Summary of LC-HRMS condition for AF determination.
\\n\\n
IntechOpen was founded by scientists, for scientists, in order to make book publishing accessible around the globe. Over the last two decades, this has driven Open Access (OA) book publishing whilst levelling the playing field for global academics. Through our innovative publishing model and the support of the research community, we have now published over 5,700 Open Access books and are visited online by over three million academics every month. These researchers are increasingly working in broad technology-based subjects, driving multidisciplinary academic endeavours into human health, environment, and technology.
\\n\\nBy listening to our community, and in order to serve these rapidly growing areas which lie at the core of IntechOpen's expertise, we are launching a portfolio of Open Science journals:
\\n\\nAll three journals will publish under an Open Access model and embrace Open Science policies to help support the changing needs of academics in these fast-moving research areas. There will be direct links to preprint servers and data repositories, allowing full reproducibility and rapid dissemination of published papers to help accelerate the pace of research. Each journal has renowned Editors in Chief who will work alongside a global Editorial Board, delivering robust single-blind peer review. Supported by our internal editorial teams, this will ensure our authors will receive a quick, user-friendly, and personalised publishing experience.
\\n\\n"By launching our journals portfolio we are introducing new, dedicated homes for interdisciplinary technology-focused researchers to publish their work, whilst embracing Open Science and creating a unique global home for academics to disseminate their work. We are taking a leap toward Open Science continuing and expanding our fundamental commitment to openly sharing scientific research across the world, making it available for the benefit of all." Dr. Sara Uhac, IntechOpen CEO
\\n\\n"Our aim is to promote and create better science for a better world by increasing access to information and the latest scientific developments to all scientists, innovators, entrepreneurs and students and give them the opportunity to learn, observe and contribute to knowledge creation. Open Science promotes a swifter path from research to innovation to produce new products and services." Alex Lazinica, IntechOpen founder
\\n\\nIn conclusion, Natalia Reinic Babic, Head of Journal Publishing and Open Science at IntechOpen adds:
\\n\\n“On behalf of the journal team I’d like to thank all our Editors in Chief, Editorial Boards, internal supporting teams, and our scientific community for their continuous support in making this portfolio a reality - we couldn’t have done it without you! With your support in place, we are confident these journals will become as impactful and successful as our book publishing program and bring us closer to a more open (science) future.”
\\n\\nWe invite you to visit the journals homepage and learn more about the journal’s Editorial Boards, scope and vision as all three journals are now open for submissions.
\\n\\nFeel free to share this news on social media and help us mark this memorable moment!
\\n\\n\\n"}]',published:!0,mainMedia:{caption:"",originalUrl:"/media/original/237"}},components:[{type:"htmlEditorComponent",content:'
After years of being acknowledged as the world's leading publisher of Open Access books, today, we are proud to announce we’ve successfully launched a portfolio of Open Science journals covering rapidly expanding areas of interdisciplinary research.
\n\n\n\nIntechOpen was founded by scientists, for scientists, in order to make book publishing accessible around the globe. Over the last two decades, this has driven Open Access (OA) book publishing whilst levelling the playing field for global academics. Through our innovative publishing model and the support of the research community, we have now published over 5,700 Open Access books and are visited online by over three million academics every month. These researchers are increasingly working in broad technology-based subjects, driving multidisciplinary academic endeavours into human health, environment, and technology.
\n\nBy listening to our community, and in order to serve these rapidly growing areas which lie at the core of IntechOpen's expertise, we are launching a portfolio of Open Science journals:
\n\nAll three journals will publish under an Open Access model and embrace Open Science policies to help support the changing needs of academics in these fast-moving research areas. There will be direct links to preprint servers and data repositories, allowing full reproducibility and rapid dissemination of published papers to help accelerate the pace of research. Each journal has renowned Editors in Chief who will work alongside a global Editorial Board, delivering robust single-blind peer review. Supported by our internal editorial teams, this will ensure our authors will receive a quick, user-friendly, and personalised publishing experience.
\n\n"By launching our journals portfolio we are introducing new, dedicated homes for interdisciplinary technology-focused researchers to publish their work, whilst embracing Open Science and creating a unique global home for academics to disseminate their work. We are taking a leap toward Open Science continuing and expanding our fundamental commitment to openly sharing scientific research across the world, making it available for the benefit of all." Dr. Sara Uhac, IntechOpen CEO
\n\n"Our aim is to promote and create better science for a better world by increasing access to information and the latest scientific developments to all scientists, innovators, entrepreneurs and students and give them the opportunity to learn, observe and contribute to knowledge creation. Open Science promotes a swifter path from research to innovation to produce new products and services." Alex Lazinica, IntechOpen founder
\n\nIn conclusion, Natalia Reinic Babic, Head of Journal Publishing and Open Science at IntechOpen adds:
\n\n“On behalf of the journal team I’d like to thank all our Editors in Chief, Editorial Boards, internal supporting teams, and our scientific community for their continuous support in making this portfolio a reality - we couldn’t have done it without you! With your support in place, we are confident these journals will become as impactful and successful as our book publishing program and bring us closer to a more open (science) future.”
\n\nWe invite you to visit the journals homepage and learn more about the journal’s Editorial Boards, scope and vision as all three journals are now open for submissions.
\n\nFeel free to share this news on social media and help us mark this memorable moment!
\n\n\n'}],latestNews:[{slug:"intechopen-supports-asapbio-s-new-initiative-publish-your-reviews-20220729",title:"IntechOpen Supports ASAPbio’s New Initiative Publish Your Reviews"},{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"}]},book:{item:{type:"book",id:"1304",leadTitle:null,fullTitle:"New Techniques in Gastrointestinal Endoscopy",title:"New Techniques in Gastrointestinal Endoscopy",subtitle:null,reviewType:"peer-reviewed",abstract:"As result of progress, endoscopy has became more complex, using more sophisticated devices and has claimed a special form. In this moment, the gastroenterologist performing endoscopy has to be an expert in macroscopic view of the lesions in the gut, with good skills for using standard endoscopes, with good experience in ultrasound (for performing endoscopic ultrasound), with pathology experience for confocal examination. It is compulsory to get experience and to have patience and attention for the follow-up of thousands of images transmitted during capsule endoscopy or to have knowledge in physics necessary for autofluorescence imaging endoscopy.\nTherefore, the idea of an endoscopist has changed. Examinations mentioned need a special formation, a superior level of instruction, accessible to those who have already gained enough experience in basic diagnostic endoscopy. This is the reason for what these new issues of endoscopy are presented in this book of New techniques in Gastrointestinal Endoscopy.",isbn:null,printIsbn:"978-953-307-777-2",pdfIsbn:"978-953-51-6509-5",doi:"10.5772/1773",price:139,priceEur:155,priceUsd:179,slug:"new-techniques-in-gastrointestinal-endoscopy",numberOfPages:324,isOpenForSubmission:!1,isInWos:null,isInBkci:!1,hash:"e108f32556a501bd10550b95901980b2",bookSignature:"Oliviu Pascu and Andrada Seicean",publishedDate:"October 3rd 2011",coverURL:"https://cdn.intechopen.com/books/images_new/1304.jpg",numberOfDownloads:55213,numberOfWosCitations:24,numberOfCrossrefCitations:15,numberOfCrossrefCitationsByBook:1,numberOfDimensionsCitations:32,numberOfDimensionsCitationsByBook:1,hasAltmetrics:0,numberOfTotalCitations:71,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"November 16th 2010",dateEndSecondStepPublish:"December 14th 2010",dateEndThirdStepPublish:"April 20th 2011",dateEndFourthStepPublish:"May 20th 2011",dateEndFifthStepPublish:"July 19th 2011",currentStepOfPublishingProcess:5,indexedIn:"1,2,3,4,5,6",editedByType:"Edited by",kuFlag:!1,featuredMarkup:null,editors:[{id:"62220",title:"Prof.",name:"Oliviu",middleName:null,surname:"Pascu",slug:"oliviu-pascu",fullName:"Oliviu Pascu",profilePictureURL:"https://mts.intechopen.com/storage/users/62220/images/1872_n.jpg",biography:"Dr. Oliviu Pascu graduated at the Faculty of General Medicine, Institute of Medicine & Pharmacy in 1963. He became professor of internal medicine and gastroenterology at the IIIrd Medical Clinic, University of Medicine and Pharmacy, Cluj-Napoca, Romania in 1990. From 1990 until 2000 he was dean (Faculty of Medicine) and then rector of the University of Medicine and Pharmacy in Cluj. Until 2009 he acted as president of the Romanian Society of Digestive Endoscopy and is presently honorary president. His first monograph on digestive endoscopy was published in Romania in 1982. while his first textbook on gastroenterology 1996-1997. He has practiced digestive endoscopy since 1969 and introduced hemostasis and polypectomy in Romania (1975). More than 150 of his articles have been published in Romanian and international medical journals. Dr. Pascu is a member of the Academy of Medical Sciences Romania and many Romanian and European scientific societies.",institutionString:null,position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"0",totalChapterViews:"0",totalEditedBooks:"3",institution:null}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,coeditorOne:{id:"68858",title:"Dr.",name:"Andrada",middleName:null,surname:"Seicean",slug:"andrada-seicean",fullName:"Andrada Seicean",profilePictureURL:"https://mts.intechopen.com/storage/users/68858/images/system/68858.jpg",biography:"Andrada Seicean graduated from the University of Medicine and Pharmacy ”Iuliu Hatieganu” in her native town in 1995, as head of her class. Since 2016 she has become professor of Gastroenterology at the same University, as well as a consulting gastroenterologist at the Regional Institute of Gastroenterology and Hepatology, Cluj-Napoca. \nHer interest has been focused on pancreatic and inflammatory bowel diseases, endoscopic ultrasound therapeutic techniques, molecular biology in pancreatic diseases and colorectal cancer. More than 70 of her articles were published in international journals. She is the author of about 70 articles in the international journals, seven national books and more than 50 chapters in different medical books.",institutionString:"Iuliu Hațieganu University of Medicine and Pharmacy",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"3",totalChapterViews:"0",totalEditedBooks:"1",institution:{name:"Iuliu Hațieganu University of Medicine and Pharmacy",institutionURL:null,country:{name:"Romania"}}},coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"1021",title:"Hepatology",slug:"gastroenterology-hepatology"}],chapters:[{id:"21070",title:"EUS Staging of Luminal Cancers in the Upper GI Tract",doi:"10.5772/23001",slug:"eus-staging-of-luminal-cancers-in-the-upper-gi-tract",totalDownloads:2565,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:null,signatures:"Juan Carlos Bucobo and Jonathan M. 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Aflatoxins (AFs) are highly toxic secondary metabolites produced by fungi belonging to several
Chemical structure of the most important AFs and their derivatives.
