Compositions of Graphite/Epoxy composites Biosensor.
\\n\\n
Released this past November, the list is based on data collected from the Web of Science and highlights some of the world’s most influential scientific minds by naming the researchers whose publications over the previous decade have included a high number of Highly Cited Papers placing them among the top 1% most-cited.
\\n\\nWe wish to congratulate all of the researchers named and especially our authors on this amazing accomplishment! We are happy and proud to share in their success!
Note: Edited in March 2021
\\n"}]',published:!0,mainMedia:{caption:"Highly Cited",originalUrl:"/media/original/117"}},components:[{type:"htmlEditorComponent",content:'IntechOpen is proud to announce that 191 of our authors have made the Clarivate™ Highly Cited Researchers List for 2020, ranking them among the top 1% most-cited.
\n\nThroughout the years, the list has named a total of 261 IntechOpen authors as Highly Cited. Of those researchers, 69 have been featured on the list multiple times.
\n\n\n\nReleased this past November, the list is based on data collected from the Web of Science and highlights some of the world’s most influential scientific minds by naming the researchers whose publications over the previous decade have included a high number of Highly Cited Papers placing them among the top 1% most-cited.
\n\nWe wish to congratulate all of the researchers named and especially our authors on this amazing accomplishment! We are happy and proud to share in their success!
Note: Edited in March 2021
\n'}],latestNews:[{slug:"webinar-introduction-to-open-science-wednesday-18-may-1-pm-cest-20220518",title:"Webinar: Introduction to Open Science | Wednesday 18 May, 1 PM CEST"},{slug:"step-in-the-right-direction-intechopen-launches-a-portfolio-of-open-science-journals-20220414",title:"Step in the Right Direction: IntechOpen Launches a Portfolio of Open Science Journals"},{slug:"let-s-meet-at-london-book-fair-5-7-april-2022-olympia-london-20220321",title:"Let’s meet at London Book Fair, 5-7 April 2022, Olympia London"},{slug:"50-books-published-as-part-of-intechopen-and-knowledge-unlatched-ku-collaboration-20220316",title:"50 Books published as part of IntechOpen and Knowledge Unlatched (KU) Collaboration"},{slug:"intechopen-joins-the-united-nations-sustainable-development-goals-publishers-compact-20221702",title:"IntechOpen joins the United Nations Sustainable Development Goals Publishers Compact"},{slug:"intechopen-signs-exclusive-representation-agreement-with-lsr-libros-servicios-y-representaciones-s-a-de-c-v-20211123",title:"IntechOpen Signs Exclusive Representation Agreement with LSR Libros Servicios y Representaciones S.A. de C.V"},{slug:"intechopen-expands-partnership-with-research4life-20211110",title:"IntechOpen Expands Partnership with Research4Life"},{slug:"introducing-intechopen-book-series-a-new-publishing-format-for-oa-books-20210915",title:"Introducing IntechOpen Book Series - A New Publishing Format for OA Books"}]},book:{item:{type:"book",id:"6979",leadTitle:null,fullTitle:"Parasites and Parasitic Diseases",title:"Parasites and Parasitic Diseases",subtitle:null,reviewType:"peer-reviewed",abstract:"Parasitic diseases are considered nowadays as an important public health problem due to the high morbidity and mortality rates registered in the world. These diseases result in more severe consequences for the social order of tropical and subtropical countries because many of them have low economic income that makes it even more difficult to design and implement health control programs. This situation opens the door to the emergence and reemergence of these diseases; therefore, it is convenient, necessary, and essential to study and update the epidemiological behavior of tropical diseases with the objective of offering official health professionals and institutions current information for decision-making in this area to ensure social welfare.",isbn:"978-1-83880-128-1",printIsbn:"978-1-83880-127-4",pdfIsbn:"978-1-83962-140-6",doi:"10.5772/intechopen.73726",price:100,priceEur:109,priceUsd:129,slug:"parasites-and-parasitic-diseases",numberOfPages:98,isOpenForSubmission:!1,isInWos:null,isInBkci:!1,hash:"f55304c8bd1d92268e33689c368f9e33",bookSignature:"Gilberto Bastidas",publishedDate:"April 24th 2019",coverURL:"https://cdn.intechopen.com/books/images_new/6979.jpg",numberOfDownloads:6658,numberOfWosCitations:2,numberOfCrossrefCitations:6,numberOfCrossrefCitationsByBook:0,numberOfDimensionsCitations:14,numberOfDimensionsCitationsByBook:0,hasAltmetrics:1,numberOfTotalCitations:22,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"March 27th 2018",dateEndSecondStepPublish:"April 17th 2018",dateEndThirdStepPublish:"June 16th 2018",dateEndFourthStepPublish:"September 4th 2018",dateEndFifthStepPublish:"November 3rd 2018",currentStepOfPublishingProcess:5,indexedIn:"1,2,3,4,5,6",editedByType:"Edited by",kuFlag:!1,featuredMarkup:null,editors:[{id:"238219",title:"Dr.",name:"Gilberto Antonio",middleName:null,surname:"Bastidas Pacheco",slug:"gilberto-antonio-bastidas-pacheco",fullName:"Gilberto Antonio Bastidas Pacheco",profilePictureURL:"https://mts.intechopen.com/storage/users/238219/images/system/238219.jpeg",biography:"Prof. Gilberto Antonio Bastidas Pacheco is a physician with degrees in Pre-hospital Emergency Care, Executive Direction for Senior Management in Health, and Occupational Health and Safety. Along with a Health Management Course equivalent to the Public Health Middle Course, Magister Scientiae in Education Management and also in Protozoology, Prof. Bastidas holds a Ph.D. in Parasitology. He is a full professor at the Faculty of Health Sciences, Department of Public Health, University of Carabobo, Valencia, Venezuela. He has authored several articles published in national and international journal, and is also an arbitrator of scientific articles, member of the editorial committees of several journals, and a textbook writer and lecturer.",institutionString:"University of Carabobo",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"1",totalChapterViews:"0",totalEditedBooks:"2",institution:{name:"University of Carabobo",institutionURL:null,country:{name:"Venezuela"}}}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"909",title:"Parasitology",slug:"parasitology"}],chapters:[{id:"66212",title:"Introductory Chapter: Parasitology and Parasitism Areas of Knowledge That Must Be Constantly Studied",doi:"10.5772/intechopen.85181",slug:"introductory-chapter-parasitology-and-parasitism-areas-of-knowledge-that-must-be-constantly-studied",totalDownloads:2323,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:null,signatures:"Bastidas Gilberto",downloadPdfUrl:"/chapter/pdf-download/66212",previewPdfUrl:"/chapter/pdf-preview/66212",authors:[{id:"238219",title:"Dr.",name:"Gilberto Antonio",surname:"Bastidas Pacheco",slug:"gilberto-antonio-bastidas-pacheco",fullName:"Gilberto Antonio Bastidas Pacheco"}],corrections:null},{id:"63084",title:"Organ Pathology and Associated IFN-γ and IL-10 Variations in Mice Infected with Toxoplasma gondii Isolate from Kenya",doi:"10.5772/intechopen.79700",slug:"organ-pathology-and-associated-ifn-and-il-10-variations-in-mice-infected-with-toxoplasma-gondii-isol",totalDownloads:860,totalCrossrefCites:1,totalDimensionsCites:1,hasAltmetrics:0,abstract:"Toxoplasma gondii is an important foodborne opportunistic pathogen that causes a severe disease in immunocompromised patients. The pathology and immune responses associated with the ensuing disease have not been well described in strains from different parts of the world. The aim of the present study is to determine the IFN-γ and IL-10 variations and organ pathology in immunocompetent and immunocompromised mice infected with T. gondii isolated from a Kenyan chicken. Two groups of BALB/c mice were infected with T. gondii cysts and administered with dexamethasone (DXM) in drinking water. Other two groups: infected untreated and uninfected mice were kept as controls. The mice were euthanized at various time points: blood collected for serum and assayed for IFN-γ and IL-10 variations. After infection, significant (p<0.05) elevated levels of IFN-γ and IL-10 were observed. A significant decline in IFN-γ and IL-10 levels (p<0.05) was observed after dexamethasone treatment. Histological sections in the liver, heart, and spleen of the mice administered with DXM revealed various degrees of inflammation characterized by infiltration of inflammatory cells. The dexamethasone-treated mice presented with progressively increased (p<0.001) inflammatory responses is compared with the infected untreated mice.",signatures:"John Mokua Mose, David Muchina Kamau,\nJohn Maina Kagira, Naomi Maina, Maina Ngotho,\nLucy Mutharia and Simon Muturi Karanja",downloadPdfUrl:"/chapter/pdf-download/63084",previewPdfUrl:"/chapter/pdf-preview/63084",authors:[{id:"255664",title:"Dr.",name:"John",surname:"Kagira",slug:"john-kagira",fullName:"John Kagira"},{id:"255668",title:"Dr.",name:"John",surname:"Mokua",slug:"john-mokua",fullName:"John Mokua"},{id:"255669",title:"Dr.",name:"David",surname:"Kamau",slug:"david-kamau",fullName:"David Kamau"},{id:"255670",title:"Prof.",name:"Naomi",surname:"Maina",slug:"naomi-maina",fullName:"Naomi Maina"},{id:"255671",title:"Dr.",name:"Maina",surname:"Ngotho",slug:"maina-ngotho",fullName:"Maina Ngotho"},{id:"255672",title:"Ms.",name:"Adele",surname:"Njuguna",slug:"adele-njuguna",fullName:"Adele Njuguna"},{id:"255673",title:"Prof.",name:"Simon",surname:"Karanja",slug:"simon-karanja",fullName:"Simon Karanja"},{id:"265279",title:"Dr.",name:"Lucy",surname:"Mutharia",slug:"lucy-mutharia",fullName:"Lucy Mutharia"}],corrections:null},{id:"62896",title:"Malaria Pathophysiology as a Syndrome: Focus on Glucose Homeostasis in Severe Malaria and Phytotherapeutics Management of the Disease",doi:"10.5772/intechopen.79698",slug:"malaria-pathophysiology-as-a-syndrome-focus-on-glucose-homeostasis-in-severe-malaria-and-phytotherap",totalDownloads:1254,totalCrossrefCites:3,totalDimensionsCites:5,hasAltmetrics:1,abstract:"Severe malaria presents with varied pathophysiological manifestations to include derangement in glucose homeostasis. The changes in glucose management by the infected human host emanate from both Plasmodium parasitic and host factors and/or influences which are aimed at creating a proliferative advantage to the parasite. This also includes morphological changes that that take place to both infected and uninfected cells as structural alterations occur on the cell membranes to allow for increased nutrients (glucose) transportation into the cells. Without the availability, effective and efficient intervention there is a high cost incurred by the human host. Hyperglycaemia, hypoglycaemia and hyperinsulinemia are critical aspects displayed in severe malaria. Conventional treatment to malaria renders itself hostile to the host with negative glucose metabolism changes experiences in the young, pregnant women and malaria naïve individuals. In malaria, therefore, host effects, parasite imperatives and treatment regimens play a pivotal role in the return to wellness of the patient. Phytotherapeutics are emerging as treatment alternatives that ameliorate glucose homeostasis alternations as well as combat malaria parasitaemia. The phytochemicals e.g. triterpenes, have been shown to alleviate the “disease” and “parasitic” aspects of malaria pointing at key aspects in ameliorating malaria glucose homeostasis fallings-out that are experienced in malaria.",signatures:"Greanious Alfred Mavondo, Joy Mavondo, Wisdom Peresuh, Mary\nDlodlo and Obadiah Moyo",downloadPdfUrl:"/chapter/pdf-download/62896",previewPdfUrl:"/chapter/pdf-preview/62896",authors:[{id:"202805",title:"Prof.",name:"Alfred Mavondo-Nyajena Mukuwa",surname:"Greanious",slug:"alfred-mavondo-nyajena-mukuwa-greanious",fullName:"Alfred Mavondo-Nyajena Mukuwa Greanious"},{id:"263433",title:"Dr.",name:"Obadiah",surname:"Moyo",slug:"obadiah-moyo",fullName:"Obadiah Moyo"},{id:"263434",title:"Mrs.",name:"Joy",surname:"Mavondo",slug:"joy-mavondo",fullName:"Joy Mavondo"},{id:"263435",title:"Ms.",name:"Mary",surname:"Dlodlo",slug:"mary-dlodlo",fullName:"Mary Dlodlo"},{id:"263436",title:"Mr.",name:"Wisdom",surname:"Peresu",slug:"wisdom-peresu",fullName:"Wisdom Peresu"}],corrections:null},{id:"62893",title:"Prevalence and Intensity of Intestinal Parasites and Malaria in Pregnant Women at Abobo District in Abidjan, Côte d’Ivoire",doi:"10.5772/intechopen.79699",slug:"prevalence-and-intensity-of-intestinal-parasites-and-malaria-in-pregnant-women-at-abobo-district-in-",totalDownloads:866,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"A prospective study was carried out from 2010 to 2012 at the Hôpital Général d’Abobo (HGA) in Abidjan, in order to determine the impact of infectious and parasitic diseases on child cognitive development. Blood samples were examined by means of drop thick and blood smear, as for stool by direct examination and concentration by formalin-ether method. We evaluated the prevalence and the parasite load of malaria and gastrointestinal parasites and then investigated the risk factors for these disorders. Overall, 331 pregnant women in the last trimester of their pregnancy were enrolled. The plasmodic index was 3.9% with an infestation specific rate for P. falciparum of 100%. Concerning digestive protozoa, it has been observed 71.3% of nonpathogenic, against 9.7% of pathogens, either an overall prevalence of 51.4% of digestive parasites. The calculated average parasitic loads revealed 3089.2 tpz/μl of blood (95% CI, 591.1–5587.3) for malaria, 6.5 eggs per gram of stool (95% CI, 0.4–13.4) for intestinal helminths, and one (1) parasite by microscopic field for protozoa (common infestation). It has been shown that the occurrence of malaria has been linked to the nonuse of impregnated mosquito nets (χ2 = 0.012, p = 0.018) to age. No link could be established between the presence of digestive parasites and the age of pregnant women or socioeconomic conditions (level of education, profession, type of toilet). Malaria is less common in pregnant women, while the rate of digestive parasites remains high.",signatures:"Gaoussou Coulibaly, Kouassi Patrick Yao, Mathurin Koffi, Bernardin\nAhouty Ahouty, Laurent Kouassi Louhourignon, Monsan N’Cho and\nEliézer Kouakou N’Goran",downloadPdfUrl:"/chapter/pdf-download/62893",previewPdfUrl:"/chapter/pdf-preview/62893",authors:[{id:"254981",title:"Ph.D. Student",name:"Gaoussou",surname:"Coulibaly",slug:"gaoussou-coulibaly",fullName:"Gaoussou Coulibaly"}],corrections:null},{id:"63554",title:"Current Aspects in Trichinellosis",doi:"10.5772/intechopen.80372",slug:"current-aspects-in-trichinellosis",totalDownloads:1355,totalCrossrefCites:2,totalDimensionsCites:8,hasAltmetrics:0,abstract:"Currently, it is estimated that more than 11 million humans in the world are infected by helminth parasites of Trichinella species, mainly by Trichinella spiralis (T. spiralis), responsible for causing Trichinellosis disease in both animals and humans. Trichinellosis is a cosmopolitan parasitic zoonotic disease, which has direct relevance to human and animal health, because it presents a constant and important challenge to the host’s immune system, especially through the intestinal tract. Currently, there is an intense investigation of new strategies in pharmacotherapy and immunotherapy against infection by Trichinella spiralis. In this chapter, we will present the most current aspects of biology, epidemiology, immunology, clinicopathology, pharmacotherapy and immunotherapy in Trichinellosis.",signatures:"José Luis Muñoz-Carrillo, Claudia Maldonado-Tapia, Argelia López-\nLuna, José Jesús Muñoz-Escobedo, Juan Armando Flores-De La\nTorre and Alejandra Moreno-García",downloadPdfUrl:"/chapter/pdf-download/63554",previewPdfUrl:"/chapter/pdf-preview/63554",authors:[{id:"214236",title:"Dr.",name:"Jose Luis",surname:"Muñoz-Carrillo",slug:"jose-luis-munoz-carrillo",fullName:"Jose Luis Muñoz-Carrillo"},{id:"216080",title:"Dr.",name:"Alejandra",surname:"Moreno-García",slug:"alejandra-moreno-garcia",fullName:"Alejandra Moreno-García"},{id:"254888",title:"Dr.",name:"Juan Armando",surname:"Flores-De La Torre",slug:"juan-armando-flores-de-la-torre",fullName:"Juan Armando Flores-De La Torre"},{id:"254889",title:"Dr.",name:"José Jesús",surname:"Muñoz-Escobedo",slug:"jose-jesus-munoz-escobedo",fullName:"José Jesús Muñoz-Escobedo"},{id:"254890",title:"Dr.",name:"Argelia",surname:"López-Luna",slug:"argelia-lopez-luna",fullName:"Argelia López-Luna"},{id:"254891",title:"Dr.",name:"Claudia",surname:"Maldonado-Tapia",slug:"claudia-maldonado-tapia",fullName:"Claudia Maldonado-Tapia"}],corrections:null}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},subseries:null,tags:null},relatedBooks:[{type:"book",id:"9025",title:"Parasitology and Microbiology Research",subtitle:null,isOpenForSubmission:!1,hash:"d9a211396d44f07d2748e147786a2c8b",slug:"parasitology-and-microbiology-research",bookSignature:"Gilberto Antonio Bastidas Pacheco and Asghar Ali Kamboh",coverURL:"https://cdn.intechopen.com/books/images_new/9025.jpg",editedByType:"Edited by",editors:[{id:"238219",title:"Dr.",name:"Gilberto Antonio",surname:"Bastidas Pacheco",slug:"gilberto-antonio-bastidas-pacheco",fullName:"Gilberto Antonio Bastidas Pacheco"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"1692",title:"Parasitology",subtitle:null,isOpenForSubmission:!1,hash:"b2110e81c765897e4ffdfbd340495e25",slug:"parasitology",bookSignature:"Mohammad Manjur Shah",coverURL:"https://cdn.intechopen.com/books/images_new/1692.jpg",editedByType:"Edited by",editors:[{id:"94128",title:"Dr.",name:"Mohammad Manjur",surname:"Shah",slug:"mohammad-manjur-shah",fullName:"Mohammad Manjur Shah"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"5527",title:"Natural Remedies in the Fight Against Parasites",subtitle:null,isOpenForSubmission:!1,hash:"d705be119e74a50305952521b2b5ece0",slug:"natural-remedies-in-the-fight-against-parasites",bookSignature:"Hanem Khater, M. 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Ashraf",coverURL:"https://cdn.intechopen.com/books/images_new/8268.jpg",licenceType:"CC BY 3.0",editedByType:"Edited by",editors:[{id:"199287",title:"Dr.",name:"Ghulam Md",middleName:null,surname:"Ashraf",slug:"ghulam-md-ashraf",fullName:"Ghulam Md Ashraf"}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"}}},ofsBook:{item:{type:"book",id:"11814",leadTitle:null,title:"Liposomes - Recent Advances, New Perspectives and Applications",subtitle:null,reviewType:"peer-reviewed",abstract:"\r\n\tLiposome, a versatile lipid vesicle, is licensed to use for effective delivery of drugs, candidate vaccines, and nucleic acids. Also, drug-loaded ligand anchored liposomes are used to induce selective apoptosis in cells of monocyte and macrophage lineage. Therefore this book aims to cover all spheres of drug and antigen delivery for developing therapeutic interventions and vaccines against infectious diseases and beyond.
