microchips as well as other foreseen trends in CE analysis of foods is discussed. Papers that were published during the period June 2002-June 2005 are included following the previous review by Frazier and Papadopoulou (Electrophoresis 2003, 24, 4095-4105).     

Chromatographic determination of amino acids in foods

Amino acids in foods exist in a free form or bound in peptides, proteins, or nonpeptide bonded polymers. Naturally occurring L-amino acids are required for protein synthesis and are precursors for essential molecules, such as co-enzymes and nucleic acids. Nonprotein amino acids may also occur in animal tissues as metabolic intermediates or have other important functions. The development of bacterially derived food proteins, genetically modified foods, and new methods of food processing; the production of amino acids for food fortification; and the introduction of new plant food sources have meant that protein amino acids and amino acid enantiomers in foods can have both nutritional and safety implications for humans. There is, therefore, a need for the rapid and accurate determination of amino acids in foods. Determination of the total amino acid content of foods requires protein hydrolysis by various means that must take into account variations in stability of individual amino acids and resistance of different peptide bonds to the hydrolysis procedures. Modern methods for separation and quantitation of free amino acids either before or after protein hydrolysis include ion exchange chromatography, high performance liquid chromatography (LC), gas chromatography, and capillary electrophoresis. Chemical derivatization of amino acids may be required to change them into forms amenable to separation by the various chromatographic methods or to create derivatives with properties, such as fluorescence, that improve their detection. Official methods for hydrolysis and analysis of amino acids in foods for nutritional purposes have been established. LC is currently the most widely used analytical technique, although there is a need for collaborative testing of methods available. Newer developments in chromatographic methodology and detector technology have reduced sample and reagent requirements and improved identification, resolution, and sensitivity of amino acid analyses of food samples.     

Simple and rapid characterization of soybean cultivars by perfusion reversed-phase HPLC: application to the estimation of the 11S and 7S globulin contents

A perfusion RP HPLC method enabling the separation of soybean proteins in an analysis time lower than 3 min has been used to obtain the chromatographic profiles of different soybean cultivars. The chromatograms obtained for each soybean variety presented clear differences that justified the potential use of this method for cultivar characterization. The area percentages obtained were employed as variables for cluster and principal components analysis of these soybeans. The application of these multivariate methods enabled the grouping of the soybeans in different categories. The protein fractions obtained from these soybeans by the application of a fractionation method were also analyzed. The chromatographic profiles obtained enabled the assignment of peaks to the main soybean proteins (7S and 11S globulins). These data were used for the estimation, for the first time, of the 7S and 11S globulin contents in soybean cultivars.