Raw materials usually used for human food and animal feed are contaminated by this type of fungi and their metabolites. Cereals (maize, wheat, rice, barley, soy, etc.), dried fruits, nuts, coffee and other foods could be contaminated during plant growth or postharvest, depending on different factors such as temperature, humidity, water activity, concurrent mycobiota, physical damage, and other storage conditions [2]. AFs are very stable and may resist cooking processes, resulting a problem in processed foods. Human exposure to AFs can result directly from ingestion of contaminated foods, or indirectly from consumption of animal foods previously exposed to contaminated feeds. AFs have a great risk for human health, especially by their carcinogenic potential [3]. Degradation or enzymatic transformation of mycotoxins led to the appearance of modified mycotoxins, usually lesser toxic than the parent compounds. Thus, aflatoxin M1 (AFM1) is formed from the hydroxylation of AFB1 and eliminated in the milk of animals that consumed feed contaminated with this mycotoxin [4].
Therefore, it is important to develop reliable methods for the determination of AFs and their derivatives in foods and feeds, as well as toxicokinetic and toxicodynamic studies for assessment of human or animal exposure. The target and non-target qualitative and quantitative analysis using high resolution mass spectrometry (HRMS) instruments, such as time-of-flight (TOF) and Orbitrap, brings great challenges for screening of AFs [5]. Main advantages include high sensitivity, accurate mass measurement, and retrospective data analysis, allowing both the target determination of AFs and the non-targeted screening of modified AFs or unknown metabolites.
AFs are potent carcinogenic, mutagenic, teratogenic, and immunosuppressive agents. Their carcinogenicity has mainly been associated with liver and kidney, although the effect of AFs has also been reported in pancreas, bladder, bone, viscera or central nervous by some epidemiological and animal studies [6]. Their inhalation and direct contact could also cause lung and skin [7, 8] occupational cancers, respectively. In addition, feeds contaminated by AFs can involve high susceptibility to diseases, low productivity and low reproductive performance in animals [9].
Among AFs, AFB1 is considered the highest risk. The Scientific Committee on Food has established that AFs are genotoxic carcinogens [10, 11], being the order of toxicity as follows: AFB1 > AFG1 > AFB2 > AFG2. Indeed, AFB1 has been shown to be carcinogenic in all experimental animals and has been classified since 1988 by the World Health Organization (WHO) as a human carcinogen. Consequently, the International Agency for Research on Cancer (IARC) [12] has classified AFB1 within the category of Group 1 substances based on the existence of sufficient evidence about its carcinogenicity to humans, both alone and in natural mixtures with the other AFs [13, 14].
The most common route of entry of AFs into the human body is the ingestion. In the case of AFB1, the best studied aflatoxin, is absorbed in the gastrointestinal tract, due to its liposolubility, and transported by red blood cells and plasma proteins to the liver. In the liver, it is metabolized producing intermediate metabolites that have been related with the toxic and carcinogenic effects of AFs [15]. Specifically, AFB1 is biotransformed in the liver by microsomal enzymes of the cytochrome superfamily P450. Microsomal biotransformation can result in the hydrolyzation of aflatoxin B1, producing less toxic metabolites such as AFM1, aflatoxin Q1 (AFQ1), aflatoxin P1 (AFP1) and aflatoxin B2a (AFB2a). In addition, AFB1 can produce aflatoxicol (AFL) via NADPH reductase. The formation of these compounds is considered a detoxification process although the protein binding of some of them can lead to additional toxicities [16]. They are excreted in urine and feces, although AFM1 is also commonly detected in breast milk.
The action of CYP450 enzymes can also metabolize AFB1 resulting in the appearance of a reactive intermediate metabolite, AFB1–8,9-epoxide (AFBO), which has two isomers (
The interaction AFB1-DNA causes AFB1-N7-guanine adduct, which is chemically unstable and undergoes rapid urinary excretion resulting in an aputinic (AP) site on the DNA backbone [16]. Alternatively, the adduct AFB-N7-guanine may be stabilized by rearranging to a ring-opened formamidopyramidine structure (AFB1-FAPy) [17]. Both AP and AFB1-FAPy can produce mutagenesis. Figure 2 summarizes the action mechanism of AFB1.
Metabolic pathway of AFB1.
From the action mechanism of AFB1 it can be deduced that AFB1-lys, AFB1-N7-guanine, AFB1-mercapturic acid or the hydroxylated forms (AFM1, AFQ1, AFP1, AFL and AFB2a) could be effective biomarkers for assessing AF exposure.
Due to the high lesions produced by AFs, especially cancer, the European Union has established maximum permitted levels of these contaminants in various foods through Regulation No. 1881/2006 [19]. Specifically, the maximum contents for AFB1, AFB2, AFG1, AFG2 and AFM1 in nuts, cereals, milk and baby foods are included in this regulation the maximum contents are between 4 and 15 μg kg−1.
In the field of animal nutrition, the specifications regarding the presence of mycotoxins in feed are reflected in Directive 2002/32/EC [20]. Only AFB1 has been legislated. The maximum levels ranged between 5 and 50 μg kg−1. The lower limit was set for feed intended for milk-producing animals (5 μg kg−1).
Liquid chromatography (LC)-HRMS is a powerful tool for metabolomic approaches, allowing simultaneous quantitative and qualitative analysis of a wide variety of mycotoxins, as well as the search of related metabolites derived from mycotoxin biotransformation or degradation, enabling the detection and identification of unknown compounds. In addition, HRMS offers the ability to work in various modes, such as target analysis and non-target screening, or retrospective analysis. The relative incompatibility of HRMS ion sources with the continuous liquid flow of LC limited the progress of LC-HRMS coupling for years, but the development of interfaces such as electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI), where LC effluent is de-solvated, has allowed the proposal of a high number of LC-HRMS methods. Thus, complex mixtures can be separated in the chromatographic system and their components are unequivocally detected by HRMS with high sensitivity.
For AFs determination, quadrupole (Q)-TOF, Orbitrap and its hybrid Q-Orbitrap are the mass analysers most widely used. Comparing both instruments, Orbitrap shows better resolution and accuracy (Q-TOF: 60,000 full width at half maximum (FWHM) and between 1 and 10 ppm; Orbitrap: 240,000 FWHM and less than 1 ppm), and a greater range of
ESI or its variant heated ESI (HESI) working in positive mode are the best options for AF determination. Although it should be noted that many of the methods described in this chapter are multiclass methods, i.e., they determine a greater number of mycotoxins, not just AFs, and in this case, authors usually prefer two independent runs using both positive and negative polarities.
Regarding LC instruments, ultra-high-performance LC (UHPLC) is normally coupled to HRMS. Although NanoLC coupled to Q-Orbitrap for the determination of AFB1-lys in human plasma [26] and high performance LC (HPLC)-TOF for the determination of AFB1 in beer [24] have also been proposed. In addition, Qi et al. used a multiple heart-cutting two-dimensional liquid chromatography (Heart-cutting 2D-LC) coupled to Q-Orbitrap for simultaneous determination of AFs and ochratoxin A in snus [27]. The utilization of Heart-cutting 2D-LC enables to reduce matrix effect, leading to better precision of the AF contents. The mobile phase is normally a mixture of water and methanol (MeOH) or acetonitrile (ACN). Formic acid (FA), acetic acid (AA), ammonium formate or ammonium acetate are used as additives. The stationary phase was mainly C18 although C8 has been also proposed [28]. Slobodchikova et al. [22] also used a pentafluorophenyl (PFP) column whereas Qi et al. [27] combined both C18 and PFP columns in the Heart-cutting 2D-LC system.
The analysis of biological samples focused on the monitoring of the four most important AFs (AFB1, AFB2, AFG1 and AFG2), although their adducts due to the interaction of AFB1 with proteins or DNA, AFB1-N7-guanine and AFB1-lys, respectively, as well as its hydrolysed derivative AFM1 have also been determined. In food samples, besides the four main AFs, some metabolites such as AFM1, AFM2, or AFL were also detected.
Both data dependent (dd-MS2) and data independent (DIA) acquisition have been proposed. For the analysis of biological samples, Full MS combined with dd-MS2 by inclusion of a list of accurate masses of target or suspect compounds was more frequent. Although, Ogawa et al. [29] also proposed a dd-MS2 by fixing an ion intensity threshold. Regarding food analysis, authors normally prefer Full MS and DIA, specifically, all-ion fragmentation (AIF) mode, where no precursor ion isolation is carried out. Other modes such as simple Full MS, selected ion monitoring (SIM) or parallel reaction monitoring mode (PRM) have also been investigated. Renaud et al. [30] compared three acquisition modes: dd-MS2 with inclusion list, AIF and AIF using targeted high energy collision dissociation (HCD) events across a mass range for MS/MS. Good linearity was achieved by AIF with different HCD events at low concentrations, demonstrating that the limits of detection (LODs) are much higher without a Q mass filtering. The potential of AIF with different HCD events has also been studied for the determination of five AFs in nutraceutical obtained from green tea [31].