\r\n\r\n\tThis book will shed light on various methods for the preparation and characterization of liposomes and their engineered versions. Technological advancements have enabled us to develop newer methods of formulating liposomes. Over time, liposomes have been modified to a larger extent and engineered to meet our growing needs for sustained and controlled delivery for developing therapeutic interventions vaccines. Further, this book will throw light on the various methods of preparation and characterization of liposomes, and discuss several biochemical and indirect methods to understand the biological and physicochemical mechanisms of action of liposomes that decide their efficiency in vivo.
\r\n\tDespite their limitations such as encapsulation efficiency, liposomes are a well-established choice for a number of unconventional and conventional biological applications. The versatility of these lipid-based vesicles presents the importance of these nanoparticles in the future applications of nanotechnology besides targeted drug delivery. Overall, this book provides the necessary and relevant information about various aspects of liposomes and their use in nanomedicine.
\r\n\t
The basic principle of detection of a biosensor is based on the specific interaction between the analyte of interest and the recognition element. As a result of this specific interaction, changes are produced one or several physical-chemical properties (pH, electron transference, heat transfer, change of potential or mass, variation of optical properties, etc.). These changes are detected and can be measured by a specific transductor (Thévenot et al., 2001; Wang, 2004). Electrochemical biosensors are based on the electrochemical species consumed and /or generated during a biochemical interaction process of a biological active substance and analyte. Electrochemical biosensors, especially the amperometrics, have an important position among the biosensors. Since 2000s until the completion of this review, intensive research activity has been devoted to the development of amperometric biosensors. The statistic in ScienceDirect search showed 6950 items found for publication made with pub-date > 1999 and keywords Electrochemical biosensor. Of which about 38% with application to detect ethanol, 47 % for glucose and 15% to phenolic compound.
\n\t\t\tThe electrochemical biosensor usually consists of three phases: receptors phase, transducer phase and a signal amplifier phase. The receptor phase incorporates a biological or biomimetic recognition element (nucleic acid, enzyme, anti-body, tissue, organelles or whole cells). The most important phase in an amperometry biosensor is the receptor biomolecule by the selectivity of the device to a given analyte or condition. The transducer is the conductive phase, which converts a biochemical signal into a reading or measurement. And the amplifier is the computerized apparatus/software using to boosts/readout of signal.
\n\t\t\tThe role of the transducer in a biosensor is to generate a measurable signal of the analyte interacts with the biological molecule associated with the transducer surface. So, in the case of the optical transducer, it generates a signal measured as a light intensity proportional to the concentration of analyte in the sample; this may be an inverse relationship. The composite-films of the surface optical biosensor has an important role in the process in which changes in optical characteristics such as absorption, rotation, refractive index, bio/chemiluminescence, and fluorescence are related to the analyte concentration (Koncki et al., 2001). Martin (2002) showed how the fiber optical immunosensors based on long-period gratings that have limited sensitivity at the refractive index of ordinary aqueous solutions (~1.33). And using composite such as films of titanium dioxide, for example, can raise the local refractive index of the sensor (~1.42), thus increasing sensitivity. Titanium dioxide is commercially available and has been extensively used in the paint industry because of its ability to scatter visible light efficiently.
\n\t\t\tElectrochemical transducers generate a current or voltage in proportion to the analyte being measured as result of the electron transference; again this may be an inverse relationship. There are numerous components to any biosensor configuration. Through the years, a great many combinations have been proposed and demonstrated, although far fewer have been commercial successes. Figure 1 presents the two most popular platforms for electrochemical biosensor including: carbon paste electrode for discrete or continuous measurements and disposable screen-printed electrode (SPE) for low-cost and single-use field applications.
\n\t\t\tElectrochemical composite biosensor for analyte detection
In the literature different procedures have been developed to manufacture electrochemical transducers. These conducting composite materials could be considered to be random assemblies of minielectrodes, in which “edge effects” contribute significantly to the Faradic current. Several transducer composite show electrochemical advantages over those built using a single conductive material (platinum, gold, silver, carbon, mercury, graphite, etc.). The conductive parts of the arrays have different sizes and shapes, and it is this randomness that prevents theoretical models based on homogeneously distributed discs, to be used to describe the composite electrode behaviour (Barsan et al., 2009). Using different types of carbon, entrapped in a suitable, normally polymeric, binder, various composite electrodes have been extensively used in electroanalytical measurements.
\n\t\t\tThe desirable characteristics of composite-biosensors usually cited are: i) high versatility in shape and size; ii) lower cost; iii) easy fabrication; iv) higher signal-to-noise ratio; v) surface regeneration; vi) provide suitable mechanic and conducting characteristics, vii) possibility to incorporate other components in the bulk of composite (mediators, cofactors or other bio-molecules) to enhance selectivity or sensitivity, and viii) a long-term stability and lifetime. Conducting composites are interesting alternatives for the construction of electrochemical biosensors. The capability of integrating various materials is one of their main advantages. Thus, the composites used for the construction of biosensors, that have been proposed by several researchers and are used efficiently in different types of analytical determinations of various electroactive species, showing the robustness and sensitivity (Kress-Rogers & Brimelow, 2003; Ahammad et al., 2009; Lojou & Bianco,2006; Bojorge et al., 2009). Several materials have been used to construct electrochemical composite biosensor. Among them, carbon-based matrices are the most applied due to costs and their electronic conductivity.
\n\t\t\tAspects related with the protocols for bio-molecules immobilization on the composite support have been widely proposed for many researchers (Caramori & Fernandes, 2004; Ikeda et al., 2002; Shan et al., 2007; Mateo et al., 2000). Difficulties still pose problems such as being reproducible, simple to use, and stable immobilization of the biological component. For example, to obtain a fast response time and a reliable reading a thin layer of immobilized bio-molecule is desirable and shelf life and operational stability demand a high value of immobilized enzymatic activity. Adsorption to adequate surfaces including a metal electrode layer yields relatively unstable systems (Mateo et al., 2006).
\n\t\t\tThe immobilization of bio-molecules on composite surface, the composite supports should be designed to permit a minimum desorption of native proteins, which it is possible through the preparation and selection of arrays that generate for a very strong ionic adsorption of enzymes. In this aspect, Montes et al., 2006 shows as the surface of the enzyme penicillin G acylase was chemically modified under controlled conditions: chemical amination of the protein surface of carboxylic groups (using soluble carbodiimide and ethylendiamine) and chemical succinylation (using succinic anhydride) of amino groups. The full chemical modification produced some negative effects on enzyme stability and activity, although partial modification (mainly succinylation) presented negligible effects on both enzyme features. The chemical amination of the protein surface permitted the immobilization of the enzyme on carboxymethyl and dextran sulphate – coated support, while the chemical succinylation permitted the enzyme immobilization on DEAE-agarose and polyethylenimine coated supports. Immobilization was very strong on these supports, mainly in the polymeric ones, and dependent of the degree of modification, although the enzymes still can be desorbed or inactivated after incubation under drastic conditions.
\n\t\t\tThe immobilization on ionic polymeric beds allows a significant increase in enzyme stability against the inactivation and inhibitory effects of organic solvents, very likely by the promotion of a certain partition of the organic solvent out of the enzyme environment. These results suggest that the enrichment of the surface of proteins with ionic groups may be a good strategy to take advantage of the immobilization of enzymes via ionic exchange on ionic polymeric beds. For example, one should mention the work of Hentze & Antonietti (2002) that describe conventional and modern techniques of porous organic polymers synthesis. A great variety of polymer architectures and functions can be gained by foaming, phase separation, imprinting or templating approaches. Several applications of porous polymers are discussed, focusing on biotechnological and biomedical applications, such as chromatography, protein synthesis, drug carrier systems, tissue engineering and others.
\n\t\t\tThis work describes several approaches for the construction of device and rigid-composites application for detection health and environmental target analyte, such as ethanol and phenol. The first approach is based on a matrix of graphite-epoxy as electrochemical transducer where the immobilization of HRP enzyme for detection of phenol. The second approach is based on matrixes of graphite-epoxy and graphite-Teflon, which are incorporated with HRP/TYR enzymes for the detection of phenol. These bio-composites offer several potential advantages over more traditional to the electrodes based on a modified surface phase driving. The ability to integrate different materials into one is its main advantage, besides the improved electrochemical properties. The different properties of these materials are discussed and compared. The third approach is based on composite Graphite / Epoxy / Pani. Here the polyaniline polymer (PANI) was selected because of its ease of preparation and economic good environmental stability, and conductivity that occur after doping. Several arrays of composite were prepared with different fractions of PANI and were characterized by their morphological, electrical and mechanical. Ahuja et al. (2007) reviewed the state of the art of the use conducting polymers to construct biosensors, studying the different aspects of biomolecules immobilization techniques. Electrochemical biosensors based on polyaniline immobilized with different recognizing biomolecules like as oxide-reduction enzymes, nucleic acids, antibodies, were just constructed and reported (Wei & Ivaska, 2006; Dhand et al., 2011). The term polyaniline is employed to refer a class of polymers with repeated units of ‘ring-N’ and the base form is [Ping et al., 1997]:
\n\t\t\tShematic base form
The value of y in (I) can varied from unity, the completely reduced polymer with amine form for all nitrogen atoms, to zero, which is the completely oxidized form with imines form for all nitrogen atoms. The degree of protonation of the polymeric base depends on its oxidation state and on the pH of the environment. The terms leucoemeraldine (LE), emeraldine (EB) and perniganiline refer to the different oxidation states of the polymer where y = 1, 0.5 and 0, respectively. The emeraldine half-oxidised and protonated form is the conductive polymer (Abdiryim et al., 2005).
\n\t\tComposite biosensors are made from two or more constituent materials, conductive material and nonconducting-binding material, with significantly different physical or chemical properties which remain separate and distinct at the macroscopic or microscopic scale within the finished structure. The conductive materials involve the use of nature different of carbon-based matrices, such as: glassy carbon, graphite and nano-carbon and or also by mix of a polymer with electrical conductivity property. They are of highest purity grade and used as delivered from commercial sources (Aldrich, Sigma-Aldrich, Fluka). The main purpose of the conducting phase in the composite is to supply the electrical conductivity needed for the conduction of the electrical biosignal. Another material consists of agglutinating agents (insulator materials), such as: epoxy resins, silicone, polyurethane, metacrylate resin, Teflon, etc. The main purpose of agglutinating agents is mechanical consistence assuring the durability and resistance to the electrolytic working medium.
\n\t\t\tThe conductivity is defined by the connectivity of the conducting particles in the midst of the polymer. When a biocomposite is prepared, it is imperative that its calibration curve is studied to define the composition that guarantees a proper electrical conductivity without losing the mechanical and physical rigidity of the biocomposite. Electrochemical composites can be classified according to how the phases are distributed within the composite matrix. For example, a conducting composite electrode surface can be prepared as an ordered array or as a random arrangement (ensemble) of conducting regions separated by an insulator. The random composite mixtures are classified according to the distribution of the conductor within the composite matrix. If the conductor particles are distributed randomly within the composite matrix the composite is said to be of the dispersed type. If the conductor extends throughout the composite in a random fashion with regions of pure insulator and pure conductor that do not intermix, the composite is said to be of the consolidated type (Ates & Sarac, 2009).
\n\t\t\tCarbon is an ideal electrode substrate due to its wide anodic potential range, low residual current, chemical inertness, low cost, fast response time, and ease for fabrication in different configuration and size and suitability for various sensing and detection applications (Wang, 2001). Carbon electrodes allow scans to more negative potentials than platinum or gold, as well as good anodic potential windows. Carbon is observed in several variants such as, glassy carbon, graphite, fullerene, graphene and carbon nanotubes (Ates & Sarac, 2009).
\n\t\t\tGraphite is a dark gray, soft and porous material with adsorption capacity and is naturally abundant and highly conductive fillers (with an electrical conductivity of 104 S/cm at ambient temperature) for conducting polymer composites (Du et al., 2004) and electrical resistivity is 50 μ.Ω.m (Sengupta et al. 2010) and its density is 1.95-2.3 g/cm3. In graphite, the carbon atoms are only bonded in two dimensions. Bond angle in graphite is 120º. Each carbon atom in graphite is sp2 hybridized and with a distance between basal planes of 3.35 Å. Three out of four valence electrons of each carbon atom are used in bond formation with three other carbon atoms while the fourth electron of each carbon atom forms delocalized π-bonds which spreads uniformly over all carbon atoms. The carbon atoms are arranged hexagonally in a planar condensed ring. Also, the layers are stacked parallel to each other, with the atoms within the rings bonded covalently, whereas the layers are loosely bonded together by Van der Waal’s forces. The anisotropic nature of graphite is the result of the two types of bonding acting in different crystallographic directions. The ability of graphite to form a solid film lubricant may be attributed to these two contrasting chemical bonds. The weak Van der Waal’s forces govern the bonding between the individual layers and also there are no bonds between the layers, the layers can easily slip off one to another, making it an ideal lubricant, and resulting in a reduced coefficient of friction and, hence, wear, and by this same reason, the graphite is a good material for pencils - layers come off and get left on the paper as you write.
\n\t\t\t\tOn the other hand, given the good electro-catalytic properties, the graphite has been used as an electrode material in the construction of disposable amperometric biosensors for the detection of various analytes such as ethanol, phenol and glucose, and many others (Kirgözet al., 2006; Llopis et al., 2005; Mailley et al., 2003). By this, the aim of this study was to show the potential use of graphite in biosensor, based on the direct electronic transfers between the enzyme and mediator conducting salt, which are contained in a polymeric matrix of epoxy resin and graphite powder or Teflon and graphite powder. These devices combine the advantages of the biosensors based on solid composites and the electro-catalytic properties of an organic conducting salt, such as PANI.
\n\t\t\tAnother common form of carbon electrode material is the glassy carbon (GC), which is relatively expensive and difficult to build. Glassy carbon also called vitreous carbon, is an advanced material of pure carbon combining glassy and ceramic properties with these of graphite. GC is mechanically hard material, negligible porosity. The higher electrical resistance (0.48 and 0.09 vs. 0.28 and 0.18 Ω-cm at 325 and 725º C, respectively) and its linear temperature dependence in the annealed sample are attributed to formation of graphite crystalline nuclei in the previously homogeneous and amorphous glassy carbon (Barykin et al., 1976). GC is a class of non-graphitizing carbon that is widely used as an electrode material in electrochemistry and for high-temperature crucibles. It is prepared by subjecting the organic precursors to a series of heat treatments at temperatures up to 3000º C (Kinoshita et al., 1988; Jenkins et al., 1972, 1976). Unlike many non-graphitizing carbons, it is impermeable to gases and chemically extremely inert, especially when prepared at very high temperatures. It has been demonstrated that the rates of oxidation of glassy carbon in oxygen, carbon dioxide, or water vapour are lower than those of any other carbon (Harris, 2004). It is also highly resistant to attack by acids. Thus, whereas normal graphite is reduced to a powder by a mixture of concentrated sulphuric and nitric acids at room temperature, glassy carbon is unaffected by such treatment, even after several months. The structure of glassy carbon has been the subject of research since it was first produced in the early 1960s. However, the properties which make glassy carbon so valuable in these applications are poorly understood, since its detailed atomic structure is not known. A model for the structure of glassy carbon put forward many years ago has gained wide acceptance, but appears to suffer from serious shortcomings. In particular, it fails to account for the chemical inertness of the carbon, and for its high proportion of closed porosity (Harris, 2004).
\n\t\t\tThe carbon paste (CP) is used for construction of carbon paste electrodes (CPE´s) for voltammetric determination, which are made usually of high purity graphite powder or another type of carbon, as carbon nanotubes, dispersed in a non-conductive mineral oil such as Nujol or silicone oil to form a paste. Common types of carbon pastes are soft and non-compact, and have to be kept in special bodies. A holder for carbon pastes can be realized as a well drilled into a short Teflon rod (Peng et al., 1993) a glass tube (Pei et al., 1991) or a polyethylene syringe (Švancara et al., 2005) filled with a paste, which is electrically contacted via a conducting wire. Exhaustive reviews on CP-based sensors have been published the last two decades, where various types of biosensors (e.g., those for amino acids, ethanol, fructose, galactose, glucose, glycerol, lactate, xanthine, etc.) based on related oxidases and dehydrogenases, whole cells, and plant tissues are reviewed. A major advantage of CP-based biosensors are very low background current and are the feasibility of bulk modification of the electrode material with biocatalyst as well as with other components essential for their effective functioning. Renewable or disposable surfaces so that each measurement can be performed on the new surface and not be affected by the residuals from the previous measurement (Bard & Rubinstein, 1996).
\n\t\t\t\tThe constructions of CP biosensor are very simple; however, there is one aspect which makes them not very convenient for practical use and this is the necessity of refilling the carbon paste in experiments requiring a regular removal of the electrode surface layer. Another smart construction circumventing this time-consuming procedure were proposed by Švancara et al., 2005 and Kalcher et al., 2009 who proposed piston-driven electrode holders where the desired amount of the used paste could simply be extruded from the electrode body and smoothed away or cut off.
\n\t\t\t\tAnother advantage of the CP is that has no need for the sample to have a high electrical conductivity because the conduction is mainly accomplished by the graphite. The currents obtained are within a suitable range due to the small sample amounts. This means that the signal resolution is improved. The use of CP composite biosensor helps in investigating substances in small sample amount and the electrochemical reactions proceed at the surface of the paste electrode. At which, an ion transfers between the solid sample and the electrolyte solution is possible.
\n\t\t\t\tThe properties of the CP depend on the specific components employed the manner of preparation and maintenance. The properties of CP-based biosensor can be partially improved by the incorporation of additives into the paste, e.g., polyethylenimine, acetylenic polymers, polyaniline, chitosan, glutamate, cationic antibiotics, silica and carbon nanotubes to prevent the leakage of mediator, covalent binding of the mediator to a polymer matrix (Mailley et al., 2003; Anik & Çevik, 2009; Tingry et al., 2006). Recently a novel carbon paste ion selective electrode for determination of trace amount of holmium was prepared by Ganjali M. R. et al., 2009. The authors used multi-walled carbon nanotubes and nanosilica for improvement of a holmium carbon paste sensor response. The approaches to improve the properties of CP mentioned above have certain limitations and deficiencies. On one hand, it is still unclear how additives might affect the biosensor performance at the molecular level, thus making the search for suitable additives for each specific biosensor rather difficult and more random than systematic. On the other hand, covalently bound mediators exhibit modified electrochemical properties and reduced mobility, which affect the reaction rate with enzymes.
\n\t\t\t\tThe main disadvantages of the CP are the fragile surface or weak mechanical properties due to their creamy texture, which can easily lead to disintegration of the system; and the solubility of the pasting liquid in organic solvents, which often show voltammograms with a higher irreversibility than in cases where no organic binders are used. Therefore, several carbon composite electrodes based on carbon nanotubes and a solid matrix have been proposed. Dues to its insulating nature efficiency in the presence of a solid matrix decreases electrode reaction rates more than a pasting liquid does unless an active carbon surface is exposed (Bard,1996; Kalcher et al., 2006).
\n\t\t\tCarbon nanotubes (CNTs) represent one of the best examples of novel nanostructures derived by bottom-up chemical synthesis approaches. CNTs are molecular-scale tubes of graphitic carbon with outstanding properties. They are among the stiffest and strongest fibres known, and have remarkable electronic properties and many other unique characteristics. CNTs has received a great deal of attention as an electrode material, because these have good electrocatalytic properties (Wang, 2005). Merkoçi, 2006 showed an interesting review paper on CNTs in analytical sciences covers the full calendar from their discovery in 1991 until 2005 and treat analytical aspects of interest in the coupling of CNTs to enzymes, DNA, proteins and, finally, the use of CNTs for several applications such as in chromatography, sensors and biosensors, nanoprobes, etc. Commercial applications have been rather slow to develop, however, primarily because of the high production costs of the best quality nanotubes. For these reasons they have attracted huge academic and industrial interest, with several articles on nanotubes being published the lasts years (Ruoff & Lorents, 1995; Gouveia-Caridade et al., 2008; Yadav et al., 2011).
\n\t\t\t\tCarbon nanotubes have received considerable attention in the field of electrochemical sensing, due to their unique structural 1D nature (Javey, 2009), for instance, unique tubular nanostructure, due to their superior mechanical (Schadler et al., 1998) and electronic and chemical properties (Saito et al.,1998), large specific surface, excellent conductivity, superior to 100 Scm-1 (Yao et al., 2000), modifiable sidewall, good biocompatibility, and so on. Solubilization and biological functionalization of carbon nanotubes have greatly increased the usage of carbon nanotubes in biomedical applications such as biosensors and nanoprobes (Teker, 2008).
\n\t\t\t\tThe structure of CNTs is like a sheet of graphite rolled up into a tube of diameter 1-10 nm, and hence forms hollow tubules of a single layer of carbon atoms (Wang, 2005). CNTs include both singlewalled (SWNT or SWCNT) and multiwalled (MWNTS) structures (Baughman et al., 2002). CNTs have very high surface-to-volume ratios and, therefore, promise depending on the direction of hexagons, nanotubes can be classified as either zigzag, armchair or chiral. When scientists make nanotubes, they tend to get a mixture of several types due to different types of nanotubes have different properties. Today, a major challenge in nanoscience is finding a way to make just one type of nanotube. In this aspect, an interesting study made by Safarova et al., 2007, who shows how parameters of SWCNTs, specially a diameter and length of one nanotube or a bundle of nanotubes and a number of nanotubes in the bundle, can be determinated using the techniques as Transmission Electron Microscopy (TEM), Scanning Electron Microscopy (SEM) and Atomic Force Microscopy (AFM).
\n\t\t\tAccording to Céspedes & Alegret (2000), rigid carbon composites are ideal for the construction of electrochemical sensors. The plastic nature of these materials makes them modifiable, permitting the incorporation of fillers before they are cured. A large number of biological materials can be immobilised by blending them with these composites to form new biocomposite materials, showing robustness and sensitivity. These biocomposites not only act as reservoirs of the biological material but may also contain catalysts, mediators and cofactors that improve the response of the resulting electrochemical biosensors. The carbon-based matrices are the most applied due to their high conductivity, whereas epoxy resins, silicone, polyurethane, metacrylate resin, Teflon, etc, can be employed as agglutinating agents (insulator materials). The main characteristic of these composites is their rigidity, resulting in a high mechanical stability over time. This type of composite biosensor offers additional important advantages: the sensing surface can be renewed by a simple polishing procedure. The proximity of the redox centers of the biological material and the conducting sites on the sensing surface favours the transfer of electrons between electroactive species. So, for example, a universal affinity platform for electrochemical genosensing can be easily achieved by modifying the graphite-epoxy composite with avidin to obtain an avidin biocomposite, where biotinylated DNA can be rapidly single-point attached (Pividori et al., 2005). Thus, this often allows the regeneration of the biological component without using co-substrates and mediators, it can be customized easily according to biocatalytical and/or electrochemical requirements; it presents low background currents, favoring a high signal-to-noise ratio, and lower detection limits and it offers a new active layer after removal of the outer surface, extending the lifetime of such electrodes.
\n\t\t\t\tGraphite and epoxy resins are employed to construct rigids composites that can be used in aqueous or nonaqueous media. Serra et al., 2005 showed a comparison study of the behavior of different rigid composite matrices for the construction of amperometric tyrosinase biosensors, widely used for the detection of phenolics compounds. So, they showed that the great advantage of Graphite-epoxy composite (GEC) composite over carbon paste composite is their tolerance to organic solvents, due to a better reproducibility of the amperometric measurements both with and without regeneration of the electrode surface by polishing.
\n\t\t\t\t\t\n\t\t\t\t\t\tKırgöz et al., 2006 showed the modification of a GEC electrode with bacterial cells to detection of xenobiotic in waste water samples. Pseudomonas putida DSM 50026 was used as a biological component and the measurement was based on the respiratory activity of the cells. This study the combination of microorganisms with GEC composite electrodes provided economic and practical disposable biosensors.
\n\t\t\t\t\tThe physical and chemical effects of water on graphite/epoxy composite were investigated in an interesting work of Zhou and Luke (1995). This paper demonstrates by diffusion data that the time for the onset of non-Fickian behavior is inversely related to the increase in exposure temperature. So, if a relatively low temperature of exposure there is no dissolution of the surface of the material or physical damage and the behaviour profile of weight gain is Fickian. Pointing out that Fick\'s laws are differential equations that describe the flow of a substance and the concentration versus time and position. And with the temperature increase, cracks, voids, surface peeling and dissolution occur. An explanation for this effect is that the cracks retain water which contributes to the behavior of absorption higher than the theoretical Fickian diffusion curve or the epoxy resins used in manufacturing GEC are capable of undergoing a significant and irreversible thermal oxidative degradation at high temperatures after submission or within a narrow range of its maximum temperature of superior service. When exposed to temperatures high enough to cause degradation of resin, these materials experience a drop in glass transition temperature which effectively increases service temperature and significantly reduces the room-temperature mechanical strength properties of the composite. Below a certain threshold of exposure, these compounds are visually and microscopically damaged, embrittlement and cracking of the surface causes a loss in resistance the impact of the material. Therefore, composite GEC exposed to conditions of overheating may suffer irreversible damage and catastrophic in a very short time period.
\n\t\t\t\t\tFrom the viewpoint of the effects of electrochemical of the GEC electrode have an anion response close to the Nernstian behavior, 1-3 potential salt and are more sensitive to OH¯. This response of the transducer is due to an electroreduction of the adsorbed oxygen on the graphite surface in such way that an increase in the electrode potential takes place due to a change in the oxidation state (Rodriguez-Huerta et al., 2006).
\n\t\t\t\t\n\t\t\t\t\tLi & Shi (2011), published a review about the electrochemical applications of composites prepared with chemically converted graphenes (CCGs) and conducting polymers as polyaniline (PANI), polypirrole (PPy), polythiophene (PTh), to construct sensors and, consequently, biosensors. The electrochemical activity and sensing of the composites prepared with SPANI (acid doped PANI) and CGC were improved and tested with cyclic voltammetric studies. The electropolymerization of polyaniline on the carbon surface was discussed and described by Ates and Sarac (2009) in order to improve the proprieties of the conducting polymers and the application to construct sensors, biosensors, capacitors and batteries. Gómez et al., 2011, reported a chemical precipitation technique to synthesized graphene-polyaniline nanocomposite for electrodes and electroanalysis, and the morphology was characterized by using scanning electron microscopy, transmission electron microscopy, Fourier transform infrared spectroscopy and Raman and cyclic voltammetry. Tung at al., 2011, prepared a nanocomposite with selenious acid, doped polyaniline and graphite nanoplatelet (GNP), and characterized by using SEM, FTIR, X-ray diffraction, and electrical conductivity measurement. A crystalline structure was shown and the electrical conductivity increases with the increase of the GNP content although lower than the HCl doped one. Kim and Park (2011), used multi-walled carbon nanotube and grapheme to prepare a nanocomposite with polyaniline (M-GR/PANI) and higher sensitive CVs were observed when compared with a graphite-polyaniline composite CV. A bienzymatic biosensor was designed for glucose oxidase and horseradish peroxidase based on covalent attachment onto carboxylic-derived multi-walled carbon nanotube for the deposition electroactive polyaniline by Sheng and Zheng (2009), and the linear range for glucose detection was 0.05 mM – 8.0 mM. Zhan et al., 2009, proposed a new strategy for a highly sensitive amperometric biosensor immobilizing tyrosinase on the surface of the polyaniline-ionic liquid-carbon nanofiber composite for phenols detection. A linear range from 4.0 x 10-10 to 2.1 x 10-6 mM was obtained for catechol detection.
\n\t\t\tIn this section, the protocols for construction to make the maximum use of composites-transducers are described. Especially the behavior of graphite to transfer electronic device promoted the electrochemical sensor. Rigid matrices based on epoxy/graphite, and the use of thermoplastic resins such as Teflon are very useful tools in building composite electrode for its flexibility in shape and size, allowing easy adaptation to a variety of electrode configurations (conventional flow-through, screenprinted, etc.).
\n\t\t\tCarbon paste electrodes are the most popular electrodes in electroanalytical chemistry and bioelectrochemistry due to that exhibit low background current and are easily prepared. The types of carbon pastes are soft and non-compact, and have to be kept in special bodies. A holder for carbon pastes can be realized as a well drilled into a short Teflon rod, a glass tube or a polyethylene syringe filled with a paste, which is electrically contacted via a conducting wire. Carbon pastes usually employed for the fabrication of CPE are usually prepared as follow: (i) Carbon paste are prepared by mixing graphite powder with Nujol oil or paraffin oil in a mortar until it was uniformly wetted using a graphite/Nujol. The ratio varies for each biosensor proposed in the literature: of 4/1 w/w; 50:50 % m/m; 75:25% m/m. These ratios are employed as it provides convenient analytical properties. (ii) The enzymes are incorporated in the carbon paste using an enzymatic ratio up to 10% w/w. (iii) Sometimes, redoxmediators substances such as quinones and ferrocenes, are incorporated to facilitate electron transfer between the electrode and enzyme employed with ratios of mediators ranging from 1 to 6% w/w. (iv) After blending, the pastes were packed into a Teflon electrode holder (geometric surface area of about 0.07 to 3 cm2) with electrical wire. (v) The electrode surface was smoothed on a paper to produce a reproducible working surface.
\n\t\t\t\tThe use of CNTs also has recently been reported to build nanotubes paste electrode (MWCNTPE). Patrascu et al., 2011 showed the use of MWCNTPEs as voltammetric sensors to selectively detect dopamine in the presence of serotonin. Which, it was prepared by thoroughly hand-mixing the multi-wall carbon nanotubes powder with paraffin oil (60:40 w/w) in a mortar. The paste is packed into 1.0 mL polyethylene disposable syringes with a copper wire being used for electrical contact. The surfaces of the electrodes were smoothed by polishing with emery paper.
\n\t\t\tThe epoxy composites filled with graphite are prepared by solution intercalation method, whereby graphite is added to the mixtures of epoxy resin and curing agent. The materials were mechanically stirred, in order for the epoxy resin to intercalate inside the conductive fillers, especially into the graphite interlayers and pores. After the conductive fillers were mixed with epoxy resins, solvents presents in the mixture are evaporated with continuous stirring. The polymer matrix system consists of epoxy resin and a cure agent, and generally all the components are commercial products, and are used as received, without purification.
\n\t\t\t\tEpoxy resins, aromatic or aliphatic, have similar ether linkages as that of the Nafion® membranes commonly used in fuel cells. So it is reasonable to select epoxy resins as the polymer matrix in the composite bipolar plates. One of the most popularly used in composites is formed by curing diglycidal ether of bisphenol A (DGEBA). It was used under its commercial designation (Dow Chemical Company; DER 332) whit triethyltetramine (TETA). The basic reaction involves an amine reacting with the epoxy ring. Resulting in simple chain extension and cross-linking. The former reaction generates a new secondary amine that can also react with DGEBA. This is often referred to as branching. The reaction also produces a hydroxyl group that has long been considered as being ideally located to aid (catalyze) the branching reaction.