Most authors who carried out a non-targeted acquisition opted for target processing, in order to quantify and/or confirm AFs. Only Jia et al. [31] carried out a non-targeted processing consisting of: non-target fourier peak picking, (ii) spectra automated componentization, (iii) suspicion spectral library searching, and (iv) marked fragments filtering. Finally, compounds were confirmed with reference standard. In addition, Castaldo et al. [32] and Renaud et al. [30] carried out a non-targeted processing using spectral library for the tentative identification of other fungal metabolites, although they processed AFs following a targeted approach.
Table 1 summarizes the separation and detection condition of HRMS methods for AF monitorization.
AF studied | Matrix | Instrument | LC conditions | Acquisition | Ref. |
---|---|---|---|---|---|
AFB1, AFB2, AFG1, AFG2, AFM1 | Breast milk | UHPLC–Orbitrap HESI + | Hypersil GOLD C18 (100 × 2.1 mm, 1.9 μm) H2O/MeOH with FA and HCOONH4 | Full MS ( | [33] |
AFB1, AFB2, AFG1, AFG2 | Isoflavone supplements | UHPLC-Orbitrap HESI + | Hypersil Gold C18 (100 × 2.1 mm, 1.9 μm) H2O/MeOH with FA and HCOONH4 | Full MS ( ( | [34] |
AFB1, AFB2, AFG1, AFG2 | Ginkgo biloba supplements | UHPLC-Orbitrap HESI + | Hypersil Gold C18 (100 × 2.1 mm, 1.9 μm) H2O/MeOH with FA and HCOONH4 | Full MS and AIF ( | [35] |
AFB1, AFB2, AFG1, AFG2 | Green tea and royal jelly supplements | UHPLC-Orbitrap HESI + | Hypersil Gold C18 (100 × 2.1 mm, 1.9 μm) H2O/MeOH with FA and HCOONH4 | Full MS and AIF ( | [36] |
AFB1, AFB2, AFG1, AFG2, AFM1, AFL | Coix seed | UHPLC-Orbitrap HESI + | C18 (100 × 2.1 mm, 1.6 μm) H2O/MeOH with FA and CH3COONH4 | Full MS ( | [37] |
AFB1, AFB2, AFG1, AFG2 | Feed | UHPLC-Orbitrap HESI + | Hypersil Gold C18 (100 × 2.1mm,1.9 μm) H2O/MeOH/ACN with AA | Full MS and AIF ( | [38] |
AFB1, AFB1-lys | Human serum | UHPLC-Q-Orbitrap HESI + | C18 (100 × 2.1 mm, 1.7 μm) H2O/MeOH with FA and HCOONH4 | Full MS ( | [39] |
AFB1, AFM1, AFB1-N7-guanine | Human urine | UHPLC-Q-Orbitrap HESI + | Acquity BEH C18 (100 × 2.1 mm, 1.7 μm) H2O/MeOH with FA and HCOONH4 | Full MS (no range) and dd-MS2 (list dependent) | [40] |
AFB1, AFM1 | Milk | UHPLC-Q-Orbitrap HESI + | Luna Omega C18 (50 × 2.1 mm, 1.6 μm) H2O/MeOH with FA and HCOONH4 | Full MS and AIF ( | [41] |
AFB1, AFB2, AFG1, AFG2, AFM1, AFM2 | Milk | UHPLC-Q-Orbitrap HESI + | Accucore C18 (150 x 2.1 mm, 2.6 μm) H2O/ACN with FA and CH3COONH4 | Full MS ( | [42] |
AFB1, AFG1, AFG2 | Maize | UHPLC-Q-Orbitrap HESI + | Zorbax Eclipse Plus RRHD C18 column (50 × 2.1 mm, 1.8 μm) | Full MS, dd-MS2 (list dependent), AIF and AIF with HCD events | [30] |
AFB1, AFB2, AFG1, AFG2 | Cashew nut | UHPLC-Q-Orbitrap HESI + | HSS T3 (100 x 2.1 mm, 1.8 μm) H2O/MeOH with FA and HCOONH4 | PRM | [43] |
AFB1, AFB2, AFG1, AFG2 | Cereals | UHPLC-Q-Orbitrap HESI + | Kinetex C18 (50 × 3 mm, 1.7 μm H2O/MeOH with AA and CH3COONH4 | Full MS and AIF ( | [44] |
AFB1 | Durum wheat pasta and baby food pasta | UHPLC-Q-Orbitrap HESI + | Accucore C18 (100 × 2.1 mm 2.6 μm) H2O/MeOH with FA and HCOONH4 | Full MS ( | [45] |
AFB1, AFB2, AFG1, AFG2, AFM1 | Green tea supplements | UHPLC-Q-Orbitrap HESI + | Hypersil Gold C18 (100 × 2.1 mm, 1.9 μm) H2O/MeOH with FA and HCOONH4 | Full MS ( | [31] |
AFB1, AFB2, AFG1, AFG2 | Medicinal herbs | UHPLC-Q-Orbitrap HESI + | Kinetex C18 column (100 × 2.1 mm, 1.7 μm) H2O/MeOH with FA | SIM | [46] |
AFB1, AFB2, AFG1, AFG2 | Pet foods | UHPLC-Q-Orbitrap HESI + | Luna Omega C18 (50 × 2.1 mm, 1.6 μm) H2O/MeOH with FA and HCOONH4 | Full MS and AIF ( | [32] |
AFB1, AFB2, AFG1, AFG2 | Waters | UHPLC-Q-Orbitrap HESI + | C18 (125 × 2 mm, 5 μm) H2O/ ACN with FA | Full MS (no data) and dd-MS2 (list dependent) | [47] |
AFB1-lys | Human plasma | NanoLC–Q-Orbitrap HESI + | Acclaim C18 (15 cm, 75 μm) H2O/ACN with FA | Full MS ( | [26] |
AFB1, AFB2, AFG1, AFG2 | Snus | Heart-cutting 2D-LC-Q-Orbitrap HESI + | Hypersil Gold C18 (100 × 0.5 mm, 3 μm) and ACQUITY HSS PFP (100 × 2.1 mm, 1.7 μm) H2O/MeOH/ACN with FA and HCOONH4 | PRM | [27] |
AFB1, AFB2, AFG1, AFG2 | Human plasma | UHPLC–IT-Orbitrap HESI + | PFP (50 × 2.1 mm, 2.6 μm) H2O/MeOH with AA | Full MS ( | [22] |
AFB1, AFB2, AFG1, AFG2 | Beer | UHPLC-IT-Orbitrap HESI + | Gemini C18 (150 × 2 mm, 5 μm) H2O/MeOH with FA and HCOONH4 | Full MS ( | [23] |
AFB1 | Beer | HPLC-TOF ESI + | Kinetex C18 (50 × 3 mm, 1.7 μm) H2O/MeOH with FA and CH3COONH4 | Full MS ( | [24] |
AFB1 | Beer | UHPLC-TOF ESI + | Kinetex C18 (50 × 3 mm, 1.7 μm) H2O/ACN with FA and CH3COONH4 | Full MS ( | [25] |
AFB1, AFB2, AFG1, AFG2 | Human plasma | UHPLC–Q-TOF ESI + | ODS C18 (150 × 1.5 mm, 5.0 μm) H2O/MeOH with HCOONH4 | Full MS (no data) and dd-MS2 (Ion intensity-dependent) | [29] |
AFB1-lys | Human serum | UHPLC-Q-TOF ESI + | Acquity BEH C18 (50 × 2.1 mm, 1.7 μm) H2O/ACN with FA | Full MS ( | [48] |
AFB1, AFM1 | Plasma, urine, feces (pig, broiler) | UHPLC-Q-TOF ESI + | Acquity HSS T3 C18 (100 × 2.1 mm, 1.8 μm) H2O/MeOH with FA and HCOONH4 | Full MS and AIF (mass range | [49] |
AFB1 | Seeds, milk, flour, beer | UHPLC-Q-TOF ESI + | Eclipse Plus C8 RRHD (50 × 2.1 mm, 1.8 μm) H2O/ACN with FA | No data | [28] |
AFB1, AFB2, AFG1, AFG2 | Corn | UHPLC-Q-TOF ESI + | Hypersil Gold C18 (100 × 2.1mm, 1.9 μm) H2O/MeOH with FA and CH3COONH4 | Full MS ( | [50] |
Summary of LC-HRMS condition for AF determination.
AFs can grow on many foods, mainly peanuts, maize and cottonseed, although they have also been found in all types of nuts, copra, cereals, sunflower and soya beans, unrefined vegetable oils, spices, dried fruits, coffee, cocoa and animal feed [1, 16].
Because occurrence of mycotoxins in
A simple and rapid multi-mycotoxin method for the determination of 17 mycotoxins simultaneously is described on
Mycotoxins are frequently present in
A
Modern MS detectors can be used not only as detectors, but also as a “separation” tool, due to significant advances in HRMS, achieving greater sensitivity and selectivity. Thus,
Mycotoxins can be present in their parent forms and also in other forms, as
The determination of mycotoxin exposure of human populations is difficult due to the heterogeneous distribution of mycotoxins in foods and the time lag between toxin intake and the development of chronic disease. Therefore, a more reliable and relevant indication of individual exposure could be provided by biomarkers measured in biological fluids.