\n\t\t\t\tThe criterion of the selection between the type epoxy resin and the cure agent observed in the majority of the publishing paper is in order to adjust the viscosity of the solution during mixing and provide flexibility to the cured products. However, usually the curing processes of these materials need high temperatures, curing at 150º C, 80% of epoxide groups were converted (Merad et al.,2007; Laza et al., 2005), that would be fatal for the biological component. So that, should be reduced the curing temperature and prolong the curing time. These precautions can be extended to other chemical modifiers added to the biocomposite such as redoxmediators, catalysts, etc. that are quite sensitive to wide temperature variations.
\n\t\t\t\tRigid phenyl rings are particularly useful to provide resins with improved heat and thermal resistance and char formation. Undesirably, too rigid backbone structures also drastically reduce the processability of a resin due to viscosity increase, and produce a relatively still but brittle material.
\n\t\t\t\tThe table 1 show different ratios of the composition of rigid composites based in GEC resins, which isn´t standard. So, when to same resin is used, the curing conditions can vary and should be optimised depending on the nature of the fillers present in the biocomposite. For example, a study proposed by Fatibello et al., 2007 show the effect of graphite powder varying from 30 to 80% (m/m), epoxy resin from 5 to 60% (m/m), and tissue from 5 to 20% (m/m) on the biosensor response using 0.025 mol L-1 catechol solution at pH 7. The best composition obtained was in function of the remarkable robustness and sensitivity of the sensor.
\n\t\t\t\t\n\t\t\t\t\t\t\t\t | \n\t\t\t\t\t\t\t\n\t\t\t\t\t\t\t\t | \n\t\t\t\t\t\t\t\n\t\t\t\t\t\t\t\t | \n\t\t\t\t\t\t\t\n\t\t\t\t\t\t\t\t | \n\t\t\t\t\t\t\t\n\t\t\t\t\t\t\t\t | \n\t\t\t\t\t\t\t\n\t\t\t\t\t\t\t\t | \n\t\t\t\t\t\t\t\n\t\t\t\t\t\t\t\t | \n\t\t\t\t\t\t
\n\t\t\t\t\t\t\t\t | \n\t\t\t\t\t\t\t1 part wt | \n\t\t\t\t\t\t\ta part wt | \n\t\t\t\t\t\t\t- | \n\t\t\t\t\t\t\t- | \n\t\t\t\t\t\t\t55 ºC for 2 h | \n\t\t\t\t\t\t\tWang J. et al., 1989 | \n\t\t\t\t\t\t
\n\t\t\t\t\t\t\t\t | \n\t\t\t\t\t\t\t16 part wt | \n\t\t\t\t\t\t\t- | \n\t\t\t\t\t\t\t15 mg GOD/g composite | \n\t\t\t\t\t\t\tone part wt | \n\t\t\t\t\t\t\t35 – 40ºC, for a week | \n\t\t\t\t\t\t\tCéspede et al., 1993 | \n\t\t\t\t\t\t
\n\t\t\t\t\t\t\t\t | \n\t\t\t\t\t\t\t4 parts wt | \n\t\t\t\t\t\t\t- | \n\t\t\t\t\t\t\t20mg HRP /g graphite | \n\t\t\t\t\t\t\t- | \n\t\t\t\t\t\t\t40°C for 4 days | \n\t\t\t\t\t\t\tMorales et al., 1996 | \n\t\t\t\t\t\t
\n\t\t\t\t\t\t\t\t | \n\t\t\t\t\t\t\t7 1% | \n\t\t\t\t\t\t\t- | \n\t\t\t\t\t\t\t2% | \n\t\t\t\t\t\t\t9% | \n\t\t\t\t\t\t\t- | \n\t\t\t\t\t\t\tMartorell et al., 1997 | \n\t\t\t\t\t\t
\n\t\t\t\t\t\t\t\t | \n\t\t\t\t\t\t\t4 parts wt | \n\t\t\t\t\t\t\t- | \n\t\t\t\t\t\t\tg 0.9% (w/w) RIgG | \n\t\t\t\t\t\t\t- | \n\t\t\t\t\t\t\t40 °C for 1 week | \n\t\t\t\t\t\t\tSantandreu et al, 1997 | \n\t\t\t\t\t\t
\n\t\t\t\t\t\t\t\t | \n\t\t\t\t\t\t\t40% | \n\t\t\t\t\t\t\t40% | \n\t\t\t\t\t\t\t5% | \n\t\t\t\t\t\t\t5% | \n\t\t\t\t\t\t\t72 h at 28 ºC | \n\t\t\t\t\t\t\tDutra et al., 2000 | \n\t\t\t\t\t\t
\n\t\t\t\t\t\t\t\t | \n\t\t\t\t\t\t\t1 part | \n\t\t\t\t\t\t\t0.4 part | \n\t\t\t\t\t\t\t0.15 g | \n\t\t\t\t\t\t\t- | \n\t\t\t\t\t\t\t60°C for 24 h | \n\t\t\t\t\t\t\tPuig-Lleixà et al., 2001 | \n\t\t\t\t\t\t
\n\t\t\t\t\t\t\t\t | \n\t\t\t\t\t\t\t4 part wt | \n\t\t\t\t\t\t\t- | \n\t\t\t\t\t\t\t150 μl | \n\t\t\t\t\t\t\t- | \n\t\t\t\t\t\t\t40 °C for 1 week | \n\t\t\t\t\t\t\tPividori et al., 2003 | \n\t\t\t\t\t\t
\n\t\t\t\t\t\t\t\t | \n\t\t\t\t\t\t\t76% | \n\t\t\t\t\t\t\t- | \n\t\t\t\t\t\t\t5% GOD | \n\t\t\t\t\t\t\t9.5% | \n\t\t\t\t\t\t\t40 °C for 1 week | \n\t\t\t\t\t\t\tLlopis et al., 2005 | \n\t\t\t\t\t\t
\n\t\t\t\t\t\t\t\t | \n\t\t\t\t\t\t\t10% | \n\t\t\t\t\t\t\t- | \n\t\t\t\t\t\t\t15% | \n\t\t\t\t\t\t\t- | \n\t\t\t\t\t\t\t- | \n\t\t\t\t\t\t\tLupetti et al., 2006 | \n\t\t\t\t\t\t
\n\t\t\t\t\t\t\t\t | \n\t\t\t\t\t\t\t60% | \n\t\t\t\t\t\t\t40% | \n\t\t\t\t\t\t\t10mg IgG-HRP | \n\t\t\t\t\t\t\t- | \n\t\t\t\t\t\t\t25ºC for 96 h | \n\t\t\t\t\t\t\tBojorge et al., 2007 | \n\t\t\t\t\t\t
\n\t\t\t\t\t\t\t\t | \n\t\t\t\t\t\t\t70% | \n\t\t\t\t\t\t\t30% | \n\t\t\t\t\t\t\t- | \n\t\t\t\t\t\t\t- | \n\t\t\t\t\t\t\t- | \n\t\t\t\t\t\t\tPauliukaite et al. 2009 | \n\t\t\t\t\t\t
Compositions of Graphite/Epoxy composites Biosensor.
The same as epoxy resin, Teflon (polytetrafluoroethylene, PTFE) is other the nonconducting binder used to construct rigid composite transducers with electrochemical sensing applications (Peña et al., 2001; Wang & Musameh, 2003]. The main advantage of the incorporation of enzymes into graphite-Teflon composite matrixes is the compatibility of these electrodes with organic or predominantly organic solvents.
\n\t\t\t\tThe most popular procedure is summarized as: Teflon powder is dissolved in hexane by gentle mixing. Then, the hexane is evaporated under vacuum. Hexane is used to precipitate any asphaltenes presents. Then, an appropriate amount of graphite powder is added to the Teflon and mixed with the enzyme in a mortar for a time to incubate. Finally, the mixture is pressed in into pellets, by means of a Carver pellet press at 10,000 kg cm−2 for 10 min. From this mother pellet, several cylindrical portions of each pellet are bored, and each portion is press-fitted into a Teflon holder. Electrical contact is made through a stainless steel screw (Carralero et al., 2006).
\n\t\t\tOne of the most important steps of building a biosensor is to immobilize the biomolecules. A successful matrix-transducer-composite should immobilize or integrate biomolecules stably at a transducer surface and efficiently maintain the functionality of the biomolecules, while providing accessibility towards the target analyte and an intimate contact with the transducer surface.
\n\t\t\tThe bases immobilization proposed here are: Enzymes are hydrophobically adsorbed on the supports of high ionic strength. There is a covalent “intermolecular” reaction between the adsorbed absorbed protein and the media. The immobilized protein is incubated at alkaline pH to increase multipoint covalent attachment to stabilize the enzyme. The hydrophobic surface of hydrophylized support is the reaction of other groups of amino acids in order to reduce unfavourable interactions enzyme-support hydrophobic. This strategy produced a significant increase in stability, for example, immobilization of enzymes such as HRP, TYR and AOX, which has been extensively used in determination of analyte-target different such as ethanol and phenol and, in comparison with the stability achieved using conventional protocols. The development of a good biocompatible matrix for the immobilization of biomolecules is very crucial to improve the analytical performance of biosensor. A total of four electrochemical sensors were made using different composites: carbon paste, GEC and Teflon/Graphite, to which different enzymes were added to the bulk mixture (see Figure 3). The composition of the composites were selected among successful combinations in the literature (Bojorge et al., 2007; Carralero et al., 2006), and according to preliminary exploratory experiments in our laboratory.
\n\t\t\tRigid-Composite electrodes based on graphite used in the array. a) Initial assembly with electrical contact in a Teflon-tube; b) preparation of the composite paste mixture: an excess of the mixture is placed on the tip of the electrode body during the curing time; c) final aspect after curing and polishing, which is carefully performed to ensure a smooth and flat surface area of 0.015 ± 0.005 cm2.
The bi-enzymatic composite was initially prepared by homogenizing the mass from 0.150 to 0.200 g of graphite powder (Fluka, Cat. No. 50870), previously treated with H2O2 [Bojorge et al., 2007] and a specific mass of 0.050g of Tyrosinase (Sigma) and HRP (Toyobo-Brazil) in a mortar for at least 20 min. After that, epoxy resin at a ratio of 1: 4 wt/wt was subsequently added to this mixture in a mortar and mixed for at least 20 min to produce the final biocomposite paste, that was packed (1000 mg) into the tip of depth of 3mm of a cylindrical Teflon sleeve body (1.5 mm I.D) and a copper wire was used to provide the external electric contact. All composites were prepared at room temperature.
\n\t\t\t\tComposite enzyme electrodes were fabricated in the form of cylindrical pellets, as follows. Graphite (Fluka, Cat. No. 50870), 0.10 g horseradish peroxidase (E.C. 1.11.1.7, 270 I.U. mg-1; Toyobo-Brazil).), 0.014 g and mushroom Tyrosinase (EC.1.14.18.1, Sigma T3824, Sigma-Aldrich Inc.), were accurately weighed and thoroughly mixed by mechanic stirring for 1 h in a 0.4mL suspension of a 0.1 mol L-1 phosphate buffer solution of pH 7.2 at 4º C. Next, put it desiccator at room temperature to evaporate the water. Next, Teflon powder (Aldrich) was added in at proportion of 70% wt. and mixed thoroughly by hand. The mixture was pressed into pellets by using a Carver pellets press at 10000kg cm-2 for 10min. The diameter of these pellets was 1.3 cm and their thickness 0.4 cm, approximately. Several 3.0-mm diameter cylindrical portions of the pellet were bored, and each portion was press-fitted into a Teflon tube. Electrical contact was made through a stainless steel flat-tip screw.
\n\t\t\t\tThe powder graphite received a previous treatment with oxygen peroxide and dried in desiccators. The chemical modified graphite was mixed with the emeraldine polyaniline (Sigma) in a proportion of 30:70 (percentage, w/w) and epoxy resin added (60:40, w/w). The paste was used to fill the cylindrical Teflon sleeve body (1.5 mm ID) electrode and rest 2 hours at 60º C to dry. After that, the composite was well polished to immobilize the horseradish peroxidase (Toyobo-Brazil) with glutaraldehyde (2.5%, v/v) as the agent for the covalent linkage.
\n\t\t\t\tThe chemically modified graphite-emeraldine polyaniline composite (30:70, w/w) was mixed with horseradish peroxidase (Toyobo of Brazil) and alcohol oxidase (E.C.1.1.3.13, Sigma-Aldrich Inc.) adjusting the corrects UI/mg composite values. After that, the epoxy resin was added to the composite and the moisture rested for a few minutes at environment temperature for aggregation. The bi-enzymatic immobilized paste was used to fill the cylindrical Teflon electrode (Serra et al., 2003).
\n\t\t\t\t0.3 g graphite powder was added to the equivalent of 0.2 g of Nujol mineral oil (density = 0.838 g/mL) and then was added 0.01 g of HRP dispersing it slowly for about 10 minutes until a homogeneous paste. The resulting paste is inserted into a pipe end polymer (made from the body of a commercial syringe) into the tip of depth of 3mm of a cylindrical Teflon sleeve body (1.5 mm I.D) and a copper wire was used as electrical contact. The surface is gently polished with A4 paper or tissue paper to achieve a smooth flat surface. Rinsed the surface of the electrode with deionized water.
\n\t\t\t\tElectrical resistivity measurements of the all composites pellets based on graphite and were performed according to the ASTM D 257 standard testing method with a Keithley 6517A electrometer (Cleveland, OH, USA) as the source. From these measurements and the geometric dimensions of the sample composites electrodes, both the volume resistivity and surface electrical insulating materials and can be calculated as well as the corresponding conductivities.
\n\t\t\tMorphology studies of all composites pellets were carried out with SEM. All the composites pellets were mounted directly onto aluminum specimen stubs with two-sided adhesive carbon tabs (Pelco, Redding, CA) and coated with gold for approximately 45 s at 20 lA and 75 mTorr. Samples were analyzed in a JEOL JSM-6460 emission scanning electron microscope dotted with detector electron back-scattered diffraction pattern and system integrated analysis station software used to interface with the SEM and EDS detector operating at an accelerating voltage of 30 KV. Representative micrographs were chosen to illustrate each composite´s pellet.
\n\t\t\tThe electrodes were cycled for 3–5 times in distilled water in order to get stable voltammetric responses before performing the measurements with analyte samples. Cyclic Voltammetry measurements were taken using a potentiostat Autolab/PGSTAT12 (Ecochemie, Netherlands, http://www.ecochemie.nl/). The following parameters where fixed: First potential: −1.0 V; Second Potential: 1.0 V; Scan rate: 0.1 V s−1 and Step potential: 0.00244 V. All experiments were carried out without any oxygen removal from the sample and with no physical surface regeneration of the working electrodes after each measure.
\n\t\t\t\tIn order to obtain reliable measurements, and to prevent the accumulative effect of impurities on the working electrode surfaces, an electrochemical cleaning stage was considered between measures. This stage was performed by applying a conditioning potential of +1.5 V for 30 s after each experiment, in a cell containing 5 ml of distilled water.
\n\t\t\tChemical compositions of the materials of different composites obtained, the variation of the electrical conductivities are presented in Figure 5. To single filler epoxy composite as a function of the graphite loading concentration the electrical conductivity of graphite/epoxy resin increase gradually with increasing graphite filler. This phenomenon shows that the graphite acts as the transfer medium for electrons hence the electrical conductivity of the composite would be increased. However, the electrical conductivity of the graphite composite is still low, reaching only 0.0078 S/cm at loading concentration of 80 w/w %. This suggests that composites filled with graphite only are not suitable for the achievement of high electrical conductivity. A low resistivity is desirable when high currents are flowing through the electrode body in order to avoid the iR drop of the applied potential. However, when the composite are constituted of graphite/Teflon the performance of conductivity is different. The electrode resistivity was lower when working with high ratios of graphite, but an inverse tendency for mechanical strength was observed. This means that when graphite is fully intercalated with Teflon, the material is a rather poor electrical conductor, so, the change in Teflon composition fraction could create variations in conductivity probably due to an induces defects in the graphite structure with the intercalation with Teflon. In relation to the conductivity of the PANI- Graphite/epoxy composite (GEC-PANI), it can be see that the conductivity of composites increased with increasing graphite content. The conductivity found for neat polyaniline was 0.00249 Scm-1. However after addition of 1, 2 and 3 w/w % of graphite the electrical conductivity of composites increased slightly and for pellets prepared using 10-30 w/w % of graphite the electrical conductivity of composites increased significantly showing a maximum rate conductivity of 28 µScm-1 for GEC-PANI 30 w/w % sample as shown in Figure 3, indicating that, for the electrical properties, the PANI-Graphite/epoxy 30 w/w% present the better compromise between filler and polymeric matrix.