Aflatoxins can bioaccumulate in the organs and tissues of animals and humans or be excreted by biological fluids or feces [39, 40]. Sensitive analytical procedures are required for the determination of AFs in biological samples due to the very low concentrations involved. Most of the analytical methods proposed are based on LC coupled to different detection systems such as spectrophotometry, fluorescence, MS or MS/MS. Recently, HRMS, including TOF and Orbitrap, resulted an excellent technique for target analysis of AFs as well as for identifying and screening of non-target compounds in metabolomic strategies for studies concerning bioaccumulation, toxicokinetics and excretion of AFs and their metabolites.
Most of the studies are related with
In addition,
In comparison to other biological fluids such as blood, plasma and urine, the database on multi-mycotoxin levels in
Several studies propose to analyze
Figure 3 shows a distribution of the type of food and biological samples for which LC-HRMS methods have been applied in AF determination.
Type of samples more frequently analyzed. The number of published articles dealing with each matrix is indicated.
Method accuracy and precision are strongly conditioned by the effectiveness and robustness of the sample treatment stage. Both physicochemical properties of the AFs and the sample matrix composition need to be considered in the extraction procedure selection, which should ideally isolate and concentrate the analytes, eliminate interferences, and provide extracts compatible with the analytical technique to be used. For example, AFB1 was extracted in acidified ethyl acetate (EtAc) from serum previously submitted to enzymatic digestion (ED) and lipid removing by liquid–liquid extraction (LLE) with hexane. Nevertheless, this extraction medium was unable to isolate the hydrophilic metabolite AFB1-lys from the same sample and, a salting-out step with a quick, easy, cheap, effective, rugged and safe (QuEChERS) mixture was applied [39]. The objective conditions the adoption of a more or less selective sample treatment. Thus, non-selective extractions are applied for non-targeted strategies, allowing retrospective analysis of any potential compound, whereas for targeted analysis, clean and concentrated extracts are required.
Solid samples are generally homogenized by grinding [37, 38, 43, 44, 46], in order to obtain representative sample aliquots before being submitted to a solid–liquid extraction (SLE) stage. Freeze-drying of feces has also been proposed for eliminating any variations due to different moisture contents [49]. For SLE, aqueous mixtures of polar organic solvents such as MeOH, acetone or ACN have been used for AFs isolation from biological [49] and food [37, 43, 44, 46, 52] samples, being the mixtures mechanically shaken. The application of external energy in SLE procedures is sometimes proposed in order to enhance analyte recoveries in low times. Ultrasound assisted extraction (UAE) [30, 32, 50, 55] and microwave assisted extraction (MAE) [28] have efficiently extract AFs from food matrices.
On the other hand, even though food and biological liquid samples could be directly analyzed HRMS or LC combined with HRMS, previous steps are generally applied to minimize matrix effects. Thus, the addition of organic solvents such as ACN or MeOH allowed deproteinization of plasma [26, 29, 49] and milk [42] samples. Enzymatic reactions have also proven to be effective for human serum deproteinization [39, 48].
The removing of non-polar sample components, such as phospholipids, has been proposed by LLE with hexane [39, 46], by solid-phase extraction (SPE) using SPE-phospholipid cartridges for pig feces [49] and well-plates for chicken plasma [49].
The isolation of AFB1, AFB2, AFG1, AFG2 and AFM1 by LLE using EtAc has been applied for urine [49] and serum [22, 39], being recommended though a three-step LLE by Slobodchikova et al. [22] which resulted in better recoveries than those provided by SPE or protein precipitation (PP) procedures in a multi-mycotoxin method.
The simultaneous sample matrix purification and AF isolation is accomplished by many of the applied treatments in a single step. Thus, an on-line SPE device allowed the simultaneous isolation of 12 mycotoxins, including AFB1, and matrix purification of beer samples. Although SPE is more commonly applied under off- line mode, as used by Rubert et al. for isolation of AFB1, AFB2, AFG1, AFG2 in a multi-mycotoxin method proposed for beer [23]. SLE extracts obtained from solid food matrices have also been submitted to SPE [43, 44]. Polymeric sorbents are used in SPE isolation of AFs in their free forms. Whereas mixed-mode SPE-sorbents are selected for the retention of AFB1-lys adduct. Thus, a modified extraction procedure involving a PP step before enzymatic digestion (ED) with Pronase® and SPE-clean-up using strong ion mixed-mode-SPE has allowed both metabolic profiling and AFB1-lys adduct quantification in serum samples [26, 48].
Specific antibody-analyte binding is exploited as clean-up procedure in IACs, reporting interesting applications in AF analysis. Thus, gel suspensions of monoclonal antibody specific for AFs have allowed the purification of AFB1, AFB2, AFG1 and AFG2 from urine [40]. IAC have also been proposed for multi-mycotoxin studies, including AFs, for the analysis of functional and medicinal herbs [46]. In both articles reviewed dealing with IAC, AFs were eluted with MeOH after a washing step for impurities elimination using water [40] of aqueous buffer solutions [46].
QuEChERS methodology has been applied for AFs determination in both food [24, 32, 38, 41, 45, 50, 51, 53, 56] and biological samples [33, 39], using ACN as extractant solvent and a dispersive SPE (DSPE) step with the appropriate sorbents. When QuEChERS clean-up is applied omitting the use of sorbents, so that only implying organic solvent and salts, the procedure is named as simplified QuEChERS, and it has been proposed for isolation of AFB1, AFB2, AFG1, AFG2 and AFM1 from human breast milk [33] and AFB1 from durum wheat pasta [45]. The possibilities of different mixtures of solid sorbents for multi-class determinations of more than 250 compounds, pesticides and mycotoxins, including AFB1, AFB2, AFG1 and AFG2, applied to nutraceutical products have been studied by Martínez-Domínguez et al. [34, 35, 36] and compared with SLE, called in this case as “dilute and shoot” procedure. Best results have been reported by the latter procedure followed by a clean-up step using a mixture of sorbents in a DSPE mode [35, 36] or cartridge packed [34], this clean-up stage applied in order to enhance analyte recoveries and/or maintain the equipment performance for longer periods of times. When “dilute and shoot” procedure was compared to IAC for AFs in urine sample, lower LODs were achieved by the latter, because cleaner extracts with higher AF concentrations were obtained, being therefore selected [40]. An on-line automated sample preparation procedure is developed by Jia et al. [31] for multiple mycotoxin screening in nutraceutical products involving SLE, using aqueous acid solution and ACN, and the obtained supernatant being transferred to a disposable pipette extraction containing salt previously to the application of a clean-up step based on DSPE.
In the last years, nanoparticles (NPs) have received great attention in analytical chemistry due to the high surface area to volume ratio if compared to particles of higher dimensions, thus leading to very efficient extractions in lower times. A mass of 2 mg of zirconia NPs dispersed in the aqueous extract obtained by MAE from food samples allowed the DSPE isolation of AFB1 in 2 min, being then submitted to a desorption step in acidified chloroform [28]. When magnetized NPs are used, the collection of the enriched NPs is easily achieved by applying an external magnetic field, avoiding the centrifugation step. Under this named magnetic dispersive solid-phase extraction (MDSPE) methodology, AFB1, AFB2, AFG1, AFG2, AFM1 and AFM2, in a multiclass mycotoxin analysis method, have been preconcentrated with multi-walled carbon nanotubes (MWCNTs) modified with polyethylene glycol [42]. Although not implying HRMS detection for LC, it is noteworthy that AFB1 has also been preconcentrated with amino-modified magnetic MWCNTs [57].
With the aim to enhance sensitivity and clean-up purposes, the AF extracts finally obtained by applying the selected isolation procedure, are generally evaporated to dryness and reconstituted in low volumes of solvents compatible with the instrumental measurement step.
The data provided in this review demonstrate that most of the studies dealing with AF determination by LC-HRMS are focused on food, feed and biological samples analysis. In fact, only two manuscripts dealing with other matrices, waters [47] and snus [27], were found. A triple-stage SPE, consisting of a hand-made cartridge, packed with porous graphitized carbon and modified styrene-divinylbenzene polymer, coupled to a commercial HLB plus cartridge, allowed the isolation of natural toxins of different polarities. Thus, a screening method is proposed for the tentative identification of mycotoxins, cyanotoxins and plant toxins in surface waters [47]. The use of multiple Heart-cutting 2D-LC-Q-Orbitrap for AF separation and detection, respectively, has probably allowed to apply a very simple procedure in the treatment of snus samples. Thus, LLE in acidified EtAc provided similar analyte recoveries than QuEChERS method [27].
As can be appreciated in Figure 4, sample treatments for AF determination by LC-HRMS have been proposed both applying a unique methodology or through the combination of different procedures generally sequentially applied, for both food and biological samples.
Sample treatments for AF determination by LC-HRMS.
A summary of the sample treatments involved in the reviewed LC-HRMS methods appearing in the literature for AF determination in different matrices is provided as well in Table 2.