\n\t\t\t\t\tElectrical conductivity of composites in variation graphite composition
The SEM is a very good technique for investigating the morphology of composites based on graphite, due to provide information about the structure, size and distribution of the graphite particles in the matrix of the composite. Figure 4 shows scanning electron micrographs of carbon paste, GEC, Graphite-PTFE and GCE-PANI composites pre-treated as described in the experimental section. As can be seen, carbon paste composite made of graphite powder mixed and Nujol oil (Figure 5a) the electrode surface has a rough surface morphology and is heterogeneous with lighter areas, associated with the Nujol oil, and darker areas corresponding to the graphite conducting micro-structures. Figure 5b shows the SEM of GEC surface, which is less rough than CP. However, cluster appears also, which it appears agglomerated in random areas. This is due to the graphite particles randomly distributed and randomly oriented in the epoxy resin. Another important characteristic of the graphite-epoxy composite is the rather flat fracture surface indicating the nature of brittle fracture. Figure 5c show SEM image of the worn surface for Graphite-Teflon, SEM examinations of Graphite-Teflon show slightly ribbed i.e., less creation and development of the cracks occurred on the surfaces. This can be seen with the naked eye during the preparation, whose surface is brighter and sharper with Teflon than epoxy, because developing a uniform transfer layer on the composite-surface. Figure 5d shows SEM image for graphite-PANI-epoxy that revealed also the uniform morphology of the structures in which the outer layers were PANI and the inner layer consisted of graphite. Such a uniform morphology is desirable because it enables a material with high ionic conductivity to achieve fast charge/discharge rates. In the present study, however, the morphology of PANI was strictly granular and it coalesced, making the surface rough with no uniformity.
\n\t\t\t\t\tElectronic microscopic images of composites: a) GP (Graphite Paste) electrode; b) GEC (graphite epoxy composite) electrode; c) GEC (Graphite Teflon composite) electrode and d) GEC-PANI. All electrode surfaces were polished as explained in the text. The same accelerated voltage (20 KV) and resolution (5µm) were used.
\n\t\t\t\t\t\tFigure 6 shows the cyclic voltammetric behaviors of different composites based on graphite with potential sweep rates of 100 and 50 mV/s between –0.1 and 1 V in solution containing 1 mM K4[Fe(CN)6] and were recorded on the five cycle of a repetitive potential sweep program. As it can be seen, the voltammograms that show the well-defined redox peaks is the GEC, which it is related to the intrinsic characterization of graphite. However, for the voltammogram of the PANI-graphite composites show a nearly rectangular-shaped voltammogram, typical of capacitive systems. This characteristic is attributed mainly to the enhancement of the conductivity due to the graphite particles added to the polyaniline chains. The anodic peak, occurred at potential of ~ 0.2 V, is connected to doping of chloride anions indicating transition of leucoemeraldine form of PANI to emeraldine salt. Further increase in potential, above ~ 0.50 V, refers to transition of emeraldine salt to perningraniline salt. Therefore, it can be observed that the electrochemical kinetics of redox transition of PANi-graphite composites has been performed.
\n\t\t\t\t\tCyclic voltammograms for 10 mM potassium ferricyanide employing the composites sensors: a) using GP composite, b) using Graphite-Teflon composite, c) Graphite-Epoxy composite, d) Polyaniline Graphite-Epoxy composite. Scan rate 100 mV/s
In agreement with this, the reaction kinetics of the electroactive species in concentrated solutions may be slowed due to the slow mobility of the molecules. In the Figure 6d, the degradation of the PANI-perningraniline salt electrode almost was not observed for that potential region. The possible explanation could be to the fact that some forms of PANI are potential dependent and pH dependent [Tawde et al., 2001] or the fractal dimensions dependent [Ghanbari et al., 2006] evaluated from the cyclic voltammetry. Hence, at such low pH, extent of the degradation products was negligible and practically had no influence on charge/discharge characteristics of the PANI electrode during initial cyclization. However, this work is still in progress, in further studies it would be beneficial to investigate the influence of pH and anions on degradation of PANI during the cyclization. Though, attention must be paid to each measurement made with a composite film as accurate reproducibility of the arrays and polymeric composites is very difficult. The results indicated that the electrochemical methods could be used as a simple tool for analyzing the structure of conducting polymers and their composites.
\n\t\t\t\tThe electrochemical performance of the composite electrodes is strongly affected by the presence of the insulator matrix of the polymeric binder, which coats the graphite particles, at the same time increasing electrode resistivity which is influenced by the distance between the particles. Since it is known that a low rate of evaporation of the solvents provides the smoothest surfaces, the material was dried at low temperatures of ≈20 to 30º C.
\n\t\t\tDifferent morphologies are expected for different composite compositions. We can conclude that the nature of the surface film plays a key role for the electrode stability or to capacity decrease always related to an increase of the electrode itself impedance
\n\t\tThe Authors thanks for the resources received from Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (Proc. No. E26/150.997/2006) and Conselho Nacional de Pesquisa e Desenvolvimento – CNPq (Edital Universal, Process number 476654/2008-4). and in special to Professor Belkis Valdman, our inspiration, and to Students João Paulo Barros Guimarães Mendes, Erica Ferreira Southgate, Leonardo Ivar Gomes Jaldin, Gleice Santos Lima Magalhães by theirs participations in the LABSENS by the Scientific initiation program - PIBIC - UFRJ, to which we encourage to continue in this way.
\n\t\tBuckwheat is an annual herbaceous plant that botanically belongs to the order
Buckwheat grains are the main form of consumption of this pseudocereal. Hulled grains are mostly used for human consumption in the form of breakfast cereals or as flour for the production of various bakery products (bread, cakes, snacks) and noodles enriched with buckwheat flour (0.3–60%), buckwheat-improved non-bakery products (tea, honey, and tarhana) [5] and products made of buckwheat husks such as pillows, quilts, mattresses, collars, eye masks, and children’s toys [6]. In addition to flour and groats, buckwheat sprouts are increasingly used to improve bakery products [5, 7].
Since buckwheat is gluten-free, these products can be included in a gluten-free diet for patients with gluten intolerance [8, 9].
The addition of buckwheat into bakery products is of particular importance. This pseudocereal is gaining increasing attention as potentially functional food [4, 10]. Namely, buckwheat is recognized as a good source of nutritious proteins, lipids, dietary fiber and minerals and, in combination with other components that have a positive impact on health such as phenolic components and sterols, it is attracting increasing attention as a functional food.
Buckwheat is the only pseudocereal that contains rutin, which has shown anti-inflammatory, anticancer, antiatherogenic, and antioxidant activity [4, 12]. Buckwheat protein extracts are associated with anticancer and cholesterol-lowering effects in animals [13, 14]. Apparently, the incorporation of buckwheat into bread results in significantly lower blood glucose and insulin responses compared to white wheat bread [15]. Buckwheat grain contains very rare D-
Buckwheat proteins have a high biological value thanks to a well-balanced amino acid composition. The protein content of buckwheat is relatively lower than the protein content of legumes. However, the amino acid score of buckwheat protein is 100 and the content of essential amino acids corresponds to the recommended amino acid intake for children and adults [17].
They are rich in lysine, which is the first limiting amino acid of plant proteins, and arginine [18, 19]. However, the content of glutamine and proline is much lower compared to wheat [20], and threonine and methionine are the first and second limiting acids in buckwheat. Furthermore, Giménez-Bastida et al. [5] stated that buckwheat proteins are rich in albumin and globulin and that they are very poor in prolamin and gluten. Therefore, buckwheat flour is suitable for use in the diet of people with celiac disease due to its low non-toxic prolamin content [21]. The protein content in buckwheat flour is significantly higher compared to rice, wheat, corn, millet, and sorghum flour. While it is lower only compared to the protein content of oat flour [19]. Guo et al. [22] pointed out that the average protein content in buckwheat is 12.94%.
The digestibility of buckwheat proteins is about 80%, which is lower compared to proteins of animal origin such as hemoglobin and ovalbumin. However, it is higher than cereal proteins (e.g., sorghum 55–59%; corn 66–75%) and has a value approximate to rice bran (89%) and wheat germ (77–93%). Despite the balanced composition of essential amino acids, the bioavailability of buckwheat protein after digestion is not complete. Relatively low digestibility is attributed to the molecular structure of buckwheat protein and the presence of antinutritive factors in flour and protein isolates [17].
Buckwheat, along with other pseudocereals such as quinoa and amaranth, is recommended for use in creating new value-added bakery products because it can provide high levels of essential amino acids in the human diet [23].
The literature states that buckwheat proteins have many unique physiological functions, such as treating chronic diseases in humans, reducing serum cholesterol, suppressing gallstones and tumors, inhibiting angiotensin I-converting enzyme (ACE), and so on [17, 24, 25]. An ACE inhibitory tripeptide (Gly-Pro-Pro) was isolated and identified from common buckwheat [26]. In humans, buckwheat consumption has also been associated with a lower prevalence of hyperglycemia and improved glucose tolerance in people with diabetes [27]. Since many health benefits of buckwheat are inherently related to peptide radical binding activity from digested proteins, it is hypothesized that buckwheat protein hydrolysis may release peptide fragments capable of stabilizing reactive oxygen kinds and inhibiting lipid oxidation. By in vitro digestion of buckwheat protein six peptide fractions were obtained, whereas LC-MS/MS identified Trp-Pro-Leu, Val-Pro-Trp, and Val-Phe-Pro-Trp (IV), Pro-Trp (V), and tryptophan (VI) as the prominent peptides/amino acid in these fractions [28]. Six peptides DVWY (H-Asp-Val-Trp-Tyr-OH), FDART (H-Phe-Asp-Ala-Arg-Thr-OH), FQ (H-Phe-Gln-OH), VAE (H-Val-Ala-Glu-OH), VVG (H-Val-Val-Gly-OH), and WTFR (H-Trp-Thr-Phe-Arg-OH) identified from buckwheat sprouts fermented with
Many health benefits of buckwheat are attributed to the high content of phenolic components and high antioxidant activity [18]. Whole grain buckwheat was found to contain 2–5 times more phenolic components than barley and oats, while the husk and bran of buckwheat have 2–7 times higher antioxidant activity compared to barley, triticale, and oats [32, 33]. The research by Begić et al. [34] showed that Tartary buckwheat contains about 20 times more total phenol content and that it shows antioxidant activity nine times higher than common buckwheat.
Among the polyphenolic components present in buckwheat, those from the group of flavonoids, and among them rutin, are the most important ones.
The presence and amount of flavonoids in buckwheat grain make it specific compared to cereals, which contain small amounts of flavonoids. This group of polyphenolic components is the basic antioxidant of buckwheat [35, 36]. Buckwheat is considered to be one of the best dietary sources of rutin [36]. The content and composition of flavonoids are different in different types of buckwheat. In general, the flavonoid content in
Flavonoids demonstrate a protective effect in lipid oxidation in vitro as “scavengers” of free radicals and metal chelators [39]. They generally occur as O-glycosides in which one or more hydroxyl groups are bound to sugars.
Six flavonoids were isolated and identified from whole buckwheat grains: rutin, quercetin, orientin, isoorientin, vitexin, and isovitexin. The presence of rutin and isovitexin was found in hulled grain while buckwheat husk contained all 6 flavonoids [40, 41]. Buckwheat is the only pseudocereal that contains rutin and is, therefore, a useful source of this flavonoid [25]. Except in buckwheat, rutin has not been detected in cereals and pseudocereals [41, 42]. Rutin (quercetin-3-O-β-rutinoside), a secondary metabolite present in buckwheat, is the best-known glycoside derived from flavonol quercetin. Buckwheat is considered the best source of dietary rutin. Buckwheat grains (groats and husk) and sprouts are important sources of rutin and their content depends on the type and conditions of growth [43, 44]. It is important to develop new well-adaptive varieties with a high content of rutin, and improved biological value of the proteins [45].
Tartary buckwheat groats contain more rutin—80.94 mg/g dry matter (DM) than common buckwheat groats—0.20 mg/g DM [46, 47] while Tartary buckwheat sprouts have 2,2 times more rutin than common buckwheat sprouts [48]. Li et al. [49] stated that Tartary buckwheat can contain up to 100 times more rutin than common buckwheat.
Rutin has attracted much attention mainly because of its many health benefits observed in vitro and in vivo: anti-inflammatory, antidiabetic, hypocholesterolemic, antiatherogenic, antiatherosclerotic, and anticancer ones [4, 12, 36, 50, 51, 52, 53] and its activity are related to antioxidant capacity [54]. Furthermore, rutin may be effective in preventing the toxic effects of methotrexate on the kidneys [55].
Rutin has relaxing effects on smooth muscles and is effective in preventing capillary apoplexy and retinal bleeding, lowers high blood pressure and shows antioxidant activity and lipid peroxidation activity. It also has lipid-lowering activity by reducing dietary cholesterol absorption, as well as reducing plasma and liver cholesterol [56, 57]. In addition, possibilities of rutin as a new strategy for the prevention of type 2 diabetes are noted [58]. Alkaline luminol chemiluminescence and electron spin resonance analysis revealed the formation of the rutin-ovalbumin complex which significantly increases the radical-binding activity in rutin. Rutin has also demonstrated antioxidant activity against hydroxyl radicals in a DNA protection test [59].
Quercetin (quercetin-3-ramnoside) is another glycoside present in buckwheat in concentrations ranging from 0.01 to 0.05% DM in Tartary and from 0.54 to 1.80% DM in common buckwheat [46, 60]. Isoquercetin (quercetin-3-glucoside) is present in buckwheat hypocotyl (1.4 μM/g DM) [61] and has been shown to exhibit antidiabetic and anticancer activity [36, 49, 62, 63]. Quercetin, an aglycone of rutin, is present in hulled grain (semolina) of buckwheat (0.001 mg/g DM) and husk (0.009–0.029 mg/g DM) in lower concentrations than rutin [18, 47]. Quercetin is the most studied flavonoid, primarily due to its pronounced antioxidant activity, as well as significant absorption in the digestive tract. It is predominantly in the form of glycoside as rutin (quercetin-3-O-beta-rutinoside). In addition to this, kaempferol-3-O-rutinoside and quercetin 3-O-rutinoside-3’-O-β glucopyranoside have been found in buckwheat seeds [49, 64].
Three flavonoids from Tartary buckwheat bran: quercetin, isoquercetin and rutin were evaluated as R-glucosidase inhibitors (controlling blood glucose) using fluorescence spectroscopy and enzyme kinetics. The R-glucosidase activity was clearly influenced by extractives (mostly rutin) and their hydrolysis products (a mixture of quercetin, isoquercetin, and rutin) from buckwheat bran [65].
Recent research relating to the examination of the antiviral activity of rutin in the treatment of patients with COVID-19 symptoms have been topical [66, 67].
In addition to rutin, catechins, the antioxidant activity of which is higher than the antioxidant activity of rutin, were isolated from ethanol extracts of buckwheat groats. Four catechins were isolated and their structures were determined as: (−)-epicatechin, (+)-catechin-7-O-β-D-glucopyranoside, (−)-epicatechin 3-O-p-hydroxybenzoate, and (−)-epicatechin 3-O-(3,4-di-O-methyl)gallate [68]. The following components from the catechin group were identified in buckwheat: catechin, epicatechin, catechin glucoside (A or B isomers), catechin gallate, epicatechin gallate, epicatechin-O-3,4-dimethylgallate, epiaphzelchin-(4─8)-epicatechin-3,4-O- dimethylgallate, while catechin-3,4-O- dimethylgallate was identified in thermally treated buckwheat and epiaphzelchin- (4─6) -epicatechin was identified in green buckwheat [69].
These ingredients in plant tissue are influenced by numerous environmental factors such as ultraviolet (UV) radiation, harvest time and damage caused by pests, and genetic and aging-related factors. Studies have shown significant positive correlations between the mean altitude of the growth site and the amount of individual phenolic antioxidants [70, 71]. Buckwheat, as a source of rutin, can be successfully grown in Mediterranean conditions, too [72]. Flavon-3-glycosides present in buckwheat (vitexin, isovitexin, orientin, and homoorientin), anthocyanin and proanthocyanin content [61] and the presence of squalene, epicatechin, and vitamin E [73] make buckwheat a good antioxidant source in the human diet.
Phenolic acids in buckwheat also contribute to its antioxidant activity. In the grain of different varieties of Tartary buckwheat, p-hydroxybenzoic, ferulic and protocatechuic stand out, and other acids, including p-coumaric, gallic, caffeic, vanillic, and syringic acid, were also detected [74]. Several phenolic acids have been described during the flowering of different varieties of buckwheat: chlorogenic, p-anisic, salicylic, and methoxycinnamic [75].
Buckwheat is also an important source of vitamins, especially those of the B group. The total content of B vitamins, including B1 (thiamine, 2.2–3.3 μg/g DM), B2 (riboflavin, 10.6 μg/g DM), B3 (niacin, 18 μg/g), B5 (pantothenic acid, 11 μg/g) and B6 (pyridoxine, 1.5 μg/g) is higher in Tartary buckwheat compared to common buckwheat. The levels of vitamin C indicated for it in the literature go as high as 50 μg/g DM while its content, as well as the total amount of vitamins B1 and B6, increases by germination of buckwheat and, consequently, the content of vitamin C in buckwheat sprouts reaches 250 μg/g DM [4, 54, 76, 77]. Vitamin B1 is found in thiamine-binding proteins of buckwheat grains, which, according to Li and Zhang [37] increases the availability of vitamin B1 and improves its stability during storage. The content of vitamin E (tocopherols) in buckwheat is higher compared to wheat, barley, oats, and rye [18]. The most common tocopherol in buckwheat is γ-tocopherol. In addition to γ-tocopherol, α- and δ-tocopherol have also been identified in buckwheat [78]. The concentration of total tocopherols in buckwheat grains ranges from 14.3 to 21.7 mg/kg [79]. Tocopherols, along with the other components mentioned above, make buckwheat a good antioxidant source in the human diet. Tocotrienols were not detected in buckwheat [80, 81], while Piironen et al. [82] identified traces of tocotrienols in whole buckwheat grains. High levels of vitamin E intake are associated with a reduction in cardiovascular disease, a reduction in the risk of Alzheimer’s disease, and an improvement in the immune system [73].
Squalene is an isoprenoid component that has six isoprene units and antioxidant activity and is widely produced in buckwheat plants. Squalene protects cells from radicals, strengthens the immune system, and reduces the risk of various types of cancer. There are some differences between buckwheat species, especially in the content of squalene and rutin [73].
Buckwheat grains contain a very rare D-
So far, several studies have described the role of D-
Buckwheat is also significant for its phytosterol content. Phytosterols present in buckwheat, although in low concentrations, also show a positive effect in lowering blood cholesterol levels. In addition, phytosterol intake significantly reduces in vivo cholesterol absorption. Buckwheat phytosterols are present throughout the grain, but their content varies by grain parts [37]. The most abundant phytosterol in buckwheat flour is β-sitosterol (0.86 mg/g DM) and makes up about 70% of total sterols, followed by campesterol (0.11 mg/g DM) and stigmasterol (0.02 mg/g DM) [94].
According to the research of Dziedzic et al. [95]. the sterols content in Tartary buckwheat whole grains was 15,398 mg/kg of lipids and the most prevalent was the β-sitosterol (10,944 mg/kg of lipids).
D-fagomin is a minor component from the group of iminosugars detected in the dehulled grain of common buckwheat which shows a glucose-lowering effect [96, 97].
Similar to D-fagomin, other imino sugars such as 1-deoxynojirimycin (DNJ) are intestinal glucosidase inhibitors, associated with a reduced risk of developing insulin resistance, gaining weight, and suffering from excess potentially pathogenic bacteria [97, 98]. Anthraquinone emodin is present in buckwheat concentrations between 1.72 and 2.71 mg/kg DM [99]. Due to the wide range of biological activities that emodin exhibits, it can be considered an important bioactive factor in buckwheat [100].
Along with vitamins, other components were detected, such as glutathione (1.10 mmol/g DM in buckwheat groats), phytic acid (35–38 mg/g DM in bran), carotenoids (2.10 mg/g DM in grain), and melatonin (470 pg/g DM in groats). These components may contribute to the antioxidant activity of buckwheat [4, 47].
Both types of buckwheat, common and Tartary, have a high tannin content (1.76 and 1.54%). Tannins isolated from buckwheat showed a relatively high level of activity against
It has also been found that γ-aminobutyric acid (GABA) and 2″-hydroxynicothianamine (2HN) serve as functional components in buckwheat. Grains and sprouts contain GABA, while 2HN was identified in flour. Literature states that these components lower blood pressure in humans and inhibit the activity of angiotensin I-converting enzyme (ACE) [102, 103, 104]. Suzuki et al. [105] quantified GABA and 2HN concentrations in common and Tartary buckwheat leaves 14, 28, and 42 days after sowing (DAS). The concentration of GABA reached a peak at 42 DAS, while the concentration of 2HN decreased with the age of the plant.
It is well known that processing can cause chemical changes in food products. Therefore, it is important to consider the effects on bioactive components in buckwheat. Today, there are several technological processes related to buckwheat, which will be presented below.
Milling is one of the technological processes that is inevitable during the processing of buckwheat into flour. During the processing of buckwheat grains into white flour, the husk and outer layers are separated, which lowers the ratio of fibers, minerals, and polyphenolic components.
Hung and Morita [106] explored the possibility of improving the functionality of buckwheat flour by successively milling buckwheat and they found that in 16 different fractions of flour the content of ferulic acid and rutin increases with an increased ratio of outer grain layers. The same authors found that the antiradical activity on DPPH extracts of free and bound polyphenolic components of buckwheat, the fraction of the successive milling of buckwheat is highest for fractions containing external grain parts. Additionally, better antiradical activity on DPPH was registered for extracts of free polyphenolic components compared to extracts of bound polyphenolic components in buckwheat grain.
Inglett et al. [107] examined the antioxidant activity of ethanolic extracts of four types of commercial buckwheat flour and found the highest antiradical activity on DPPH in buckwheat flour containing a high ratio of husk and aleurone layer, while the lowest antiradical activity was registered in white flour consisting exclusively of the endosperm. The highest content of total polyphenolic components and total flavonoids was registered in whole buckwheat flour. Gallardo et al. [108] established that the content of rutin in buckwheat flour is 0.7 mg/100 g and 11.2 mg/100 g in buckwheat husk.
A recent study found that the buckwheat protein contents decreased from the exterior to the interior parts of the groats [109]. Significantly higher content of amino acids, fatty acids, polyphenols, and flavonoids was found in the bran of Tartary buckwheat, compared to the flour [110].
It should be noted that the milling conditions should be adapted to the type of buckwheat. The granulation composition of common and Tartary buckwheat flour differed under the same milling conditions and affected the physical characteristics of the obtained flour fractions. Tartary buckwheat flour contained larger fractions compared to common buckwheat flour under the same milling conditions [111]. By adjusting the grinding and knowing the content of different components in the fractions of Tartary buckwheat, it is possible to obtain products of different nutritional value [45, 111].
The number of studies researching the effects of heat treatment on buckwheat foods has increased significantly. Today, many new thermal techniques are used in the food industry to improve the quality of functional buckwheat food. The extrusion process has become important in the production of pasta, ready-to-eat cereals, snacks, animal feeds, and textured plant proteins. Microwave heating has gained popularity in food processing due to the ability of this technique to achieve high heating rates, significantly reduce cooking time, provide more uniform heating and safe handling. This technique could change the taste and nutritional properties of food to a lesser extent, as opposed to conventional heating during the cooking process [112]. However, data on the effects of heat treatments on the antioxidant capacity of buckwheat and its products are still limited. In general, most studies are aimed at determining the effect of heat treatment on the content of total phenols and flavonoids due to their role in the management of human health and diseases.
It was established that the heat treatment of buckwheat causes changes in its chemical composition and, above all, that it affects the functional properties of the selected bioactive components. The results published so far on the effects of the heat treatment on buckwheat grain and processed flour are contradictory.
One of the first studies was conducted by Dietrych-Szostak and Oleszek [40] who examined the effect of heat processing on flavonoid content in hulled grains and buckwheat husks by removing the husk using heat. Removing the husk from buckwheat grain by applying heat treatment resulted in a product that was both visually and chemically different. The peeling process removed primarily the multitude of tannins and crude fibers that are naturally present in the husk. As for the concentration of total flavonoids, dehulling process with different temperature treatments caused a drastic reduction of the total flavonoid concentration in the grain (by 75% of the control) and smaller but significant (15–20%) reduction in the hulls.
Kreft et al. [12] compared the content of rutin in buckwheat products with its content in the raw materials used to obtain these products. Noodles prepared with 70% of integral buckwheat flour contained much less rutin (78 mg/kg DM) compared to the integral buckwheat flour (218 mg/kg DM) out of which they were produced. As a possible explanation for this reduction in rutin content in the product, the authors cited the presence and activity of an enzyme that degrades rutin, flavonol 3-glucosidase, during dough mixing. The presence of this enzyme in buckwheat was confirmed by Suzuki et al. [113]. In raw (uncooked) hulled buckwheat grain (raw buckwheat semolina) the rutin content was 230 mg/kg DM, while in pre-cooked hulled buckwheat grain its content was 88 mg/kg DM. The aforementioned authors explained the established reduction of rutin content in hulled buckwheat grain due to hydrothermal treatment by possible degradation of rutin molecules or its combination with some other molecules, in such a way that it becomes insoluble in the applied solvent. A similar reduction in rutin content was observed during bread production in different combinations of wheat and Tartary buckwheat flour (100:0; 70:30; 50:50; and 0:100) where the effect of making bread and baking on the content of rutin, quercetin, and polyphenols and the antioxidant activity of said loaves was examined. After baking, rutin (0.47 mg/g) was present in bread which is made of 100% Tartary buckwheat flour, together with quercetin (4.83 mg/g). The dough that this bread was made of contained a lower concentration of rutin and greater concentration of quercetin compared to flour used to prepare it; wherein 0.0175 mmol of rutin degraded with the addition of water and yeast to Tartary buckwheat flour, and 0.0149 mmol of quercetin was obtained at the same time. This indicates that 85% of the rutin was converted to quercetin by adding water and yeast to the flour [114].
Degradation of rutin can be the result of activities of the rutin-degrading enzyme found in buckwheat. This enzyme is stable and active at pH 5–7 and below 40°C. Based on the comparison of the level of concentration, it appears that quercetin is more stable compared to rutin in the process of proofing and baking bread. There were no significant differences in the content of rutin and quercetin between the bread crumb and crust. Additionally, the results showed a reduction in the total polyphenol content in all samples of bread as a result of the heat treatment in the baking process [112].
The obtained results were consistent with the results of the authors Alvarez-Jubete et al. [21] which showed a significant decrease in the concentration of total polyphenols, particularly phenolic acids in bread made of common buckwheat flour (0.65 mg GAE/g) compared to the concentrations of these components in buckwheat grain (3.23 mg GAE/g). During the process of mixing and proofing the bread, there was a modest increase in the concentration of total polyphenols in bread samples made with 100% wheat flour and 100% Tartary buckwheat flour, and a slight reduction in the other samples, containing a combination of both kinds of flour (70:30 and 50:50).
Reductions in polyphenol content and antioxidant activity were also reported during baking of bread samples prepared in different combinations (90:10; 80:20; and 70:30) of rice and buckwheat flour (wholemeal and white) relative to their content in flours. It was noticed that the baking process resulted in a higher percentage of reduction in total polyphenols in bread samples made white buckwheat flour, while only minor or insignificant changes were observed in lower percentages (10 and 20%) in samples with wholemeal buckwheat flour, and only the sample with 30% of wholemeal buckwheat flour had a decrease of about 17%. In addition, the decrease in antioxidant activity was more pronounced in bread samples prepared with white buckwheat flour. During baking, there was a loss in rutin content in bread samples relative to its assumed (calculated) content, and this loss increased with increasing the proportion of buckwheat flour, both types (wholemeal and white), in the range of 4.57–40.4%. The opposite trend was observed in the quercetin content, which increased from 1.5 to 7 times, probably due to the hydrolysis of rutin into quercetin [115].
Similar results were shown in bread samples produced with the addition of buckwheat in the amount of 15 g/100 g and 30 g/100 g. A decrease in the content of total phenols, total flavonoids, and antioxidant activity in bread samples relative to their content in flour was found. The content of total flavonoids in bread samples was 2 to 4 times lower compared to its content in flour [116].
The thermal treatment of Tartary buckwheat bran and flour significantly reduced the content of fatty acids, polysaccharides, and polyphenols. As for the content of amino acids and total flavonoids, their content in bran after heat treatment decreased, while increased in the flour of Tartary buckwheat [110].
In addition to bread and cakes, a decrease in the content of bioactive components was observed in the production and cooking of other products with the addition of buckwheat such as spaghetti, pasta, noodles, etc. A decrease in the content of free (about 74.5%) and bound (about 80%) phenolic components “farm to table”, i.e., from flour to cooked spaghetti with buckwheat was found. Regarding the content of total phenolic components, the spaghetti production process (mixing, extrusion, and drying) caused a loss of 45.9%, which the authors explain by the increase in temperature during the extrusion process and the high temperature (about 95°C) reached during drying. Further degradation of phenolic components was found after cooking spaghetti. The boiling process caused the degradation of 52.9% of the total phenolic components. This degradation was significantly different (p < 0.05) compared to post-production degradation. This effect can be attributed to the solubility of phenolic components in boiled water. Of the total phenolic components that were present in the spaghetti after the drying process, 11.6% were dissolved in water after cooking [117].
Biney and Beta [118] also reported that the production and cooking process led to a reduction in phenol content and antioxidant activity in spaghetti enriched with buckwheat flour and bran. The production process did not cause statistically (p < 0.05) significant changes in the content of total phenols between flour mixtures and uncooked products. However, cooking significantly reduced total phenols in all spaghetti samples. Although the addition of buckwheat flour resulted in a significantly higher content of these components in all spaghetti samples, the average percentage of decrease in total cooking phenols due to cooking was higher in samples containing buckwheat flour or bran, compared to control samples prepared from semolina. The production and cooking process also led to a significant reduction in the content of rutin and total flavonoids in spaghetti samples. The higher the proportion of buckwheat in the spaghetti formulation, the greater the losses in rutin content.
The results of similar research (pasta enriched with buckwheat flour in the amount of 20%) showed a reduction of about 44% of total phenolic components after cooking compared to their content in pasta after drying; 8.37% of total phenols from dried pasta was present in the water in which it was boiled, and 35.63% was degraded. The cooking process reduced the rutin content by about 8.50%. During cooking, rutin was converted from its bound form to quercetin, which is shown in the increase of quercetin content by about 20%. The results also showed that catechin showed a minimum tolerance to the cooking process, with a loss of about 57% [119].
Furthermore, the autoclaving of buckwheat grains caused a decrease in free and an increase in bound phenolic forms in flour. Similarly, this was found in noodles produced by adding this flour to the formulation with wheat flour, compared to the content of these components in noodles produced in the same way with flour obtained from untreated buckwheat grains. Although autoclaving caused an initial reduction in rutin in treated grain flours, it prevented further degradation and conversion of rutin to quercetin in uncooked and cooked samples obtained from these flours, causing a possible improvement in the sensory acceptability of noodles. The loss of phenolic components in noodle samples with added buckwheat flour during cooking (48.1–61.1%) was at the same level as in the control sample with wheat flour only, indicating that buckwheat-containing pasta can maintain the quality during cooking [120].
Cho and Lee [121] examined the thermal stability of rutin in wheat instant fried noodles fortified with rutin-enriched material (REM) from buckwheat milling fractions. The noodles were fried at different temperatures (150, 170, and 190°C) during different periods of time (1, 2, and 3 minutes). Also, noodles were placed in boiling water at different periods (0, 3, and 6 minutes) to examine the effect of cooking on rutin content. The results showed that different temperatures and frying times did not negatively affect the rutin content, while a marked loss of rutin was observed after cooking the noodles.
However, the results of another study showed a reduction in bioactive components in buckwheat products during various heat treatment processes and reported an increase in the total antioxidant activity in buckwheat sprouts and shoots after autoclaving treatment. Furthermore, an increase of 20% and a reduction of 7% of total phenols were observed in buckwheat sprouts and shoot respectively [122].
Contradictory results were also found by Zieliński et al. [123] during extrusion of buckwheat, which showed a decrease in antioxidant capacity accompanied by a reduction in rutin and isovitexin, but at the same time an increase in free phenolic acids and those freed from ester bonds. The authors stated that the reported increase in phenolic acids could be due to the increased release of these bioactive components from the matrix, making them available for extraction. The same authors report that, although the extrusion caused a marked reduction in antioxidant content in hulled buckwheat grain, the amount of bioactive component in hulled buckwheat grain after thermal treatment was still significant, resulting in a decrease in antioxidant activity of only 10%.
Hes et al. [124] also reported contradictory results when testing the impact of cooking in water on the antioxidant properties and dietary fiber of hulled buckwheat grain. It was shown that cooking in water for 30 minutes in a ratio of 2:1 (water: grain) has no negative effects on the nutritional characteristics of the hulled buckwheat grain. Extracts of cooked hulled grain showed a significantly higher content of polyphenols and total dietary fiber compared to raw grain. The detected higher content of polyphenols in cooked hulled grain is explained by the authors as a result of their partial release from the bound form of the protein as a result of cooking. Additionally, phenols can also be associated with other components such as carbohydrates. In terms of individual phenolic components, a significantly higher content of catechin particularly stands out, and, in contrast to that, a considerably lower content of p-coumaric acid in the extracts of cooked buckwheat grain compared to the extracts of the raw buckwheat grain. Cooking did not cause any changes in rutin content.
It has been recognized that the possible beneficial effects of phytochemicals present in buckwheat may be related to the inherent antioxidant capacity of these components. Therefore, during the last decade, the relationship between antioxidant capacity and these components after heat treatment has been exposed. The antioxidant capacity of buckwheat products is linked to flavonoid concentrations after hydrothermal treatment [125]. Kreft et al. [12] described significant correlations between rutin content and antioxidant activity of buckwheat grain and buckwheat food products. Chlopicka et al. [116] found positive and significant correlations between total phenols and antioxidant activity in buckwheat bread samples, as well as between total phenols and antioxidant activity of buckwheat bread samples, and, finally, between antioxidant activities themselves. Zhang et al. [126] reported that the baking, heating under steam pressure, and microwave heating of integral buckwheat flour had a statistically significant (P < 0.05) effect on the decrease in total flavonoids and antioxidant activity of flour, while the decrease in total phenols in buckwheat flour was less pronounced for all three applied treatments. As a possible explanation, the authors cited the creation of Maillard reaction products, which react with Folin-Ciocalteu reagent, resulting in masking the actual decrease in polyphenol content.