Matrix | Sample treatment | Ref. |
---|---|---|
Human serum | ED of 0.5 mL sample with Pronase®, LLE degreasing with hexane and: LLE with acidified EtAc (for AFB1), QuEChERS (for FB1-lys) | [39] |
Human serum | ED of 0.25 mL sample with Pronase® and mixed-mode SPE. Elution with acidified MeOH | [48] |
Human plasma | PP of 0.23 mL sample with MeOH/water, supernatant ED with Pronase® and mixed-mode SPE. Elution with acidified MeOH | [26] |
Human plasma | PP of 0.1 mL sample with EtOH/ACN | [29] |
Human plasma | 3-step LLE of 0.1 mL sample with EtAc | [22] |
Human urine | IAC for 2 mL sample. Elution with MeOH | [40] |
Human breast milk | Simplified QuEChERS | [33] |
Pig plasma, urine, and feces. Broiler chicken plasma and excreta | Pig plasma: PP of 0.25 mL sample with ACN. Pig urine: LLE of 0.5 mL sample with EtAc. Pig feces: SLE of 0.25 g sample with acetone and SPE for phospholipid removal. Chicken plasma: PP of 0.15 mL sample with ACN and well-plates. Chicken excreta: SLE of 0.25 g sample with ACN | [49] |
Beer | QuEChERS for 5 mL sample | [24] |
Beer | On-line SPE for 0.5 mL sample. Elution with acidified ACN/CH3COONH4 | [25] |
Beer | SPE for 10 mL sample. Elution with ACN/MeOH | [23] |
Milk | PP of 4 g sample with ACN and MDSPE with 10 mg MNPs. Desorption with acidified EtAc | [42] |
Milk | QuEChERS for 10 mL sample | [41] |
Peach seed, milk powder, corn flour and beer | MAE of solid samples (0.2 g) in MeOH/water and DSPE with 2 mg zirconia NPs. Desorption with MeOH | [28] |
Maize | UAE of 0.7 g sample in acidified MeOH/dichloromethane/EtAc | [55] |
Maize | UAE of 0.5 g sample with acidified ACN | [30] |
Corn | UAE of 2 g sample in ACN/water and QuEChERS | [50] |
Cereal foods (flours and bread) | SLE of 10 g sample with ACN/water and SPE. Elution with MeOH | [44] |
Durum wheat pasta and baby food pasta | Simplified QuEChERS for 4 g sample | [45] |
Cashew nut | SLE of 1 g sample in MeOH/water and SPE. Elution with MeOH | [43] |
Coix seed | SLE of 5 g sample with acidified ACN | [37] |
Isoflavone supplements | SLE of 2.5 g sample with acidified ACN and clean-up by SPE | [34] |
Ginkgo biloba nutraceuticals | SLE of 2.5 g sample with acidified ACN and clean-up by DSPE | [35] |
Green tea and royal jelly supplements | SLE of 2.5 g sample with acidified ACN and clean-up by DSPE | [36] |
Green tea nutraceuticals | SLE of 1 g sample with acidified ACN and clean-up by DSPE | [31] |
Functional and medicinal herbs | SLE of 2 g sample with PBS, LLE degreasing with hexane and IAC. Elution with MeOH | [46] |
Pet foods | UAE of 2 g sample in acidified ACN and QuEChERS | [32] |
Feed | Simplified QuEChERS | [38] |
Surface and drinking waters | SPE for 100 mL sample. Elution with MeOH/water/acetone | [47] |
Snus | LLE with acidified EtAc | [27] |
Summary of the sample treatments used in the AF determination by LC-HRMS.
AFs are synthesized via multiple intermediates by a complex pathway in several species of the
As regards mycotoxin degradation, decontamination techniques for AFs in food and feed attract continuous interest due to their adverse health effects and large economic losses for producers. In this sense, physical, chemical and biological strategies have been proposed. Thus, ABF1 degradation products by electron beam irradiation have been identified, as well as the possible pathway, using UHPLC-Q-TOF-MS [61, 62]. High-voltage atmospheric cold plasma (HVACP) is other physical strategy applied for AFB1 decontamination, providing a 76% efficiency when the non-thermal treatment was applied for 5 min in air containing 40% relative humidity. Thus, molecular formulas of six degradation products were elucidated by HPLC-TOF and their structures were further studied by Orbitrap MS. Two of the detected degradation compounds were ozonolysis products of AFB1, and the other four indicated the action of other reactive species besides ozone, generated during HVACP treatment [63]. The proven degradation power of ultrasounds for AFB1 aqueous solutions allows to perceive this physical detoxification technology as promising for food industry. An ultrasound exposure of 80 min degraded AFB1 by 85.1%, being eight main reaction products identified by UHPLC-Q-Orbitrap [64]. The study of degradation pathways and structural identification of photodegradation products of AFB1 in aqueous medium [65], ACN [66] and on peanut surface [67], has been carried out using UHPLC-Q-TOF after ultraviolet irradiation of different intensities.
Biological degradation, mainly caused by bacterial and fungal enzymes, appears as a strategy for AFB1 removal, with inherent advantages over physical and chemical strategies such as being friendly to the environment. LC-Q-TOF has been applied in the monitorization of AFB1 degradation products, and the obtained results lead the authors to propose bacterial strain
AFs are secondary toxic metabolites which may be present mainly in contaminated food and biological samples at very low levels. Among them, AFB1 is considered the most toxic, being classified as a human carcinogen. The analysis of food and biological samples is very complex and includes different steps as extraction, clean-up, separation, and detection approaches. This chapter reports the main analytical procedures developed for the AF determination by LC-HRMS. Different sample preparation techniques have been proposed, being QuEChERS, SPE and SLE the more frequently used. New nanomaterials including magnetic nanoparticles have been recently applied as adsorbents, increasing extraction efficiency and specificity. Separation of AFs is usually performed using HPLC, which performance was improved when using UHPLC. Different detectors are proposed, being MS or MS/MS widely applied, ensuring a specific confirmation for targeted analysis. However, the toxicological pathway of AFs in biological samples leads to the appearance of modified or masked mycotoxins, whose structures must be accurately established, making their detection difficult using routine analytical methods. On the other hand, the lack of commercial analytical standards results a great challenge for accurate identification and quantitation of modified AFs. In this field, HRMS has proven to be a very effective tool to enable the rapid determination of both parent and modified AFs. The use of metabolomic platforms combined with HRMS is nowadays considered the most appropriate way to study the toxicokinetic behavior of AFs in order to establish, when possible, maximum tolerable intakes and to investigate whether they have any relationship with certain clinical pathologies and cancer processes.
The authors acknowledge the financial support of the Comunidad Autónoma de la Región de Murcia (CARM, Fundación Séneca, Project 19888/GERM/15), the Spanish MICINN (PGC2018-098363-B-100), the University of Murcia (R-987/2020) and the European Commission (FEDER).
AA | acetic acid |
ACN | acetonitrile |
AFAR | aflatoxin aldehyde reductase |
AFL | Aflatoxicol |
AFs | aflatoxins |
AFB1 | aflatoxin B1 |
AFB2 | aflatoxin B2 |
AFBO | aflatoxin-8,9-epoxide |
AFG1 | aflatoxin G1 |
AFG1 | aflatoxin G2 |
AFM1 | aflatoxin M1 |
AFM2 | aflatoxin M2 |
AFP1 | aflatoxin P1 |
AFQ1 | aflatoxin Q1 |
AIF | all-ion fragmentation |
AP | aputinic |
DART | direct analysis in real time |
2D | Two-dimensional |
dd-MS2 | data dependent |
DIA | data independent |
DMSPE | dispersive magnetic solid-phase extraction |
DPEP | dipeptidase |
DSPE | dispersive solid-phase extraction |
ED | enzymatic digestion |
ESI | electrospray ionization |
EtAc | ethyl acetate |
EtOH | ethanol |
FA | formic acid |
FAPy | formamidopyramidine |
FI | flow injection |
GGT | γ-glutamyltransferase |
GSH | glutathione |
GST | glutathione S-transferase |
HCD | high energy collision dissociation |
HESI | heated electrospray ionization |
HRMS | high resolution mass spectrometry |
HVACP | high-voltage atmospheric cold plasma |
IACs | immunoaffinity columns |
IARC | International Agency for Research on Cancer |
IDMS | isotope dilution mass spectrometry |
IT | ion trap |
LC | liquid chromatography |
LLE | liquid–liquid extraction |
LOD | limit of detection |
LOQ | limit of quantification |
lys | lysine |
MAE | microwave assisted extraction |
MDSPE | magnetic dispersed solid-phase extraction |
MeOH | methanol |
MS | mass spectrometry |
MS/MS | tandem mass spectrometry |
MWCNT | multi-walled carbon nanotubes |
NAT | N-acetyltransferase |
NPs | nanoparticles |
PBS | phosphate-buffered solution |
PFP | pentafluorophenyl |
PP | protein precipitation |
PRM | parallel reaction monitoring mode |
Q | quadrupole |
QqQ | triple quadrupole |
QuEChERS | quick easy cheap effective rugged and safe |
SIM | selected ion monitoring |
SLE | solid–liquid extraction |
SPE | solid-phase extraction |
TOF | time-of-flight |
TOF-SIMS | time-of-flight secondary ion mass spectrometry |
UAE | ultrasound assisted extraction |
UHPLC | ultra-high performance liquid chromatography |
WHO | World Health Organization. |
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\\n"}]'},components:[{type:"htmlEditorComponent",content:'Copyright is the term used to describe the rights related to the publication and distribution of original Works. Most importantly from a publisher's perspective, copyright governs how Authors, publishers and the general public can use, publish, and distribute publications.
\n\nIntechOpen only publishes manuscripts for which it has publishing rights. This is governed by a publication agreement between the Author and IntechOpen. This agreement is accepted by the Author when the manuscript is submitted and deals with both the rights of the publisher and Author, as well as any obligations concerning a particular manuscript. However, in accepting this agreement, Authors continue to retain significant rights to use and share their publications.