Similar conclusions regarding the formation of Maillard reaction products were reached by the authors Constantini et al. [127] during the production of bread with the addition of Tartary buckwheat flour, where a loss in the total antioxidant capacity and content of total polyphenols and flavonoids was observed, relative to their values in flour mixtures. The aforementioned authors pointed out that it is possible that the real reduction is greater than what was found in this study. As an explanation, they stated that heat treatment of cereals and pseudocereals, such as during baking, can also result in the synthesis of substances with antioxidant properties, including certain products of the Maillard reaction that occur in the crust of bread. These syntheses can mask the actual decrease in the content of total phenols and flavonoids (which are able to react with Folin-Ciocalteu reagent), as well as any loss in total antioxidant capacity.
Aside from phenols, other components, such as proteins, appear to be involved in the formation of the antioxidant activity of buckwheat products. The frying hulled buckwheat grain, in addition to reducing antioxidant activity, also resulted in a decrease in protein content and quality, while heat treatment did not show an effect on whole grain proteins [125]. In addition, during thermal treatment, Maillard components are generated due to a chemical reaction between the free amino groups of lysine and the carbonyl groups of reducing sugars [128]. It was observed the formation of Maillard products was caused by heat treatment of both whole and hulled buckwheat grains. Although Maillard components may be harmful to health, they may contribute to an increase in antioxidant activity, masking the actual decrease in total phenolic components, as highlighted in the above studies [125, 126, 127]. In addition, it has been suggested that antioxidant capacity may increase as a result of dissociation (separation) of phenolic forms and release of phenols bound to cell walls due to heat treatment followed by polymerization/oxidation of phenolic constituents or by-product generation [122].
The influence of baking on the content of tocopherols in buckwheat bread was investigated. Vitamin E loss was found to be about 30%. Smaller losses were observed in bread samples of 100% buckwheat flour compared to samples in which the share of buckwheat flour was 50% [81]. A significant reduction in vitamin E content (about 63%) in buckwheat was also found during the extrusion process [123].
The importance of common and Tartary buckwheat is generally recognized. However, one should also keep in mind some disadvantages of their application in the bakery in terms of sensory impression. This primarily refers to the particle size and the proportion of bran that can negatively affect the rheological properties of the dough and result in an inappropriate texture of bakery products. In addition, the finished products with Tatary buckwheat may appear a slightly bitter taste [6].
Based on all of the above, it is indicative that the contradictory results obtained so far greatly emphasize the importance of determining the exact composition and ratio of bioactive components. In addition, more studies are needed to identify the effect of heat treatment on the functional components, including proteins and phenolic components, of buckwheat products, in order to ultimately obtain buckwheat of consumption quality. Therefore, processing conditions, such as time and temperature, need to be optimized to preserve the functionality of bioactive components.
High pressure has been shown to be a viable alternative to heat treatment, with no adverse effects such as forming an off flavor, loss of vitamins and phytochemical properties, and discoloration [129]. The effect of high hydrostatic pressure treatment (200 MPa at 2, 4, and 9 minutes) on total antioxidant capacity (TAC), reducing capacity (RC), and rutin content of raw and roasted buckwheat groats were examined. After high-pressure treatment, the content of TAC and rutin differed in the case of raw and fried semolina. The TAC of raw and fried semolina subjected to high-pressure treatment was 16–20% and 12.5–17% lower, respectively, compared to the TAC of untreated semolina. Hydrophilic antioxidants were the main components contributing to the TAC of raw and fried semolina subjected to high-pressure treatment. RC decreased in the case of raw buckwheat (raw semolina), while the rutin content dropped in a shorter time compared to fried semolina. In contrast, overpressure in fried semolina increased the RC formed by hydrophilic antioxidants by 18% when measured by cyclic voltammetry on average and decreased the concentration of rutin after treatment [130].
The results of Zhou et al. [131] suggested that treatment under high pressure at 45 °C improves the nutritional properties of buckwheat compared to untreated and treated under high pressure at room temperature.
Radiation is a method of treating food to make it safer to eat and to extend its shelf life. Traditionally, this process is used to control surface microorganisms on vegetables and fruits without affecting nutritional quality. Hayashi et al. [132] reduced the microbial load to a lower level by exposing buckwheat grains to soft-electrons without affecting their quality. Chun and Song [133] conducted a study in which aqueous chlorine dioxide, fumaric acid, modified packaging atmosphere enriched with CO2, and ultraviolet radiation (UV) were combined in the treatment of buckwheat sprouts to improve microbiological quality. A decrease in total aerobic bacteria, yeasts and moulds, and enterobacteria to low levels was observed without affecting sensory quality. However, after the treatment, there was an increase in the concentration of rutin. A comparative study by Orsák et al. [134] studied the effects of UV, microwave, and γ-radiation on three buckwheat samples. Different effects were observed depending on the radiation system and the applied dose on the content of polyphenols and rutin. In addition, it has been described that the content of rutin and flavone C-glycosides is improved in sprouts after exposure to LED (light-emitting diodes) [135].
Therefore, radiation could be offered as a way to increase the half-life of food, maintain sensory quality, improve microbiological quality and increase nutritional value due to bioactive components in buckwheat products. Although public knowledge about radiation remains limited, interest in buying “safe—radiation-enhanced food” is increasing, especially after obtaining information about the potential benefits and risks.
A detailed review of the literature showed that the data on the stability of the most important bioactive components in all types of buckwheat products (bakery and others) during storage is quite limited. Unlike stability during processing, which can still be stated to have been the subject of a significant number of studies in recent years, and that the number of studies is constantly increasing, stability during storage is still almost unexplored. One of the studies that could be related to a certain extent to the mentioned topic is the one conducted by the authors Cho and Lee [121].
Namely, these authors established experimental extraction procedures for preparing rutin-enriched material (REM) from buckwheat milling fractions. Then the REM was used for the fortification of wheat instant fried noodles with rutin. After frying, the noodle samples were stored for 14 days at 60°C and its peroxide value was measured every other day in order to examine the effect of REM on the oxidative stability of the noodles. The monitoring showed that the peroxide number of noodle samples tended to increase with increasing storage time. However, it was noted that the rate of increase in peroxide value was markedly lower in noodle samples with incorporated REM compared to the control sample which did not contain REM. This indicates that the oxidizing improvement of instant fried noodles in storage was reduced using REM. These results can be expected from the high level of rutin in REM, since rutin has strong antioxidant activity [32].
Tabaković et al. [136] found that the most suitable way to store buckwheat seeds for a long period is paper material in order to retain their physiological and morphological properties to the greatest extent.
Buckwheat, a pseudocereal belonging to the
When it comes to processing, there are several processes associated with buckwheat, the first and inevitable one being the milling process. Milling is the processing (conversion) of buckwheat grains into flour, the primary raw material for the production of bakery products. During the processing of buckwheat grains into white flour, the husk and outer layers are separated, which removes the ratio of fibers, minerals, and polyphenolic components. Then there is the process of heat treatment, and baking, which belongs to the category of heat treatment, is also inevitable during the production of bakery products. In general, when it comes to the effects of heat treatment on bioactive components in buckwheat, a considerable amount of research has been conducted so far. However, the results of the conducted studies are contradictory. Although in most cases heat treatment resulted in a decrease in bioactive components, on the other hand, there are studies in which heat treatment did not have a significant effect on bioactive components. Therefore, it is necessary to conduct more studies with an emphasis on optimizing the conditions of heat treatment in order to ultimately obtain buckwheat and buckwheat-based products of appropriate quality for use. Recently, high-pressure treatments and the application of radiation are becoming increasingly important. However, although knowledge about radiation remains limited, there is an increased interest in buying “safe—radiation-enhanced food”, especially after obtaining information about the potential benefits and risks.
Although a considerable number of studies have been conducted on the effects of processing, and primarily on the effects of heat treatment, there are still significant gaps in this area. They are primarily related to the fact that most of the conducted studies are aimed at determining the effects of processing on the content of phenolic components, especially flavonoids, due to their recognized role in health benefits. Therefore more research that focuses on the effects of processing on all other bioactive components in buckwheat is needed in the future.
When it comes to the effects of storage, this topic is still almost unexplored. A review of the literature revealed that there is a small number of studies dealing with this topic, and therefore further research is needed to identify the effect of storage on bioactive components of buckwheat products, and ultimately to preserve their quality for as long as possible.
This is a brief overview of the main steps involved in publishing with IntechOpen Compacts, Monographs and Edited Books. Once you submit your proposal you will be appointed a Author Service Manager who will be your single point of contact and lead you through all the described steps below.
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A new type of memristors with a floating photogate based on biocompatible graphene and other 2D crystals with extremely low power consumption and footprint is considered. The photocatalytic oxidation of graphene is proposed as an effective method of creating synapse-like 2D memristive devices with photoresistive switching for nonvolatile electronic memory of ultrahigh density. 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This chapter will discuss the state-of-the-art research trend on neuromorphic computing with memristors as electronic synapses. Furthermore, a novel three-dimensional (3D) neuromorphic computing architecture combining memristor and monolithic 3D integration technology would be introduced; such computing architecture has capabilities to reduce the system power consumption, provide high connectivity, resolve the routing congestion issues, and offer the massively parallel data processing. 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His research interests involve understanding the molecular mechanisms of host defense during human viral infections and developing new predictive, preventive, and therapeutic strategies for them using Japanese encephalitis virus (JEV), HIV, and emerging viruses as a model via stem cell and cell culture technologies. His research work has been published in various high-impact factor journals (Science, PNAS, Nature Medicine) with a high number of citations. He has received many awards and honors in India and abroad including various Young Scientist Awards, BBSRC India Partnering Award, and Dr. JC Bose National Award of Department of Biotechnology, Min. of Science and Technology, Govt. of India. 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She graduated from Gazi University Faculty of Dentistry, Ankara, Turkey in 2000. \r\nLater she received her Ph.D. degree from the Oral Diagnosis and Radiology Department; which was recently renamed as Oral and Dentomaxillofacial Radiology, from the same university. \r\nShe is working as a full-time Associate Professor and is a lecturer and an academic researcher. \r\nHer expertise areas are dental caries, cancer, dental fear and anxiety, gag reflex in dentistry, oral medicine, and dentomaxillofacial radiology.",institutionString:"Gazi University",institution:{name:"Gazi University",institutionURL:null,country:{name:"Turkey"}}}]},{type:"book",id:"7139",title:"Current Approaches in Orthodontics",subtitle:null,coverURL:"https://cdn.intechopen.com/books/images_new/7139.jpg",slug:"current-approaches-in-orthodontics",publishedDate:"April 10th 2019",editedByType:"Edited by",bookSignature:"Belma Işık Aslan and Fatma Deniz Uzuner",hash:"2c77384eeb748cf05a898d65b9dcb48a",volumeInSeries:2,fullTitle:"Current Approaches in Orthodontics",editors:[{id:"42847",title:"Dr.",name:"Belma",middleName:null,surname:"Işik Aslan",slug:"belma-isik-aslan",fullName:"Belma Işik Aslan",profilePictureURL:"https://mts.intechopen.com/storage/users/42847/images/system/42847.jpg",biography:"Dr. Belma IşIk Aslan was born in 1976 in Ankara-TURKEY. 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Having been personally trained by Dr. Robert Kerstein on this method, Dr. Gözler has been lecturing on the T-Scan Occlusion Analysis Method in conferences both in Turkey and abroad. Dr. Gözler has various articles and presentations on Digital Occlusion Analysis methods. 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Dr. Al Ostwani is an assistant professor and faculty member at IUST University since 2014. \nDuring his academic experience, he has received several awards including the scientific research award from the Union of Arab Universities, the Syrian gold medal and the international gold medal for invention and creativity. Dr. Al Ostwani is a Member of the International Association of Dental Traumatology and the Syrian Society for Research and Preventive Dentistry since 2017. 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He also obtained an MSc in Molecular and Genetic Medicine, and a Ph.D. in Clinical Immunology and Human Genetics from the University of Sheffield, UK. He also completed a short-term fellowship in Pediatric Clinical Immunology and Bone Marrow Transplantation at Newcastle General Hospital, England. Dr. Rezaei is a Full Professor of Immunology and Vice Dean of International Affairs and Research, at the School of Medicine, Tehran University of Medical Sciences, and the co-founder and head of the Research Center for Immunodeficiencies. He is also the founding president of the Universal Scientific Education and Research Network (USERN). Dr. Rezaei has directed more than 100 research projects and has designed and participated in several international collaborative projects. He is an editor, editorial assistant, or editorial board member of more than forty international journals. He has edited more than 50 international books, presented more than 500 lectures/posters in congresses/meetings, and published more than 1,100 scientific papers in international journals.",institutionString:"Tehran University of Medical Sciences",institution:{name:"Tehran University of Medical Sciences",country:{name:"Iran"}}},{id:"180733",title:"Dr.",name:"Jean",middleName:null,surname:"Engohang-Ndong",slug:"jean-engohang-ndong",fullName:"Jean Engohang-Ndong",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/180733/images/system/180733.png",biography:"Dr. Jean Engohang-Ndong was born and raised in Gabon. After obtaining his Associate Degree of Science at the University of Science and Technology of Masuku, Gabon, he continued his education in France where he obtained his BS, MS, and Ph.D. in Medical Microbiology. He worked as a post-doctoral fellow at the Public Health Research Institute (PHRI), Newark, NJ for four years before accepting a three-year faculty position at Brigham Young University-Hawaii. Dr. Engohang-Ndong is a tenured faculty member with the academic rank of Full Professor at Kent State University, Ohio, where he teaches a wide range of biological science courses and pursues his research in medical and environmental microbiology. Recently, he expanded his research interest to epidemiology and biostatistics of chronic diseases in Gabon.",institutionString:"Kent State University",institution:{name:"Kent State University",country:{name:"United States of America"}}},{id:"188773",title:"Prof.",name:"Emmanuel",middleName:null,surname:"Drouet",slug:"emmanuel-drouet",fullName:"Emmanuel Drouet",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/188773/images/system/188773.png",biography:"Emmanuel Drouet, PharmD, is a Professor of Virology at the Faculty of Pharmacy, the University Grenoble-Alpes, France. As a head scientist at the Institute of Structural Biology in Grenoble, Dr. Drouet’s research investigates persisting viruses in humans (RNA and DNA viruses) and the balance with our host immune system. He focuses on these viruses’ effects on humans (both their impact on pathology and their symbiotic relationships in humans). He has an excellent track record in the herpesvirus field, and his group is engaged in clinical research in the field of Epstein-Barr virus diseases. He is the editor of the online Encyclopedia of Environment and he coordinates the Universal Health Coverage education program for the BioHealth Computing Schools of the European Institute of Science.",institutionString:null,institution:{name:"Grenoble Alpes University",country:{name:"France"}}},{id:"131400",title:"Prof.",name:"Alfonso J.",middleName:null,surname:"Rodriguez-Morales",slug:"alfonso-j.-rodriguez-morales",fullName:"Alfonso J. Rodriguez-Morales",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/131400/images/system/131400.png",biography:"Dr. Rodriguez-Morales is an expert in tropical and emerging diseases, particularly zoonotic and vector-borne diseases (especially arboviral diseases). He is the president of the Travel Medicine Committee of the Pan-American Infectious Diseases Association (API), as well as the president of the Colombian Association of Infectious Diseases (ACIN). He is a member of the Committee on Tropical Medicine, Zoonoses, and Travel Medicine of ACIN. He is a vice-president of the Latin American Society for Travel Medicine (SLAMVI) and a Member of the Council of the International Society for Infectious Diseases (ISID). Since 2014, he has been recognized as a Senior Researcher, at the Ministry of Science of Colombia. He is a professor at the Faculty of Medicine of the Fundacion Universitaria Autonoma de las Americas, in Pereira, Risaralda, Colombia. He is an External Professor, Master in Research on Tropical Medicine and International Health, Universitat de Barcelona, Spain. He is also a professor at the Master in Clinical Epidemiology and Biostatistics, Universidad Científica del Sur, Lima, Peru. In 2021 he has been awarded the “Raul Isturiz Award” Medal of the API. Also, in 2021, he was awarded with the “Jose Felix Patiño” Asclepius Staff Medal of the Colombian Medical College, due to his scientific contributions to COVID-19 during the pandemic. He is currently the Editor in Chief of the journal Travel Medicine and Infectious Diseases. His Scopus H index is 47 (Google Scholar H index, 68).",institutionString:"Institución Universitaria Visión de las Américas, Colombia",institution:null},{id:"332819",title:"Dr.",name:"Chukwudi Michael",middleName:"Michael",surname:"Egbuche",slug:"chukwudi-michael-egbuche",fullName:"Chukwudi Michael Egbuche",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/332819/images/14624_n.jpg",biography:"I an Dr. Chukwudi Michael Egbuche. I am a Senior Lecturer in the Department of Parasitology and Entomology, Nnamdi Azikiwe University, Awka.",institutionString:null,institution:{name:"Nnamdi Azikiwe University",country:{name:"Nigeria"}}},{id:"284232",title:"Mr.",name:"Nikunj",middleName:"U",surname:"Tandel",slug:"nikunj-tandel",fullName:"Nikunj Tandel",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/284232/images/8275_n.jpg",biography:'Mr. Nikunj Tandel has completed his Master\'s degree in Biotechnology from VIT University, India in the year of 2012. He is having 8 years of research experience especially in the field of malaria epidemiology, immunology, and nanoparticle-based drug delivery system against the infectious diseases, autoimmune disorders and cancer. He has worked for the NIH funded-International Center of Excellence in Malaria Research project "Center for the study of complex malaria in India (CSCMi)" in collaboration with New York University. The preliminary objectives of the study are to understand and develop the evidence-based tools and interventions for the control and prevention of malaria in different sites of the INDIA. Alongside, with the help of next-generation genomics study, the team has studied the antimalarial drug resistance in India. Further, he has extended his research in the development of Humanized mice for the study of liver-stage malaria and identification of molecular marker(s) for the Artemisinin resistance. At present, his research focuses on understanding the role of B cells in the activation of CD8+ T cells in malaria. Received the CSIR-SRF (Senior Research Fellow) award-2018, FIMSA (Federation of Immunological Societies of Asia-Oceania) Travel Bursary award to attend the IUIS-IIS-FIMSA Immunology course-2019',institutionString:"Nirma University",institution:{name:"Nirma University",country:{name:"India"}}},{id:"334383",title:"Ph.D.",name:"Simone",middleName:"Ulrich",surname:"Ulrich Picoli",slug:"simone-ulrich-picoli",fullName:"Simone Ulrich Picoli",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/334383/images/15919_n.jpg",biography:"Graduated in Pharmacy from Universidade Luterana do Brasil (1999), Master in Agricultural and Environmental Microbiology from Federal University of Rio Grande do Sul (2002), Specialization in Clinical Microbiology from Universidade de São Paulo, USP (2007) and PhD in Sciences in Gastroenterology and Hepatology (2012). She is currently an Adjunct Professor at Feevale University in Medicine and Biomedicine courses and a permanent professor of the Academic Master\\'s Degree in Virology. She has experience in the field of Microbiology, with an emphasis on Bacteriology, working mainly on the following topics: bacteriophages, bacterial resistance, clinical microbiology and food microbiology.",institutionString:null,institution:{name:"Universidade Feevale",country:{name:"Brazil"}}},{id:"229220",title:"Dr.",name:"Amjad",middleName:"Islam",surname:"Aqib",slug:"amjad-aqib",fullName:"Amjad Aqib",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/229220/images/system/229220.png",biography:"Dr. Amjad Islam Aqib obtained a DVM and MSc (Hons) from University of Agriculture Faisalabad (UAF), Pakistan, and a PhD from the University of Veterinary and Animal Sciences Lahore, Pakistan. Dr. Aqib joined the Department of Clinical Medicine and Surgery at UAF for one year as an assistant professor where he developed a research laboratory designated for pathogenic bacteria. Since 2018, he has been Assistant Professor/Officer in-charge, Department of Medicine, Manager Research Operations and Development-ORIC, and President One Health Club at Cholistan University of Veterinary and Animal Sciences, Bahawalpur, Pakistan. He has nearly 100 publications to his credit. His research interests include epidemiological patterns and molecular analysis of antimicrobial resistance and modulation and vaccine development against animal pathogens of public health concern.",institutionString:"Cholistan University of Veterinary and Animal Sciences",institution:null},{id:"62900",title:"Prof.",name:"Fethi",middleName:null,surname:"Derbel",slug:"fethi-derbel",fullName:"Fethi Derbel",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/62900/images/system/62900.jpeg",biography:"Professor Fethi Derbel was born in 1960 in Tunisia. He received his medical degree from the Sousse Faculty of Medicine at Sousse, University of Sousse, Tunisia. He completed his surgical residency in General Surgery at the University Hospital Farhat Hached of Sousse and was a member of the Unit of Liver Transplantation in the University of Rennes, France. He then worked in the Department of Surgery at the Sahloul University Hospital in Sousse. Professor Derbel is presently working at the Clinique les Oliviers, Sousse, Tunisia. His hospital activities are mostly concerned with laparoscopic, colorectal, pancreatic, hepatobiliary, and gastric surgery. He is also very interested in hernia surgery and performs ventral hernia repairs and inguinal hernia repairs. He has been a member of the GREPA and Tunisian Hernia Society (THS). During his residency, he managed patients suffering from diabetic foot, and he was very interested in this pathology. For this reason, he decided to coordinate a book project dealing with the diabetic foot. Professor Derbel has published many articles in journals and collaborates intensively with IntechOpen Access Publisher as an editor.",institutionString:"Clinique les Oliviers",institution:null},{id:"300144",title:"Dr.",name:"Meriem",middleName:null,surname:"Braiki",slug:"meriem-braiki",fullName:"Meriem Braiki",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/300144/images/system/300144.jpg",biography:"Dr. Meriem Braiki is a specialist in pediatric surgeon from Tunisia. She was born in 1985. She received her medical degree from the University of Medicine at Sousse, Tunisia. She achieved her surgical residency training periods in Pediatric Surgery departments at University Hospitals in Monastir, Tunis and France.\r\nShe is currently working at the Pediatric surgery department, Sidi Bouzid Hospital, Tunisia. Her hospital activities are mostly concerned with laparoscopic, parietal, urological and digestive surgery. She has published several articles in diffrent journals.",institutionString:"Sidi Bouzid Regional Hospital",institution:null},{id:"229481",title:"Dr.",name:"Erika M.",middleName:"Martins",surname:"de Carvalho",slug:"erika-m.-de-carvalho",fullName:"Erika M. de Carvalho",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/229481/images/6397_n.jpg",biography:null,institutionString:null,institution:{name:"Oswaldo Cruz Foundation",country:{name:"Brazil"}}},{id:"186537",title:"Prof.",name:"Tonay",middleName:null,surname:"Inceboz",slug:"tonay-inceboz",fullName:"Tonay Inceboz",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/186537/images/system/186537.jfif",biography:"I was graduated from Ege University of Medical Faculty (Turkey) in 1988 and completed his Med. PhD degree in Medical Parasitology at the same university. I became an Associate Professor in 2008 and Professor in 2014. I am currently working as a Professor at the Department of Medical Parasitology at Dokuz Eylul University, Izmir, Turkey.\n\nI have given many lectures, presentations in different academic meetings. I have more than 60 articles in peer-reviewed journals, 18 book chapters, 1 book editorship.\n\nMy research interests are Echinococcus granulosus, Echinococcus multilocularis (diagnosis, life cycle, in vitro and in vivo cultivation), and Trichomonas vaginalis (diagnosis, PCR, and in vitro cultivation).",institutionString:"Dokuz Eylül University",institution:{name:"Dokuz Eylül University",country:{name:"Turkey"}}},{id:"71812",title:"Prof.",name:"Hanem Fathy",middleName:"Fathy",surname:"Khater",slug:"hanem-fathy-khater",fullName:"Hanem Fathy Khater",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/71812/images/1167_n.jpg",biography:"Prof. Khater is a Professor of Parasitology at Benha University, Egypt. She studied for her doctoral degree, at the Department of Entomology, College of Agriculture, Food and Natural Resources, University of Missouri, Columbia, USA. She has completed her Ph.D. degrees in Parasitology in Egypt, from where she got the award for “the best scientific Ph.D. dissertation”. She worked at the School of Biological Sciences, Bristol, England, the UK in controlling insects of medical and veterinary importance as a grant from Newton Mosharafa, the British Council. Her research is focused on searching of pesticides against mosquitoes, house flies, lice, green bottle fly, camel nasal botfly, soft and hard ticks, mites, and the diamondback moth as well as control of several parasites using safe and natural materials to avoid drug resistances and environmental contamination.",institutionString:null,institution:{name:"Banha University",country:{name:"Egypt"}}},{id:"99780",title:"Prof.",name:"Omolade",middleName:"Olayinka",surname:"Okwa",slug:"omolade-okwa",fullName:"Omolade Okwa",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/99780/images/system/99780.jpg",biography:"Omolade Olayinka Okwa is presently a Professor of Parasitology at Lagos State University, Nigeria. She has a PhD in Parasitology (1997), an MSc in Cellular Parasitology (1992), and a BSc (Hons) Zoology (1990) all from the University of Ibadan, Nigeria. She teaches parasitology at the undergraduate and postgraduate levels. She was a recipient of a Commonwealth fellowship supported by British Council tenable at the Centre for Entomology and Parasitology (CAEP), Keele University, United Kingdom between 2004 and 2005. She was awarded an Honorary Visiting Research Fellow at the same university from 2005 to 2007. \nShe has been an external examiner to the Department of Veterinary Microbiology and Parasitology, University of Ibadan, MSc programme between 2010 and 2012. She is a member of the Nigerian Society of Experimental Biology (NISEB), Parasitology and Public Health Society of Nigeria (PPSN), Science Association of Nigeria (SAN), Zoological Society of Nigeria (ZSN), and is Vice Chairperson of the Organisation of Women in Science (OWSG), LASU chapter. She served as Head of Department of Zoology and Environmental Biology, Lagos State University from 2007 to 2010 and 2014 to 2016. She is a reviewer for several local and international journals such as Unilag Journal of Science, Libyan Journal of Medicine, Journal of Medicine and Medical Sciences, and Annual Research and Review in Science. \nShe has authored 45 scientific research publications in local and international journals, 8 scientific reviews, 4 books, and 3 book chapters, which includes the books “Malaria Parasites” and “Malaria” which are IntechOpen access publications.",institutionString:"Lagos State University",institution:{name:"Lagos State University",country:{name:"Nigeria"}}},{id:"273100",title:"Dr.",name:"Vijay",middleName:null,surname:"Gayam",slug:"vijay-gayam",fullName:"Vijay Gayam",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/273100/images/system/273100.jpeg",biography:"Dr. Vijay Bhaskar Reddy Gayam is currently practicing as an internist at Interfaith Medical Center in Brooklyn, New York, USA. He is also a Clinical Assistant Professor at the SUNY Downstate University Hospital and Adjunct Professor of Medicine at the American University of Antigua. He is a holder of an M.B.B.S. degree bestowed to him by Osmania Medical College and received his M.D. at Interfaith Medical Center. His career goals thus far have heavily focused on direct patient care, medical education, and clinical research. He currently serves in two leadership capacities; Assistant Program Director of Medicine at Interfaith Medical Center and as a Councilor for the American\r\nFederation for Medical Research. As a true academician and researcher, he has more than 50 papers indexed in international peer-reviewed journals. He has also presented numerous papers in multiple national and international scientific conferences. His areas of research interest include general internal medicine, gastroenterology and hepatology. He serves as an editor, editorial board member and reviewer for multiple international journals. His research on Hepatitis C has been very successful and has led to multiple research awards, including the 'Equity in Prevention and Treatment Award” from the New York Department of Health Viral Hepatitis Symposium (2018) and the 'Presidential Poster Award” awarded to him by the American College of Gastroenterology (2018). He was also awarded 'Outstanding Clinician in General Medicine” by Venus International Foundation for his extensive research expertise and services, perform over and above the standard expected in the advancement of healthcare, patient safety and quality of care.",institutionString:"Interfaith Medical Center",institution:{name:"Interfaith Medical Center",country:{name:"United States of America"}}},{id:"93517",title:"Dr.",name:"Clement",middleName:"Adebajo",surname:"Meseko",slug:"clement-meseko",fullName:"Clement Meseko",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/93517/images/system/93517.jpg",biography:"Dr. Clement Meseko obtained DVM and PhD degree in Veterinary Medicine and Virology respectively. He has worked for over 20 years in both private and public sectors including the academia, contributing to knowledge and control of infectious disease. Through the application of epidemiological skill, classical and molecular virological skills, he investigates viruses of economic and public health importance for the mitigation of the negative impact on people, animal and the environment in the context of Onehealth. \r\nDr. Meseko’s field experience on animal and zoonotic diseases and pathogen dynamics at the human-animal interface over the years shaped his carrier in research and scientific inquiries. He has been part of the investigation of Highly Pathogenic Avian Influenza incursions in sub Saharan Africa and monitors swine Influenza (Pandemic influenza Virus) agro-ecology and potential for interspecies transmission. He has authored and reviewed a number of journal articles and book chapters.",institutionString:"National Veterinary Research Institute",institution:{name:"National Veterinary Research Institute",country:{name:"Nigeria"}}},{id:"158026",title:"Prof.",name:"Shailendra K.",middleName:null,surname:"Saxena",slug:"shailendra-k.-saxena",fullName:"Shailendra K. Saxena",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRET3QAO/Profile_Picture_2022-05-10T10:10:26.jpeg",biography:"Professor Dr. Shailendra K. Saxena is a vice dean and professor at King George's Medical University, Lucknow, India. His research interests involve understanding the molecular mechanisms of host defense during human viral infections and developing new predictive, preventive, and therapeutic strategies for them using Japanese encephalitis virus (JEV), HIV, and emerging viruses as a model via stem cell and cell culture technologies. His research work has been published in various high-impact factor journals (Science, PNAS, Nature Medicine) with a high number of citations. He has received many awards and honors in India and abroad including various Young Scientist Awards, BBSRC India Partnering Award, and Dr. JC Bose National Award of Department of Biotechnology, Min. of Science and Technology, Govt. of India. Dr. Saxena is a fellow of various international societies/academies including the Royal College of Pathologists, United Kingdom; Royal Society of Medicine, London; Royal Society of Biology, United Kingdom; Royal Society of Chemistry, London; and Academy of Translational Medicine Professionals, Austria. He was named a Global Leader in Science by The Scientist. 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She is a professor in the Stomatology Faculty, St. Petersburg State University. She has expertise in the development and evaluation of a wide range of live mucosal vaccines against influenza and bacterial complications. 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Govindarajan completed his BSc degree in Zoology at Government Arts College (Autonomous), Kumbakonam, and MSc, MPhil, and PhD degrees at Annamalai University, Annamalai Nagar, Tamil Nadu, India. He is serving as an assistant professor at the Department of Zoology, Annamalai University. His research interests include isolation, identification, and characterization of biologically active molecules from plants and microbes. He has identified more than 20 pure compounds with high mosquitocidal activity and also conducted high-quality research on photochemistry and nanosynthesis. He has published more than 150 studies in journals with impact factor and 2 books in Lambert Academic Publishing, Germany. He serves as an editorial board member in various national and international scientific journals.",institutionString:null,institution:null},{id:"274660",title:"Dr.",name:"Damodar",middleName:null,surname:"Paudel",slug:"damodar-paudel",fullName:"Damodar Paudel",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/274660/images/8176_n.jpg",biography:"I am DrDamodar Paudel,currently working as consultant Physician in Nepal police Hospital.",institutionString:null,institution:null},{id:"241562",title:"Dr.",name:"Melvin",middleName:null,surname:"Sanicas",slug:"melvin-sanicas",fullName:"Melvin Sanicas",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/241562/images/6699_n.jpg",biography:null,institutionString:null,institution:null},{id:"337446",title:"Dr.",name:"Maria",middleName:null,surname:"Zavala-Colon",slug:"maria-zavala-colon",fullName:"Maria Zavala-Colon",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Puerto Rico, Medical Sciences Campus",country:{name:"United States of America"}}},{id:"338856",title:"Mrs.",name:"Nur Alvira",middleName:null,surname:"Pascawati",slug:"nur-alvira-pascawati",fullName:"Nur Alvira Pascawati",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Universitas Respati Yogyakarta",country:{name:"Indonesia"}}},{id:"441116",title:"Dr.",name:"Jovanka M.",middleName:null,surname:"Voyich",slug:"jovanka-m.-voyich",fullName:"Jovanka M. Voyich",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Montana State University",country:{name:"United States of America"}}},{id:"330412",title:"Dr.",name:"Muhammad",middleName:null,surname:"Farhab",slug:"muhammad-farhab",fullName:"Muhammad Farhab",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Agriculture Faisalabad",country:{name:"Pakistan"}}},{id:"349495",title:"Dr.",name:"Muhammad",middleName:null,surname:"Ijaz",slug:"muhammad-ijaz",fullName:"Muhammad Ijaz",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Veterinary and Animal Sciences",country:{name:"Pakistan"}}}]}},subseries:{item:{id:"95",type:"subseries",title:"Urban Planning and Environmental Management",keywords:"Circular economy, Contingency planning and response to disasters, Ecosystem services, Integrated urban water management, Nature-based solutions, Sustainable urban development, Urban green spaces",scope:"