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On September, 29th 2006 he has won a post PhD fellowship from the university of Bologna (from October 2006 to October 2008), at the competitive examination he was ranked first in the industrial engineering area. He extensively served as referee for several international journals. He is author/coauthor of more than 100 research papers. He has been involved in some projects supported by MURST and European Community. His research interests include pattern recognition, bioinformatics, and biometric systems (fingerprint classification and recognition, signature verification, face recognition).",institutionString:null,institution:null},{id:"496",title:"Dr.",name:"Carlos",middleName:null,surname:"Leon",slug:"carlos-leon",fullName:"Carlos Leon",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Seville",country:{name:"Spain"}}},{id:"512",title:"Dr.",name:"Dayang",middleName:null,surname:"Jawawi",slug:"dayang-jawawi",fullName:"Dayang Jawawi",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Technology Malaysia",country:{name:"Malaysia"}}},{id:"528",title:"Dr.",name:"Kresimir",middleName:null,surname:"Delac",slug:"kresimir-delac",fullName:"Kresimir Delac",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/528/images/system/528.jpg",biography:"K. 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Insect pests are one of the main causes for losses in agriculture production, and current control technologies based on pesticide application or the use of transgenic crops expressing Bacillus thuringiensis toxin proteins are facing efficacy challenges. Novel approaches to control pests are urgently necessary. RNA interference (RNAi) is a gene silencing mechanism triggered by providing double-stranded RNA (dsRNA), that when ingested into insects can lead to death or affect the viability of the target pest. Transgenic plants expressing dsRNA version of insect specific target genes are the new generation of resistant plants. However, the RNAi mechanism is not conserved among insect orders, and its elucidation is the key to develop commercial RNAi crops. In this chapter, we review the core RNAi pathway in insects and the dsRNA uptake, amplification, and spread of systemic silencing signals in some key insect species. We also highlight some of the experimental steps before developing an insect-pest-resistant “RNAi plant”. Lastly, we review some of the most recent development studies to control agricultural insect pests by RNAi transgenic plants.",book:{id:"5090",slug:"rna-interference",title:"RNA Interference",fullTitle:"RNA Interference"},signatures:"Thais Barros Rodrigues and Antonio Figueira",authors:[{id:"176770",title:"Dr.",name:"Thais B.",middleName:null,surname:"Rodrigues",slug:"thais-b.-rodrigues",fullName:"Thais B. 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Histologically, collagen decreases, and the dermis is replaced by abnormal elastic fibers as a cause of wrinkle formation through the loss of skin elasticity. There have been numerous studies of skin aging performed to elucidate the underlying molecular mechanisms and to develop various antiaging therapeutics and preventive strategies. We summarized the molecular mechanisms and treatments of skin aging. Mainly UV radiation induces ROS formation and DNA damage, leading to increased production of MMPs and decreased production of collagen in keratinocytes and fibroblasts, which reflect the central aspects of skin aging. Besides UV radiation exposure, extrinsic factors including tobacco smoking, exposure to environmental pollutants, infrared radiation, and heat contribute to premature skin aging. Like UV radiation, these factors cause ROS formation and increase expression of MMPs, thus accelerating skin aging by inducing extracellular matrix (ECM) degradation. Accumulated collagen fibrils inhibit the new collagen synthesis and account for the further degradation of the ECM through this positive feedback loop. Accumulating evidence for molecular mechanisms of skin aging should provide clinicians with an expanding spectrum of therapeutic targets in the treatment of skin aging.",book:{id:"5258",slug:"molecular-mechanisms-of-the-aging-process-and-rejuvenation",title:"Molecular Mechanisms of the Aging Process and Rejuvenation",fullTitle:"Molecular Mechanisms of the Aging Process and Rejuvenation"},signatures:"Miri Kim and Hyun Jeong Park",authors:[{id:"47695",title:"Prof.",name:"Hyun Jeong",middleName:null,surname:"Park",slug:"hyun-jeong-park",fullName:"Hyun Jeong Park"},{id:"185767",title:"Prof.",name:"Miri",middleName:null,surname:"Kim",slug:"miri-kim",fullName:"Miri Kim"}]},{id:"49637",title:"RNA Interference Technology — Applications and Limitations",slug:"rna-interference-technology-applications-and-limitations",totalDownloads:4120,totalCrossrefCites:1,totalDimensionsCites:1,abstract:"RNA interference (RNAi), an evolutionarily conserved mechanism triggered by double-stranded RNA (dsRNA), causes gene silencing in a sequence-specific manner. RNAi evolved naturally to mediate protection from both endogenous and exogenous pathogenic nucleic acids and to modulate gene expression. Multiple technological advancements and precision in gene targeting have allowed a plethora of potential applications, ranging from targeting infections in crop plants to improving health in human patients, which have been reviewed in this chapter.",book:{id:"5090",slug:"rna-interference",title:"RNA Interference",fullTitle:"RNA Interference"},signatures:"Devi Singh, Sarika Chaudhary, Rajendra Kumar, Preeti Sirohi,\nKamiya Mehla, Anil Sirohi, Shashi Kumar, Pooran Chand and Pankaj\nKumar Singh",authors:[{id:"176625",title:"Prof.",name:"Devi",middleName:null,surname:"Singh",slug:"devi-singh",fullName:"Devi Singh"},{id:"176744",title:"Ms.",name:"Preeti",middleName:null,surname:"Sirohi",slug:"preeti-sirohi",fullName:"Preeti Sirohi"},{id:"176745",title:"Dr.",name:"Rajendra",middleName:null,surname:"Kumar",slug:"rajendra-kumar",fullName:"Rajendra Kumar"},{id:"176746",title:"Mrs.",name:"Sarika",middleName:null,surname:"Chaudhary",slug:"sarika-chaudhary",fullName:"Sarika Chaudhary"},{id:"176747",title:"Dr.",name:"Kamiya",middleName:null,surname:"Mehla",slug:"kamiya-mehla",fullName:"Kamiya Mehla"},{id:"176748",title:"Dr.",name:"Pankaj Kumar",middleName:null,surname:"Singh",slug:"pankaj-kumar-singh",fullName:"Pankaj Kumar Singh"},{id:"176749",title:"Dr.",name:"Shashi",middleName:null,surname:"Kumar",slug:"shashi-kumar",fullName:"Shashi Kumar"},{id:"176809",title:"Dr.",name:"Pooran",middleName:null,surname:"Chand",slug:"pooran-chand",fullName:"Pooran Chand"}]},{id:"43280",title:"Gene Therapy for Diabetic Retinopathy – Targeting the Renin-Angiotensin System",slug:"gene-therapy-for-diabetic-retinopathy-targeting-the-renin-angiotensin-system",totalDownloads:2482,totalCrossrefCites:2,totalDimensionsCites:2,abstract:null,book:{id:"3509",slug:"gene-therapy-tools-and-potential-applications",title:"Gene Therapy",fullTitle:"Gene Therapy - Tools and Potential Applications"},signatures:"Qiuhong Li, Amrisha Verma, Ping Zhu, Bo Lei, Yiguo Qiu, Takahiko Nakagawa, Mohan K Raizada and William W Hauswirth",authors:[{id:"155578",title:"Dr.",name:"Qiuhong",middleName:null,surname:"Li",slug:"qiuhong-li",fullName:"Qiuhong Li"},{id:"360660",title:"Dr.",name:"Amrisha",middleName:null,surname:"Verma",slug:"amrisha-verma",fullName:"Amrisha Verma"},{id:"360661",title:"Dr.",name:"Ping",middleName:null,surname:"Zhu",slug:"ping-zhu",fullName:"Ping Zhu"},{id:"360662",title:"Dr.",name:"Bo",middleName:null,surname:"Lei",slug:"bo-lei",fullName:"Bo Lei"},{id:"360663",title:"Dr.",name:"Yiguo",middleName:null,surname:"Qiu",slug:"yiguo-qiu",fullName:"Yiguo Qiu"},{id:"360664",title:"Dr.",name:"Takahiko",middleName:null,surname:"Nakagawa",slug:"takahiko-nakagawa",fullName:"Takahiko Nakagawa"},{id:"360665",title:"Dr.",name:"Mohan K",middleName:null,surname:"Raizada",slug:"mohan-k-raizada",fullName:"Mohan K Raizada"},{id:"360666",title:"Dr.",name:"William W",middleName:null,surname:"Hauswirth",slug:"william-w-hauswirth",fullName:"William W Hauswirth"}]},{id:"49416",title:"Microinjection-Based RNA Interference Method in the Water Flea, Daphnia pulex and Daphnia magna",slug:"microinjection-based-rna-interference-method-in-the-water-flea-daphnia-pulex-and-daphnia-magna",totalDownloads:2209,totalCrossrefCites:5,totalDimensionsCites:8,abstract:"It is well known that most daphnid species have several attractive life history characteristics such as cyclical parthenogenesis, environmental sex determination, and predator-induced defense formation. Recent advances in high-throughput omics technologies make it easy to obtain a huge number of potential candidate factors involved in environmental stimuli-triggered phenotypic alterations. Furthermore, our group has developed a microinjection system to introduce foreign materials such as nucleotides and chemicals into the early-stage (one-cell stage) egg of Daphnia pulex and Daphnia magna. Consequently, we established a microinjection-based RNAi system that allows arbitrary gene functions to be investigated. However, this microinjection system does not seem to have pervaded in the daphnid research community due to its low throughput and high level of skills required. In this chapter, we review the microinjection method and its RNAi system in water fleas, D. pulex and D. magna, providing some technical tips and making challenging proposals for the development of novel high-throughput RNAi methods. Finally, we provide an overview of recently developed gene functional analysis methods such as overexpression and genome-editing systems.",book:{id:"5090",slug:"rna-interference",title:"RNA Interference",fullTitle:"RNA Interference"},signatures:"Kenji Toyota, Shinichi Miyagawa, Yukiko Ogino and Taisen Iguchi",authors:[{id:"92826",title:"Dr.",name:"Taisen",middleName:null,surname:"Iguchi",slug:"taisen-iguchi",fullName:"Taisen Iguchi"},{id:"176835",title:"Dr.",name:"Kenji",middleName:null,surname:"Toyota",slug:"kenji-toyota",fullName:"Kenji Toyota"}]},{id:"55603",title:"RNA‐seq: Applications and Best Practices",slug:"rna-seq-applications-and-best-practices",totalDownloads:3782,totalCrossrefCites:7,totalDimensionsCites:8,abstract:"RNA‐sequencing (RNA‐seq) is the state‐of‐the‐art technique for transcriptome analysis that takes advantage of high‐throughput next‐generation sequencing. Although being a powerful approach, RNA‐seq imposes major challenges throughout its steps with numerous caveats. There are currently many experimental options available, and a complete comprehension of each step is critical to make right decisions and avoid getting into inconclusive results. A complete workflow consists of: (1) experimental design; (2) sample and library preparation; (3) sequencing; and (4) data analysis. RNA‐seq enables a wide range of applications such as the discovery of novel genes, gene/transcript quantification, and differential expression and functional analysis. This chapter will encompass the main aspects from sample preparation to downstream data analysis. It will be discussed how to obtain high‐quality samples, replicates amount, library preparation, sequencing platforms and coverage, focusing on best recommended practices based on specialized literature. Basic techniques and well‐known algorithms are presented and discussed, guiding both beginners and experienced users in the implementation of reliable experiments.",book:{id:"5944",slug:"applications-of-rna-seq-and-omics-strategies-from-microorganisms-to-human-health",title:"Applications of RNA-Seq and Omics Strategies",fullTitle:"Applications of RNA-Seq and Omics Strategies - From Microorganisms to Human Health"},signatures:"Michele Araújo Pereira, Eddie Luidy Imada and Rafael Lucas Muniz\nGuedes",authors:[{id:"202103",title:"Ph.D. Student",name:"Michele",middleName:"Araújo",surname:"Pereira",slug:"michele-pereira",fullName:"Michele Pereira"},{id:"202456",title:"MSc.",name:"Eddie Luidy",middleName:null,surname:"Imada",slug:"eddie-luidy-imada",fullName:"Eddie Luidy Imada"},{id:"202460",title:"Dr.",name:"Rafael",middleName:null,surname:"Guedes",slug:"rafael-guedes",fullName:"Rafael Guedes"}]}],onlineFirstChaptersFilter:{topicId:"419",limit:6,offset:0},onlineFirstChaptersCollection:[],onlineFirstChaptersTotal:0},preDownload:{success:null,errors:{}},subscriptionForm:{success:null,errors:{}},aboutIntechopen:{},privacyPolicy:{},peerReviewing:{},howOpenAccessPublishingWithIntechopenWorks:{},sponsorshipBooks:{sponsorshipBooks:[],offset:0,limit:8,total:null},allSeries:{pteSeriesList:[{id:"14",title:"Artificial Intelligence",numberOfPublishedBooks:9,numberOfPublishedChapters:90,numberOfOpenTopics:6,numberOfUpcomingTopics:0,issn:"2633-1403",doi:"10.5772/intechopen.79920",isOpenForSubmission:!0},{id:"7",title:"Biomedical Engineering",numberOfPublishedBooks:12,numberOfPublishedChapters:108,numberOfOpenTopics:3,numberOfUpcomingTopics:0,issn:"2631-5343",doi:"10.5772/intechopen.71985",isOpenForSubmission:!0}],lsSeriesList:[{id:"11",title:"Biochemistry",numberOfPublishedBooks:33,numberOfPublishedChapters:330,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2632-0983",doi:"10.5772/intechopen.72877",isOpenForSubmission:!0},{id:"25",title:"Environmental Sciences",numberOfPublishedBooks:1,numberOfPublishedChapters:19,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2754-6713",doi:"10.5772/intechopen.100362",isOpenForSubmission:!0},{id:"10",title:"Physiology",numberOfPublishedBooks:14,numberOfPublishedChapters:145,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2631-8261",doi:"10.5772/intechopen.72796",isOpenForSubmission:!0}],hsSeriesList:[{id:"3",title:"Dentistry",numberOfPublishedBooks:9,numberOfPublishedChapters:141,numberOfOpenTopics:2,numberOfUpcomingTopics:0,issn:"2631-6218",doi:"10.5772/intechopen.71199",isOpenForSubmission:!0},{id:"6",title:"Infectious Diseases",numberOfPublishedBooks:13,numberOfPublishedChapters:123,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2631-6188",doi:"10.5772/intechopen.71852",isOpenForSubmission:!0},{id:"13",title:"Veterinary Medicine and Science",numberOfPublishedBooks:11,numberOfPublishedChapters:112,numberOfOpenTopics:3,numberOfUpcomingTopics:0,issn:"2632-0517",doi:"10.5772/intechopen.73681",isOpenForSubmission:!0}],sshSeriesList:[{id:"22",title:"Business, Management and Economics",numberOfPublishedBooks:1,numberOfPublishedChapters:22,numberOfOpenTopics:3,numberOfUpcomingTopics:0,issn:"2753-894X",doi:"10.5772/intechopen.100359",isOpenForSubmission:!0},{id:"23",title:"Education and Human Development",numberOfPublishedBooks:0,numberOfPublishedChapters:11,numberOfOpenTopics:1,numberOfUpcomingTopics:1,issn:null,doi:"10.5772/intechopen.100360",isOpenForSubmission:!0},{id:"24",title:"Sustainable Development",numberOfPublishedBooks:1,numberOfPublishedChapters:19,numberOfOpenTopics:5,numberOfUpcomingTopics:0,issn:"2753-6580",doi:"10.5772/intechopen.100361",isOpenForSubmission:!0}],testimonialsList:[{id:"6",text:"It is great to work with the IntechOpen to produce a worthwhile collection of research that also becomes a great educational resource and guide for future research endeavors.",author:{id:"259298",name:"Edward",surname:"Narayan",institutionString:null,profilePictureURL:"https://mts.intechopen.com/storage/users/259298/images/system/259298.jpeg",slug:"edward-narayan",institution:{id:"3",name:"University of Queensland",country:{id:null,name:"Australia"}}}},{id:"13",text:"The collaboration with and support of the technical staff of IntechOpen is fantastic. 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",coverUrl:"https://cdn.intechopen.com/series/covers/3.jpg",latestPublicationDate:"August 14th, 2022",hasOnlineFirst:!0,numberOfPublishedBooks:9,editor:{id:"419588",title:"Ph.D.",name:"Sergio",middleName:"Alexandre",surname:"Gehrke",slug:"sergio-gehrke",fullName:"Sergio Gehrke",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y000038WgMKQA0/Profile_Picture_2022-06-02T11:44:20.jpg",biography:"Dr. Sergio Alexandre Gehrke is a doctorate holder in two fields. The first is a Ph.D. in Cellular and Molecular Biology from the Pontificia Catholic University, Porto Alegre, Brazil, in 2010 and the other is an International Ph.D. in Bioengineering from the Universidad Miguel Hernandez, Elche/Alicante, Spain, obtained in 2020. In 2018, he completed a postdoctoral fellowship in Materials Engineering in the NUCLEMAT of the Pontificia Catholic University, Porto Alegre, Brazil. He is currently the Director of the Postgraduate Program in Implantology of the Bioface/UCAM/PgO (Montevideo, Uruguay), Director of the Cathedra of Biotechnology of the Catholic University of Murcia (Murcia, Spain), an Extraordinary Full Professor of the Catholic University of Murcia (Murcia, Spain) as well as the Director of the private center of research Biotecnos – Technology and Science (Montevideo, Uruguay). Applied biomaterials, cellular and molecular biology, and dental implants are among his research interests. He has published several original papers in renowned journals. In addition, he is also a Collaborating Professor in several Postgraduate programs at different universities all over the world.",institutionString:null,institution:{name:"Universidad Católica San Antonio de Murcia",institutionURL:null,country:{name:"Spain"}}},editorTwo:null,editorThree:null},subseries:{paginationCount:4,paginationItems:[{id:"14",title:"Cell and Molecular Biology",coverUrl:"https://cdn.intechopen.com/series_topics/covers/14.jpg",isOpenForSubmission:!0,editor:{id:"165627",title:"Dr.",name:"Rosa María",middleName:null,surname:"Martínez-Espinosa",slug:"rosa-maria-martinez-espinosa",fullName:"Rosa María Martínez-Espinosa",profilePictureURL:"https://mts.intechopen.com/storage/users/165627/images/system/165627.jpeg",biography:"Dr. Rosa María Martínez-Espinosa has been a Spanish Full Professor since 2020 (Biochemistry and Molecular Biology) and is currently Vice-President of International Relations and Cooperation development and leader of the research group 'Applied Biochemistry” (University of Alicante, Spain). Other positions she has held at the university include Vice-Dean of Master Programs, Vice-Dean of the Degree in Biology and Vice-Dean for Mobility and Enterprise and Engagement at the Faculty of Science (University of Alicante). She received her Bachelor in Biology in 1998 (University of Alicante) and her PhD in 2003 (Biochemistry, University of Alicante). She undertook post-doctoral research at the University of East Anglia (Norwich, U.K. 2004-2005; 2007-2008).\nHer multidisciplinary research focuses on investigating archaea and their potential applications in biotechnology. She has an H-index of 21. She has authored one patent and has published more than 70 indexed papers and around 60 book chapters.\nShe has contributed to more than 150 national and international meetings during the last 15 years. Her research interests include archaea metabolism, enzymes purification and characterization, gene regulation, carotenoids and bioplastics production, antioxidant\ncompounds, waste water treatments, and brines bioremediation.\nRosa María’s other roles include editorial board member for several journals related\nto biochemistry, reviewer for more than 60 journals (biochemistry, molecular biology, biotechnology, chemistry and microbiology) and president of several organizing committees in international meetings related to the N-cycle or respiratory processes.",institutionString:null,institution:{name:"University of Alicante",institutionURL:null,country:{name:"Spain"}}},editorTwo:null,editorThree:null},{id:"15",title:"Chemical Biology",coverUrl:"https://cdn.intechopen.com/series_topics/covers/15.jpg",isOpenForSubmission:!0,editor:{id:"441442",title:"Dr.",name:"Şükrü",middleName:null,surname:"Beydemir",slug:"sukru-beydemir",fullName:"Şükrü Beydemir",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y00003GsUoIQAV/Profile_Picture_1634557147521",biography:"Dr. Şükrü Beydemir obtained a BSc in Chemistry in 1995 from Yüzüncü Yıl University, MSc in Biochemistry in 1998, and PhD in Biochemistry in 2002 from Atatürk University, Turkey. He performed post-doctoral studies at Max-Planck Institute, Germany, and University of Florence, Italy in addition to making several scientific visits abroad. He currently works as a Full Professor of Biochemistry in the Faculty of Pharmacy, Anadolu University, Turkey. Dr. Beydemir has published over a hundred scientific papers spanning protein biochemistry, enzymology and medicinal chemistry, reviews, book chapters and presented several conferences to scientists worldwide. He has received numerous publication awards from various international scientific councils. He serves in the Editorial Board of several international journals. Dr. Beydemir is also Rector of Bilecik Şeyh Edebali University, Turkey.",institutionString:null,institution:{name:"Anadolu University",institutionURL:null,country:{name:"Turkey"}}},editorTwo:{id:"13652",title:"Prof.",name:"Deniz",middleName:null,surname:"Ekinci",slug:"deniz-ekinci",fullName:"Deniz Ekinci",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002aYLT1QAO/Profile_Picture_1634557223079",biography:"Dr. Deniz Ekinci obtained a BSc in Chemistry in 2004, MSc in Biochemistry in 2006, and PhD in Biochemistry in 2009 from Atatürk University, Turkey. He studied at Stetson University, USA, in 2007-2008 and at the Max Planck Institute of Molecular Cell Biology and Genetics, Germany, in 2009-2010. Dr. Ekinci currently works as a Full Professor of Biochemistry in the Faculty of Agriculture and is the Head of the Enzyme and Microbial Biotechnology Division, Ondokuz Mayıs University, Turkey. He is a member of the Turkish Biochemical Society, American Chemical Society, and German Genetics society. Dr. Ekinci published around ninety scientific papers, reviews and book chapters, and presented several conferences to scientists. He has received numerous publication awards from several scientific councils. Dr. Ekinci serves as the Editor in Chief of four international books and is involved in the Editorial Board of several international journals.",institutionString:null,institution:{name:"Ondokuz Mayıs University",institutionURL:null,country:{name:"Turkey"}}},editorThree:null},{id:"17",title:"Metabolism",coverUrl:"https://cdn.intechopen.com/series_topics/covers/17.jpg",isOpenForSubmission:!0,editor:{id:"138626",title:"Dr.",name:"Yannis",middleName:null,surname:"Karamanos",slug:"yannis-karamanos",fullName:"Yannis Karamanos",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002g6Jv2QAE/Profile_Picture_1629356660984",biography:"Yannis Karamanos, born in Greece in 1953, completed his pre-graduate studies at the Université Pierre et Marie Curie, Paris, then his Masters and Doctoral degree at the Université de Lille (1983). He was associate professor at the University of Limoges (1987) before becoming full professor of biochemistry at the Université d’Artois (1996). He worked on the structure-function relationships of glycoconjugates and his main project was the investigations on the biological roles of the de-N-glycosylation enzymes (Endo-N-acetyl-β-D-glucosaminidase and peptide-N4-(N-acetyl-β-glucosaminyl) asparagine amidase). From 2002 he contributes to the understanding of the Blood-brain barrier functioning using proteomics approaches. He has published more than 70 papers. His teaching areas are energy metabolism and regulation, integration and organ specialization and metabolic adaptation.",institutionString:null,institution:{name:"Artois University",institutionURL:null,country:{name:"France"}}},editorTwo:null,editorThree:null},{id:"18",title:"Proteomics",coverUrl:"https://cdn.intechopen.com/series_topics/covers/18.jpg",isOpenForSubmission:!0,editor:{id:"200689",title:"Prof.",name:"Paolo",middleName:null,surname:"Iadarola",slug:"paolo-iadarola",fullName:"Paolo Iadarola",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bSCl8QAG/Profile_Picture_1623568118342",biography:"Paolo Iadarola graduated with a degree in Chemistry from the University of Pavia (Italy) in July 1972. He then worked as an Assistant Professor at the Faculty of Science of the same University until 1984. In 1985, Prof. Iadarola became Associate Professor at the Department of Biology and Biotechnologies of the University of Pavia and retired in October 2017. Since then, he has been working as an Adjunct Professor in the same Department at the University of Pavia. His research activity during the first years was primarily focused on the purification and structural characterization of enzymes from animal and plant sources. During this period, Prof. Iadarola familiarized himself with the conventional techniques used in column chromatography, spectrophotometry, manual Edman degradation, and electrophoresis). Since 1995, he has been working on: i) the determination in biological fluids (serum, urine, bronchoalveolar lavage, sputum) of proteolytic activities involved in the degradation processes of connective tissue matrix, and ii) on the identification of biological markers of lung diseases. In this context, he has developed and validated new methodologies (e.g., Capillary Electrophoresis coupled to Laser-Induced Fluorescence, CE-LIF) whose application enabled him to determine both the amounts of biochemical markers (Desmosines) in urine/serum of patients affected by Chronic Obstructive Pulmonary Disease (COPD) and the activity of proteolytic enzymes (Human Neutrophil Elastase, Cathepsin G, Pseudomonas aeruginosa elastase) in sputa of these patients. More recently, Prof. Iadarola was involved in developing techniques such as two-dimensional electrophoresis coupled to liquid chromatography/mass spectrometry (2DE-LC/MS) for the proteomic analysis of biological fluids aimed at the identification of potential biomarkers of different lung diseases. He is the author of about 150 publications (According to Scopus: H-Index: 23; Total citations: 1568- According to WOS: H-Index: 20; Total Citations: 1296) of peer-reviewed international journals. He is a Consultant Reviewer for several journals, including the Journal of Chromatography A, Journal of Chromatography B, Plos ONE, Proteomes, International Journal of Molecular Science, Biotech, Electrophoresis, and others. He is also Associate Editor of Biotech.",institutionString:null,institution:{name:"University of Pavia",institutionURL:null,country:{name:"Italy"}}},editorTwo:{id:"201414",title:"Dr.",name:"Simona",middleName:null,surname:"Viglio",slug:"simona-viglio",fullName:"Simona Viglio",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRKDHQA4/Profile_Picture_1630402531487",biography:"Simona Viglio is an Associate Professor of Biochemistry at the Department of Molecular Medicine at the University of Pavia. She has been working since 1995 on the determination of proteolytic enzymes involved in the degradation process of connective tissue matrix and on the identification of biological markers of lung diseases. She gained considerable experience in developing and validating new methodologies whose applications allowed her to determine both the amount of biomarkers (Desmosine and Isodesmosine) in the urine of patients affected by COPD, and the activity of proteolytic enzymes (HNE, Cathepsin G, Pseudomonas aeruginosa elastase) in the sputa of these patients. Simona Viglio was also involved in research dealing with the supplementation of amino acids in patients with brain injury and chronic heart failure. She is presently engaged in the development of 2-DE and LC-MS techniques for the study of proteomics in biological fluids. The aim of this research is the identification of potential biomarkers of lung diseases. She is an author of about 90 publications (According to Scopus: H-Index: 23; According to WOS: H-Index: 20) on peer-reviewed journals, a member of the “Società Italiana di Biochimica e Biologia Molecolare,“ and a Consultant Reviewer for International Journal of Molecular Science, Journal of Chromatography A, COPD, Plos ONE and Nutritional Neuroscience.",institutionString:null,institution:{name:"University of Pavia",institutionURL:null,country:{name:"Italy"}}},editorThree:null}]},overviewPageOFChapters:{paginationCount:42,paginationItems:[{id:"82914",title:"Glance on the Critical Role of IL-23 Receptor Gene Variations in Inflammation-Induced Carcinogenesis",doi:"10.5772/intechopen.105049",signatures:"Mohammed El-Gedamy",slug:"glance-on-the-critical-role-of-il-23-receptor-gene-variations-in-inflammation-induced-carcinogenesis",totalDownloads:15,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"Chemokines Updates",coverURL:"https://cdn.intechopen.com/books/images_new/11672.jpg",subseries:{id:"18",title:"Proteomics"}}},{id:"82875",title:"Lipidomics as a Tool in the Diagnosis and Clinical Therapy",doi:"10.5772/intechopen.105857",signatures:"María Elizbeth Alvarez Sánchez, Erick Nolasco Ontiveros, Rodrigo Arreola, Adriana Montserrat Espinosa González, Ana María García Bores, Roberto Eduardo López Urrutia, Ignacio Peñalosa Castro, María del Socorro Sánchez Correa and Edgar Antonio Estrella Parra",slug:"lipidomics-as-a-tool-in-the-diagnosis-and-clinical-therapy",totalDownloads:7,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"Fatty Acids - Recent Advances",coverURL:"https://cdn.intechopen.com/books/images_new/11669.jpg",subseries:{id:"17",title:"Metabolism"}}},{id:"82440",title:"Lipid Metabolism and Associated Molecular Signaling Events in Autoimmune Disease",doi:"10.5772/intechopen.105746",signatures:"Mohan Vanditha, Sonu Das and Mathew John",slug:"lipid-metabolism-and-associated-molecular-signaling-events-in-autoimmune-disease",totalDownloads:17,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"Fatty Acids - Recent Advances",coverURL:"https://cdn.intechopen.com/books/images_new/11669.jpg",subseries:{id:"17",title:"Metabolism"}}},{id:"82483",title:"Oxidative Stress in Cardiovascular Diseases",doi:"10.5772/intechopen.105891",signatures:"Laura Mourino-Alvarez, Tamara Sastre-Oliva, Nerea Corbacho-Alonso and Maria G. 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